Handbook of PDF

Food Science and Technology
Handbook of Functional
Nutraceutical
11
Science
and
Technology
Nutraceutical Science
and Technology
Series Editor: Fereidoon Shahidi
11
Beverages and Human Health
Handbook of
Handbook of Functional Beverages

Handbook of Functional Beverages and Human Health provides potential appli-
cations and new developments in functional beverages, nutraceuticals, and health
foods. In addition to serving as a reference manual, it summarizes the current state
of knowledge in key research areas and contains novel ideas for future research and Functional
Beverages and
development. Additionally, it provides an easy-to-read text suitable for teaching senior
undergraduate and postgraduate students in the relevant areas.
and Human Health

Human Health
The book is divided into seven major sections: Section I covers market trends,
global regulations, flavor challenges, chemistry, and health with specific reference
to cancer chemoprevention and the prevention of postprandial metabolic stress due
to the consumption of functional beverages. Section II, by far the largest part of
the book, has 39 chapters on the most popular fruit juices (apple juice, lemon juice,
pomegranate juice, watermelon juice, etc.). Section III reports on herbal and vegetable
juices (carrot juice, Chinese medicinal herbs and root-based beverages, tomato juice,
and vegetable-containing juices).
Edited by
Section IV details caffeinated beverages, including different varieties of tea (green,
black, oolong, and herbal teas), coffee (coffee and beverages from green coffee beans), Fereidoon Shahidi
and cocoa and chocolate. Section V is on dairy and soy beverages, while Section VI is
on alcoholic beverages (wine) and water (maple water). Finally, Section VII describes
Cesarettin Alasalvar
fermented (kefir, koumiss, and ayran) and fortified functional beverages (applications
of plant sterols and stanols in functional beverages, beverages fortified with omega-3
fatty acids, dietary fiber, minerals and vitamins, probiotics and prebiotics in functional
beverages, functional beverages in weight management, fortified sports drinks, and
peptide-enriched functional beverages).
Shahidi
Alasalvar
K20775
Handbook of
Functional
Beverages and
Human Health
NUTRACEUTICAL SCIENCE AND TECHNOLOGY
Series Editor
FEREIDOON SHAHIDI
Ph.D., FACS, FAOCS, FCIC, FCIFST, FIAFoST, FIFT, FRSC
University Research Professor
Department of Biochemistry
Memorial University of Newfoundland
St. John’s, Newfoundland, Canada
1. Phytosterols as Functional Food Components and Nutraceuticals

edited by Paresh C. Dutta
2. Bioprocesses and Biotechnology for Functional Foods and Nutraceuticals
edited by Jean-Richard Neeser and Bruce J. German
3. Asian Functional Foods
John Shi, Chi-Tang Ho, and Fereidoon Shahidi
4. Nutraceutical Proteins and Peptides in Health and Disease
edited by Yoshinori Mine and Fereidoon Shahidi
5. Nutraceutical and Specialty Lipids and Their Co-Products
edited by Fereidoon Shahidi
6. Anti-Angiogenic Functional and Medicinal Foods
edited by Jack N. Losso, Fereidoon Shahidi, and Debasis Bagchi
7. Marine Nutraceuticals and Functional Foods
edited by Colin Barrow and Fereidoon Shahidi
8. Tea and Tea Products: Chemistry and Health-Promoting Properties
edited by Chi-Tang Ho, Jen-Kun Lin, and Fereidoon Shahidi
9. Tree Nuts: Composition, Phytochemicals, and Health Effects
edited by Cesarettin Alasalvar and Fereidoon Shahidi
10. Functional Foods of the East
edited by John Shi, Chi-Tang Ho, and Fereidoon Shahidi
11. Handbook of Functional Beverages and Human Health
edited by Fereidoon Shahidi and Cesarettin Alasalvar
Handbook of
Functional
Beverages and
Human Health
Edited by
Fereidoon Shahidi
Boca Raton London New York
CRC Press is an imprint of the

Taylor & Francis Group, an informa business
CRC Press
Taylor & Francis Group
6000 Broken Sound Parkway NW, Suite 300
Boca Raton, FL 33487-2742
© 2016 by Taylor & Francis Group, LLC
CRC Press is an imprint of Taylor & Francis Group, an Informa business
No claim to original U.S. Government works

Version Date: 20160120
International Standard Book Number-13: 978-1-4665-9642-9 (eBook - PDF)
This book contains information obtained from authentic and highly regarded sources. Reasonable efforts have been
made to publish reliable data and information, but the author and publisher cannot assume responsibility for the valid-
ity of all materials or the consequences of their use. The authors and publishers have attempted to trace the copyright
holders of all material reproduced in this publication and apologize to copyright holders if permission to publish in this
form has not been obtained. If any copyright material has not been acknowledged please write and let us know so we may
rectify in any future reprint.
Except as permitted under U.S. Copyright Law, no part of this book may be reprinted, reproduced, transmitted, or uti-
lized in any form by any electronic, mechanical, or other means, now known or hereafter invented, including photocopy-
ing, microfilming, and recording, or in any information storage or retrieval system, without written permission from the
publishers.
For permission to photocopy or use material electronically from this work, please access www.copyright.com (http://
www.copyright.com/) or contact the Copyright Clearance Center, Inc. (CCC), 222 Rosewood Drive, Danvers, MA 01923,
978-750-8400. CCC is a not-for-profit organization that provides licenses and registration for a variety of users. For
organizations that have been granted a photocopy license by the CCC, a separate system of payment has been arranged.
Trademark Notice: Product or corporate names may be trademarks or registered trademarks, and are used only for
identification and explanation without intent to infringe.
Visit the Taylor & Francis Web site at
http://www.taylorandfrancis.com
and the CRC Press Web site at
http://www.crcpress.com
Contents
Preface....................................................................................................................................................... xi
Editors......................................................................................................................................................xiii
Contributors.............................................................................................................................................. xv
Section I Market Trends, Regulations, Chemistry, and Health Aspects

1. Functional Beverages: Market Trends and Market-Oriented New Product Designs................ 3
Joe Bogue and Amy Jane Troy
2. Global Nutraceutical Regulations for Functional Beverages.......................................................17

Anand Swaroop, Manashi Bagchi, and Debasis Bagchi
3. Flavor Challenges in Functional Beverages.................................................................................. 27

Keith R. Cadwallader
4. Chemistry of Functional Beverages.............................................................................................. 35

Shiming Li, Fereidoon Shahidi, and Chi-Tang Ho
5. Cancer Chemopreventive Effects of Selected Fruit Juices......................................................... 47

Joydeb Kumar Kundu, Kyung-Soo Chun, and Juthika Kundu
6. Fruit Juices and the Prevention of Postprandial Metabolic Stress in Humans........................ 69

Giuseppa Morabito and Mauro Serafini
Section II Fruit Juices

7. Acerola Juice.................................................................................................................................... 85
Delia B. Rodriguez-Amaya
8. Apple Juice....................................................................................................................................... 93
H.P. Vasantha Rupasinghe and Surangi Thilakarathna
9. Apricot Juice/Nectar..................................................................................................................... 107

Emine Aytunga Arık Kibar and Hatice İmge Oktay Başeğmez
10. Aronia Juice....................................................................................................................................119

Maria Glibetić and Aleksandra Konić-Ristić
11. Blackberry Juice.............................................................................................................................135

Mirela Kopjar and Vlasta Piližota
12. Black Currant Juice.......................................................................................................................147

Bradley W. Bolling, Derek A. Martin, Ruisong Pei, Liyang Xie, and Diana M. DiMarco
v
vi Contents
13. Blueberry Juice...............................................................................................................................163

William L. Kerr
14. Cherry Juice...................................................................................................................................175

Gamze Toydemir, Dilek Boyacioglu, Jules Beekwilder, Robert D. Hall, and Esra Capanoglu
15. Cherry Laurel Syrup (Pekmez)....................................................................................................187

16. Coconut Juice................................................................................................................................. 193

Melinda Phang and Manohar Garg
17. Cranberry Juice............................................................................................................................ 205

Monique Lacroix and Khanh Dang Vu
18. Date Syrup......................................................................................................................................217

Sami Fattouch, Karima Dhaouadi, and Manel Belkhir
19. Dragon Fruit Juice.........................................................................................................................231

Lee-Fong Siow
20. Goji Berry Juice............................................................................................................................ 239

Patricia Navarro, Luis Noguera-Artiaga, Santiago López-Miranda,
Ángel A. Carbonell-Barrachina, and Antonio J. Pérez-López
21. Golden Berry and Selected Tropical (Açai, Acerola, and Maqui) Juices.................................251
Coralia Osorio, Maria Elisa Schreckinger, Prerna Bhargava, Woo Young Bang,
Daniel A. Jacobo-Velázquez, and Luis Cisneros-Zevallos
22. Grape Juice.................................................................................................................................... 271

Gian Carlo Tenore
23. Grapefruit Juice............................................................................................................................ 281

İncinur Hasbay
24. Guava Juice.................................................................................................................................... 297

İncinur Hasbay and Emine Aytunga Arık Kibar
25. Hawthorn Juice..............................................................................................................................311

Petras Rimantas Venskutonis
26. Indian Gooseberry (Amla) Juice..................................................................................................321

Neelima Garg and Pushpa Chethan Kumar
27. Kiwifruit Juice................................................................................................................................331

Asim K. Duttaroy
28. Lemon Juice................................................................................................................................... 339

Lucia Maria Jaeger de Carvalho, Lara de Azevedo Sarmet Moreira Smiderle,
Ediane Maria Gomes Ribeiro, and José Luiz Viana de Carvalho
Contents vii
29. Lime Juice...................................................................................................................................... 349

Lucia Maria Jaeger de Carvalho, Gisela Maria Dellamora-Ortiz,
and José Luiz Viana de Carvalho
30. Mango Juice................................................................................................................................... 359

Sui Kiat Chang and Amin Ismail
31. Mangosteen Juice.......................................................................................................................... 373

Mark L. Failla, Fabiola Gutierrez-Orozco, Chureeporn Chitchumroonchokchai,
and Florian Diekmann
32. Melon Juice.................................................................................................................................... 385

Ayse Karadag and Banu Bayram
33. Mulberry Juice.............................................................................................................................. 399

Meltem Türkyılmaz and Mehmet Özkan
34. Noni Fruit Juice............................................................................................................................. 409

Johannes Westendorf
35. Orange Juice.................................................................................................................................. 423

Rita Maria Velázquez-Estrada, José Afid Chávez-Ocegueda,
Ma. Manuela Hernández-Herrero, and Artur Xavier Roig-Sagués
36. Papaya Juice................................................................................................................................... 439

Sui Kiat Chang and Cesarettin Alasalvar
37. Passion Fruit Juice.........................................................................................................................455

Chin-Kun Wang
38. Peach Juice..................................................................................................................................... 463

Emilio Alvarez-Parrilla, Laura A. de la Rosa, Joaquín Rodrigo-García,
Gustavo A. González-Aguilar, and Jesús F. Ayala-Zavala
39. Pear Juice........................................................................................................................................475

Dulcineia Ferreira Wessel, Elisabete Coelho, and Manuel A. Coimbra
40. Pineapple Juice.............................................................................................................................. 489

Nauman Khalid, Hafiz Ansar Rasul Suleria, and Iftikhar Ahmed
41. Plum, Prune, and Ume Juices...................................................................................................... 501

Kent Fanning, Roger Stanley, Bruce Topp, Dougal Russell, and Michael Netzel
42. Pomegranate Juice.........................................................................................................................513

Tao Yuan and Navindra P. Seeram
43. Raspberry Juice............................................................................................................................. 527

Bradley W. Bolling, Diana M. DiMarco, Katherine Lainas, and Sarah Kranz
viii Contents
44. Strawberry Juice............................................................................................................................541

Rong Tsao and Hongyan Li
45. Watermelon Juice...........................................................................................................................553

Beraat Ozcelik and Merve Yavuz
Section III Herbal and Vegetable Juices

46. Carrot Juice................................................................................................................................... 565
Ralf Martin Schweiggert and Reinhold Carle
47. Chinese Medicinal Herbs and Root-Based Beverages............................................................... 583

Chin-Lin Hsu, Chi-Cheng Lu, and Gow-Chin Yen
48. Tomato Juice.................................................................................................................................. 593

María Jesús Periago and Francisco-Javier García-Alonso
49. Vegetable-Containing Juices (Carrot, Kale, and Sprout)......................................................... 609

Daniel A. Jacobo-Velázquez, Erika Ortega-Hernández, and Luis Cisneros-Zevallos
Section IV Caffeinated Beverages: Tea, Coffee, and Cocoa/Chocolate

50. Teas (Green, Oolong, and Black)................................................................................................. 629
Rui Jiao, Jingnan Chen, Yu Huang, and Zhen-Yu Chen
51. Herbal Teas.................................................................................................................................... 645

Sha Li, Shu-Ke Li, Dong-Ping Xu, An-Na Li, and Hua-Bin Li
52. Coffee...............................................................................................................................................661
Iziar A. Ludwig, Michael N. Clifford, Michael E.J. Lean, and Alan Crozier
53. Beverages from Green Coffee Beans........................................................................................... 677

Yuanyuan Ma and Ronald B. Pegg
54. Cocoa and Hot Chocolate............................................................................................................. 687

Beatriz Sarriá, Raquel Mateos, and Laura Bravo
Section V Dairy and Soy Beverages

55. Dairy Beverages............................................................................................................................. 707
Ranjan Sharma
56. Soybean Beverages........................................................................................................................ 725

Tzou-Chi Huang and Chi-Tang Ho
Contents ix
Section VI Alcoholic Beverages and Water

57. Wine................................................................................................................................................ 739
Andrew L. Waterhouse, Rosa M. Lamuela-Raventós, Paola Quifer-Rada,
and Creina S. Stockley
58. Maple Water...................................................................................................................................757

Section VII Fermented and Fortified Functional Beverages

59. Fermented Functional Beverages (Kefir, Koumiss, and Ayran)................................................767
Frank Sherkat, Kambiz Shamsi, and Amir Arjmand
60. Applications of Plant Sterols and Stanols in Functional Beverages......................................... 785

Jerzy Zawistowski
61. Beverages Fortified with Omega-3 Fatty Acids, Dietary Fiber, Minerals, and Vitamins...... 801
Fereidoon Shahidi and Priyatharini Ambigaipalan
62. Probiotics and Prebiotics in Functional Beverages....................................................................815

Koen Venema
63. Functional Beverages in Weight Management........................................................................... 829

Debasis Bagchi, Anand Swaroop, and Manashi Bagchi
64. Fortified Sports Drinks................................................................................................................. 839

Hernan Brice Kenmogne-Domguia and Cesarettin Alasalvar
65. Peptide-Enriched Functional Beverages..................................................................................... 853

Kenji Sato and Tamami Kiyono
Index........................................................................................................................................................861
Preface
The market for functional beverages represents the largest and fastest growing segment of the functional
foods sector, with an annual growth rate of almost 20% in the United States. The production and consump-
tion of functional beverages has gained much importance due to their major contribution to health pro-
motion and disease risk reduction. They constitute an excellent delivery means for nutrients and bioactive
compounds, including vitamins, minerals, antioxidants, omega-3 fatty acids, plant extracts, sterols/stanols,
dietary fiber, amino acids and biopeptides, prebiotics, and probiotics, among others. There have been con-
tinuous innovations in functional beverages and their associated market over the last decade as consumers
seek novelty and health benefits from their beverages. The market for new functional beverages with added
bioactive ingredients with health benefits has grown rapidly with positioning strategies linked to energy, ath-
letic performance, digestion, aging, satiety, cognitive ability, hydration, weight management, cardiovascular
health, cancer, diabetes, bone and joint health, and fatigue and stamina, among others.
This handbook consists of 65 chapters divided into 7 sections. Section I includes six chapters on
market trends, global regulations, flavor challenges, chemistry, health with specific reference to cancer
chemoprevention, and the prevention of postprandial metabolic stress due to consumption of functional
beverages. Section II, by far the largest part of the book, has 39 chapters on the most popular fruit
juices (acerola juice, apple juice, apricot juice/nectar, aronia juice, blackberry juice, black currant juice,
blueberry juice, cherry juice, cherry laurel syrup [pekmez], coconut juice, cranberry juice, date syrup,
dragon fruit juice, goji berry juice, golden berry and selected tropical juices [açai, acerola, and maqui],
grape juice, grapefruit juice, guava juice, hawthorn juice, Indian gooseberry [amla] juice, kiwifruit juice,
lemon juice, lime juice, mango juice, mangosteen juice, melon juice, mulberry juice, noni fruit juice,
orange juice, papaya juice, passion fruit juice, peach juice, pear juice, pineapple juice, plum, prune, and
ume juices, pomegranate juice, raspberry juice, strawberry juice, and watermelon juice). Section III
reports on herbal and vegetable juices (carrot juice, Chinese medicinal herbs and root-based beverages,
tomato juice, and vegetable-containing juices [carrot, kale, and sprout]). Section IV details caffeinated
beverages, including different varieties of tea (green, black, oolong, and herbal teas), coffee (coffee
and beverages from green coffee beans), and cocoa and chocolate. Section V is on dairy and soy bever-
ages, while Section VI is on alcoholic beverage (wine) and water (maple water). Finally, Section VII
describes fermented (kefir, koumiss, and ayran) and fortified functional beverages (applications of plant
sterols and stanols in functional beverages, beverages fortified with omega-3 fatty acids, dietary fiber,
minerals, vitamins, probiotics, and prebiotics in functional beverages, functional beverages in weight
management, fortified sports drinks, and peptide-enriched functional beverages).
We are most grateful to the contributors to this handbook, who are internationally renowned research-
ers, for their comprehensive account of the global perspective on the issues of concern related to nutri-
tional characteristics, bioactive and antioxidant efficacy, phytochemicals, and health effects of beverages.
The book will serve as a major resource for those interested in the potential applications and new devel-
opments in functional beverages, nutraceuticals, and health foods. Biochemists, chemists, food scien-
tists/technologists, nutritionists, and health professionals from academia, government laboratories, and
beverage industries will find the contents of this handbook of much interest. Although this book serves
primarily as a reference manual, it also summarizes the current state of knowledge in key research areas
and contains novel ideas for future research and development. In addition, it provides easy-to-read text
suitable for teaching senior undergraduate and postgraduate students in the relevant areas. Finally, we
trust that this handbook paves the way for better appreciation of the concepts, products, and opportuni-
ties in the field for professionals, regulators, processors, and consumers.
Fereidoon Shahidi
xi
Editors
Fereidoon Shahidi, Ph.D., FACS, FAGFD-ACS, FAOCS, FCIC, FCIFST, FIAoFST, FIFT, FRSC,
is a university research professor, the highest rank the university gives for research, in the Department
of Biochemistry at the Memorial University of Newfoundland (MUN) in Canada. He is cross-appointed
to the Department of Biology, the Department of Ocean Sciences, and the Aquaculture Program. He is a
chair professor at National Chung Hsing University in Taiwan, an honorary professor at the Chung Shan
Medical University, also in Taiwan, a visiting professor at Jiangnan University, and Dalian Polytechnic
University in P.R. China. He collaborates with many other universities in countries such as Brazil,
France, Korea, Japan, Poland, Thailand, Turkey, the United States, and elsewhere around the globe.
He is also an advisor to the Chinese Academy of Agricultural Sciences for special projects on cereals
and oilseeds. Dr. Shahidi has made numerous outstanding and innovative quality contributions to both
the basic and applied areas of food and nutraceutical science and technology and antioxidant phenolics
and omega-3 oils in health and disease. He is the only Canadian on the ISI list of top 10 (3rd to 8th)
most highly cited scientists in agricultural sciences, first recognized as the most highly cited (top 15)
individual and the most productive scientist in the area of food, nutrition, and agricultural science for
the 1991–2001 period and 3rd in citations for 2001–2011, and is now in 6th place. He has received
numerous awards from different societies and organizations for his pioneering scientific achievements.
Dr. Shahidi’s work has led to the publication of more than 760 research articles in the form of peer-
reviewed journals and book chapters. He is also the editor/author of some 64 books and holds 10 patents.
These publications, along with his extensive list of presentations, have led to the advancement of the
discipline of food science at both the national and international levels. Dr. Shahidi has trained more
than 100 Ph.D. and M.Sc. students and research assistants/associates, postdoctoral fellows, and visiting
professors and scholars, and has educated the future generation of scientists. His former students, now
his colleagues, occupy key positions as faculty members, government workers, and industry leaders in
more than a dozen countries on five continents.
Cesarettin Alasalvar, Ph.D., FIFT, is the director of the Food Institute at TÜBİTAK Marmara
Research Centre (MRC) in Turkey and is also an associate professor of food science and engineering.
He received his Ph.D. in food science and technology in 1994 from the University of Lincoln (United
Kingdom) and conducted postdoctoral research at the same university (1995–1997). Dr. Alasalvar is
a recipient of a fellowship award from the Japanese Science and Technology Agency (1997–1998).
He was then appointed as a senior research fellow/lecturer both at the Food Research Centre and the
Department of Food Science and Technology at the University of Lincoln (1998–2005). He has been
working at different positions (chief research scientist, deputy director, and director) at the Food Institute
of TÜBİTAK MRC since 2006. Dr. Alasalvar is a leading international researcher in bioactive compo-
nents from marine resources and plant materials, especially hazelnuts. He is recognized for his impact
in identifying bioactives and phytochemicals present in foods and plant-based products. He has coedited
5 books, published more than 60 scientific articles in peer-reviewed journals and 25 book chapters,
given more than 100 presentations at different international scientific conferences, and holds a patent.
He has delivered invited lectures, served as a session chairperson and poster-award chair for various
international congresses, and has organized international congresses, seminars, and brokerage events.
Dr. Alasalvar has been active in the Institute of Food Technologists (IFT) programs for many years
and has played a leadership role in the Nutraceuticals and Functional Foods Division. He served as a
past chair of the division and serves as an editorial board member of Food Chemistry and the Journal
of Functional Foods. He also served as local chair of the International Society for Nutraceuticals and
Functional Foods (ISNFF) 2014 Annual Conference and Exhibition and is currently the chair-elect of
xiii
xiv Editors
ISNFF (2014–2016). Dr. Alasalvar serves on the expert advisory board of the Turkish Goverment and
Higher Education on R&D projects and as a panelist for projects funded by the European Union (EU). He
coordinates two major EU-funded projects, EU-FP7 (NutraHEALTH) and EU-IPA (INNOFOOD), and
has received a number of international prestigious awards, including the IFT-Fellow Award (2012), the
TÜBİTAK MRC–Most Successful Researcher Award (2012), the ISNFF Merit Award for Outstanding
Contributions to the Nutraceuticals and Functional Foods Discipline and Service to the ISNFF (2014),
and the Sabri Ülker International Science Award on Public Health and Nutrition (2015) in recognition of
his pioneering scientific achievements.
Contributors
Iftikhar Ahmed Manashi Bagchi

National Institute for Genomics and Advanced Departments of Pharmacology and Toxicology
Biotechnology Cepham Research Center
National Agricultural Research Centre Piscataway, New Jersey
Islamabad, Pakistan
Woo Young Bang
Cesarettin Alasalvar National Institute of Biological Resources
Food Institute Environmental Research Complex
TÜBİTAK Marmara Research Center Incheon, South Korea
Gebze-Kocaeli, Turkey
Hatice İmge Oktay Başeğmez
Emilio Alvarez-Parrilla Food Institute
Department of Chemical and Biological Sciences TÜBİTAK Marmara Research Center
Autonomous University of Ciudad Juarez Gebze-Kocaeli, Turkey
Ciudad Juarez, Mexico
Banu Bayram
Food Institute
Priyatharini Ambigaipalan
TÜBİTAK Marmara Research Center
Department of Biochemistry
Gebze-Kocaeli, Turkey
St. John’s, Newfoundland and Labrador, Canada
Jules Beekwilder
BU Bioscience
Amir Arjmand Plant Research International
School of Science Wageningen University and Research Centre
RMIT University Wageningen, The Netherlands
Melbourne, Victoria, Australia
Manel Belkhir
Jesús F. Ayala-Zavala Faculty of Sciences of Tunis
Department of Technology of Food of Plant University El Manar
Origin Tunis, Tunisia
Research Center for Food & Development
(CIAD) Prerna Bhargava
Hermosillo, Mexico Department of Horticultural Sciences
Texas A&M University
Debasis Bagchi College Station, Texas
Departments of Pharmacology and Toxicology
Cepham Research Center Joe Bogue
Piscataway, New Jersey Department of Food Business and Development
University College Cork
and
Cork, Ireland
Department of Pharmacological and
Pharmaceutical Sciences Bradley W. Bolling
College of Pharmacy Department of Food Science
University of Houston University of Wisconsin–Madison
Houston, Texas Madison, Wisconsin
xv
xvi Contributors
Dilek Boyacioglu Jingnan Chen

Department of Food Engineering School of Food Science and Technology
Istanbul Technical University Henan University of Technology
Istanbul, Turkey Zhengzhou, Henan, People’s Republic of China
Laura Bravo Zhen-Yu Chen

Department of Metabolism and Nutrition School of Life Sciences
Institute of Food Science, Technology, and The Chinese University of Hong Kong
Nutrition (ICTAN–CSIC) Shatin, Hong Kong, People’s Republic of China
Madrid, Spain
Chureeporn Chitchumroonchokchai
Keith R. Cadwallader Department of Human Sciences
Department of Food Science and Human The Ohio State University
Nutrition Columbus, Ohio
University of Illinois at Urbana–Champaign
Urbana, Illinois Kyung-Soo Chun
College of Pharmacy
Keimyung University
Esra Capanoglu
Daegu, South Korea
Department of Food Engineering
Istanbul Technical University
Luis Cisneros-Zevallos
Istanbul, Turkey
Department of Horticultural Sciences
Texas A&M University
Ángel A. Carbonell-Barrachina
College Station, Texas
Department of Agro-Food Technology
Miguel Hernández University
Michael N. Clifford
Alicante, Spain
School of Biosciences and Medicine
University of Surrey
Reinhold Carle
Guildford, United Kingdom
Institute of Food Science and Biotechnology
University of Hohenheim
Stuttgart, Germany Elisabete Coelho
Department of Chemistry
José Luiz Viana de Carvalho University of Aveiro
Embrapa Food Technology Aveiro, Portugal
Rio de Janeiro, Brazil
Manuel A. Coimbra
Lucia Maria Jaeger de Carvalho Department of Chemistry
Department of Natural Products and Food University of Aveiro
School of Pharmacy Aveiro, Portugal
Federal University of Rio de Janeiro
Alan Crozier
Sui Kiat Chang Department of Nutrition
Department of Nutrition and Dietetics University of California–Davis
Universiti Putra Malaysia Davis, California
Selangor, Malaysia
Laura A. de la Rosa
José Afid Chávez-Ocegueda Department of Chemical and Biological
Integral Laboratory of Food Research Sciences
Tepic Institute of Technology Autonomous University of Ciudad Juarez
Tepic, Mexico Ciudad Juarez, Mexico
Contributors xvii
Gisela Maria Dellamora-Ortiz Francisco-Javier García-Alonso

Department of Natural Products and Food Department of Food Science and Nutrition
School of Pharmacy Catholic University of Murcia
Federal University of Rio de Janeiro Murcia, Spain
Manohar Garg
School of Biomedical Sciences and Pharmacy
Karima Dhaouadi The University of Newcastle
National Institute of Applied Sciences and Callaghan, New South Wales, Australia
Technology
University of Carthage
Tunis, Tunisia Neelima Garg
Department of Post Harvest Management
ICAR–Central Institute for Subtropical
Florian Diekmann Horticulture
Food, Agricultural, and Environmental Sciences Lucknow, India
Library
The Ohio State University Maria Glibetić
Columbus, Ohio Centre of Research Excellence in Nutrition and
Metabolism
Institute for Medical Research
Diana M. DiMarco University of Belgrade
Department of Nutritional Sciences Belgrade, Serbia
University of Connecticut
Storrs, Connecticut Gustavo A. González-Aguilar
Department of Food Technology of Plant Origin
Research Center for Food and Development
Asim K. Duttaroy
(CIAD)
Department of Nutrition
Hermosillo, Mexico
Faculty of Medicine
University of Oslo
Oslo, Norway Fabiola Gutierrez-Orozco
Interdisciplinary Ph.D. Program in Nutrition
The Ohio State University
Mark L. Failla Columbus, Ohio
Department of Human Sciences
and Robert D. Hall
Interdisciplinary Ph.D. Program in Nutrition Department of Bioscience
The Ohio State University Plant Research International
Columbus, Ohio Wageningen University and Research Centre
Wageningen, The Netherlands
Kent Fanning and
Department of Agriculture and Fisheries
Agri-Science Queensland Laboratory of Plant Physiology
Coopers Plains, Queensland, Australia Wageningen University
Wageningen, The Netherlands
Sami Fattouch
National Institute of Applied Sciences and İncinur Hasbay
Technology Food Institute
University of Carthage TÜBİTAK Marmara Research Center
Tunis, Tunisia Gebze-Kocaeli, Turkey
xviii Contributors
Ma. Manuela Hernández-Herrero Ayse Karadag

Special Research Centre–Food Technology Plant Food Institute
(CERPTA) TÜBİTAK Marmara Research Center
XaRTA Gebze-Kocaeli, Turkey
Tecnio, Malta
and Hernan Brice Kenmogne-Domguia
Department of Animal and Food Science Food Institute
School of Veterinary Medicine TÜBİTAK Marmara Research Center
Autonomous University of Barcelona Gebze-Kocaeli, Turkey
Barcelona, Spain
Chi-Tang Ho William L. Kerr

Department of Food Science Department of Food Science and Technology
Rutgers University The University of Georgia
New Brunswick, New Jersey Athens, Georgia
Chin-Lin Hsu
Nauman Khalid
School of Nutrition
Graduate School of Agricultural and Life
Chung Shan Medical University
Sciences
and
The University of Tokyo
Tokyo, Japan
Chung Shan Medical University Hospital
Taichung, Taiwan, People’s Republic of China
Emine Aytunga Arık Kibar
Tzou-Chi Huang Food Institute
Department of Biological Sciences and TÜBİTAK Marmara Research Center
Technology Gebze-Kocaeli, Turkey
National Pingtung University of Science and
Technology Tamami Kiyono
Pingtung, Taiwan, People’s Republic of China Division of Applied Life Sciences
Graduate School of Life and Environmental
Yu Huang Sciences
School of Biomedical Sciences Kyoto Prefectural University
The Chinese University of Hong Kong Kyoto, Japan
Shatin, Hong Kong, People’s Republic of China
Amin Ismail Aleksandra Konić-Ristić

Department of Nutrition and Dietetics Centre of Research Excellence in Nutrition and
and Metabolism
Halal Products Research Institute Institute for Medical Research
Universiti Putra Malaysia University of Belgrade
Selangor, Malaysia Belgrade, Serbia
Daniel A. Jacobo-Velázquez
Mirela Kopjar
Biotechnology Center–FEMSA Monterrey
Department of Food Technologies
Institute of Technology
Josip Juraj Strossmayer University of Osijek
Monterrey, Mexico
Osijek, Croatia
Rui Jiao
Department of Food Science and Engineering Sarah Kranz
Jinan University Department of Nutritional Sciences
Guangzhou, Guangdong, People’s Republic University of Connecticut
of China Storrs, Connecticut
Contributors xix
Pushpa Chethan Kumar Hongyan Li

Department of Post Harvest Management State Key Lab of Food Science and Technology
ICAR–Central Institute for Subtropical Institute for Advanced Study
Horticulture Nanchang University
Lucknow, India Jiangxi, Xinjian, People’s Republic of China
Joydeb Kumar Kundu Hua-Bin Li

College of Pharmacy Department of Nutrition
Keimyung University School of Public Health
Daegu, South Korea Sun Yat-Sen University
Guangzhou, Guangdong, People’s Republic
Juthika Kundu of China
College of Pharmacy
Keimyung University Sha Li
Daegu, South Korea Department of Nutrition
School of Public Health
Monique Lacroix Sun Yat-Sen University
Research Laboratories in Sciences Applied Guangzhou, Guangdong, People’s Republic
to Food of China
Canadian Irradiation Centre
Institut National de la Recherche Scientifique–
Shiming Li
Institut Armand-Frappier
College of Life Sciences
Université du Québec
Huanggang Normal University
Laval, Québec, Canada
Huanggang, Hubei, People’s Republic of China
Katherine Lainas and
Department of Nutritional Sciences
Department of Food Science
University of Connecticut
Rutgers University
Storrs, Connecticut
New Brunswick, New Jersey
Rosa M. Lamuela-Raventós
Department of Nutrition and Food Science Shu-Ke Li
School of Pharmacy Department of Nutrition
University of Barcelona School of Public Health
Barcelona, Spain Sun Yat-Sen University
Guangzhou, Guangdong, People’s Republic
and
of China
The Spanish Biomedical Research Centre
Physiopathology of Obesity and Nutrition Santiago López-Miranda
Health Institute of Carlos III Department of Food Science and Nutrition
Madrid, Spain Catholic University of Murcia
Murcia, Spain
Michael E.J. Lean
College of Medical, Veterinary, and Life Sciences
University of Glasgow Chi-Cheng Lu
Glasgow, United Kingdom Department of Food Science and Biotechnology
National Chung Hsing University
An-Na Li Taichung, Taiwan, People’s Republic of China
School of Public Health Iziar A. Ludwig
Sun Yat-Sen University Department of Food Technology
Guangzhou, Guangdong, People’s Republic University of Lleida
of China Lleida, Spain
xx Contributors
Yuanyuan Ma Mehmet Özkan

Department of Food Science and Technology Department of Food Engineering
The University of Georgia Ankara University
Athens, Georgia Ankara, Turkey
Derek A. Martin Ronald B. Pegg

Department of Food Science Department of Food Science and Technology
University of Wisconsin–Madison The University of Georgia
Madison, Wisconsin Athens, Georgia
Raquel Mateos Ruisong Pei

Department of Metabolism and Nutrition Department of Food Science
Institute of Food Science, Technology, and University of Wisconsin–Madison
Nutrition (ICTAN–CSIC) Madison, Wisconsin
Madrid, Spain
Antonio J. Pérez-López
Giuseppa Morabito Department of Food Science and Nutrition
Functional Food and Metabolic Stress Prevention Catholic University of Murcia
Laboratory Murcia, Spain
Center for Food and Nutrition, CREA
Rome, Italy María Jesús Periago
Department of Food Science and Nutrition
Patricia Navarro Catholic University of Murcia
Department of Food Science and Nutrition Murcia, Spain
Catholic University of Murcia
Murcia, Spain Melinda Phang
Women’s and Children’s Health Research
Michael Netzel Institute
Centre for Nutrition and Food Sciences Women’s and Children’s Hospital
Queensland Alliance for Agriculture and Food North Adelaide, South Australia, Australia
Innovation
The University of Queensland Vlasta Piližota
Brisbane, Queensland, Australia Department of Food Technologies
Josip Juraj Strossmayer University of Osijek
Luis Noguera-Artiaga Osijek, Croatia
Department of Agro-Food Technology
Miguel Hernández University Paola Quifer-Rada
Alicante, Spain Department of Nutrition and Food Science
School of Pharmacy
Erika Ortega-Hernández University of Barcelona
Biotechnology Center–FEMSA Monterrey Barcelona, Spain
Institute of Technology
Monterrey, Mexico and
The Spanish Biomedical Research Centre
Coralia Osorio Physiopathology of Obesity and Nutrition
Department of Chemistry Health Institute of Carlos III
National University of Colombia Madrid, Spain
Bogotá, Colombia
Ediane Maria Gomes Ribeiro
Beraat Ozcelik Department of Natural Products and Food
Department of Food Engineering School of Pharmacy
Istanbul Technical University Federal University of Rio de Janeiro
Istanbul, Turkey Rio de Janeiro, Brazil
Contributors xxi
Joaquín Rodrigo-García Navindra P. Seeram

Department of Chemical and Biological Sciences Bioactive Botanical Research Laboratory
Autonomous University of Ciudad Juarez Department of Biomedical and Pharmaceutical
Ciudad Juarez, Mexico Sciences
University of Rhode Island
Delia B. Rodriguez-Amaya Kingston, Rhode Island
Federal University of the Southern Frontier
Laranjeiras do Sul Campus
Parana, Brazil Mauro Serafini
Functional Food and Metabolic Stress Prevention
Artur Xavier Roig-Sagués Laboratory
Special Research Centre–Food Technology Plant Center for Food and Nutrition, CREA
(CERPTA) Rome, Italy
XaRTA
Tecnio, Malta
Fereidoon Shahidi
and Department of Biochemistry
Department of Animal and Food Science
St. John’s, Newfoundland and Labrador, Canada
School of Veterinary Medicine
Autonomous University of Barcelona
Barcelona, Spain Kambiz Shamsi
School of Science
H.P. Vasantha Rupasinghe RMIT University
Department of Environmental Sciences Melbourne, Victoria, Australia
Dalhousie University
Truro, Nova Scotia, Canada
Ranjan Sharma
Dougal Russell Dairy Innovation Australia Limited
Department of Agriculture and Fisheries Werribee, Victoria, Australia
Agri-Science Queensland
Nambour, Queensland, Australia
Frank Sherkat
Beatriz Sarriá School of Science
Department of Metabolism and Nutrition RMIT University
Institute of Food Science, Technology, and Melbourne, Victoria, Australia
Nutrition (ICTAN–CSIC)
Madrid, Spain Lee-Fong Siow
School of Science
Kenji Sato Monash University Malaysia
Division of Applied Biosciences Selangor, Malaysia
Graduate School of Agriculture
Kyoto University
Kyoto, Japan Lara de Azevedo Sarmet Moreira Smiderle
Department of Natural Products and Food
Maria Elisa Schreckinger School of Pharmacy
Department of Horticultural Sciences Federal University of Rio de Janeiro
Texas A&M University Rio de Janeiro, Brazil
College Station, Texas
Ralf Martin Schweiggert Roger Stanley

Institute of Food Science and Biotechnology Centre for Food Innovation
University of Hohenheim University of Tasmania
Stuttgart, Germany Launceston, Tasmania, Australia
xxii Contributors
Creina S. Stockley Koen Venema

The Australian Wine Research Institute Beneficial Microbes Consultancy
Urrbrae, South Australia, Australia Wageningen, The Netherlands
Hafiz Ansar Rasul Suleria and

School of Agriculture and Food Sciences Department of Human Biology
The University of Queensland Maastricht University
Brisbane, Queensland, Australia Venlo, The Netherlands
Anand Swaroop Petras Rimantas Venskutonis
Departments of Pharmacology and Toxicology Department of Food Science and Technology
Cepham Research Center Kaunas University of Technology
Piscataway, New Jersey Kaunas, Lithuania
Gian Carlo Tenore Khanh Dang Vu
Department of Pharmacy Research Laboratories in Sciences Applied
University of Naples “Federico II” to Food
Naples, Italy Canadian Irradiation Centre
Institut National de la Recherche Scientifique–
Surangi Thilakarathna
Institut Armand–Frappier
Department of Environmental Sciences
Université du Québec
Dalhousie University
Laval, Québec, Canada
Truro, Nova Scotia, Canada
Chin-Kun Wang
Bruce Topp School of Nutrition
Centre for Plant Science Chung Shan Medical University
Queensland Alliance for Agriculture and Food Taichung, Taiwan, People’s Republic of China
Innovation
The University of Queensland Andrew L. Waterhouse
Nambour, Queensland, Australia Department of Viticulture and Enology
University of California–Davis
Gamze Toydemir Davis, California
Department of Food Engineering
Okan University Dulcineia Ferreira Wessel
Istanbul, Turkey Department of Food Industry
Agrarian School
Amy Jane Troy Polytechnic Institute of Viseu
Department of Food Business and Development Viseu, Portugal
University College Cork Johannes Westendorf
Cork, Ireland Institute of Experimental Pharmacology and
Toxicology
Rong Tsao
University Clinic Hamburg–Eppendorf
Guelph Food Research Centre
Hamburg, Germany
Agriculture and Agri-Food Canada
Guelph, Ontario, Canada Liyang Xie
Department of Nutritional Sciences
Meltem Türkyılmaz University of Connecticut
Institute of Food Safety Storrs, Connecticut
Ankara University
Ankara, Turkey Dong-Ping Xu
Rita María Velázquez-Estrada School of Public Health
Integral Laboratory of Food Research Sun Yat-Sen University
Tepic Institute of Technology Guangzhou, Guangdong, People’s Republic
Tepic, Mexico of China
Contributors xxiii
Merve Yavuz Tao Yuan

Department of Food Engineering Bioactive Botanical Research Laboratory
Istanbul Technical University Department of Biomedical and Pharmaceutical
Istanbul, Turkey Sciences
University of Rhode Island
Kingston, Rhode Island
Gow-Chin Yen Jerzy Zawistowski
Department of Food Science and Biotechnology Department of Food, Nutrition, and Health
National Chung Hsing University The University of British Columbia
Taichung, Taiwan, People’s Republic of China Vancouver, British Columbia, Canada
Section I
Market Trends, Regulations,

Chemistry, and Health Aspects
1
Functional Beverages:
Market Trends and Market-Oriented
New Product Designs
Joe Bogue and Amy Jane Troy
CONTENTS
1.1 Introduction....................................................................................................................................... 3
1.2 Global Functional Beverage Market................................................................................................. 4
1.3 Consumer-Oriented New Product Development Case Study: Functional Beverages....................... 5
1.4 Semiotic Approach to Market-Oriented Product Concept Optimization......................................... 5
1.5 Consumer Insights on Functional Beverages.................................................................................... 6
1.6 Semiotics and Functional Beverages................................................................................................ 7
1.6.1 Stage 1: The Semiotic Sort................................................................................................... 8
1.6.2 Stage 2: Interpretation of Package Signs by Respondents................................................... 8
1.7 Semiotic Results................................................................................................................................ 9
1.7.1 Stage 1: Results..................................................................................................................... 9
1.7.2 Stage 2: Results....................................................................................................................11
1.8 Preferences and Purchase Habits toward Functional Beverages.....................................................11
1.9 Semiotics and New Product Development...................................................................................... 12
1.10 Lessons from the Case Study.......................................................................................................... 13
1.11 Conclusion........................................................................................................................................14
References..................................................................................................................................................14
1.1 Introduction
There have been many changes and innovations in the beverage market over the past years as consum-
ers seek new benefits from their beverages. One of the most important benefits sought by consumers
is health and wellness. The market for new functional beverages with added ingredients and related
health benefits has grown rapidly with positioning strategies linked to energy, digestion, aging, satiety,
cognitive ability, hydration, weight management, and fatigue, among others. While the opportunity for
developing functional beverages is high, manufacturers often struggle to achieve market success, and
the challenges new functional beverage developers face include the technological challenge of develop-
ing and marketing new products with new ingredients; differentiating brands in ultracompetitive mar-
kets; identifying the most appropriate positioning platforms of convenience; and marketing science and
technology to consumers, as well as health, natural, and legal obstacles. The high failure rates suggest
an inability to understand consumer preferences and choice motives in relation to the purchase of func-
tional beverages.
This chapter examines the main trends in the functional beverage market and identifies the key issues
related to the consumer and the functional beverage market. Following this, a case study is introduced
that focuses on developing market-oriented functional beverages based on consumer insights. It exam-
ines the importance of market-oriented approaches in developing new functional beverages and views
3
4 Handbook of Functional Beverages and Human Health
how firms can use the voice of the consumer information to design products that more closely meet con-
sumer needs. The case study examines utilizing qualitative research techniques to generate information
at the early stages of the new product development (NPD) process. In particular, it looks at using semiot-
ics to generate information on beverage packaging as it can greatly influence consumers’ first purchase
of a new product and repeat purchases.
1.2 Global Functional Beverage Market

By 2017, the global market for beverages will be worth approximately US$1347 billion. Market growth
drivers include urbanization, expansion in the middle-class population, and an increase in double income
families [1]. Moreover, there are also significant opportunities for functional food and beverages in
emerging markets, such as in China, Brazil, Asia Pacific, and Latin America, where companies can
successfully innovate by observing closely studied trends, understanding local consumers, and differen-
tiating their products from their competitors [2]. Global trends that will impact the development of new
functional food and beverages include age complexity, gender complexity, life stage complexity, income
complexity, convenience, health, sensory, comfort, and individualism [3].
The global retail value of health and wellness beverages reached US$274 billion in 2011. This was
approximately 44% of the retail value sales of nonalcoholic beverages [4]. The growth of health and
wellness beverages was set to outperform the wider soft and hot beverage industry over the period 2012–
2016 [5]. In addition, concerns continue to grow around the link between carbonated beverages and
specific health issues such as obesity, diabetes, and coronary heart disease [6,7]. Global market trends
indicate that demand for health beverages, specifically the low or no calories ones, has shown signs of
decline, while there is an increased demand for beverages that help address health conditions by includ-
ing natural ingredients and those focused on specific health benefits, such as digestive or heart health
[4,8]. The inclusion of functional foods and beverages into consumers’ diets can also be viewed as part of
future public health prevention strategies that aim at reducing expenditure on health care [9].
Often, consumers associated low-calorie beverages with artificial ingredients, and this finding created
the opportunity for functional beverages to be considered as naturally healthy [10]. In addition, 48% of
consumers believed a natural product was one that was produced in adherence with strict regulations,
while 46% of consumers aligned natural products closely with organic products [5]. Consumers believed
ingredients were key to the concept of a natural beverage with beverages labeled as 100% juice, all natu-
ral, and containing no artificial colors, perceived as healthier beverages.
Drivers of innovation in the beverage industry over the period 2012–2017 include new functional ingre-
dients, advances in technology in relation to taste masking and encapsulation, reduction of sugar content,
and wider channels of distribution [11]. Specifically, in relation to future functional beverage innovation,
the focus will be on beverages targeted at specific parts of the body such as bones, joints, eyes, as well as
sleep improvement, weight management, cholesterol management, maintenance of healthy teeth, energy,
the elderly market segment, and beverages with omega-3 fatty acids [12]. This innovation focus will lead
to market opportunities across demographic groups, as beverages are developed that are tailored more
and more to the rapidly changing needs of individuals.
It has been suggested that beverage developers should follow a product development strategy that
combines health, convenience, modern packaging, and affordable prices [5]. Firms can use differ-
ent strategies, such as using natural ingredients, or novel ingredients, or adding natural and/or low-
calorie sweeteners to beverages [13]. Alternatively, they can incorporate ingredients into beverages such
as omega-3 oils, fiber, or probiotics. However, products should be built on what consumers believe is
inherent nutritional functionality, and foods or beverages should be developed based on what is cultur-
ally relevant as a delivery medium [14]. In addition, consumers in developed regions made the switch
to low-calorie soft beverages a number of years ago, and as these markets had entered into maturity,
consumers were actively seeking healthier soft beverages [5]. This offers opportunities to develop added
value through functionality, in the form of energy beverages and those with associated health claims.
Increased use of vitamins, in the energy beverage sector, indicated the shift in consumer attitudes toward
products that had more natural appeal [15].
Functional Beverages 5
Consumption of superfruit juice is also on the rise globally, and with increasing consumer awareness,
there are opportunities to appeal to more mass-market consumers [5]. China leads in the consumption of
superfruit juice at 1587 million liters per annum, followed by the United States and Japan. As superfruit
juices move to a wider range of beverage categories, more novel flavors and ingredients are likely to be
available to consumers. Blueberry, pomegranate, and aloe vera are consumer favorites worldwide, but
more unusual varieties will be used in future beverage manufacture such as acai berry, baobab, mango-
steen, goji berry, and sea buckthorn [5].
A key part of developing novel beverages with new ingredients will be consumer acceptance of new
ingredients, their knowledge of the potential health benefits of such ingredients, and how the specific
science and technology is marketed to consumers. In addition, consumers’ interest in health is related
to what health benefits are relevant to them and their lifestyles and, thus, are highly individualized [16].
This suggests that beverage developers need to generate deep insights into consumers’ perceptions and
how these beverages fit in with their lifestyles.
1.3 Consumer-Oriented New Product Development

Case Study: Functional Beverages
The success of food and beverage firms is dependent on their ability to develop and market new products
that provide consumers with superior value to that of their competitors. However, the driver of innovation
within the functional food sector is often based on R&D within the food firm, rather than the consumer.
This provides a science push rather than a consumer pull focus to the innovation and has often been
attributed to failures within the functional beverage sector, where new products frequently do not meet
consumer needs or expectations [17]. Reasons for functional food failures, which are relevant to func-
tional beverages, include too many benefits from a single brand, benefits that are often not relevant to the
consumer, relying on the selling power of the ingredient rather than the benefit, and using a nonrelevant
carrier [16]. For example, many omega-3 fatty acid–fortified products have had little impact on the global
market as they provide benefits that consumers cannot quickly see or feel [16].
There is a strong positive relationship between market orientation and the NPD activities of firms
[18,19]. Market-oriented NPD entails generating information on consumers’ needs and choice motives,
integrating this information with the early stages of the NPD process, and developing an optimal prod-
uct with attributes that maximize consumer acceptance. Information gathered for functional beverage
development may include consumer perceptions of functional beverages; identifying consumer segments
that are functionally driven, for example, consumers that will pay a premium for beverages with added
functional benefits; understanding consumer knowledge of the health benefits associated with functional
ingredients and health claims; identifying the key intrinsic (texture, mouth feel, and flavor) and extrinsic
(packaging, brand, and health claim) product attributes that will influence consumer acceptance; and
how consumers interpret the message (through images, colors, and icons) communicated by functional
beverage packaging and the type of delivery method (shots, stick packs, and ready to drink).
Generation of consumer information is an important part of the NPD process that helps identify new
ideas, defines the target market more explicitly, and then aids in the design of specific marketing strate-
gies that position new products on markets [20]. Qualitative techniques are important for information
generation at the early stages of the NPD process, and many studies acknowledge that both in-depth
interviews and focus groups are particularly good at exploring concepts, generating ideas, and eliciting
opinions on packaging [19,21–23]. They facilitate the integration of consumer insights at the earliest
stages of the NPD process, and thus consumers act as codesigners in NPD.
1.4 Semiotic Approach to Market-Oriented Product Concept Optimization

Differentiation of a product or brand is seen as one of the key factors for developing successful func-
tional foods. One of the ways of achieving this differentiation is through packaging design [16].
Semiotics can play an important role in the process of designing differentiated product packaging for
functional beverages. Semiotics is an abductive method of analyzing meanings by examining signs that

communicate information [24,25].
Research estimates that approximately 70% of purchase decisions are usually made at point of sale,
and that for approximately 40 weeks of the year, packaging is the main attribute that leads to the sale
of a product [26,27]. For low-involvement purchases, the package is the product, particularly because
impressions formed during initial contact can have lasting effects. Beyond providing pure information,
the emotional aspects of packaging graphics are more subliminal, which evolve from the styling of
various graphical elements, including logo styling, symbols, icons, colors, textures, photography, and
illustrations [28].
Semiotics can be used to understand the important role of packaging design in motivating consumer
purchase of functional beverages. This entails an analysis of consumer perceptions of all icons, colors,
and images on the package and how these combined might influence the purchase decision. Good pack-
aging can support a brand in highlighting its difference in the marketplace [16].
In the following case study, some key activities at the early design stages of the NPD process are out-
lined in terms of the use of qualitative techniques to understand consumer requirements for functional
beverages.
1.5 Consumer Insights on Functional Beverages

In this first step of developing new functional beverage concepts, a total of 12 in-depth interviews and
3 focus groups were conducted to generate consumer information on functional beverages. A convenient
sampling technique was used to select participants for the in-depth interviews and focus groups [29].
Majority of the samples were middle-aged, educated to third level, in full-time employment, and lived
in Cork City, Ireland. Both interview and focus group guides consisted of semistructured questions, and
participants were rewarded with €30 for their time and effort in line with best practice [30]. Results were
tape-recorded and the data were analyzed using the QSR N6 software package [31].
Focus group participants felt that the concept of a functional beverage was relatively new to the market
and were positive toward the product idea. Five functional beverage concepts were generated from the
participants. Most participants had a positive attitude toward a symbiotic beverage that consisted of the
combined benefit of fiber and a probiotic culture in a functional beverage. Focus Group 2, which con-
sisted of younger females, was very interested in the health benefits of such beverages. These consumers
were familiar with the fermentation process, which they felt was traditional and were positive toward the
concept of fermented functional beverages. They were also familiar with many other fermented food and
drink products that they consumed on a daily basis. However, there were mixed attitudes toward certain
functional beverage concepts, where they were unsure of the health benefits of certain ingredients and
also the high retail prices charged for such beverages. In addition, the ingredient carrier, whether juice or
dairy, for example, was seen as having an important influence on the purchase of functional beverages.
Some interviewees felt that a functional beverage could be marketed as a health drink and positioned
as such in the marketplace. These interviewees felt that since some functional beverages were lactose-
free, they could also be marketed as a nondairy probiotic category and had the potential to rival soy and
probiotic dairy beverages. Focus group participants also suggested that functional beverages could be
targeted at the lactose-intolerant consumer segment. Participants’ comments included the following:
I think functional beverages are suitable for those who do not like dairy products or are allergic
to lactose. (Focus Group 1, male, 18–24 years)
I know that there are many probiotic beverages on the market and most of them are dairy-based.
I think a non-dairy probiotic beverage may be of interest to consumers. (Interviewee 9, male,
35–44 years)
Some young participants, across both focus groups and interviews, suggested that functional beverages
could be positioned as healthy meal replacements or, more particularly, breakfast meal replacements.
They felt that those with busy lifestyles could consume functional beverages at the breakfast meal occa-
sion as they were convenient and could be consumed on the go based on the method of delivery. These
products could contain the nutritional and vitamin profile of a healthy breakfast and be positioned as an
on-the-go beverage. Moreover, they suggested that functional beverages could also be marketed to dif-
ferent demographic groups as healthy alternatives to carbonated sports or energy beverages with a natu-
ral positioning strategy. However, these consumers were also concerned about the high calorie content
of functional beverages. They might not consume a functional beverage as a meal component due to its
perceived high calorie content. Examples of participants’ comments were as follows:
A meal replacement at breakfast is an option and would be welcomed by people with busy life-
styles. (Focus Group 1, male, 18–24 years)
I think it is a good idea that you can take one (functional beverage) in the morning instead of
breakfast, but I would not have it with a meal because of its high calorie content. (Interviewee
11, female, 45–54 years)
The health benefits of functional beverages were considered valuable marketing cues by both focus
group participants and interviewees. The most important health benefits that would encourage purchase
of functional beverages were enhancing the immune system, aiding the digestion, lowering cholesterol,
and having high fiber and reduced sugar. Older focus group participants were interested in disease-
preventing benefits, such as cholesterol reduction or cancer prevention. Generally, males were more
interested in the specific health claims of the product. However, female participants focused more on
their knowledge of functional ingredients and their health benefits. From a marketing perspective, this
illustrates the significance of correctly identifying a suitable target market and then positioning an opti-
mal functional beverage toward the target market.
Many participants also indicated that product information had a strong influence on their willingness
to purchase functional beverages. They suggested that product descriptions on functional beverage pack-
aging should contain information on the functional ingredients, the health benefits, the suggested daily
dosage, and the reference daily intake. The following was a typical comment:
I want to know whether I have to be careful of the dosage I consume daily. (Focus Group 1,
female, over 55 years)
These participants also mentioned that the brand name, logo, images, and color on the packaging would
influence their motivation to purchase functional beverages. In addition, many of the respondents per-
ceived that beverages that utilized a single-serve packaging format were healthier than other formats.
Fifteen product attributes were generated from consumer interviews that would strongly influence par-
ticipants’ purchase of functional beverages. It also revealed that consumers’ purchase decision in relation
to functional foods was complex and influenced by many intrinsic and extrinsic attributes such as taste,
the added ingredients to the products, health benefits, price, product volume, packaging (color, images,
typeface, and product shape), brand, and label information. Label information and packaging were seen
by many participants as being central to the purchase decision particularly with respect to reduced risk
in relation to new ingredients, new products, or new brands. Packaging has frequently been mentioned
as a direct aid for consumers for evaluating product quality and, therefore, should be carefully designed
to effectively convey product attributes to the consumer.
1.6 Semiotics and Functional Beverages

The next stage in the process was to conduct a semiotic analysis of functional beverages to see how the
information can be used to inform decisions on packaging design. Semiotics, a market-oriented meth-
odology, consisted of two separate stages and complemented the information generated from the focus
groups and interviews in the first part of this case study.
1.6.1 Stage 1: The Semiotic Sort

Stage 1, the semiotic sort, was conducted and classified information attributes (brand, color, and logo) not
only by things they had in common but also by any shared meanings [32]. An electronic sort consisted
of an electronic collection of the most prominent functional beverages on the market. Any functional
beverage that was positioned on the basis of health was included in the sort. A sort is a gathering of
digital images and products, in this case, of functional beverages. Signs most prevalent on these pack-
ages were recorded such as color, brand, health/nutritional claims, iconic symbols, and images. From
this, the main sign system for the semiotic analysis of functional beverages was developed and can be
seen in Table 1.1.
The images gathered in the sort were utilized in a primary semiotic analysis of functional beverage
packaging. The primary semiotic analysis required the researcher to interpret signs and encoded mean-
ings from all functional beverage packages collected during the sort. All signs and encoded meanings
were decoded based on the existing literature. Existing literature included manufacturers’ design speci-
fications, manufacturers’ intended meanings, and interpretation of color and images. This information
was readily available through semiotic journals, corporate reports, and corporate websites. Literature
focused on color interpretation, shape and typography, and consumer perceptions of various package
designs and types. This first stage identified suitable visual stimuli that would be presented to consumers
in the second stage of the study in conjunction with an in-depth interview guide.
1.6.2 Stage 2: Interpretation of Package Signs by Respondents

To ensure the methodology remained market oriented, Stage 2 focused on the interpretation of package
signs by respondents. Twelve respondents, five males and seven females, were recruited through the use
of convenience sampling [33]. Information was generated that focused on consumers’ attitudes toward
functional beverages, the purchase experience when buying functional beverages, and functional bever-
age packaging. A semistructured interview was utilized to gather the information [34].
Thirty-six visual stimuli were presented to respondents through a PowerPoint presentation at the inter-
view. The stimuli chosen were informed by the findings of the focus group and interviews conducted
in the earlier part of the case study, in addition to the results of the semiotic sort. This ensured that the
research followed a market-oriented methodology. The 36 visual stimuli consisted of 12 full packaging
concepts with all external stimuli removed, 12 brands and logos separate from the packaging concepts,
6 images found on packaging concepts, and 6 full packaging concepts complete with all external stimuli.
The PowerPoint presentation ensured that all the images were displayed on a neutral background and
were presented to consumers in the same sequence. This process illustrated how the signified concept
of an individual sign (a brand logo) might change due to its paradigmatic relationship with other signs
(how the brand logo might interact with other colors and icons on the packaging). Studies have shown
TABLE 1.1
Main Sign System for Semiotic Analysis of Functional Beverages
Main Sign
System Headings Examples
Brand Innocent, Actimel, and Red Bull.
Signifier Any material thing used to signify something: use of a wheat kernel and heart symbol to signify
health.
Signified The concept that the signifier refers to: healthy ingredients, energy, and naturalness.
Code A set of conventions understood in a given society: modern lifestyles, healthiness, and wellness.
Metaphor Expressing the unfamiliar in terms of the familiar: the use of phrases or images to imply that
something is healthy and natural such as the use of a halo to imply pure.
Connotation The cultural meaning of signs: the use of the color green to mean organic or natural.
Imagery The use of graphics to convey a meaning: a picture of an active person to convey the meaning that
the product is for active people.
that graphic elicitation, such as PowerPoint presentations, encouraged contributions from consumers that
were difficult to obtain by other means and this facilitated a more market-oriented methodology [35–37].
The semiotic interviews were transcribed from the audiotapes and analyzed using the software pack-
age QSR N6 [31]. Various codes were allocated and assigned to key segments of information within the
data [30]. A detailed analysis of the codes identified signs that when incorporated onto a product package
could motivate the purchase of new or existing functional beverages.
1.7 Semiotic Results

1.7.1 Stage 1: Results
The codes present from the interaction of multiple signs allowed for an initial semiotic analysis to be
performed for six selected functional beverages. The main sign system for each of these functional
beverages is outlined in Tables 1.2 through 1.7.
The individual signs prevalent on functional beverages were then analyzed. The semiotic sort revealed
that the colors white and green were synonymous with certain functional beverages. The use of the color
white was used primarily on dairy-based functional beverages such as Actimel and Yakult. The color
green was primarily used by juice-based functional beverages such as Naked and Tropicana. It was
found that the colors blue and yellow were most utilized for products that were trying to convey the ben-
efit of energy. Transparent packaging was common for a large number of beverages, such as Innocent,
Naked, Ribena, and Yakult. The transparent packaging allowed for the beverage color to be seen by
the purchaser and so generally denoted the flavor of the beverage. Iconic imagery was frequently used
to convey the ingredients used within the beverage. These images were often used in conjunction with
images associated with nature such as sun, grass, leaves, or growing crops.
TABLE 1.2
Semiotic Analysis of Functional Beverage 1
Beverage Brand Innocent Pure Fruit Smoothie
Signifier Brand and image of a head with a halo.
Signified Healthy ingredients, no artificial ingredients.
Code Wellness, dietary behavior, convenience, and modern lifestyles.
Metaphor The signs transfer the qualities of the signified for another, thus creating a metaphorical sign that
offers the meaning that the beverage can easily be incorporated into the everyday diet to enhance
its healthiness.
Connotation The use of the white label, the brand name, and the image with a halo suggests that the beverage
is healthy to the consumer and does not contain negative ingredients.
Imagery Face with a halo to portray the perception of healthiness.
TABLE 1.3
Beverage Brand Naked Juice Smoothie
Signifier Brand and logo, dominant use of the ingredient color (green).
Signified Naturally healthy juice, energy giving.
Code Natural, organic, and healthy.
offers the meaning that the beverage is a natural product with natural health-enhancing benefits.
Connotation The dominant use of the color green offers a connotative relationship between the green
ingredients and the green color utilized in the packaging.
Imagery Fruit and vegetables to represent the ingredients used within the product and the use of leaves to
convey the perception of naturalness.
TABLE 1.4
Beverage Brand Actimel Original
Signifier Brand and image of rising sun.
Signified Fun, energy giving, and natural.
Code Wellness, naturalness, health, and convenience.
offers the meaning that consumption of this beverage would improve the healthiness of the diet.
Connotation The dominant use of the color white offers a connotative relationship between the dairy carrier used
and the perception of natural ingredients.
Imagery Rising sun over a green pasture offers the perception of energy, freshness, and naturalness.
TABLE 1.5
Beverage Brand Ribena Plus Summer Fruits
Signifier Brand and descriptor: plus.
Signified Rich in fruits, high in vitamins and antioxidants.
Code Healthy and thirst quenching.
offers the meaning that the juice is high in fruits and therefore high in vitamins.
Connotation The use of primary color in conjunction with the blue sky and clouds offers the perception of a
refreshing drink.
Imagery Medallion illustrating the calcium content and fruits reflecting the ingredients of the product.
TABLE 1.6
Beverage Brand Yakult
Signifier Brand: dominant use of transparent packaging with red typeface for the brand.
Signified Small and powerful.
Code Convenience, innovation, wellness, simple, and effective.
offers the meaning that the beverage is novel, healthy, and powerful.
Connotation The use of the red typeface for the brand on the transparent packaging offers the idea that the
beverage is simple and effective.
Imagery Not applicable.
TABLE 1.7
Beverage Brand Tropicana Essentials Orange Juice and Omega 3
Signifier Brand and descriptor (essentials) in green typeface.
Signified Necessary for healthy living and natural.
Code Convenience, health, and wellness.
offers the meaning that the beverage is essential to everyday health.
Connotation The use of the green typeface for the brand on the white packaging gives the perception of a
natural beverage with no artificial ingredients.
Imagery Faded image of male and female on the center of the package illustrating the product is essential
for every person. Iconic images of oranges reaffirming the flavor and content of the beverage.
1.7.2 Stage 2: Results

Respondents were first asked a variety of questions related to their functional beverage purchase habits
and preferences. They then viewed 36 images and were asked questions on each image to ascertain what
messages they decoded from the information given to them.
1.8 Preferences and Purchase Habits toward Functional Beverages

The interviews revealed that the packaging color played a significant part in consumer acceptance of
functional beverages. For dairy beverages, packaging that was primarily white and contained iconic
images of the ingredients were perceived as most healthy by respondents. For juices, packaging that was
transparent was most preferred by respondents. Most interviewees admitted they made presumptions
about the flavor of a beverage based on the color of the base ingredient. These presumptions varied based
on whether the beverage was a dairy-based product or a juice-based product. Respondents felt that the
carrier had a very important influence on product purchase, whether the carrier was juice, water, cereal,
or dairy based. In addition, noncarbonated beverages with more natural ingredients were perceived as
healthier by respondents.
For dairy-based products, light pink was associated with strawberry, light yellow was associated with
banana, purple was associated with berry, and white was mostly associated with a vanilla-type flavor.
For juice-based beverages, pink was associated with grapefruit, and yellow and orange colors were
used interchangeably to describe the flavor for orange juice, although tropical juice was also mentioned.
Purple was used to denote black currant and green was mostly associated with apple flavors. Most
females indicated they preferred strawberry or vanilla flavors, while males did not have any particular
preferences. Some females, who regularly purchased functional juice beverages for health benefits, men-
tioned the sugar content and indicated they paid particular attention to the label to ensure they chose
the healthiest product available. In addition, respondents believed that both juice-based and dairy-based
products, which contained images of fruits, were healthiest.
The packaging on functional beverages in the retail environment was also mentioned by respondents
as an important influence on purchase. In addition, the location where the beverage was merchandised in
the retail outlet also impacted on the purchase decision. If a beverage was available from the refrigerated
section of the retail outlet, respondents were more likely to purchase the product. They also mentioned
the need for healthy on-the-go beverages, explained by some as “those beverages that keep you going.”
These beverages were described as those that had extra caffeine, or other ingredients, such as ginseng
or guarana, that provided a boost. These products were associated with strong and bright colors, in par-
ticular orange and red. There was a strong interest in noncarbonated energy beverages with more natural
ingredients.
Dairy-based functional beverages, such as Danone Actimel and Danone Activia, were the most fre-
quently purchased brands mentioned by interviewees. However, it was evident that a large proportion
of respondents tried to minimize the quantity of dairy consumed in their diets, due to either health or
weight concerns. These respondents frequently mentioned fruit juices, such as Innocent, as healthy bev-
erages. Juice-based carriers gave the perception of a healthy beverage. A small number of respondents
purchased own-brand functional beverages, such as Tesco yogurt or Tesco juice beverages, with a wide
variety of added vitamins.
The consumption occasion also had a significant influence on the beverage brand purchased. A number
of respondents were willing to purchase own-brand juice beverages for breakfast consumption. However,
when beverages were consumed outside the home, a brand name was nearly always preferred.
I just get the Tesco orange juice with added calcium but I always buy Actimel to take to work.
(R2, male, 41–50 years)
A majority of interviewees gave significant consideration to the preferences of the overall family unit
in the purchase of healthy beverages. Interestingly, a large number of female consumers indicated they
would purchase additional single-serve beverages to meet their own specific health and wellness needs.
These beverages were nearly always a popular branded product that reflected a premium price.
I will buy a multi-vitamin juice for everyone so they are getting all the vitamins they need but I
will have my own shot every morning. My kids would not drink those. (R10, female, 31–40 years)
The packaging type was also of particular importance to consumers. The majority of respondents indi-
cated that a resealable package that was easy to open and close was essential, in addition to being light-
weight and rectangular in shape, so that it could easily fit on the refrigerator shelf. For products that were
positioned on the basis of single-serve portions, a number of consumers mentioned the need for compact
packaging so that a number of portions could fit in the fridge. A small number of female consumers
also mentioned how these products should also have durable packaging for on-the-go consumption. The
importance of on-the-go consumption to the purchase decision was evident across many of the demo-
graphic groups.
The font size of nutritional and labeling information on packaging was an important issue for respon-
dents. They noted that food firms used smaller font sizes on the back of pack labeling, and, in some cases,
the additional information provided to support a health claim or logo. Importantly, these respondents
were reluctant to try any new products that displayed difficult to read text.
I find that the writing explaining ingredients has become very small and I hate having to look for
my glasses. So I will just pick up another one [beverage] instead. (R11, male, 61+ years)
Most respondents believed that juice and dairy beverages they regularly purchased were healthy and
positively contributed to the overall wellness of their diet. In particular, dairy was perceived to be a
naturally healthy product category, and older consumers mentioned a number of health benefits including
calcium, phosphorus, and protein. However, younger respondents were more positive toward water and
juice-based beverages particularly with energy, hydration, and beauty benefits.
Respondents were unsure how cholesterol reduction could be effectively conveyed through an image.
The majority of respondents revealed that a heart image would contribute to the perception of reduced
cholesterol levels, while images that conveyed the idea of natural ingredients and active individuals were
also deemed appropriate. In addition, a smaller number of respondents emphasized the need for key
words to accompany the image in order to avoid confusion at the retail point of purchase for the con-
sumer. Similar responses were found for the health benefits of omega-3 fatty acids. Respondents identi-
fied the image of a heart as appropriate to describe the benefits of this ingredient, and they stressed the
need for text that clearly outlined omega-3 benefits. Typical comments included the following:
I would like to see an active person and the healthy heart. (R2, male, 41–50 years)
Natural ingredients and a heart like before, but I would have to see the words Omega-3 to know
for sure. (R3, female, 51–60 years)
The image of a wheat shaft or grain kernel was suggested as a symbol for fiber in functional beverages.
Respondents also associated these images with whole grain. The most common health benefit for this
type of product was gut health and digestion. The use of images of leaves and grass on a beverage were
associated with fresh organic products. Although not a specific health benefit, respondents indicated that
these images were successful in attracting their attention to products in the retail environment.
1.9 Semiotics and New Product Development

The semiotics revealed that consumers preferred partially transparent packaging for beverages they were
unfamiliar with or for beverages that claimed a certain health and wellness benefit. This allowed con-
sumers to make assumptions about the ingredients and flavors of the product they were unfamiliar with.
Moreover, opaque packaging was associated with very familiar beverages that made no specific health
and wellness claims. Transparent packaging has been associated with reduction of the uncertainty or
purchase risk associated with novel products [38].
Typographical elements are often used to intentionally signify specific things, namely, the subcultural
context, the strategy of the food manufacturer, and the target group of consumers [39]. This was evident
for a number of beverage packages that were examined in the semiotic analysis. It was clear the respon-
dents held a preference for typeface that was simple and clearly legible for beverages that claimed to
have a functional benefit or positioned as a healthier product. Overemphasis on the depth and curvature
of the type gave the perception of a cheaper quality product for the vast majority of respondents. For this
reason, functional beverage packaging needs to incorporate plain, possibly Antigua-style text, as this text
is mostly associated with healthier products [26].
The consumer interviews also provided an insight into how the presence of a claim could interact with
other aspects of packaging to influence consumers’ attitudes and purchase intentions. The strategic use of
color is regarded as a fundamental tool in corporate marketing strategies and provides a means of product
and brand differentiation [40,41]. Importantly, this case study revealed that color was the primary sign
that attracted consumers to a brand on the market that they were unfamiliar with. In addition, consumers
made assumptions about the taste of the product based on the primary color used in the packaging.
It was also evident that the use of certain colors in smaller amounts on product packaging, that is, other
than the main package color, encouraged the consumer to interpret metonymic relationships. The most
common of which was the use of the colors green, white, and yellow: green to portray the meanings of
natural, healthy, organic, and fresh; white to portray the meanings of natural, fresh, and free-from; and
yellow to portray the meanings of sunlight, morning, energy, and fresh. In addition, the use of pictures
and images also attracted consumer attention to brands they were unfamiliar with on the market [37].
1.10 Lessons from the Case Study

The aim of the case study in this chapter is to explore market-oriented design issues of new functional
beverages. Information was generated about new functional beverages from the consumers’ perspective.
In addition, a semiotic analysis was conducted to inform the design of product packaging that would
encourage first purchase, and repeat purchase, by consumers of functional beverages. A key role of infor-
mation is to reduce market uncertainties and then to create, build, and maintain competitive advantage,
through an in-depth understanding of consumers’ needs during the NPD process.
The case study illustrated the important role that consumers can play in the design and marketing of
functional beverages in terms of the development of new product concepts and associated new product
packaging and also the identification of suitable target markets. The case study showed the importance
of generating information on functional beverages in relation to how consumers perceive them and how
different attributes, such as the carrier, taste, packaging, and price, may influence purchase. It illustrated
the importance of consumer knowledge of functional beverages and how these products fit in with con-
sumers’ healthy lifestyles.
The importance of packaging to the purchase decision was clear from this case study in terms of
attracting consumer attention to functional beverages. Involving consumers at the early stages of the
NPD process to assist in codesigning product packaging can ensure that benefits of the product are com-
municated effectively through various signs, symbols, and colors. This may result in increased accep-
tance of new functional beverages and repeat purchase of existing functional beverages. Consumers’
initial interaction with novel products is through the medium of packaging. Therefore, consideration
should be given to the development of product packages that successfully communicate intrinsic attri-
butes of the product through the use of appropriate signs and codes. This case study found that colors
such as white, green, and yellow/orange were most synonymous with healthy beverages. In addition, the
inclusion of images such as wheat shafts, healthy hearts, and leaves further added to the overall percep-
tion of increased health and wellness.
The information generated in this case study can then be used with quantitative techniques, such
as conjoint analysis, to identify optimal product attributes. Conjoint analysis is a multivariate concept
optimization research technique that is used to measure consumer preferences, through utility trade-
offs, for product concepts to understand preferences for products [42]. It is premised on the idea that
consumers evaluate the value of an object by combining the separate amounts of value provided by each
attribute. This enables the development of an optimal functional beverage that creates the most value for
consumers.
1.11 Conclusion
The market for functional beverages continues to grow as consumer demand for traditional carbonated
beverages falls, in line with changing consumer health and wellness lifestyles. This market offers huge
opportunities for firms that develop market-oriented beverages, where the intrinsic and extrinsic attri-
butes are designed to closely meet consumer expectations, offering benefits as part of a healthy lifestyle.
A market-oriented approach to the development of new functional beverages incorporates the voice of
the consumer information at the early stages of the NPD process in order to increase the likelihood of
consumer acceptance of such beverages. This is particularly important in the very competitive func-
tional beverage sector where consumers are faced with new choices, innovations, and brands on a very
regular basis.
REFERENCES
1. De Angelis, A., Global beverage market, April 2013. Published online at: http://www. companies
andmarkets.com/MarketInsight/Food-and-Drink/Global-Beverage-Market/NI6953 (accessed April 16,
2014).
2. Cowland, D., Demand for functional food and drink on the rise in emerging markets, December 2012.
Published online at: http://www.nutraceuticalsworld.com/blog/marketwatch/2012-12-19/demand-for-
functional-food-drink-on-the-rise-in-emerging-markets (accessed April 16, 2014).
3. Bleiel, J., Functional foods from the perspective of the consumer: How to make it a success? Int. Dairy J.,
20(4), 303–306, 2010.
4. Cowland, D., Health and wellness beverages outperform wider drinks industry, February 2013.
Published online at: http://blog.euromonitor.com/2013/02/health-and-wellness-beverages-outperform-
wider-drinks-industry.html (accessed April 16, 2014).
5. Euromonitor, Functionality, naturalness and stevia: Key to developing beverages to fit today’s trends,
January 2013. Published online at: http://www.euromonitor.com/functionality-naturalness-and-stevia-
key-to-developing-beverages-to-fit-todays-trends/report (accessed April 16, 2014).
6. Fulgoni, L.V. and Quann, E.E., National trends in beverage consumption in the children from birth to
5 years: Analysis of NHANES across three decades. Nutr. J., 11, 92, 2012.
7. Kleiman, S., Ng, S.W., and Popkin, B., Drinking to our health: Can beverage companies cut calories
while maintaining profits? Obes. Rev., 13, 258–274, 2012.
8. Cooper, B., Functional drinks—Part II: Growth potential, investment and challenges, October 2011.
Published online at: http://www.just-drinks.com/management-briefing/growth-potential-investment-
and-challenges_id105305.aspx (accessed April 16, 2014).
9. Holub, B., Potential benefits of functional food and nutraceuticals to reduce the risks and costs of disease
in Canada, Report submitted to Agriculture and Agri-Food Canada, Food Bureau, Ottawa, Ontario,
Canada, 2002.
10. Euromonitor, Health and wellness in Ireland, November 2013. Published online at: http://www.
euromonitor.com/health-and-wellness-in-ireland/report (accessed April 16, 2014).
11. Cowland, D., Stevia going from strength to strength, August 2012. Published online at: http://www.
nutraceuticalsworld.com/blog/marketwatch/2012-08-15/stevia-going-from-strength-to-strength
(accessed April 16, 2014).
12. Pranevičius, A., Top functional beverage trends for 2013. Published online at: http://mydrinkbeverages.
com/blog/top-functional-beverage-trends-for-2013 (accessed April 16, 2014).
13. Lipp, M., Beverages at the forefront of innovation in booming functional food market, June–July 2012.
Published online at: http://www.foodsafetymagazine.com/magazine-archive1/junejuly-2012/regulatory-
report-beverages-at-the-forefront-of-innovation-in-booming-functional-food-market/ (accessed April 16,
2014).
14. Demeritt, L., 2012 trends in functional foods and beverages. Published online at: http://www.
naturalproductsinsider.com/articles/2012/11/2012-trends-in-functional-foods-and-beverages.aspx
15. Fuhrman, E., Delivering on a functional promise. Bev. Ind., 101, 60–64, 2010.
16. Mellentin, J., Failures in Functional Foods and Beverages, New Nutrition Business, Hammersmith,
London, UK, 2009.
17. Wennström, P. and Mellentin, J., The Food and Health Marketing Handbook, New Nutrition Business,
Hammersmith, London, UK, 2003.
18. Mattsson, J. and Helmersson, H., Food product development: A consumer-led text analytic approach to
generate preference structures. Br. Food J., 109, 246–259, 2007.
19. Kohn, K., Idea generation in new product development through business development through business
environmental scanning: The case of XCar. Mark. Intell. Plann., 23, 688–704, 2005.
20. van Kleef, E., Van Trijp, H.C.M., Luning, P., and Jongen, W., Consumer-oriented functional food devel-
opment: How well do functional disciplines reflect the voice of consumer? Trends Food Sci. Technol.,
13, 93–101, 2002.
21. Mitchell, K. and Branigan, P., The role of focus groups in evaluation, in Evaluating Health Promotion:
Practice and Methods, Thorogood, M. and Coombes, Y., eds., Oxford University Press, Oxford, UK,
2000.
22. van Kleef, E., van Trijp, H.C.M., and Luning, P., Consumer research in the early stages of new product
development: A critical review of methods and techniques, Food Qual. Prefer., 16, 181–201, 2005.
23. Webb, J.R., Understanding and Designing Marketing Research, The Dryden Press, London, UK, 1995.
24. Harmon, T.R., The meaning behind marketing: Semiotic-oriented research in marketing and con-
sumer research. Published online at: http://faculty.quinnipiac.edu/charm/CHARM%20proceedings/
CHARM%20article%20archive%20pdf%20format/Volume%2012%202005/144%20harmon.pdf
25. Bignell, J., Media Semiotics, Manchester University Press, Manchester, UK, 2002.
26. Anderson, T.H. and Boeriis, M., Semiotic cereals, in Systematic Functional Linguistics in Use,
Nørgaard, N., ed., Odense Working Papers in Language and Communication (OWPLC), University of
Southern Denmark, Odense, Denmark, 2008.
27. Rettie, R. and Brewer, C., The verbal and visual components of package design. J. Prod. Brand Manage.,
9, 89–105, 2000.
28. Lu, L., Gargallo, S., and Munar, M., Packaging as a Strategic Tool, School of Business and Engineering,
University of Halmstad, Halmstad, Sweden, 2007.
29. Fink, A. and Kosecoff, J., How to Conduct Surveys: A Step-by-Step Guide, 2nd edn., Sage Publications,
Thousand Oaks, CA, 1998.
30. Krueger, R.A., Focus Groups: A Practical Guide for Applied Research, 2nd edn., Sage Publications,
Thousand Oaks, CA, 1994.
31. QSR International, N6 Non-Numerical Unstructured Data Indexing Searching and Theorizing
Qualitative Data Analysis Program, Version 6.0, QSR International Pty Ltd., Melbourne, Victoria,
Australia, 2002.
32. Oswald, L., Semiotics and Sensory Marketing, Marketing Semiotics Inc., New York, 2001.
33. Anderson, D.R., Sweeney, D.J., and Williams, T.A., Essentials of Statistics for Business and Economics,
4th edn., Cengage Learning, London, UK, 2005.
34. Punch, K.F., Introduction to Social Research: Quantitative and Qualitative Approaches, Sage
Publications, London, UK, 1998.
35. Crilly, N., Blackwell, A.F., and Clarkson, J.B., Graphic elicitation using research diagrams as interview
stimuli. Qual. Res., 17, 13–26, 2006.
36. Harper, D., Talking about pictures. Visual Stud., 17, 13–26, 2002.
37. Underwood, R.L., Klein, M.M., and Burke, R.R., Packaging and communication: Attentional effects of
product imagery. J. Prod. Brand Manage., 10, 402–422, 2001.
38. Sogn-Grundvåg, G. and Østli, J., Consumer evaluation of unbranded and unlabelled food products: The
case of bacalhau. Eur. J. Mark., 43, 213–228, 2009.
39. Spitzmüller, J., Visible by design. The significance of typography in media communication, March 2007.
Published online at: http://www.ds.uzh.ch/spitzmueller/docs/pres-tokio-2007-03-02.pdf (accessed April
16, 2014).
40. Grossman, R.P. and Wisenblit, J.Z., What we know about consumers’ color choices. J. Mark. Pract.
Appl. Mark. Sci., 5, 78–88, 1999.
41. Schmitt, B.H. and Pan, Y., Managing corporate and brand identities in the Asia-Pacific region. Calif.
Manage. Rev., Summer, 32–48, 1994.
42. Hair, J.F., Black, W.C., Babin, B.J., Anderson, R.E., and Tathan R.C., Multivariate Data Analysis,
Pearson International, Upper Saddle River, NJ, 2006.
2
Global Nutraceutical Regulations
for Functional Beverages
Anand Swaroop, Manashi Bagchi, and Debasis Bagchi
CONTENTS
2.1 Introduction......................................................................................................................................17
2.2 Beverages and Regulations..............................................................................................................18
2.3 Beverages versus Liquid Dietary Supplements................................................................................18
2.4 Powdered Premix Products and Liquid Concentrations................................................................. 19
2.5 Regulatory Requirements for Ingredients in Beverages and Dietary Supplements....................... 20
2.6 Regulatory Requirements for Labeling of Beverages..................................................................... 20
2.7 International Regulatory Norms..................................................................................................... 21
2.7.1 Japan................................................................................................................................... 21
2.7.2 European Union.................................................................................................................. 22
2.7.3 China.................................................................................................................................. 22
2.7.4 Canada................................................................................................................................ 22
2.7.5 South Korea........................................................................................................................ 23
2.7.6 India.................................................................................................................................... 23
2.7.7 Australia............................................................................................................................. 23
2.7.8 New Zealand....................................................................................................................... 23
2.7.9 Israel................................................................................................................................... 24
2.8 World Market.................................................................................................................................. 24
2.9 Conclusion....................................................................................................................................... 25
References................................................................................................................................................. 25
2.1 Introduction
Phytopharmaceuticals have been used for centuries as novel prophylactic agents for the prevention and
treatment of diseases and disorders in humans and animals as well as for the improvements of chronic
degenerative conditions. Approximately more than 2500 years ago, the father of modern medicine
“Hippocrates” proclaimed the association of food with health benefits and quoted “Let food be thy medi-
cine and medicine be thy food” (Hippocrates, 460–377 BC). Nutritionists and health professionals are
continuously unveiling the beneficial health effects of diverse functional foods and nutraceuticals. This
may range from isolated nutrients as dietary supplements, herbal products, and fortified diets that can
be used as soups, cereals, fortified juices, or beverages, among others. Functional beverages are specifi-
cally designed to quench thirst and maintain healthy fluid and water levels and nutrition. Some examples
include orange juice fortified with vitamin C, calcium, and phytosterols, berry drinks with anthocyanins,
and green tea fortified with epigallocatechin gallate. It is very important that functional beverages carry
appropriate labeling information for the benefit of the consumer. It is also essential that these functional
beverages strictly follow regulatory guidelines to attract consumer confidence in the marketplace [1].
Nutraceuticals and functional foods are becoming increasingly popular around the world, and
especially the demand for functional nutraceutical beverages is on the rise. A significant number of
17
nutraceutical beverages have been introduced in the United States over the last two to three decades and
become increasingly popular. In 2010, the diverse functional beverage market including energy drinks,
sports drinks, various functional drinks, yogurt drinks, smoothies, and ready-to-drink teas and coffees
reached US$23.4 billion, and the same trend is similar globally [2]. The growth of functional beverages
is quite obvious in developing nations because of increased awareness of maintaining good health, body,
and mind. Another reason may be affordability and convenience. This chapter focuses on the intricate
aspect of nutraceutical beverage regulation process in the United States and around the world.
2.2 Beverages and Regulations

It has been observed that there is a remarkable upsurge in the marketing of beverages with a variety of
nutraceutical ingredients and intended uses. Products are being marketed as dietary supplements and/or
conventional foods. It should be pointed out that several of these products may be misbranded, because
their labeling are entirely inconsistent with the product category. However, the regulatory environment for
nutraceutical beverages is not clearly defined [3]. A significant number of functional beverages continue
to expand globally, working with domestic and international trade associations and agencies, struggle to
survive, and harmonize with the regulatory hurdles continue. While consumers purchase these bever-
ages depending on their usefulness and cost, the regulatory organizations are imposing requirements to
demonstrate broad-spectrum safety as well as to track adverse events [4]. Increasingly, the nutraceuti-
cal beverage manufacturing and trading companies are trying to comply with domestic, interstate, and
international regulatory requirements.
A number of key factors are associated to meet the requirements:
• A major emphasis has been given on the safety and appropriate labeling claims, which can be
achieved partly through the good manufacturing practice (GMP) regulations and keeping track
of adverse event reporting.
• Increased enforcement of regulations will streamline the comparatively new and fragmented
companies for regulatory compliance.
• Beverages are conventional foods that may not be marketed as dietary supplements. Under sec-
tion 201(ff)(2)(B) of the Federal Food, Drug, & Cosmetic (FD&C) Act (21 U.S.C. 321(ff)(2)(B)),
“dietary supplement” means a product that, among other requirements, “is not represented for use
as a conventional food or as a sole item of a meal or diet. On the other hand, beverages are conven-
tional foods under the FD&C Act” [4]. Sometimes, when the label of a product characterizes it as a
dietary supplement, the product may not meet the requirements of a dietary supplement. Beverages
or products in liquid form can be represented as conventional foods as a result of factors such as
their products or brand name, packaging, serving size, and recommended daily intake (RDI) or
the volume that is specified to be consumed, composition, recommendations, directions for use,
statements or graphic representations in labeling or advertising, and other marketing requirements.
2.3 Beverages versus Liquid Dietary Supplements

There are several factors that distinguish beverages from liquid dietary supplements. The most vital rep-
resentation of a product’s use is claims made for the product in its labeling and advertising. In addition
to labeling and advertising, a product’s name, packaging, serving size, RDI, recommended conditions
of use, composition, marketing practices, and representation are important determinants of whether the
product is represented as a conventional food, and, if so, it cannot be marketed as a dietary supplement.
The following are some of the salient features:
Claims: Claims are used for a product in its labeling or advertising.
Product representation: A product’s name, packaging details, labeling, serving size, RDI, other recom-
mended conditions of use, composition, and marketing practices, advertising, and representations are
Global Nutraceutical Regulations for Functional Beverages 19
important determinants whether the product is represented as a conventional food and may not be mar-
keted as a dietary supplement. However, in special circumstances, a single factor may determine whether
the product can be termed as a “conventional food,” but, in most circumstances, a combination of factors
would determine whether the product is represented as a conventional food.
Labeling: It has been outlined by Food and Drug Administration (FDA) that statements, graphics dis-
played on product labels, labeling, and advertisement details including websites and social media should
be available when the agency evaluates the intended use and its appropriate representation. It is very
important to emphasize that a product that exhibits a supplement fact panel may still be a conventional
food if it contains statements that it is intended to “refresh” or “rehydrate,” in which statements indicate
that the intended use is as a beverage (which is a conventional food). Graphic representation in terms of
symbols, vignettes, schematic, or pictorial serving suggestions is another representation of a product as a
“conventional food.” In salad dressings, one can see the advertisement or label with a picture of a liquid
product being poured onto a salad would identify the product as a salad dressing.
Product identification: The brand name or product name uses the terms “beverage,” “bottled water,”
“iced tea,” “coffee,” “apple cider,” “juice,” “orange juice,” “soda,” or “drink” on the label to represent
the products as conventional foods because “bottled water” is a terminology identifying a specific cat-
egory of conventional food, which is defined in a “food standard regulation” (see 21 Code of Federal
Regulations [CFR] 165.100).
Packaging: Packaging is a modern art, which is a great marketing tool to contain, hold, preserve, and
exhibit the aesthetic appeal of the product as well as to provide directions as to how the product is to be
used. It should also include the size, volume of liquid, shape, storage conditions, color and design of the
container, and packaging details including whether it is reclosable or to be consumed in a single serving.
These types of packaging and labeling are extensively used for common beverages. A good example is a
Coca Cola pop-top aluminum can bearing a silver strip indicating “cola supplement” that shows that the
product is cola-flavored soft drink intended to be consumed in a single serving. It is very important to
indicate that containers indicate other specifics, including serving size and RDI that need to differenti-
ate the product from a conventional food, even if the container looks like a regular beverage container.
Composition and generally recognized as safe (GRAS) status: To overcome the regulatory hurdles, the
ingredients which can be incorporated in functional beverages must be safe as demonstrated by a bat-
tery of toxicological studies and follow all regulations that are imposed by the regulatory agencies in the
country of use. In the United States, it is expected that the ingredients used in beverages are self-affirmed
or FDA-notified GRAS and qualified for the requirement need for food additives.
Recommended use: The serving size and RDI are very important criteria. Average daily drinking fluid
intake is approximately 1.2 L/day. Liquid formulations that suggest on their labels serving size and/or
RDI to consume up to three 16 ounce bottles (~1.4 L)/day that they are intended to be consumed in
amounts that provide all or a significant part of the entire daily fluid intake of an average person in the
United States are effectively represented as conventional foods [5]. It is important to note that even if
a product is not expressly represented as an alternative to a beverage, when the practical result of the
labeled serving size and/or RDI is that the product is used as a beverage or replaces beverages that serve
as ordinary sources of drinking fluid, FDA would generally consider the representation of the product
for use as a conventional food.
Sales and marketing practices: Appropriate marketing practices should be used. Appropriate labeling,
advertising, and all promotional activities should comply with the regulatory norms.
2.4 Powdered Premix Products and Liquid Concentrations

Powdered premix products intended to be consumed in water, fruit juice, or milk products have long
been used globally. If these products are properly labeled as dietary supplements, these should not be
considered as beverages. Generally, these powdered premixes were introduced in the marketplace for
their convenience or stability. The structural integrity of some or all of the active ingredients of these
powdered premixes may be unstable in aqueous solution and hence are not considered beverages for use.
2.5 Regulatory Requirements for Ingredients in Beverages and

Dietary Supplements
It is important to ensure that any beverage or liquid dietary supplement getting introduced in the market-
place complies with all applicable regulatory norms and requirements of the substances being added to
the formulation. Product compliance is an important parameter. The following important features must
be considered:
Substances intentionally added to beverages: Several nutraceuticals and functional food ingredients
are added to beverages as food additives, which require premarket approval by FDA (Section 409 of the
FD&C Act 21 U.S.C. 348 and 21 CFR Part 171). However, a product is exempt from the definition of
a food additive and exempt from premarket approval if it is identified as a GRAS by qualified experts
under the conditions of its intended use in food (21 CFR 170.30) or if the product falls under another
exception of the food additive definition in section 201(s) of the FD&C Act 21 U.S.C. 321(s).
Dietary ingredients in dietary supplements: Dietary ingredients under section 201(ff)(1) of the FD&C Act
(21 U.S.C.321[ff][1]) must not adulterate the dietary supplement to which they are added (Section 402 of the
FD&C Act [21 U.S.C. 342]). Furthermore, dietary ingredients that were not marketed in the United States
before October 15, 1994, are “new dietary ingredients” subject to the requirements of section 413 of the
FD&C Act (21 U.S.C. 350b) and 21 C.F.R. 190.6. The “dietary supplements” [6], “new dietary ingredients
in dietary supplements—background for industry” [7], and “new dietary ingredients notification process”
[8] include links to regulatory requirements and recommendations that apply to new dietary ingredients.
Substances (other than dietary ingredients) intentionally added to dietary supplements: Section 201(s)
of the FD&C Act 21 U.S.C. 321(s) exempts dietary ingredients used in dietary supplements from the
“food additive” definition. Although a dietary ingredient used in dietary supplement must not adulterate
the supplement under section 402(f) of the FD&C Act (21 U.S.C. 342[f]), it does not have to be GRAS for
its intended use in the supplement. On the contrary, other excipients such as binders, additives, diluents,
and fillers, intended for use in dietary supplements, are not exempt from the food additive definition and
must meet the same requirements as substances added to conventional foods [9,10]. Thus, nondietary
ingredients added to a dietary supplement must follow the food additive regulation or be GRAS affirmed
for their intended use unless these ingredients qualify for another exception to the food additive defini-
tion [6,7]. More detailed information is available in the FDA websites [9,10].
2.6 Regulatory Requirements for Labeling of Beverages

Beverages should comply with all applicable labeling requirements in their respective locations.
General requirements: Claims, statements, and graphics in the labeling of beverages and liquid dietary
supplements need to comply with Section 403(a)(1) of the FD&C Act (21 U.S.C. 343[a][1]), which indi-
cates that a food is misbranded if its labeling is misleading or false.
Health claims: Beverage and liquid dietary supplements may bear health claims, which exhibit relation-
ship between a food or food component and a disease or health-related condition. It is recommended to
critically review the following sections in the U.S. FDA Redbook for details:
1. Section 21 CFR 101.14(a)(1)

2. Section 403(r)(1)(B), (r)(3), (r)(4), and (r)(5) of the FD&C Act (21 U.S.C. 343[r][1][B], [r][3],
[r][4], [r][5])
3. Section 21 CFR 101.14, 21 CFR 101.70, and 21 CFR 101.72–101.83
4. Label claims [11]

5. Guidance for industry: evidence-based review system for the scientific evaluation of health
claims [12]
6. Qualified health claims [13]
7. FDA modernization act (FDAMA) claims [14]
8. Summary of qualified health claims subject to enforcement discretion [15]
Nutrient content claims: Both beverages and liquid dietary supplements, which identify the nutrient level
in a food according to 21 CFR 101.13(b). In addition, the following sections provide additional details:
1. Section 403(r)(1)(A), (r)(2), (r)(4), and (r)(5) of the FD&C Act (21 U.S.C. 343[r][1][A], [r][2],
[r][4], [r][5])
2. Section 21 CFR 101.13, 21 CFR 101.69, and 21 CFR 101.54–101.67
3. FDAMA claims [14]
4. Label claims [11]
Structure function claims for conventional foods: Conventional foods and beverages may bear certain
kinds of claims about effects on the structure or function of the body. “Food” is defined in Section
201(f) of the FD&C Act (21 U.S.C. 321[f]) as (1) articles used for food or drink for man or other animals,
(2) chewing gum, and (3) articles used for components of any such article. It is also recommended to
consult Section 201(g)(1)(C) of the FD&C Act (21 U.S.C. 321[g][1][C]).
Structure function claims for dietary supplements: Labeling of dietary supplements needs to comply
with Section 403(r)(6) of the FD&C Act (21 U.S.C. 343(r)(6) and 21 CFR 101.93). These claims are about
general well-being and benefits.
General food labeling requirements: FDA’s general food labeling requirements, including those that
apply to dietary supplements, are in 21 CFR Part 101. Labeling requirements for beverages and conven-
tional foods differ greatly from dietary supplements. Beverages need to exhibit nutrition information in
the nutrition facts format as shown in 21 CFR 101.9, while dietary supplements need to exhibit nutrition
information in the Supplement Facts format (21 CFR 101.36). A beverage or other conventional foods
should not be labeled with the FDA disclaimer, which is required for dietary supplements.
2.7 International Regulatory Norms

In addition to U.S. regulation given earlier, international regulatory norms are also discussed briefly.
2.7.1 Japan
The terminology “foods with health claims” was first incorporated for nutraceuticals and functional
foods in Japan. It is worthwhile to mention that Japanese are extremely health conscious and the second
largest consumer of nutraceuticals. There are two basic categories:
1. Foods with Nutrient Function Claims: Basically, it satisfies the minimum and maximum daily
levels of selected vitamins and micronutrients.
2. Foods for Specified Health Uses: This requires premarketing approval. This needs evalua-
tion of effectiveness and approval by the Pharmaceutical Affairs and Food Sanitation Council
and the Ministry of Health, Labour and Welfare (MHLW). Japan also established Consumer
Affairs Agency, which assumed the MHLW responsibility [1,3,16].
The new Japanese regulation reformation process is in process and will be implemented soon.
2.7.2 European Union

European Union (EU) food laws and legislations are basically based on General Food Law Regulation
178/2002; Foods (EC Regulation 172/2002); Food Supplements Directive 2002/46; Fortified Foods
Regulation 1925/2006 on the addition of vitamins, minerals, and other nutraceuticals to food; Foods
for Particular Nutritional Uses (Dietetic Foods) (Directive 2009/39); Nutrition and Health Claims
Regulation 1924/2006; Novel Foods Regulation 258/97; and Foods for Particular Nutritional Use
(PARNUTS) (Directive 89/398/EEC). From 2007, nutrition and health claims regulation is covered by
European Commission, European Food Safety Authority, and national authorities [17,18]. In the United
Kingdom, food products are regulated under the Food Safety Act 1990, and “food” is defined in Article
2 of EC Regulation 172/2002.
In Article 2 of EC Regulation 172/2002, it clearly states that “Food shall not include medicinal prod-
ucts within the meaning of Council Directive 2001/83/EC.” Food in this definition covers any food,
beverage (drink), or supplement ingested and considered to be part of the normal human diet. Products
that provide benefits beyond their traditional nutritional value, however, are not considered to be foods.
If not considered a food, these may be PARNUTS under Directive 89/398/EEC.
This directive defines PARNUTS as foods which “owing to their special composition or manufactur-
ing process are clearly distinguishable from foodstuffs for normal consumption, which are suitable for
the claimed nutritional purpose and which are marketed in such a way as to indicate such suitability.”
The EU PARNUTS directive contains rules relating to the specific compositional details and labeling
requirements of foods and six categories of permitted substances: (1) vitamins, (2) minerals, (3) amino
acids, (4) carnitine and taurine, (5) nucleotides, and (6) choline and inositol [1,17,18].
Nutraceuticals, normally derived from existing food products, however, will not fall under the Article 2
definition of food since they purport to provide a benefit beyond their traditional nutritional value. While
not a food product, nutraceuticals do not fit the PARNUTS directive definition either, as their composi-
tion does not clearly fit any of the six PARNUTS categories [17,18].
The only other basis on which nutraceuticals may be regulated is as “novel ingredients” under
Regulation (EC) No. 258/97. These ingredients have not been marketed in an EU member state before 1997.
Authorization from the relevant EU member state’s competent authority needs to be obtained [1,17,18].
2.7.3 China
The nutraceutical market in 2008 was US$6 billion in China [19] and China Health Care Association, a
government-appointed body that regulates the nutraceutical industries. Other agencies include State Food
and Drug Administration (SFDA) that regulates the nutraceutical supplements, Ministry of Health (MoH)
that oversees SFDA and monitors the approval of novel food ingredients, and Administration of Quality
Supervision Inspection and Quarantine that regulates imports and exports of nutraceuticals and func-
tional foods. However, the regulatory position for functional beverages are not transparent at all [1,3,19].
2.7.4 Canada
Vitamins, minerals, botanical herbs based on dietary supplements, traditional Chinese medicines, pro-
biotics, and enzymes are called natural health products (NHPs) and regulated under the Canadian Food
and Drugs Act. The Canadian Government Health Authority—Health Canada has approved more than
61,000 NHPs for sale in Canada since 2004. NHP has classified a three-class system based on risk,
namely, (1) Class 1, (2) Class 2, and (3) Class 3. The regulatory approval process requires evidence
requirements based on risk and health claims. Furthermore, NHP Directorate has outlined procedures for
the evaluation of multi-ingredient formulations. Postmarket activities and vigilance are also performed
by regulatory agencies [1,20]. However, nutraceutical beverage regulation has not been independently
classified.
2.7.5 South Korea

Korean Health and Welfare Committee of the National Assembly proposed the Health/Functional Food
Act (HFFA) in late 2000 and the act was established to cover nutraceuticals and functional foods in
2002. In 2004, HFFA approved 37 generic nutraceutical and functional foods including vitamins, min-
erals, essential amino acids, proteins, dietary fiber, and essential fatty acids. The Ministry of Food and
Drug Safety (MFDS) authority evaluates the specifications, process standardization, safety, and effi-
cacy data very critically. As of October 2012, approximately more than 165 functional ingredients have
been approved by MFDS. Claims including platelet aggregation, triacylglycerols, blood pressure, blood
glucose, antioxidant, skin health, fatigue, cholesterol, calcium absorption, dental caries, fat reduction,
prostate function, gastrointestinal function, cognition, physical performance, urinary function, immune
function, ocular health, antistress, memory function, joint/bone health, menopause, and liver health have
been approved for product-specific HFFs [1]. However, no clear directive is available for nutraceutical
beverages.
2.7.6 India
Nutraceutical and functional foods are regulated by Food Safety and Standards Act. Manufacturers
follow the standards of Indian Pharmacopoeia. Federation of Indian Chambers of Commerce and
Industry is somewhat associated with the improvement of regulation on nutraceutical market and func-
tional beverage [1,21]. However, no detailed information is available.
2.7.7 Australia
Botanical herbs, vitamins, minerals, nutraceutical supplements, and homeopathic and aromatherapy
preparations are referred to as “complimentary medicines” and are regulated under the Therapeutic
Goods Act (TGA) 1989 [22,23]. A complimentary medicine, including a nutraceutical, is defined as
“a therapeutic good consisting principally of one or more designated active ingredients mentioned
in Schedule 14 of the Regulations, each of which has a clearly established identity and traditional
use,” which include an amino acid; charcoal; a choline salt; an essential oil; plant or herbal extract;
homeopathic preparation; a microorganism, whole or extracted, except a vaccine; mineral; mucopoly-
saccharide; nonhuman animal material; a lipid, phospholipid, or an essential fatty acid; royal jelly;
bee pollen; propolis; a sugar; polysaccharide or carbohydrate; and a vitamin or provitamin. Australia
has a two-tiered system for the regulation of complimentary medicines: (1) higher-risk products need
to be registered on the Australian Register of Therapeutic Goods (ARTG) [22–24], which requires the
evaluation of quality, safety, and efficacy and (2) lower-risk products that contains preapproved low-risk
ingredients and has limited claims listed in ARTG. TGA’s postmarket regulatory activity of complimen-
tary medicines and adverse event reporting are standard procedures. No clear directive is available on
nutraceutical beverage.
2.7.8 New Zealand

New Zealand’s Medicines and Medical Devices Safety Authority (MEDSAFE) is responsible for thera-
peutic products available in New Zealand. The interface between therapeutic-type and food-type dietary
supplements is subject to consultation between the New Zealand Food Safety Authority and MEDSAFE.
Premarketing approval is mandatory, and postmarketing surveillance monitors the safety and adverse
events. Handling complaints and investigations and GMP auditing are routine mandatory practices. Food
& Beverage Information Project 2011 overviewed a final report on nutraceuticals and foods for health in
October 2011 [25], and basically New Zealand followed the U.S. regulations. Although the report indi-
cated foods and beverages, however, the beverage section is not extensive.
2.7.9 Israel
Israel has been termed as one of the key innovation hubs for the nutraceutical industries, and their major
revenue comes from the export of nutraceuticals and functional foods to the United States and Europe.
MoH regulates nutraceuticals and functional foods [1,3].
2.8 World Market

Table 2.1 demonstrates the key regulatory terms in the United States and around the world and the
approximate revenues in the said territories [17,20,22–26], which is very promising. However, the nutra-
ceutical beverage regulations in the international marketplace still need to be established. According
to the available information, the regulatory and judicial formalities on nutraceutical beverages are in
progress.
TABLE 2.1
Nutraceutical Supplements, Regulatory Authorities, and Estimated Annual Business
Estimated Annual
Country Regulatory Authorities Business (US$) References
United States Code of Federal Regulations ~75.9 billion in 2018 [22]
Food and Drug Administration
Federal Trade Commission
Generally Recognized as Safe
Good Manufacturing Practices
Japan Consumer Affairs Agency ~26 billion in 2006 [23]
Foods with Nutrient Function Claims
Foods for Special Dietary Uses
Food for Specified Health Use
Ministry of Health, Labour, and Welfare
European Union European Food Safety Authority ~35 billion in 2010 [17]
Foods for Particular Nutritional Use
China China Health Care Association ~6 billion in 2008 [25]
State Food and Drug Administration
Ministry of Health
Administration of Quality Supervision Inspection and
Quarantine
Canada Natural Health Products Directorate na —
Health Canada
South Korea Health/Functional Food Act na —
Ministry of Food and Drug Safety
India Food Safety and Standards Act ~4 billion in 2018 [24]
Federation of Indian Chambers of Commerce and Industry
Australia Australia New Zealand Therapeutic Products Authority ~1.5 billion each year [26]
Australian Register of Therapeutic Goods
Complementary and Alternative Medicine
Therapeutic Goods Act
New Zealand Australia New Zealand Therapeutic Products Authority ~1 billion in 2010 [20]
Medicines and Medical Devices Safety Authority
New Zealand Food Safety Authority
Israel Ministry of Health na —
Abbreviation: na, not available.
2.9 Conclusion
Innovations and marketing of nutraceuticals and functional foods are now the fastest-growing segments
for this industry. Currently, the rising costs and toxicity of some pharmaceuticals are driving the popu-
lation around the world to move forward with safe, efficacious, and less expensive nutraceuticals and
functional food supplements and beverages. Especially, there is a massive global upsurge of functional
beverages in its sales and consumption. Especially, the younger generation has a trend to using func-
tional beverages over conventional cola beverages. Although nutraceutical beverage regulation has been
defined, further clarification is required to make it more effective and safe. Functional beverages are very
popular and extensively used in Japan, South Korea, China, and Thailand. Health professionals, nutri-
tionists, and regulatory toxicologists should strategically work shoulder to shoulder to derive appropriate
regulatory standards and to provide the optimal health and therapeutic benefits to mankind globally.
REFERENCES
1. Bagchi, D., Nutraceutical and Functional Food Regulations in the United States and around the World,
2nd edn., Elsevier/Academic Press, New York, 2014.
2. Functional Beverage Market Update, Functional beverage market update. Published online at: http://
www.nutraceuticalsworld.com/issues/2012–07/view_features/functional-beverage-market-update/,
July 1, 2012 (accessed April 17, 2015).
3. Wordpress, An overview of global regulatory trends in the nutraceutical industry—April 2013. Published
online at: https://bournepartners.wordpress.com/2013/04/22/an-overview-of-global-regulatory-trends-
in-the-nutraceutical-industry-april-2013, April 22, 2013 (accessed April 16, 2015).
4. FDA, Guidance for industry: Distinguishing liquid dietary supplements from beverages. Published
online at: http://www.fda.gov/Food/GuidanceRegulation/GuidanceDocumentsRegulatoryInformation/
DietarySupplements/ucm381189.htm, April 15, 2015 (accessed April 20, 2015).
5. RDI, Recommended daily water intake. Published online at: http://www.myfooddiary.com/resources/
ask_the_expert/recommended_daily_water_intake.asp, March 2015 (accessed April 18, 2015).
6. FDA, Dietary supplements. Published online at: http://www.fda.gov/Food/DietarySupplements/default.
htm, April 28, 2015 (accessed May 1, 2015).
7. FDA, New dietary ingredients in dietary supplements—Background for industry. Published online
at: http://www.fda.gov/Food/DietarySupplements/ucm109764.htm, March 3, 2015 (accessed May 1,
2015).
8. FDA, New dietary ingredients notification process. Published online at: http://www.fda.gov/Food/
DietarySupplements/NewDietaryIngredientsNotificationProcess/default.htm, November 13, 2014 (accessed
May 1, 2015).
9. FDA, Ingredients, packaging & labeling. Published online at: http://www.fda.gov/Food/Ingredients
PackagingLabeling/default.htm, March 13, 2015 (accessed April 4, 2015).
10. FDA, Guidance for industry: Consideration regarding substances added to foods, including to bever-
ages and dietary supplements 2014. Published online at: http://www.fda.gov/Food/GuidanceRegulation/
GuidanceDocumentsRegulatoryInformation/default.htm, April 14, 2015 (accessed April 16, 2015).
11. FDA, Label claims, general information. Published online at: http://www.fda.gov//Food/Ingredients
PackagingLabeling/LabelingNutrition/ucm2006873.htm, September 12, 2014 (accessed April 4, 2015).
12. FDA, Guidance for industry: Evidence-based review system for the scientific evaluation of health
claims 2009. Published online at: http://www.fda.gov/Food/GuidanceRegulation/GuidanceDocuments
RegulatoryInformation/LabelingNutrition/ucm07332.htm, January 2009 (accessed April 6, 2015).
13. FDA, Qualified health claims. Published online at: http://www.fda.gov/Food/IngredientsPackaging
Labeling/LabelingNutrition/ucm2006877.htm, November 24, 2014 (accessed April 6, 2015).
14. FDA, FDA modernization act (FDAMA) claims. Published online at: http://www.fda.gov/Food/
IngredientsPackagingLabeling/LabelingNutrition/ucm2006874.htm, December 24, 2014 (accessed
April 6, 2015).
15. FDA, Summary of qualified health claims subject to enforcement discretion. Published online at: http://
www.fda.gov/Food/IngredientsPackagingLabeling/LabelingNutrition/ucm073992.htm, December 14,
2014 (accessed April 6, 2015).
16. ICIS, Demand for nutraceuticals in Japan continues to increase. Published online at: http://www.icis.
com/resources/news/2007/11/05/9075285/demand-for-nutraceuticals-in-japan-continues-to-increase/,
November 5, 2007 (accessed April 25, 2015).
17. Nutraceuticalsworld, Nutraceuticals regulation back on European Commission Agenda for 2013.
Published online at: http://www.nutraceuticalsworld.com/issues/2012–11/view_features/nutraceuticals-
regulation-back-on-european-commission-agenda-for-2013/, November 1, 2012 (accessed April 25, 2015).
18. MyDrink Beverages, Trends of nutraceutical functional beverages in Europe. Published online at: http://
mydrinkbeverages.com/trends-of-nutraceutical-functional-beverages-in-europe, April 2015 (accessed
April 25, 2015).
19. China’s Nutraceutical Industry, China’s nutraceutical industry. Published online at: http://www.
nutraceuticalsworld.com/issues/2008–11/view_features/china-s-nutraceutical-industry/, November 1, 2008
20. Nutraceuticals World, Canada stands at regulatory crossroads for nutraceutical products. Published
online at: http://www.nutraceuticalsworld.com/contents/view_features/1999–03–01/canada-stands-at-
regulatory-crossroads-for-nutrace/, March 1, 1999 (accessed April 25, 2015).
21. Indian Nutraceutical Market, Indian nutraceutical market is filled with promise. Published online at:
http://www.naturalproductsinsider.com/articles/2014/05/indian-nutraceutical-market-is-filled-with-
promis.aspx, May 20, 2014 (accessed April 25, 2015).
22. TGA, Therapeutic goods act 1989. Published online at: http://www.comlaw.gov.au/Series/C2004A03952,
August 14, 2015 (accessed August 25, 2015).
23. TGA, Australian register of therapeutic goods. Published online at: https://www.tga.gov.au/australian-
register-therapeutic-goods, July 5, 2013 (accessed April 25, 2015).
24. The Guardian, Vitamins take Australia. Published online at: http://www.theguardian.com/world/2013/
jun/11/vitamins-take-australia-hollywood-names, June 10, 2013 (accessed April 25, 2015).
25. Food & Beverage Information, Food & Beverage Information Project 2011, Depth sector stream—
Nutraceuticals & foods for health. Published online at: https://www.med.govt.nz/sectors-industries/food-
beverage/pdf-docs-library/information-project/nutraceuticals-2011.pdf and www.foodandbeverage.govt.
nz, February 2012 (accessed April 25, 2015).
26. Bizjournals, US nutraceuticals market outlook 2018. Published online at: http://www.bizjournals.com/
prnewswire/press_releases/2014/07/16/BR70682, April 22, 2014 (accessed April 25, 2015).
3
Flavor Challenges in Functional Beverages
Keith R. Cadwallader
CONTENTS
3.1 Introduction..................................................................................................................................... 27
3.2 Flavor Perception............................................................................................................................ 28
3.3 Off Flavors Associated with Functional Ingredients...................................................................... 28
3.3.1 Off Odors............................................................................................................................ 28
3.3.2 Bitter and Astringent Substances....................................................................................... 28
3.4 Flavor Modification Techniques..................................................................................................... 28
3.4.1 Traditional Approaches...................................................................................................... 28
3.4.1.1 Odor Masking by Mixture Suppression and Odor Synergy............................... 30
3.4.1.2 Bitterness Masking by Suppression.................................................................... 30
3.4.1.3 Taste Masking by Viscosity Modification.......................................................... 30
3.4.2 Advanced Approaches........................................................................................................ 30
3.4.2.1 Taste Masking by Inclusion Complexation......................................................... 30
3.4.2.2 Bitter-Blocking Agents........................................................................................31
3.5 Masking and Flavoring of Functional Beverages............................................................................31
3.5.1 Partnering with a Flavor Company.................................................................................... 32
3.5.2 In-House Product Development......................................................................................... 32
3.5.2.1 Flavor Considerations......................................................................................... 32
3.6 Conclusion....................................................................................................................................... 33
References................................................................................................................................................. 33
3.1 Introduction
Formulated functional beverages differ from traditional beverages in that they are produced using
ingredients with scientifically proven physiological and health benefits. These products are often based
on patents, industry trade secrets, or other type of proprietary knowledge. As with any food product, the
ultimate goal is to make a product with an acceptable flavor profile that is characterized by the imme-
diate impact of an identifying flavor (e.g., vanilla, chocolate, and strawberry), rapid development of a
balanced and full-bodied flavor, compatible mouthfeel and texture, lack of off flavors, and a minimal
(short) aftertaste. It is important that when consumers first open or taste a product, their first impression
is that of the intended, desirable flavor. Functional beverages face many of the same flavor challenges
encountered with pharmaceuticals due to the inherent off flavors associated with the ingredients used
in their formulation. Products highly fortified with vitamins, minerals, and intensely bitter functional
ingredients present a particularly difficult challenge.
Several excellent reviews provide an exhaustive overview of methods for masking off flavors in phar-
maceutical products [1–5], and techniques for reducing bitterness in functional foods have recently been
published [6]. This chapter highlights traditional and emerging technologies and discusses practical
approaches to improve the flavor characteristics of functional beverages.
27
3.2 Flavor Perception

Flavor is the integrated response to the simultaneous perception of taste, odor, trigeminal, and tactile
sensations and is often influenced by visual and auditory cues perceived during food consumption [7].
Although the peripheral sensory organs for detection of taste and smell stimuli are distinct, their signals
are integrated in the orbitofrontal and other areas of the cerebral cortex of the brain to generate the
perception of “flavor” [8]. It is the complexity of flavor perception that makes it particularly challenging
to successfully modify the inherent flavor characteristics of a functional beverage to produce a highly
acceptable product.
3.3 Off Flavors Associated with Functional Ingredients

Functional beverages often contain ingredients that cause undesirable flavors (odors and tastes), which
can ultimately impact the flavor quality and consumer acceptability of the finished product. In order
to develop an effective strategy for reducing or eliminating the perception of off flavors, it is critical
that product developers know the nature of all ingredients used in the creation of the base formulation.
Of particular importance is the flavor and off flavor potential, possible interactions (flavor binding), and
process, storage, and shelf-life limitations (stability) of each functional ingredient.
3.3.1 Off Odors

Extracts made from herbs, spices, and medical plants often contain residual volatile compounds that can
cause undesirable odors in the final product. Other sources of off odors include those caused by fortification
with minerals, vitamins, omega-3 fatty acids, or healthy proteins (e.g., soy and whey). In addition, off odors
may develop from the degradation of ingredients during manufacture (thermal processing) and storage.
3.3.2 Bitter and Astringent Substances

Many functional ingredients, especially plant extractives, have the potential to cause bitterness and
astringency. These include herbal extracts containing caffeine, such as guarana, kola nut, yerba mate,
green tea, and cocoa extract enriched in theobromine and caffeine. Polyphenolics represent the largest
group of bitter and astringent substances used in functional beverages. These are derived in the form of
extracts or concentrates from plant materials, including green tea, grape (skin and seed), berry fruits,
apple (seed), soy, and citrus (peel and seed), among others. Structures of some bitter and astringent con-
stituents of functional ingredients are shown in Figure 3.1.
The tastes elicited by polyphenols can range from mainly bitter (trans-resveratrol) to being both bitter
and astringent (e.g., (+)-catechin and (−)-epicatechin). The taste properties of some phenolics depend
upon the degree of polymerization. For example, the monomeric phenols (+)-catechin and (−)-epicatechin
are perceived as more bitter than astringent, while their dimers and trimers illicit nearly equal or greater
astringency than bitterness, respectively [9]. Not all polyphenolic compounds are bitter or astringent;
occasionally, they can be sweet (e.g., neohesperidin dihydrochalcone) or tasteless (e.g., anthocyanins), in
which case the use of these functional ingredients should not cause any bitterness issues.
3.4 Flavor Modification Techniques

3.4.1 Traditional Approaches
Several strategies are commonly employed to reduce the perception of undesirable odors, tastes, and
mouthfeel characteristics of functional beverages (Table 3.1). Most involve the use of some sort of mask-
ing technology, which acts to suppress or interfere with the perception of undesirable flavors without
actually changing their concentrations in the product.
Flavor Challenges in Functional Beverages 29
OH
OH
O
HO O O
HO
O
O O
N N
N N
O OH O
N NH HO
O N N OH
O OH
Caffeine Theobromine Naringin
(bitter) (bitter) (bitter)
OH
HO OH
OH
O
OH
OH
Resveratrol
(bitter) OH
OH
OH OH
O
OH
OH OH
HO O HO O
OH OH
OH
OH OH OH
OH OH Dimeric procyanidin B8
(+)-Catechin (2R,3S) (–)-Epicatechin (2R,3R) (4α 6 catechin-epicatechin)
(bitter/astringent) (bitter/astringent) (astringent/bitter)
FIGURE 3.1 Chemical structures of some bitter and astringent constituents of functional ingredients.
TABLE 3.1
Traditional and Emerging Methods for Improving the Flavor Characteristics of Functional Beverages
Flavor Challenge Strategy Methods
Off odor reduction Odor masking Mixture suppression by addition of complex flavorings.
Assimilation masking by addition of flavorings that complement
the odors already present in base formulation.
Matrix modification Subdue or modulate odor release (availability) by addition of fat,
fat replacers, or bulking agents.
Bitterness and Congruent or assimilation Choose flavoring that complements lingering or persistent bitter
astringency masking of bitterness and and astringent tastes (e.g., coffee, tea, or dark chocolate).
reduction astringency
Mask bitterness by suppression Suppress bitterness perception by addition of NaCl, amino acids,
sugar, or high-intensity sweeteners.
Mask bitterness and astringency Addition of thickening agents (polysaccharides or gums).
by decreasing (oral) diffusion Addition of emulsifiers (lipids or lecithin).
Mask by physical separation of Encapsulation of bitter and astringent ingredients. Addition of
bitter and astringent compounds cyclodextrins.
Blocking of bitter receptors Addition of substances (bitter-blocking agents) that suppress
bitterness by interfering with bitter receptors and/or receptor
signaling pathways.
3.4.1.1 Odor Masking by Mixture Suppression and Odor Synergy

Masking of off odors can be accomplished by addition of more complex flavorings. The effect is the
result of mixture suppression, where the perceived intensity of an odorant mixture is less than that of
the individual components [10]. Assimilation masking can also be accomplished by adding flavors that
complement those already present in the product. Often, the individual flavor ingredients have limited
masking capability, but in combination provide synergy to produce a unique odor, thus enhancing the
masking effect [11]. Off odors may also be subdued by modulating or decreasing flavor release by addi-
tion of fat, fat replacers (polydextrose), and bulking agents.
3.4.1.2 Bitterness Masking by Suppression

It is possible to suppress bitterness by the addition of sugar or salt (NaCl) [12,13]. For obvious reasons,
sugar and salt are not generally used in functional beverages developed for health conscious consumers.
Instead, nonnutritive and high-intensity sweeteners (e.g., sucralose and aspartame) have found practical
application for bitterness reduction of functional ingredients and pharmaceuticals. However, the poten-
tial of these sweeteners themselves to illicit bitter and metallic aftertastes at higher use levels should be
considered during product formulation.
Conventional taste masking strategies such as use of sweeteners, amino acids, and flavoring agents
alone or in combination are often inadequate for reducing or eliminating off flavors associated with
certain functional ingredients, especially those containing intensely bitter and astringent substances.
In these cases, it may be necessary to employ more advanced techniques such as use of bitter-blocking
agents and inclusion complexes.
3.4.1.3 Taste Masking by Viscosity Modification

It is possible to reduce both the bitterness and astringency by increasing viscosity of a functional bev-
erage. This can be accomplished by the addition of thickening agents such as natural gums or car-
bohydrates or by addition of certain lipophilic substances (lipids, lecithin, and so on). The increased
viscosity acts to slow the diffusion of bitter and astringent compounds to the surface of the tongue
and oral cavity, thus reducing the perception of these substances. Chalkiness, grittiness, and other
mouthfeel characteristics may also be modified by addition of modifiers such as gums (e.g., pectin) that
provide lubricity, creaminess, and fullness. The use of natural, plant-based gums to increase viscosity
offers the additional advantage of serving as source of dietary fiber, thus potentially increasing the
nutritional value of the product.
3.4.2 Advanced Approaches

3.4.2.1 Taste Masking by Inclusion Complexation
In addition to protecting flavors, microencapsulation can be used to help mask the unpleasant sensory
characteristics of functional ingredients such as bitter herbal extracts and fishy smelling omega-3 oils.
A multitude of encapsulation technologies exist. These vary in the materials and processes employed in
their manufacture. Most produce dry, free-flowing powders designed to release their payloads under spe-
cific conditions (e.g., hydration, heating, and shearing). The use of inclusion complexation (or molecular
encapsulation) is a particularly attractive and effective method for masking of functional beverages and
is based on the molecular inclusion of an odorant or taste substance inside the cavity of another molecule.
This prevents perception of the substance during consumption. The most often used inclusion complex-
ation systems are based on the use of cyclodextrins as the host material [14].
Cyclodextrins are cyclic oligosaccharides composed of D-glucose units. There are three types of
cyclodextrin, which contain either six (α), seven (β), and eight (γ) glucose units. β-Cyclodextrin is the
most widely used complexation material due to its ability to form inclusion complexes with a variety
of molecules, especially bitter and astringent compounds, its availability, and reasonably low cost [15].
Complexation using various types of cyclodextrins has been shown to be effective in reducing or
eliminating bitterness in a variety of foods and beverages. These include the use of β-cyclodextrin to
reduce the bitterness of naringin and limonin in citrus juice [16] and use of either β- or γ-cyclodextrin
for the reduction of bitterness of ginseng solutions [17]. In addition to reducing bitterness, cyclodextrins
can alter the sensory profile through flavor encapsulation and could interfere with the action of masking
agents or bitter-blocking agents [18].
3.4.2.2 Bitter-Blocking Agents

Bitter-blocking agents function by occupying (or blocking) bitter taste receptors or signaling pathways
without initiating a sensory perception. Certain umami substances exhibit bitter-inhibiting activity. For
example, adenosine monophosphate, a naturally occurring bitter-blocking agent with generally recog-
nized as safe (GRAS) status, suppresses bitterness by interfering with the bitter receptor signaling pro-
tein gustducin. Other naturally occurring bitter-blocking agents include phosphatidic acid and tannic
acid [19] and riboflavin-binding protein (RBP) from chicken egg [20]. RBP inhibits the binding of vari-
ous bitter substances including quinine HCl, naringin, theobromine, and caffeine.
The recent elucidation of the TAS2Rs group of about 25 bitter receptors has aided the develop-
ment of more effective bitter-blocking/bitter-masking agents. In general, bitter-blocking agents have
fairly narrow bitter-blocking capabilities since they do not block all 25 receptors, but instead they are
designed to block receptors for specific bitter substances. For example, probenecid (a uricosuric drug
used primarily for treating gout and hyperuricemia) has been shown to block bitterness of salicin by
inhibiting the activation of the subset of bitter taste receptors involved in its detection [21]. Several
proprietary or patented bitter-blocking agents have been developed by flavor companies. Givaudan
flavors offer the bitter-blocking agents (GIV3727 and GIV3616), which reportedly block the bitter and
metallic tastes associated with the consumption of high-intensity artificial sweeteners. Senomyx offers
the patented (US7939671) bitter blockers (S6821 and S7958), which effectively block the bitter tastes
associated with beverages containing soy or whey proteins, or caffeine. A potential drawback to using
taste-blocking agents is that they may cause the suppression of some other desirable or characterizing
flavors. For this reason, it might be necessary to adjust or rebalance any added flavoring agents to
compensate for this change.
3.5 Masking and Flavoring of Functional Beverages

First, it is of utmost importance to have intimate knowledge about the off flavor potential of each ingredi-
ent and of the base formulation, including any potential effects of processing or storage. Product devel-
opers must not only consider the functional or bioactive aspects of the ingredients but also any negative
sensory qualities these may possess. They must work with the inherent flavor attributes of the base
formulation and not try and create an incompatible flavor. That is, the target flavor and residual flavor of
the base formation cannot be totally incongruous.
Ideally, one would consider masking or neutralizing any off-notes before attempting to apply the target
or characterizing flavoring. Sometimes, it is necessary to adjust the ingredients in the base rather than to
rely solely on the addition of masking agents and flavors. When optimizing flavor use level, it is impor-
tant to consider both the effects of any flavor interactions (flavor interactions of functional ingredients)
and other reactions that might occur during processing or storage. Especially, problematic is flavor fade
that is often encountered in high protein (soy, whey, and so on)-containing products. Flavor fade is the
perceived loss of flavor caused by the nonspecific binding of flavor compounds to the protein. It does not
occur equally among the added flavor compounds, thus in addition to a decrease in overall flavor inten-
sity, a shift in the flavor profile or flavor imbalance may result [22].
Depending upon the company’s research and development (R&D) resources and capabilities, the
product developer may elect to partner with a flavor company or develop the technology in-house.
Both approaches have several advantages and disadvantages as discussed in the following sections.
3.5.1 Partnering with a Flavor Company

Flavor companies are well adept at providing custom solutions to solve off flavor problems. They offer
products and technologies that not only serve to mask undesirable flavors but also provide a desirable
flavor in the finished product [23]. The advantage of involving a flavor company is they have the exper-
tise, resources, and know-how needed to successfully flavor even the most difficult functional beverages.
They also have enough experience to know if a flavor or masking agent will complex or react with any of
the functional ingredients. Partnering with a flavor company is especially attractive to small companies
that possess only limited R&D capabilities. An obvious disadvantage to this approach is that intellectual
property is owned and controlled by the flavor company and the client may have no knowledge about
the composition of the masking agents, flavorings, and technology used to flavor the finished product.
3.5.2 In-House Product Development

Internal R&D of masking and flavoring solutions has the key advantage in that the intellectual property
(formulation/technology) is owned and controlled by the company. Prior to formulation of the base, the
flavor and mouthfeel attributes of the individual ingredients should be identified by descriptive sensory
analysis (DSA) using trained panelists. During the initial stages of product development, it is important
to minimize the number of functional ingredients with off flavor potential, since the taste synergy from
too many ingredients may create a base that’s nearly impossible to flavor.
Any of the aforementioned masking technologies can be applied to help neutralize the off flavors of
the base. It is also possible to obtain masking agents from flavor companies. DSA should be used to aid
in the development of the masking technologies and to identify and characterize any residual odors,
tastes, and mouthfeel properties in the finished base. Ideally, masking technologies should be applied
before overlaying the base with added flavoring (mask first, then flavor). The main advantage of flavoring
after neutralizing the base is that it helps prevent the over flavoring of the product. It is important that the
target flavor be chosen such that it complements the residual flavors in the base. Certain combinations
are impractical or nearly impossible. For example, you cannot take a bitter base and expect to make an
acceptable banana flavor. Instead, it is more practical to consider a more compatible or congruent flavor,
such as cola, citrus, dark chocolate, or coffee, in which a bitter note is expected or at least tolerated to
some extent.
It is possible that the masking ingredients might also cause the suppression or modification of the
added flavorings. Therefore, it might be necessary to later rebalance or adjust the flavoring to correct for
these changes. It is also important to consider other factors that might cause flavor changes, such as flavor
binding, thermal processing (pasteurization), and storage.
3.5.2.1 Flavor Considerations

The goal of the product developer should be to produce a functional beverage that is highly acceptable
to a wide range of consumers. Some flavors may appear healthier or more wholesome to consumers.
For example, products with citrus or berry flavors might be perceived as healthier than products with
more indulgent flavors, such as vanilla and chocolate. Flavors such as chai, exotic/tropical fruit, citrus
(lemon–lime), and berry flavors work well for energy beverages, while vanilla and chocolate flavors are
more appropriate for protein-fortified beverages. Whenever feasible, complementary (congruent) flavor-
ing strategies should be used to produce the most acceptable finished product.
It is well established that olfaction can influence taste perception in both simple and complex matrices
[24]. The result of integration is product dependent and related to food experience. For this reason, it may
have either desirable or negative consequence with respect to product quality. Labbe et al. [24] showed
that olfactory–taste interactions in a cocoa beverage caused an enhancement of bitterness induced by
the cocoa flavoring and an increase in sweetness from the vanilla flavoring. However, in caffeinated
milk, the addition of vanilla flavoring did not significantly impact sweetness, but unexpectedly bitterness
perception was enhanced. The aforementioned results highlight the need for congruency in flavoring of
functional beverages.
Use of a flavoring that complements residual or lingering or persistent aromatics and tastes is referred
to as congruent or assimilation masking. An example of this approach is the use of a coffee and dark
chocolate flavoring to complement the bitter taste and astringent mouthfeel and green, beany, and cereal
aromatics associated with soy-fortified beverages. Similarly, the earthy note of St. John’s wort blends or
assimilates well with chocolate or coffee.
3.6 Conclusion
It is the ultimate goal of the product developer to provide consumers with functional beverages that not
only deliver the intended health-promoting benefits but also taste great. Various strategies can be used
in functional beverages to decrease off odors, bitter tastes, and astringent mouthfeel characteristics.
Traditional methods can be effective, but recent advances in the development of bitter-blocking agents
offer new and potentially more effective ways to inhibit bitterness. These may have particular appeal for
the targeted blocking of intensely bitter functional ingredients, especially polyphenolics that are com-
monly used in functional beverages. The use of several approaches, for example, traditional masking
strategies combined with inclusion complexation and bitter binding agents, is the most effective option
for the effective flavoring functional beverages.
REFERENCES
1. Roy, G., Modifying Bitterness: Mechanism, Ingredients, and Applications, CRC Press LLC, Boca
Raton, FL, 1997.
2. Sohi, H., Sultana, Y., and Khar, R.K., Taste masking technologies in oral pharmaceuticals: Recent
developments and approaches. Drug Dev. Ind. Pharm., 30, 429–448, 2004.
3. Ley, J.P., Masking bitter taste by molecules. Chem. Percept., 1, 58–77, 2008.
4. Sharma, S. and Lewis, S., Taste masking technologies: A review. Int. J. Pharm. Pharm. Sci., 2, 6–13,
2010.
5. Deepak, S., Dinesh, K., Mankaran, S., Gurmeet, S., and Singh, R.M., Taste masking technologies:
A novel approach for the improvement of organoleptic property of pharmaceutical active substances.
Int. Res. J. Pharm., 3, 108–116, 2012.
6. Gaudette, N. and Pickering, G.J., Modifying bitterness in functional food systems. Crit. Rev. Food Sci.
Nutr., 53, 464–481, 2013.
7. Auvray, M. and Spence, C., The multisensory perception of flavor. Conscious. Cogn., 17, 1016–1031,
2008.
8. Chaudhari, N. and Roper, S.D., The cell biology of taste. J. Cell Biol., 190, 285–296, 2010.
9. Peleg, H., Gacon, K., Schlich, P., and Noble, A.C., Bitterness and astringency of flavon-3-ol monomers,
dimers and trimers. J. Sci. Food Agric., 79, 1123–1128, 1999.
10. Cain, W.S., Odor intensity: Mixtures and masking. Chem. Senses Flavor, 1, 339–352, 1975.
11. Liang, D.G., Perceptual odour interactions and objective mixture analyses. Food Qual. Pref., 5, 75–80,
1994.
12. Calviño, A.M., García-Medina, M.R., and Cometto-Muñiz, J.E., Interactions in caffeine-sucrose and
coffee-sucrose mixtures: Evidence of taste and flavor suppression. Chem. Senses, 15, 505–519, 1990.
13. Keast, R.S.J., Breslin, P.A.S., and Beauchamp, G.K., Suppression of bitterness with sodium salts.
Chimia, 55, 441–447, 2001.
14. Szente, L. and Szejtli, J., Cyclodextrins as food ingredients. Trends Food Sci. Technol., 15, 137–142,
2004.
15. Astray, G., Gonzalez-Barreiro, C., Mejuto, J.C., Rial-Otero, R., and Simal-Gándara, J., A review on the
use of cyclodextrins in foods. Food Hydrocolloid., 23, 1631–1640, 2009.
16. Konno, A., Misaki, M., Toda, J., Wada, T., and Yasumatsu, K., Bitterness reduction of naringin and
limonin by β-cyclodextrin. Agric. Biol. Chem., 46, 2203–2208, 1982.
17. Tamamoto, L.C., Schmidt, S.J., and Lee, S.-Y., Sensory properties of ginseng solutions modified by
masking agents. J. Food Sci., 75, S341–S347, 2010.
18. Gaudette, N.J. and Pinkering, G.J., Modifying bitterness in functional food systems. Crit. Rev. Food
Sci. Nutr., 53, 464–491, 2013.
19. Nakamura, T., Tanigake, A., Miyanaga, Y., Ogawa, T., Akiyoshi, T., Matsuyama, K., and Uchida, T.,
The effect of various substances on the suppression of the bitterness of quinine-human gustatory sensa-
tion, binding, and taste sensor studies. Chem. Pharm. Bull. (Tokyo), 50, 1589–1593, 2002.
20. Maehashi, K., Matano, M., Nonaka, M., Udaka, S., and Yamamoto, Y., Riboflavin-binding protein is a
novel bitter inhibitor. Chem. Senses, 33, 57–63, 2008.
21. Green, T.A., Alarcon, S., Thomas, A., Berdougo, E., Doranz, B.J., Breslin, P.A.S., and Rucker, J.B.,
Probenecid inhibits the human bitter taste receptor TAS2R16 and suppresses bitter perception of salicin.
PLoS One, 6, e201232011, 2011.
22. Suppavorasatit, I. and Cadwallader, K.R., Flavor-soy protein interactions, in Chemistry, Texture and
Flavor of Soy, Cadwallader, K.R. and Chang, S.K.C., eds., ACS Symposium Series 1059, American
Chemical Society, Washington, DC, 2010, pp. 339–359.
23. Brantd, L.A., Flavor masking: Strategies for success. Prepared Foods, 170, 63–66, 2001.
24. Labbe, D., Damevin, L., Vaccher, C., Morgenegg, C., and Martin, N., Modulation of perceived taste by
olfaction in familiar and unfamiliar beverages. Food Qual. Prefer., 17, 582–589, 2006.
4
Chemistry of Functional Beverages
Shiming Li, Fereidoon Shahidi, and Chi-Tang Ho
CONTENTS
4.1 Introduction..................................................................................................................................... 35
4.2 Chemistry and Bioactivities of Phytochemicals............................................................................. 36
4.2.1 Polyphenols......................................................................................................................... 36
4.2.2 Flavonoids........................................................................................................................... 36
4.2.3 Terpenoids and Carotenoids............................................................................................... 36
4.2.4 Saponins............................................................................................................................. 38
4.2.5 Phytosterols........................................................................................................................ 39
4.2.6 Polysaccharides.................................................................................................................. 39
4.2.7 Alkaloids.............................................................................................................................41
4.3 Selected Functional Beverages........................................................................................................41
4.3.1 Tea...................................................................................................................................... 42
4.3.2 Coffee................................................................................................................................. 42
4.3.3 Fruit and Vegetable Beverages........................................................................................... 43
4.3.4 Energy Drinks.................................................................................................................... 44
4.4 Conclusion....................................................................................................................................... 44
References................................................................................................................................................. 45
4.1 Introduction
Functional beverages, a subsector of the functional food industry and the fastest-growing sector of the
functional food market, have become increasingly popular among conscientious consumers due to their
perceived health benefits. Convenience and health benefits are two of the most important factors when
consumers make decisions about purchasing foods and beverages. Functional beverages claim to improve
athletic endurance, energy, and hydration, and are associated with various health benefits such as gen-
eral wellness, antioxidant activity, healthy cardiovascular system, cancer prevention, healthy digestive
system, immune defense, body weight reduction, and joint health improvement, among others [1].
Apart from water, the most popular and traditional functional beverages worldwide are tea, coffee,
and fruit juices [2]. Newly developed functional drinks sometimes contain vitamins and minerals, but in
most cases they contain functional ingredients from fruits or other parts of medicinal plants, such as açai,
pomegranate, cranberry, blueberry, and monk fruits, to name a few. Hence, the phytochemical ingredi-
ents that are the building blocks of functional beverages, which provide targeted health functionality
need to be investigated and their content summarized. Bioactive phytochemicals in functional beverages
can be classified based on their chemical structures as polyphenols, including flavonoids, terpenoids,
carotenoids, saponins, phytosterols, polysaccharides, and alkaloids, among others, even though there
are some overlaps among the aforementioned classifications. This chapter highlights the most popular
beverages and their major ingredients from an array of chemical profiles.
35
4.2 Chemistry and Bioactivities of Phytochemicals

4.2.1 Polyphenols
Polyphenols, as literally indicated by their name, can have two or more hydroxyl groups bonded to the
aromatic ring(s) in the same molecule (Figure 4.1). Hydroxyl groups on phenyl rings are also termed
phenolics and they have a strong electron-donating capability due to their conjugation with the phenyl
core moiety. Hence, the phenyl ring and phenolic groups consist of an electron-rich system, which allows
the easy loss of electrons and potential for readily being oxidized. It is often judged from the structural
chemistry theory that the more hydroxyl groups a polyphenol possesses, the stronger antioxidant activity
it has, although the antioxidant activity is more often related with the locations of hydroxyl groups on a
conjugated aromatic system. Phenolics and polyphenolics exist ubiquitously in the plant kingdom and
usually have strong antioxidant activity. Major classes of phenolics and polyphenols, as illustrated in
Figure 4.1, include monobenzone core polyphenols, hydroxylated benzoic acids, hydroxylated cinnamic
acids, stilbenoids, lignans, and various flavonoids and chalcones. Specific examples of polyphenols are
gallic acid, pyrogallol, resveratrol, pterostilbene, caffeic acid, chlorogenic acid, ferulic acid, and some
flavonoids such as catechins, hesperidins, quercetins, luteolins, and anthocyanidins, among others. Most
flavonoids exist in glycosylated forms in plants [3].
Over the past two decades or so, both academic research and food industry have gained increasing
interest in the concept of polyphenols. The main reasons for the interest include the recognition of their
ubiquitous availability and abundant resource in our diet, antioxidant activity, and perhaps pivotal role
in preventing various diseases associated with oxidative stress, such as inflammation, cancer, and car-
diovascular and neurodegenerative diseases. Medicinal plants employed worldwide, although different
among various regions, are common in at least one aspect: they are rich in polyphenol content. It has
been illustrated that polyphenols can modulate a wide range of enzyme activities. Thus, the relationship
between the health benefits of protective nutrition and polyphenol intake is gradually established [3].
4.2.2 Flavonoids
Flavonoids, a particular class of polyphenols, have a C6–C3–C6 skeleton structure and consist of several
subgroups: flavones, flavonols, flavanones, flavanols, isoflavones, chalcones, anthocyanidins, and procy-
anidins. Chalcones are the only subgroup in the flavonoid category that has two phenyl groups connected
by an acryl bond, whereas the majority of other flavonoids form a C-ring with a C3 skeleton (Figure 4.1).
Flavonoids are widely distributed in fruits and vegetables such as apples, citrus, berries, and soybeans;
in grains; and in beverages such as tea, coffee, and wine. They not only show strong antioxidant activity
but also exhibit bioactivities related to anti-inflammation, risk-lowering effect of cardiovascular disease
(CVD), cancer prevention, and antiobesity. The bioactivity of flavonoids is also assumed to originate
from their perceived antioxidant property owing to multiple phenolic groups and the hydrogen bonding
interaction between proteins and functional groups on flavonoids, such as carbonyl as hydrogen acceptor
and hydroxyl groups as hydrogen donor. Examples of flavonoids are quercetins, hesperidins, luteolins,
naringins, and tannins [4].
4.2.3 Terpenoids and Carotenoids

Terpenoids, also called isoprenoids, have C5 isoprene unit(s) in common and can be assembled in numer-
ous ways (Figure 4.2). Some examples are lycopene, β-carotene, astaxanthin, citral, menthol, and cam-
phor. Terpenoids are universally present in living organisms and play vital roles in plant physiology and
serve important functions in all cellular membranes. Some terpenoids, such as retinol, play an important
role in human health as they are good antioxidants. Some terpenes have strong anticancer activity such
as the triterpene taxol [5]. Carotenoids are a subclass of terpenoids composed of eight isoprene units and
a total of 40 carbon atoms. They are natural fat-soluble pigments that are synthesized by plants and are
responsible for the bright colors of various fruits and vegetables. There are several dozen carotenoids in
Chemistry of Functional Beverages 37
Hydroxybenzoic
Polyphenols acids:
Simple OH Stilbenes:
polyphenols:
HO R RO
OH
OH HO R
HO OH
R CO2H RO
R—
— H: Catechol R—
—OH: Gallic acid R—
—H: Resveratrol
Polyphenol R—
—OH: Pyrogallol R—
—H: Procatechuic acid R—
—Me: Pterostilbene
Hydroxycinnamic acids: Lignans:

R2
R1 HO CO2H
HO OH
O
CO2H MeO
HO O OH OMe
R1—
— OH, R2—
— H: p-Coumaric acid
R1— OH HO OH
—R2—
— OH: Caffeic acid OH
—OH, R2—
R1— — OCH3: Ferullic acid Chlorogenic acid Secoisolariciresinol
Flavonoids Flavonols: Anthocyanindins:

R5
R5
OH
Flavones: R4
+
HO O
OH R3
HO
R3
HO OH
R
OH
OH
OOH —R5—
Pelargonidine: R3— —H
OH O —
Kaempferol: R4—OH, —R5—
R3— —H Cyanidin: R3—
—OH, R5—
—OH
Apigenin: R —
—H —R4—
Quercerin: R3— — OH, R5—
—H —R5 —
Delphinidin: R3— —OH
Luteolin: R —
—OH Myricetin: R3—
—H —OH, R5—
Petunidin: R3— —OMe
—R5—
Malvidin: R3— —OMe
Flavanones: Flavanols: Procyanidins:

R4 OH OH
HO OH
R3 HO O
OH
HO O
R
OH
OH O OH OH
Naringenin: R4— OH
— OH, R3—
—H OH
Eriodictyol: R3—
—R4—
— OH Catechins: R —
—H HO O
Hesperitin: R3——OH, R4—
—OMe OH
Gallocatechins: R —
—O H
OH
OH n
OH
Isoflavanones: OH
n> = 1
OH O n = 2: Dimeric procyanidin
R O HO
R n = 3: Trimeric procyanidin
......
OH
HO O OH
Daidzein: R —
—H (–)-Epicatechin: R —
—H
Genistein: R —
—OH (–)-Epigallocatechin: R —
— OH
FIGURE 4.1 Chemical structures of polyphenols and flavonoids found in tea, coffee, soy, and fruit beverages.
Terpenoids
Monoterpene:
OH Diterpene:
OH
Isoprene
trans-retinol
Linalool Limonene
Carotenoids
β–Carotene
OH
Lutein
HO
OH
Zeaxanthin
HO
Lycopene
FIGURE 4.2 Chemical structures of terpenoids and carotenoids found in some fruit beverages.
our daily dietary intake. Carotenoids act as antioxidants and are reported to have the ability of prevent-
ing chronic diseases. β-Carotene is the most common carotenoid in food matrices and exists in carrots,
apricots, tomatoes, and pumpkins, among others. The antioxidant activity of carotenoids is believed to
be responsible for the health-promoting properties of fruits and vegetables. Furthermore, the function of
carotenoids with provitamin A activity is very important for a healthy vision [5,6]. Low-dose lycopene
intake has been reported to assist cardiovascular health [6,7] and intake in high doses has been reported
to reduce symptoms of benign prostatic hyperplasia [8].
4.2.4 Saponins
Saponin, whose name is gained from its soap foaming characteristic when suspended in water, consists
of a lipophilic triterpene (C30) or steroid (C27) and hydrophilic glycosides (Figure 4.3). The foam-
ing ability of saponins is due to the combination of a hydrophobic sapogenin and a hydrophilic sugar
portion. Sapogenin is the aglycone part of a saponin. They are a class of amphipathic compounds and
abundant in various plant species. For instance, saponins can be found in most vegetables, beans, and
herbs. Daily dietary intake of saponins is estimated at 15–240 mg. Saponins have many health benefits,
such as reduction of blood cholesterol, cancer prevention, and stimulation of the immune system. Studies
have illustrated that saponins cause cholesterol reduction by preventing their reabsorption [9]. They also
exhibit antitumor activity and can lower the risk of human cancers by inhibiting the growth of cancer
cells and may also help the immune system by protecting against viruses and bacteria; some saponins
have protective effects on bone loss. Reported examples include alfalfa saponins for decreasing lipid
Saponins
Aglycones of steroidal saponins:
O OH
OH HO
O O
OH
HO HO
H H HO
Spirostanol Furostanol Cholesterol
Tetracyclic triterpenoid saponins:

β–D-Glc1-β–D-6Glc
O
OH
H
β–D-Glc1-β–D-2Glc—O
Ginenoside Rb1
FIGURE 4.3 Chemical structures of major saponins (except cholesterol) found in some fruit beverages.
and cholesterol concentration in mouse liver; ginseng saponins for reducing hypertension by blocking
the calcium channel; and Panax notoginseng saponins for inhibiting inflammation, decreasing bleeding
time, and providing protection against cancer [9].
4.2.5 Phytosterols
Phytosterols are steroid compounds naturally occurring in plants and are structurally similar to choles-
terol. They exist widely in fruits, vegetables, berries, and nuts and are rich in vegetable oils. Good food
sources include whole grains, unrefined vegetable oils, nuts, seeds, and legumes. The daily intake of
phytosterols ranges from 150 to 450 mg or even higher in some vegetarian diet. In the human diet, the
common phytosterols are β-sitosterol, campesterol, stigmasterol, sitostanol, and campestanol [10,11].
Two major classes of phytosterols are sterols and stanols. Stanols are saturated sterols and have no double
bonds in their structural ring (Figure 4.4). Major health claims of phytosterols include reduction of
plasma total cholesterol, low-density lipoprotein (LDL) cholesterol, and perhaps triacylglycerols (TAG).
Between 1954 and 1982, phytosterol was originally used as a cholesterol-lowering drug (Cytellin) in
high doses. As a functional food additive to margarine, orange juice, and others, it was introduced to the
Finnish market in 1995 for its cholesterol-lowering functionality [11] and in 2000 with the introduction
of sterol esters under the Novel Food regulation in the EU.
4.2.6 Polysaccharides
Saccharides, also called carbohydrates, are a group of biological molecules consisting of hydrogen (H),
carbon (C), and oxygen (O) atoms. The ratio of the three atoms (H–C–O) is usually 2:1:1, but the oxygen
atom could be different. Saccharides include sugar, starch, and cellulose, whereas they can also be divided
into monosaccharides, disaccharides, oligosaccharides, and polysaccharides. Monosaccharides are simple
sugars and disaccharides consist of two monosaccharides covalently linked. Polysaccharides have polymeric
carbohydrate structures, consisting of repeating units of mono- or disaccharides covalently linked by glyco-
sidic bonds (Figure 4.5). These structures are often linear, but may be branched. Polysaccharides are often
Phytosterols
Sterols:
H H H
H H H
H H H H H H
HO HO HO
β-Sitosterol Campesterol Stigmasterol
Stanols:
Cholesterol:
H H
HO
H H
H H H H OH
HO HO
H H
β-Sitostanol Campestanol HO
FIGURE 4.4 Major phytosterols (except cholesterol) found in some fruit beverages.
Polysaccharides
Monosaccharrides: Disaccharrides:
OH OH
HO O HO O
O HO O
HO HO HO
HO
HO OH OH O OH
OH HO
OH OH OH
Glucose Fructose Sucrose
OH
O
O OH
HO
OH O OH
O
HO
OH O O
OH
HO
OH O
O
HO
OH
O
n
Starch
FIGURE 4.5 Chemical structures of polysaccharides found in functional beverages.

Alkaloids
Purine derivatives: Imidazole derivatives:
O O O
H
N N HN N N N N NH2
O N N O N N O N N HN
Caffeine Theobromine Theophylline Histamine
FIGURE 4.6 Major alkaloids found in coffee and tea beverages.
heterogeneous, containing slight modifications of the repeating unit. Depending on the specific structures
of these macromolecules, they can have distinct properties depending on their monosaccharide building
blocks. They may be amorphous or insoluble in water. Natural saccharides often contributing to sweetness
are monosaccharides or disaccharides in general, such as glucose, fructose, and sucrose [12].
Digestible polysaccharides such as starch are a common source of energy. Polysaccharides that are
indigestible may have other functionalities that affect human health. For example, cellulose, chitin, and
pectin polysaccharides cannot be broken down to monosaccharides by many organisms including human
microorganisms. However, they provide a good source of dietary fiber having the functions of enhancing
digestion and reducing the absorption of cholesterol and sugars [13,14].
4.2.7 Alkaloids
Alkaloids, a class of naturally occurring organic nitrogen-containing compounds, are produced primar-
ily in plants, which can also be found in bacteria, fungi, and animals. The name alkaloid in fact came
from alkali. More than 27,000 different types of alkaloids have, so far, been identified, with 21,000 of
them are from plants [15]. They contain one or more nitrogen atoms and can be primary, secondary, and
tertiary amines. Alkaloids are usually classified based on their nitrogen-containing structures, such as
pyrrolidines, piperidines, quinolines, isoquinolines, and indoles. Traditionally, in the structure of alka-
loids, the nitrogen atom is part of the ring system, but this is not necessarily true. When the nitrogen is in
the exocyclic position in naturally occurring nitrogen compounds, they are usually classified as amines
[15,16]. Figure 4.6 lists some examples of alkaloids.
Many alkaloids possess pharmacologic effects. Most commonly used as drugs are often alkaloids from
natural sources, such as the anticancer drug taxol. Alkaloids with biological activity in humans mostly
affect the nervous system. Popular alkaloids in beverages are often purine derivatives, particularly caf-
feine, which is a stimulant of the human central nervous system, slowing down sleepiness and restoring
alertness. It is the world’s most widely used psychoactive drug. Caffeine achieves most of its effects by
blocking the activity of adenosine, a neurotransmitter affecting almost the entire body system. It also has
other reported beneficial health properties. For instance, it is a weak bronchodilator and at low doses it
is shown to provide some improvement in lung function. It is postulated that caffeine’s regulation of the
body’s neurotransmitters may also provide health benefits such as cognitive improvement, effectiveness,
and physical activity improvement and some specific therapeutic benefits such as pain relief [16].
4.3 Selected Functional Beverages

Functional beverages are nonalcoholic drinks and include in their formulation ingredients such as herbs,
vitamins, minerals, amino acids, proteins, additional raw fruits and/or vegetables, and those that claim
to provide health benefits. In addition to water, flavored, carbonated, and alcoholic drinks are not catego-
rized as functional beverages. Examples of functional beverages include tea, coffee, fruit and vegetable
beverages (apple juice, orange juice, and soy beverages), and energy drinks, among others. Detailed
information about these beverages can be found in a separate chapter of this book. Therefore, these bev-
erages are reviewed concisely here.
4.3.1 Tea
Tea (Camellia sinensis) is cultivated worldwide, particularly in China, Sri Lanka, India, Kenya, Turkey,
and some other Asian countries. It is the most consumed flavored functional beverage in the world. As a
result of tea plant variety and different delicate manufacturing processes, there are numerous tea prod-
ucts commercially available in the global market. Generally, tea has three major types: green tea, oolong
tea, and black tea. Black tea accounts for about 78% of the total worldwide tea consumption and green
tea about 20%, whereas approximately 2% belongs to oolong tea [17]. Although all types of tea have been
gaining popularity worldwide, in regional preference, green, white, and oolong tea is dominant in China
and Japan while black tea occupies the majority of the market in Western countries.
Original tea consumption was mainly for its central nerve stimulating and soothing effects, but tea
drinking has been linked to health-promoting effects for centuries. Tea consumption is associated
with many health benefits such as antioxidant and anti-inflammatory activities, cancer prevention, and
reduced risk of coronary heart disease (CHD), among others [18]. Scientific data have demonstrated that
the health effects of tea are mainly attributed to its polyphenolic compounds (Table 4.1). Tea contains
different polyphenols in terms of content and variety [17]. Green tea polyphenols, that is, catechins, are
the most abundant polyphenols in green, white, and oolong tea, and even in most of the black tea bever-
ages on the market. Polyphenols in black tea also include theaflavins, thearubigins, and other catechin
polymeric pigments that exist in higher amounts than catechins. However, the latter are still present in
black tea and its extracts in relatively large percentages because the conversion of green tea catechins to
black tea polyphenols is always incomplete [17]. Tea polyphenols in oolong tea consist chiefly of green
tea catechins and a small percentage of black tea theaflavins and thearubigins due to limited fermenta-
tion process. The polyphenolic composition of pu-erh tea or raw pu-erh tea is the same as that of oolong
tea, whereas the fully fermented pu-erh tea mainly contains gallic acid and does not contain both green
and black tea polyphenols [17]. Epidemiological evidence shows that the intake of tea polyphenols has
a myriad of beneficial health effects including antioxidant, anticancer, anti-inflammatory, antidiabetic,
antiatherosclerotic, antihyperlipidemic, antibacterial, and antiviral activities, among others [2,18].
4.3.2 Coffee
Consumption of coffee has been reported to be positively associated with reduced risk of chronic and
degenerative diseases such as cancer, diabetes, Parkinson’s disease, inflammation, and CVD [19,20].
Coffee consumption is also associated with a lower risk of a variety of liver diseases, including liver
cirrhosis and liver cancer. Coffee is brewed by infusion and/or percolation of roasted ground coffee with
boiled water. Similar to tea, coffee contains a wide range of phytochemicals represented by caffeine and
chlorogenic acids with many potential beneficial bioactivity [19,21]. Although the biological effects of
coffee are highly dependent on plant variety and processing conditions such as in blending and brewing,
which can produce wide variations in the phytochemical compositions of the resulting beverage, the
basic fingerprint profile of coffee phytochemicals remains similar [22]. It is easy to construe that caffeine
TABLE 4.1
Major Tea Polyphenols
Polyphenols No. Name Acronym R R′
Catechins I Epicatechin EC H H
II Epigallocatechin EGC H OH
III Epicatechin gallate ECG Galloyl H
IV Epigallocatechin gallate EGCG Galloyl OH
Theaflavins V Theaflavin TF1 H H
VI Theaflavin-3-monogallate TF2a Galloyl H
VII Theaflavin-3′-monogallate TF2b H Galloyl
VIII Theaflavin-3,3′-digallate TF3 Galloyl Galloyl
is the most extensively studied compound in coffee and its bioactivity has been stated in the last section.
Less abundant alkaloids theobromine and theophylline are also present in coffee [19].
The most abundant polyphenols in coffee are chlorogenic acids and the major variant is 5-caffeoylquinic
acid [21]. The concentration of chlorogenic acids in coffee can reach as high as 840 mg/L. Apart from chlo-
rogenic acids, hydroxycinnamates, including caffeic acid, ferulic acid, and p-coumaric acid (Figure 4.1),
are some of the major polyphenols found in coffee [19,23]. The antioxidant activity of chlorogenic acids
allows them to inhibit the formation or scavenging of reactive oxygen species. Thus, they may play impor-
tant roles in the prevention of certain diseases caused by oxidative stress, such as CVD. It has been
reported that chlorogenic acids exert inhibitory effects on carcinogenesis in the large intestine, liver, and
tongue and a protective action on oxidative stress in vivo. Chlorogenic acids may also have neuropro-
tective effects. An animal feeding study with chlorogenic acids and caffeic acids found the absorption
of phenolic acids and the suppressed expression of P-selectin on mouse platelets, indicating significant
protective effect against CVD by the two phenolic acids in coffee. Chlorogenic acids were also found to
improve glucose tolerance and decrease the levels of cholesterol and TAG in rat plasma and liver [19,21].
Chlorogenic acids are one of the most abundant polyphenols in the human diet with coffee, fruits, and
vegetables as its major sources. For instance, they are an important group of nonvolatile compounds
in green coffee beans. Although 30 different species of chlorogenic acids have now been identified in
green beans, the vast majority of the compounds found belong to three classes: monocaffeoylquinic
acids at 3-, 4-, or 5-position of quinic acids, dicaffeoylquinic acids, and feruloylquinic acids. In addi-
tion to coffee, these compounds are also found at significant levels in plant foods such as apples, pears,
tomatoes, potatoes, eggplants, strawberries, pineapples, sunflowers, and blueberries. A small quantity
of free quinic acid occurs in green coffee beans. A greater quantity of quinic acid occurs as a series
of chlorogenic acids esters. They are a family of esters formed between trans-cinnamic acids (caffeic,
coumaric, and ferulic acids) and (−)-quinic acid. Chlorogenic acids are also found as a significant
component in some commonly used medicinal herbs, including chrysanthemum flower, hawthorn fruit,
artemisia leaves, epimedium leaves, artichoke leaves, burdock root, dandelion root, and echinacea root,
among others.
4.3.3 Fruit and Vegetable Beverages

Juices are often perceived as healthy drinks by consumers because they have been marketed as a
healthy, natural source of vitamins, minerals, and antioxidants. For example, the American Academy of
Pediatrics’ guidelines include the consumption of 100% fruit juice in moderate amounts, for example,
120–180 mL/day for children ages 1–6 years and 240–360 mL/day for older children [24]. Fruit juices
can help children get the nutrients they need and help them meet fruit intake recommendations. In the
United States, fruit juices can only legally be used to describe a product that consists of 100% fruit juice.
There are many phytochemicals that contribute to the health-promoting properties of fruit juices apart
from its high sugar and vitamin C content. Some examples of fruit juices are orange juice, apple juice,
and soy beverages.
Pure orange juice provides a variety of vitamins and minerals without fat and cholesterol. It is one of
the most popular healthy beverages. A few of the important well-known nutrients that make orange juice
one of the most naturally healthy beverages are its high content of vitamin C, folic acid, thiamin, cal-
cium, and potassium. In addition, polyphenolic compounds such as various flavonoids have been found
to be essential to claim orange juice as superior over other beverages in terms of nutraceutical and health-
promoting usefulness. The high content of flavonoids include hesperidin, neohesperidin, and narirutin.
Another important series of phytochemicals in orange juice is terpenoids, which include essential oils
and carotenoids [25].
Apple and apple beverages (apple juice and apple cider) are rich in antioxidants, particularly large
amount of polyphenols including flavonoids and phenolic acids. Polyphenols present include querce-
tin glycosides, procyanidins, epicatechins, chlorogenic acids, and phloretin glycosides, along with vita-
min C [26]. Other functional phytochemicals in apple and apple beverages also include gallic acid,
catechin, kaempferol, myricetin, cyanidin glycosides, coumaric acid, triterpenoids, and vitamin B,
among others [27]. Concentrations of hydroxycinnamic acids range from 57 to 259 mg/L in commercial
TABLE 4.2
Phytonutrients in Food Sources and Claimed Health Benefits
Phytonutrients Health Benefits Claimed Fruits and Vegetables
Allicin and allylic sulfides Antibacterial, antifungal, antiviral, and Chives, garlic, leeks, and onions
antioxidant activities; lowering the risk
of stomach and colon cancers.
Anthocyanidins and Strong antioxidants; maintaining Dark grapes, berries, cherries, and red wines
proanthocyanidins elasticity of capillary walls; anti-
inflammatory; inhibiting cancer cell
formation and proliferation.
Flavonoids (quercetin, Potent antioxidants; anticarcinogenic; Teas, citrus fruits, berries, cherries, apples,
kaempferol, hesperidin, anti-inflammatory; lowering the risk of grapes, papayas, cantaloupes, plums,
naringinin, CVD; decreasing fat absorption; tomatoes, apricots, beans, cocoa beans,
neohesperidin, and their increasing energy expenditure; broccolis, parsleys, celeries, onions, and soy
glycoside) neuroprotective effects. products
Carotenoids (α- and Important antiaging and antioxidants; Carrots, sweet potatoes, all berries, citrus peels,
β-carotenes, lycopene, enhancing immune function; balancing watercress, pumpkins, tomatoes, watermelons,
and lutein) blood sugars; reducing the risk of and dark green leafy vegetables
CVD and cancer.
Coumarins Antioxidant, anti-inflammatory, Blackberries, cranberries, raspberries,
antitumor, antimicrobial, and antiviral strawberries, cherries, grapes, black currants,
activities; neuroprotective effects. and apricots
Glucosinolates Reducing the risk of breast, colorectal, Cabbage family vegetables, such as broccolies,
lung, and stomach cancers. Brussels sprouts, collards, and kales
Phytosterols Blocking cholesterol uptake and thereby Most plants
preventing CHD.
Abbreviations: CVD, cardiovascular disease; CHD, coronary heart disease.
apple juice and the number is even higher for fresh apple juice. Flavonoid concentration is between 27
and 593 mg/L with fresh juice having higher values. The total polyphenols in commercial apple juice and
fresh apple juice are 110–459 mg/L and 154–970 mg/L, respectively [26,27].
Many reports about the health benefits of soy and soy products including soy milk and soy drinks
have emerged. Soy and its products are protein rich and have a high content of fat with moderate car-
bohydrates. They are rich in isoflavones (genistein, daidzein, and glycitein), saponins, β-sitosterol, and
lecithin [28,29]. Soy product consumption is associated with a reduction of TAG and LDL cholesterol,
reducing the risk of CVD [30]. Soy isoflavones help to ease menopause symptoms and reduce certain
cancer risks, including those of the breast and prostate [29]. There are many other phytochemicals in soy
products that are associated with certain health benefits [28,30].
4.3.4 Energy Drinks

Energy drinks usually have three major phytonutrients, namely, caffeine, vitamin B complex, and tau-
rine. Taurine is an amino acid that exists naturally in the body. It is found in meat, fish, and breast milk
and is also available as a dietary supplement. Some studies suggest that taurine has antioxidant prop-
erties. Its supplementation may improve athletic performance and when combined with caffeine may
improve mental performance. However, these findings remain controversial [31] (Table 4.2).
4.4 Conclusion
Functional beverages have been dramatically gaining consumer interests and market shares. Due to
the extensive research and product exploration, vast scientific data for the correlation between phyto-
chemicals and biological activities have been gathered from the combined fields of natural products,
biochemistry, molecular biology, and nutrigenomics. Phytochemicals and their beneficial health effects
have rapidly changed traditional consumption patterns. For example, ready-to-drink teas, exotic blends
of fruit juices, antioxidant-rich “superfruits,” combinations of fruit and vegetable juices, and smoothies
continue to expand market territories and to gain impressive sales. Another nontraditional trend is that
many companies are harnessing the power and potency of vegetables due to their phytochemical-rich
and less-carbohydrate contents. In addition, the evolution of drinks with vitamins, minerals, and other
functional ingredients, particularly non-water-soluble nutrients, has led to the application of nanoemul-
sion and microencapsulation, which enables the introduction of hydrophobic ingredients into the aqueous
media and hence the development of water-soluble beverages rich in vitamins A, D, and E, omega-3 fatty
acids, and CoQ10, among others. The concepts of clear protein beverage and sports drinks have also led
to the discovery of whey protein and its extraction technology.
Developing functional beverages with effective health benefits such as disease risk reduction should be
the top priority among many aspects as appearance, flavor, and stability, which are also practical chal-
lenges to satisfy health conscientious customers. Biofunctionality of functional beverages with health-
promoting properties has been focused on preventing oxidation, maintaining cardiovascular health,
improving cognitive function and nutrition, lowering LDL cholesterol, decreasing insulin resistance and
preventing diabetes, reducing body fat, nurturing healthy bone and joints, and preventing cancer. With
the unequivocal and strong science-based data support, functional beverages will certainly grow sub-
stantially in the near future.
REFERENCES
1. Adams, R., Formulating functional beverages, food product design, July 2014. Published online at: http://
www.foodproductdesign.com/blogs/formulating-foods/2014/07/formulating-functional-beverages.aspx
(accessed January 26, 2015).
2. Crespy, V. and Williamso, G., A review of the health effects of green tea catechins in in vivo animal
models. J. Nutr., 2004, 134(Suppl.), 3431S–3440S.
3. Quideau, S., Deffieux, D., Douat-Casassus, C., and Pouysegu, L., Plant polyphenols: Chemical proper-
ties, biological activities, and synthesis. Agew. Chem. Int. Ed., 50, 586–621, 2011.
4. Manach, C., Scalbert, A., Morand, C., Rémésy, C., and Jiménez. L., Polyphenols: Food sources and
bioavailability. Am. J. Clin. Nutr., 2004, 79, 727–747.
5. Gershenzon, J. and Dudareva, N., The function of terpene natural products in the natural world.
Nat. Chem. Biol., 2007, 3, 408–414.
6. Tanmay, H., Anamika, D., and Aparnathi, K.D., Lycopene: A phytochemical with nutraceutical poten-
tial. J. Food Sci. Technol., 3, 16–22, 2014.
7. Burton-Freeman, B.M. and Howard, D., Whole food versus supplement: Comparing the clinical
evidence of tomato intake and lycopene supplementation on cardiovascular risk factors. Adv. Nutr.,
5, 457–485, 2014.
8. Pagano, E., Laudato, M., Griffo, M., and Capasso, R., Phytotherapy of benign hyperplasia: A mini-
review. Phytother. Res., 28, 949–955, 2014.
9. Sparg, S.G., Light, M.E., and van Staden, J., Biological activities and distribution of plant saponins.
J. Ethnopharmacol., 94, 219–243, 2004.
10. Ågren, J.J., Tvrzicka, E., Nenonen, M.T., Helve, T., and Hänninen, O., Divergent changes in serum ste-
rols during a strict uncooked vegan diet in patients with rheumatoid arthritis. Br. J. Nutr., 85, 137–139,
2007.
11. Fransen, H.P., de Jong, N., Wolfs, M., Verhagen, H., Verschuren, W.M., Lutjohann, D., von Bergmann, K.,
Plat, J., and Mensink, R.P., Customary use of plant sterol and plant stanol enriched margarine is asso-
ciated with changes in serum sterol and stanol concentrations in the human. J. Nutr., 137, 1301–1306,
2007.
12. Fennema, O.R., Food Chemistry, 3rd edn., Marcel Dekker, New York, 1996.
13. Eastwood, M. and Kritchevsky, D., Dietary fiber: How did we get where we are? Annu. Rev. Nutr., 25,
1–8, 2005.
14. Anderson, J.W., Baird, P., Davis, R.H., Ferreri, S., Knudtson, M., Koraym, A., Waters, V., and Williams,
C.L., Health benefits of dietary fiber. Nutr. Rev., 67, 188–205, 2009.
15. Manfred, H., Alkaloids: Nature’s Curse or Blessing? Wiley-VCH, Weinheim, Germany, 2002.
16. Dewick P.M., Medicinal Natural Products, 3rd edn., Wiley-Blackwell, Oxford, UK, 2009.
17. Li, S., Lo, C.Y., Pan, M.H., Lai, C.S., and Ho, C.-T., Black tea: Chemical analysis and stability. Food
Funct., 4, 10–18, 2012.
18. Pan, M.-H., Lai, C.-S., Wang, H., Lo, C.-Y., Ho, C.-T., and Li, S., Black tea in chemo-prevention of can-
cer and other human diseases. Food Sci. Hum. Well., 2, 12–21, 2013.
19. Ludwig, I.A., Clifford, M.N., Lean, M.E.J., Ashihara, H., and Crozier A., Coffee: Biochemistry and
potential impact on health. Food Funct., 5, 1695–1717, 2014.
20. Carman, A.J., Dacks, P.A., Lane, R.F., Shineman, D.E., and Fillit, H.M., Current evidence for the use
of coffee and caffeine to prevent age-related cognitive decline and Alzheimer’s disease. J. Nutr. Health
Aging, 18, 383–392, 2014.
21. Upadhyay, R. and Rao, J.M., An outlook on chlorogenic acids: Occurrence, chemistry, technology, and
biological activities. Crit. Rev. Food Sci. Nutr., 53, 968–984, 2013.
22. Lopez-Garcia, E., van Dam, C.M., Li, T.Y., Rodriguez-Artalejo, F., and Hu, F.B., The relationship of
coffee consumption with mortality. Ann. Intern. Med., 148, 904–914, 2008.
23. Mussatto, S.I., Machado, E.M.S., Martins, S., and Teixera, J.A., Production, composition and applica-
tion of coffee and its industrial residues. Food Bioprocess Technol., 4, 661–672, 2011.
24. Baker, S.S., Cochran, W.J., Greer, F.R., Heyman, M.B., Jacobson, M.S., Jaksic, T., and Krebs, N.F., The
use and misuse of fruit juice in pediatrics. Pediatrics, 107, 1210–1213, 2001.
25. Gattuso, G., Barreca, D., Gargiulli, G., Leuzzi, U., and Caristi, C., Flavonoid composition of citrus
juices. Molecules, 12, 1641–1673, 2007.
26. Hyson, A., A comprehensive review of apples and apple components and their relationship to human
health. Adv. Nutr., 2, 408–420, 2011.
27. Gerhauser, C., Cancer chemopreventive potential of apples, apple juice, and apple components. Planta
Med., 74, 1608–1624, 2008.
28. Wu, X. and Kang, J., Phytochemicals in soy and their health effects, in Phytochemicals—Bioactivities
and Impact on Health, Rasooli, I., Ed., Intech, Rijeka, Croatia, 2011, pp. 43–76.
29. Pudenz, M. and Roth, K., Impact on the epigenome in cancer prevention. Nutrients, 6, 4218–4272, 2014.
30. Tripathi, M.K., Kumar, V., Yadav, M.K., Yadav, D., and Pandey, S., Beneficial role of soybean phytoes-
trogens. Octa J. Biosci., 1, 170–176, 2013.
31. Ripps, H. and Shen, W., Review—Taurine: A “very essential” amino acid. Mol. Vis., 18, 2673–2686,
2012.
5
Cancer Chemopreventive Effects
of Selected Fruit Juices
Joydeb Kumar Kundu, Kyung-Soo Chun, and Juthika Kundu
CONTENTS
5.1 Introduction..................................................................................................................................... 47
5.2 Combination Approach of Cancer Chemoprevention: Fruit Juice as a Gold Standard.................. 48
5.3 Cancer Chemopreventive Effects of Selected Fruit Juices............................................................. 50
5.3.1 Apple Juice..........................................................................................................................51
5.3.2 Berry Juices........................................................................................................................ 55
5.3.2.1 Chokeberry......................................................................................................... 56
5.3.2.2 Cranberry............................................................................................................ 56
5.3.2.3 Other Berry Juices.............................................................................................. 57
5.3.3 Cherry Juice........................................................................................................................ 57
5.3.4 Pomegranate Juice.............................................................................................................. 57
5.3.5 Mango Juice........................................................................................................................ 58
5.3.6 Noni Juice........................................................................................................................... 58
5.3.7 Tomato Juice....................................................................................................................... 59
5.3.8 Citrus Juices........................................................................................................................ 59
5.3.9 Miscellaneous Fruit Juices................................................................................................. 60
5.4 Conclusion........................................................................................................................................61
Acknowledgments......................................................................................................................................61
References..................................................................................................................................................61
5.1 Introduction
Despite the remarkable progress in developing a wide spectrum of anticancer therapies, cancer still
remains as one of the major global health challenges. Cancer develops through a multistep process
involving multiple gene mutations caused by various lifestyle factors, such as exposure to dietary
carcinogens and solar radiation, smoking, and increased alcohol intake, among others. The multistage
carcinogenesis apparently involves three distinct phases: initiation, promotion, and progression [1,2].
The tumor initiation phase is characterized by irreversible changes in cellular DNA by genotoxic
carcinogens that lead to the neoplastic cell transformation. Tumor promotion is a reversible process
when the initiated tumor cells undergo clonal expansion to form a benign tumor. During the promotion
stage, aberrant alterations in cellular biochemical networks result in the increased proliferation and
neovascularization of the growing tumor. The final stage of carcinogenesis is the tumor progression
phase, when cells from the localized solid tumor lose their adhesion properties and attain migratory
properties. At this stage, cancer cells gain motility and invade through the host stromal tissue and
disseminate to distant organs to form metastatic tumors [1,2]. It is now well accepted that cancer
is a preventable disease because many of the cancer-causing lifestyle factors are simply modifiable
and the biochemical changes occurring during tumor promotion are reversible. About 30%–40% of
47
cancers can be prevented by appropriate dietary habits and lifestyle modifications. Because oxidative
stress and inflammation play key roles in all phases of carcinogenesis by causing oxidative or covalent
modification of cellular macromolecules, and activation of oncogenic signal transduction pathways,
substances with antioxidative and anti-inflammatory properties are considered to be effective in pre-
venting cancer. Since the introduction of the concept of cancer prevention, termed as “chemopreven-
tion” in the 1970s [3], numerous studies have demonstrated that antioxidant and anti-inflammatory
plant constituents are effective in preventing carcinogenesis [4,5]. The biochemical basis of cancer
chemoprevention with a wide variety of structurally diverse plant metabolites, also known as phyto-
chemicals, includes inhibition of carcinogen activation and oxidative damage of cellular macromol-
ecules, suppression of inflammatory responses, induction of growth arrest and apoptosis in cancer
cells, inhibition of tumor growth by blocking angiogenesis, and the blockade of invasion, migration,
and metastasis of cancer [4,5]. This chapter highlights the potential of selected fruit juices in cancer
chemoprevention.
5.2 Combination Approach of Cancer Chemoprevention:

Fruit Juice as a Gold Standard
Multiple lines of epidemiological and preclinical studies suggest that an inverse correlation exists
between the regular consumption of fruits or fruit juices and the risk of various organ-specific cancers
[6,7]. The European Prospective Investigation into Cancer and Nutrition (EPIC) study demonstrates that
regular consumption of fruits can reduce the risk of certain cancers [8,9]. Meta-analysis of eight cohort
studies has also indicated the lung cancer–preventing effects of fruits [10]. Common fruits that exhibit
cancer chemopreventive effects in various preclinical and clinical studies include, but are not limited
to, apples, apricots, avocadoes, different types of berries, citrus fruits, pomegranate, grapes, mangoes,
mangosteens, prunes, plums, and persimmons, among others. In addition to the extensive research on
the anticancer effects of many dried fruits and their bioactive constituents [11], there has been a wide
spectrum of fascinating reports of cancer chemoprevention with different fruit juices [12–16].
Naturally, fruit juice contains a variable number of chemopreventive phytochemicals (Figure 5.1),
which often have synergistic effects. For example, pomegranate juice showed the greatest antiprolifera-
tive activity in human oral, colon, and prostate cancer cells as compared to the effects of its individual
components, such as punicalagin, ellagic acid, and total pomegranate tannins [17]. Likewise, a combi-
nation of pomegranate juice components, such as luteolin, ellagic acid, and punicic acid, reduced the
migratory and chemotactic properties of human breast cancer cells [18]. Another major constituent of
pomegranate juice is ellagitannin, which is hydrolyzed first into ellagic acid and subsequently converted
into urolithin-A through the metabolism of ellagic acid by colonic microflora. Treatment with a combi-
nation of ellagic acid and urolithin-A synergistically inhibited the proliferation and induced apoptosis
in human prostate cancer (DU-145 and PC3) cells [19]. Schaefer et al. [20] examined the effects of a
reconstituted mixture of rutin, phloridzin, chlorogenic acid, caffeic acid, and epicatechin, five major
constituents of apple juice extract, on oxidative DNA damage in human colon cancer (Caco-2 and HT-29)
cells. According to this study, the reconstituted mixture showed higher trolox equivalent antioxidant
capacity (TEAC) and was more effective in preventing menadione-induced oxidative DNA damage as
compared to the original juice extract. Likewise, the whole orange fruit juice elicited more potent free
radical and superoxide anion scavenging activity as compared to its different polyphenol fractions [21].
In a rat colon carcinogenesis model, daily consumption of cloudy apple juice or its fractions, such as
polyphenol fraction or cloud fraction or the combination of polyphenol and cloud fraction for 7 weeks
starting 1 week prior to challenge with 1,2-dimethylhydrazine (DMH), showed that DMH-induced geno-
toxicity was significantly attenuated by cloudy apple juice as a whole, but not with the polyphenol or
cloud fractions or their combinations [22]. As compared to the polyphenol or cloud fractions, drinking
of the cloudy apple juice exhibited the most significant inhibition of DMH-induced proliferation of colo-
nocytes. Moreover, cloudy apple juice, but not the fractions, reduced the number of large aberrant crypt
foci (ACF) formation in DMH-challenged rats [22].
Cancer Chemopreventive Effects of Selected Fruit Juices 49
Since cancer is caused by mutations of multiple genes and involves perturbation of diverse oncogenic
signaling pathways, the use of a combination of multitargeted naturally occurring antioxidant and anti-
inflammatory phytochemicals would be a rational approach for chemoprevention [23]. The approach
of “combination chemoprevention” proposed by Sporn [23] has gained the experimental proof through
several studies where simultaneous administration of different chemopreventive phytochemicals showed
better anticancer effects than the individual compounds. For example, treatment of mouse skin with a
combination of pomegranate fruit extract with diallyl sulfide derived from garlic showed the most potent
inhibitory effects on the inhibition of chemically induced mouse skin tumorigenesis as compared to that
elicited by treatment with pomegranate fruit extract or diallyl sulfide alone [24]. Likewise, a pomegran-
ate fruit juice component, ellagic acid, when co-treated with grape seed extract or resveratrol exhibited
maximum inhibition of chemically induced skin inflammation and tumorigenesis [25]. Zessner et al. [26]
reported the differential antioxidant and anti-inflammatory effects of apple juice constituents. According
to their study, low molecular weight (LMW) polyphenols (chlorogenic acid, flavan-3-ols, and flavonols)
and procyanidins showed 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging potential, whereas
peroxyl radical was more effectively scavenged by LMW polyphenols than procyanidins. This study
also demonstrated that among the juice constituents, quercetin aglycone was an inhibitor of carcinogen
metabolizing enzyme, cytochrome p450 (CYP)-1A, whereas phloretin and (−)-epicatechin were the most
potent inhibitors of cyclooxygenase (COX)-1. These findings suggest that fruit juices enriched with a
wide variety of polyphenolic compounds, which often work in synergy, would be the appropriate natural
formulations for multitarget-based chemoprevention of cancer.
O OH
OH HO O OH
HO
O O
HO OH
HO O OH
OH O
HO OH
OH O OH OH
Garcinone Gartanin Mangiferin
O
β-D-glucose
COO–
HO
H3C
H3C CH3 CH3 CH3
HOOC N COOH
CH3 CH3 H3C CH3
HO CH3 H
β-Cryptoxanthin Betanin
H3C CH3
CH3 CH3 CH3
OH O
CH3 CH3 CH3 O CH3
H3C CH3 HO O OH
Lycopene Mangostin
FIGURE 5.1 Chemical structures of selected chemopreventive phytochemicals present in fruit juices. (Continued)
OH
OH OH
+
O O
OH
HO OH
OH OH
O OH
HO O OH OH OH O
OH O HO OH
OH OH OH
OH OH OH OH
Epicatechin Cyanidin-3-glucoside Procyanidin B2
OH O
OH HO CO2H
HO
O O
HO O OH
HO O HO
OH OH O
OH OH
OH O OH O
Quercetin Chlorogenic acid Ellagic acid
OH
HO O
O
O O O
HO O
OH HO OH
OH OH O
Hespiridin
FIGURE 5.1 (Continued) Chemical structures of selected chemopreventive phytochemicals present in fruit juices.
5.3 Cancer Chemopreventive Effects of Selected Fruit Juices

The anticancer effects of various fruits, fruit extracts, fruit juices, and the isolated fruit-derived
phytochemicals have been extensively investigated. Many of the chemoprevention research with fruit
extracts includes the effects of organic extract of fruits, fruit peels, as well as the partially purified frac-
tions of whole fruit extracts. Moreover, molecular mechanisms of cancer chemoprevention with phyto-
chemicals commonly present in different fruits and fruit juices have been studied in detail. Since the
scope of this chapter is to focus on the chemopreventive potential of fruit juices, studies with organic
extracts or fractions of fruits have been excluded. It has been well documented that fruit juices can
elicit cancer chemoprevention activities by virtue of their antioxidant, antigenotoxic, anti-inflammatory,
antiproliferative, apoptosis inducing, and antiangiogenic properties (Figure 5.2). Several fruit juices
have been reported to reduce the genotoxicity of certain carcinogens. Platt et al. [27] evaluated the
effects of juices from 15 fruits on the genotoxicity caused by heterocyclic aromatic amines in genetically
engineered V79 hamster fibroblasts overexpressing human CYP enzymes, such as hCYP1A2 or human
N(O)-acetyltransferase (hNAT)-2*4 and human sulfotransferase (hSULT)1A1*1, which are responsible
for the metabolic activation of 2-amino-3-methylimidazo(4,5-f)quinoline (IQ) and 2-amino-1-methyl-6-
phenylimidazo(4,5-b)pyridine (PhIP). According to this study, sweet cherry juice exhibited the highest
inhibitory effect on IQ-induced genotoxicity, followed by juices from kiwi fruit, plum, and blueberry.
Juices from watermelon, blackberry, strawberry, black currant, and red delicious apple showed moderate
Antiproliferative
effects Apoptosis-
Antimutagenic/
Antigenotoxic effects inducing effects
Antioxidant Antiangiogenic
effects and
Antimetastatic
effects
Fruit
juices
FIGURE 5.2 Biochemical basis of cancer prevention with fruit juices.
suppression, whereas sour cherry, grapefruit, red currant, and pineapple juices were only weakly active
in blocking IQ-mediated genotoxic effects. On the other hand, the inhibition of PhIP-mediated genotox-
icity by these fruit juices was less prominent than their suppressive effect on the IQ-induced genotoxicity
[27]. Administration of several fruit juices has been shown to inhibit experimentally induced carcinogen-
esis in various animal models (Table 5.1). For example, drinking of pomegranate juice, cranberry juice,
and watermelon juice as 20% fruit juice preparation significantly diminished azoxymethane (AOM)-
induced ACF formation and increased the total glutathione-S-transferase (GST) activity in male Fischer
344 rats [28]. The following section will shed light on the antioxidant, anti-inflammatory, and cancer
chemopreventive effects of fruit juices and their underlying molecular mechanisms.
5.3.1 Apple Juice

In addition to their nutritive value, apple juice has been reported to prevent carcinogenesis. Apple juice
contains a number of polyphenolic compounds, such as hydroxycinnamic acids (chlorogenic acid and
p-coumaroylquinic acid), flavan-3-ols (procyanidin B2, procyanidin C1, and epicatechins), flavonols
(quercetin-3-glycoside), and dihydrochalcones (phlorizin and phloretin-2′-xyloglucoside). The total
polyphenol content of freshly prepared apple juice is much higher than the commercially available apple
juice [16,29]. Incubation of human lung epithelial (A549) cells with apple juice significantly inhibited
Cr(VI)-induced lipid peroxidation, DNA damage, and the activation of nuclear factor-kappaB (NF-κB),
suggesting the chemopreventive potential of apple juice [30]. The antioxidant activity of polyphenol-rich
apple juice is partly mediated through the activation of nuclear factor erythroid-related factor-2 (Nrf2)
and induction of a variety of cytoprotective enzymes [29]. According to a recent study, male rats were
allowed to drink polyphenol-rich or a polyphenol-free smoothie apple juice daily for 10 days followed
by a 4-day washout period, and the treatment was repeated for 4 cycles. After termination of the treat-
ment period, analysis of cytoprotective gene expression in rat colon and liver showed that the mRNA
expression of the redox-sensitive transcription factor Nrf2 and its target cytoprotective genes, such as
catalase, glutathione peroxidase (GPX)-2, and glutathione reductase (GR), was significantly induced
in colons of rats receiving clear or cloudy apple juice as compared to those receiving polyphenol-free
apple juice. Moreover, the gene expression of superoxide dismutase (SOD)-1, γ-glutamyl cysteine ligase
(GCL)-modulatory (GCLM) subunit, and NAD(P)H-quinoneoxidoreductase-1 (NQO1) was slightly
increased in the colon upon consumption of polyphenol-rich apple juice. Apple juice intake significantly
increased the liver-specific mRNA levels of GPX1 and NQO1 without affecting the expression of other
52
TABLE 5.1
Anticancer Effects of Fruit Juices
Fruit Juices Experimental Model Dose and Route Experimental Findings References
Apple Juice
Cloudy apple juice (21.5 mL/ DMH-induced ACF formation Drinking for 8 weeks; 1 week after Decreases genotoxicity, colonic epithelial cell proliferation, [13]
animal/day) and clear apple in male Fischer 344 rats juice intake, DMH was given i.p. and ACF formation; reduced COX-2 mRNA expression;
juice (22.9 mL/animal/day) once per week for 4 times and rats and increases antioxidant activity.
were sacrificed 3 weeks after the last
dose of DMH.
Cloudy apple juice, PF, or CF DMH-induced ACF formation Drinking for 8 weeks; 1 week after Only cloud juice, but not CF or PF, reduces DMH [22]
of apple juice in male Fischer 344 rats start of cloudy juice, PF, and CF, genotoxicity and DMH-induced colonic cell proliferation,
DMH was given i.p. once per week and increases antioxidant activity.
for 4 times and rats were sacrificed 3
weeks after the last AOM dose.
Procyanidin-rich fraction of SW620 colon cancer cells Incubation of cells at a concentration Inhibits cell proliferation, induces G2/M phase cell cycle [33]
apple juice of 50 μg/mL. arrest, increases caspase-3 activity, and reduces the
activity of ODC and PKC.
Rats challenged with AOM Daily drinking for 6 weeks post-AOM. Decreases AOM-induced colonic ACF formation. [33]
Berry Juices
Fruit juice plus pulp of black Rats treated with aminopyrene Daily for 3 days by gavage; AN Reduces formation of nitrosamine and inhibits liver [40]
chokeberry (AN) plus sodium nitrite in the (1 mL/100 g body weight). dystrophy.
presence or absence of AN
Polyphenol-rich chokeberry Caco-2 colon cancer cells Incubation of cells with 2% or 5% Decreases cell proliferation, and induces G2/M phase cell [41]
juice juice. cycle arrest, increases gene expression of CEACAM1 and
integrin-α2, and reduces S100A4 mRNA.
Cranberry juice concentrate Rats treated with bladder Juice concentrate (0.5 or 1 mL/rat/day) Inhibits urinary bladder hyperplasia, papilloma, and [48]
carcinogen OH-BBN given by gavage for 37 days, started carcinoma formation.
1 week after OH-BBN challenge.
Pure cranberry juice AOM-treated rats Juice (20%) in drinking water daily for Reduces the number of ACF in proximal and distal colon, [28]
17 weeks. decreases the number of crypts per ACF, and increases
liver GST activity.
(Continued)
Handbook of Functional Beverages and Human Health
TABLE 5.1 (Continued)
Freshly prepared juices of Colon cancer (Caco-2), prostate Incubation of cells, with juice 50 μL/mL Inhibits cell proliferation, increases caspase-3 activity, and [14]
raspberry, black currant, red cancer (PC3), gastric cancer for 48 h. reduces the constitutive level of cyclin-D1 and cyclin-D3
currant, white currant, goose (AGS), and breast cancer and Cdk-4 and Cdk-6.
berry, cranberry, blueberry, (MCF-7, MDA-MB-231) cells PC3 cells stimulated with or without Inhibits TNFα-induced COX-2 expression and NF-κB
and sea buckthorn TNFα were treated with juice activity.
(25 μL/mL).
Black currant juice Ehrlich tumor–bearing mice Daily for 21 days at a dose of 10 mL/kg. Inhibits tumor weight by 45%. [55]
Tart cherry fruit juice Breast cancer (MCF-7) cells Incubation with 10% or 30% (v/v) Inhibits cell proliferation and decreases bromodeoxyuridine [57]
juice. incorporation by 20%.
Pomegranate Juice
Pure pomegranate juice AOM-treated rats Juice (20%) in drinking water daily for Reduces the number of ACF in proximal and distal colon, [28]
17 weeks. decreases the number of crypts per ACF, and increases
liver GST activity.
Fresh pomegranate juice AOM-treated rats Drinking daily for 9 weeks at a dose of Decreases the number of colonic ACF; reduces [12]
57.21 mL/day (438.95 mg GAE/kg/ proliferation of colon epithelial cells; suppresses iNOS
Cancer Chemopreventive Effects of Selected Fruit Juices
day). and COX-2 protein and mRNA expression; inhibits

phosphorylation of Akt, PI3K, and mTOR; and increases
caspase activation and the level of mir126.
Colon cancer (HT29) cells Incubation with 50 μg/mL juice. Induces cytotoxicity and inhibits TNFα-induced activation [62]
of Akt and NF-κB.
Mango Juice
Whole mango juice Antioxidant activity assay; B[α] BALB/c 3T3 cells were incubated Trolox equivalents (3.41± 0.13 μM), GAE 509.1 ± 15.4 [70]
P-induced BALB/c 3T3 cell with 0.1% juice; HL-60 cells were μg/g, ascorbic acid equivalents 650.1 ± 5.3 μg/g;
transformation assay, growth incubated with 2% juice. decreases B[α]P-induced BALB/c 3T3 cell foci formation
of HL-60 cells and induces G0/G1 phase cell cycle arrest in HL-60 cells.
Noni Juice
Noni juice Lung cancer (A549) cells Incubation of cells with noni fruit Inhibits manganese chloride-induced expression of HIF-1α, [79]
juice (100 μL/mL). decreases phosphorylation of ERK1/2, JNK1, PKB and
ribosomal S6 kinase, and eIF-2α.
(Continued)
53
54

Breast tumor explant and Incubation with 5% or 10% (v/v) noni Inhibits sprouting of new blood vessels in placental vein [78]
placental vein explant fruit juice. explants, promotes degeneration of breast tumor blood
vessels, and blocks capillary initiation.
Tomato Juice
Fresh tomato juice DMBA-initiated and croton Juice given by gavage, carcinogens Decreases number of papillomas and induces hepatic GST [81]
oil–promoted mouse skin were treated topically for 12 weeks. and GPx activity.
papilloma formation and
assessment of liver antioxidant
enzymes
BBN-treated rats Juice was administered for 12 weeks Inhibits the number of urinary bladder transitional cell [82]
after BBN challenge. carcinomas.
Citrus Juices
Orange juice AOM-induced rat colon tumor Rats challenged with AOM were Decreases the colon tumor incidence by 22% and inhibits [93]
allowed to drink orange juice daily proliferation of colonic mucosa.
for 28 weeks.
DMBA-treated rat mammary Rats treated with DMBA with or Delays the onset of mammary tumor formation and [94]
tumors without high-fat diet and decreases the number of breast tumors.
supplemented with double-strength
orange juice.
Mandarin juice AOM-induced rat colon One week after AOM treatment, rats Decreases numbers of intestinal adenocarcinomas and large [95]
carcinogenesis were allowed to drink juice (7.8 mL/ bowel tumors and decreases expression of PCNA and
day/rat) for 37 weeks. cyclin D1 in tumors.
Bergamot juice Neuroblastoma (SK-N-SH and Mice treated with juice by gavage Attenuates the growth of xenograft tumor and inhibits [97]
LAN1) cells and LAN1 cell (200 μL/day) for 28 days. pulmonary metastasis of LAN1 cells.
xenograft study
Abbreviations: ACF, aberrant crypt foci; AOM, azoxymethane; AN, aronia nectar; B[a]P, benzo [α]-pyrene; BBN, N-butyl-N-(4-hydroxybutyl)-nitrosamine; Cdk, cyclin-dependent kinase;
CEACAM1, carcinoembryonic antigen-related cell adhesion molecule 1; CF, cloud fraction; COX-2, cyclooxygenase-2; DMBA, 7,12-dimethylbenz[a]anthracene; DMH,
1,2-dimethylhydrazine; ERK1/2, extracellular signal-regulated kinase-1/2; GAE, gallic acid equivalents; GPx, glutathione peroxidase; GST, glutathione-S-transferase; iNOS,
inducible nitric oxide synthase; mTOR, mammalian target of rapamycin; NF-κB, nuclear factor-kappaB; ODC, ornithine decarboxylase; OH-BBN, hydroxy-BBN; PCNA,
proliferating cell nuclear antigen; PF, polyphenol fraction; PKC, protein kinase C; TNFα, tumor necrosis factor-alpha; TPA, 12-O-tetradecanoyl phosrbol-13-acetate.
Nrf2 target genes [29]. These findings suggest that apple juice can fortify cellular antioxidant defense
by inducing Nrf2-dependent gene expression, which appears as a plausible mechanism of its cancer
chemopreventive activity.
Polyphenolic constituents of apple juice also exhibited anti-inflammatory effects. A phenolic apple
juice extract (AE04), prepared from juices of different apple varieties, significantly inhibited the expres-
sion of various inflammatory genes, such as tumor necrosis factor-α (TNFα), interleukin (IL)-β, chemo-
kine C-X-C motif ligand (CXCL)-9, CXCL-10, and COX-2 in lipopolysaccharide (LPS) plus interferon-γ
(IFNγ)-stimulated human monocytic (monoMac6) cells. AE04 was shown to contain diverse poly-
phenolic compounds, of which flavan-3-ol dimer procyanidin B2 was found to be responsible for
the anti-inflammatory activity of AE04. In addition, AE04 components dihydrochalcone aglycone
(phloretin) and the dimeric flavan-3-ol (procyanidin B1) significantly inhibited proinflammatory gene
expression and repressed NF-κB-, IFNγ-inducible protein-10 (IP-10)-, and IL-8-promoter activity in a
concentration-dependent manner [31]. Thus, the antioxidant and anti-inflammatory effects of apple juice
and/or its active constituents suggest the potential of apple juice for cancer chemoprevention.
In a pilot study, lymphocytes from female volunteers who consumed a quercetin-rich mixture of
blueberry and apple juice for 4 weeks were treated ex vivo with H2O2 or benzo(α)pyrene (B[α]P). The
juice consumption led to a decrease in H2O2-induced oxidative DNA damage and B[α]P-diol epixode
(BPDE)-DNA adduct formation. Moreover, treatment of human lymphocytes, preincubated with quer-
cetin in vitro, with H2O2 or B[α]P significantly decreased the oxidative DNA damage and BPDE-DNA
adduct formation [32]. Barth et al. [13] demonstrated that intervention with daily consumption of clear
apple juice or cloudy apple juice for 7 weeks starting 1 week prior to challenge with DMH significantly
reduced the proliferation index of colon epithelial cells. However, the cloudy apple juice, but not the clear
apple juice, reduced the number and mean size of large ACF in distal colon. A subsequent study by these
authors showed that daily consumption of cloudy apple juice itself, but not its fractions, was effective
in preventing DMH-induced genotoxicity, colonocyte proliferation, and large ACF formation [22]. The
cloudy apple juice contains a high concentration of procyanidins. Gosse et al. [33] examined the effect of
a procyanidin-rich fraction of fresh apple extract. According to their study, the procyanidin-rich extract
attenuated the proliferation of human colon cancer (SW620) cells in culture by blocking the activities of
protein kinase C (PKC) and ornithine decarboxylase (ODC). This study also demonstrated that a 6-week
intervention with procyanidin-rich apple extract significantly inhibited AOM-induced ACF formation in
rat colon [33].
The polyphenol-rich extract of commercially available apple juice inhibited the growth of human
colon cancer (HT29) cells by blocking the tyrosine kinase activity of epidermal growth factor receptor
(EGFR) and attenuating the activation of downstream mitogen-activated protein (MAP) kinases [34].
Chemical analysis of this polyphenol-rich apple juice extract revealed the presence of major phyto-
chemicals, such as proanthocyanidins B1 and B2, isoquercitrin (quercetin-3-glucoside), and hyperoside
(quercetin-3-galactoside) with substantial EGFR-inhibitory properties. However, a mixture of these
apple juice constituents showed only a marginal inhibitory effect on EGFR activation, suggesting that
the apple juice extract may contain additional chemopreventive agents and that the juice as a whole is
more effective than the isolated constituents [34]. Teller et al. [35] recently reported that the concentra-
tion of quercetin or its glycosides was too low to inhibit the EGFR activity and that the dihydrochalcones
and their glycosides diminished EGFR activity only in a cell-free system, but not in human cancer cells.
However, the fractions comprising more than 86% oligomeric procyanidins obtained from apple juice
extract inhibited the activities of EGFR and ErbB3 in cultured cancer cells [35].
5.3.2 Berry Juices

Different varieties of berry fruit juices are widely consumed. These include juices of blueberry, black-
berry, raspberry, strawberry, gooseberry, cranberry, and chokeberry, among others. A great deal of
research has been done to examine the cancer preventive potential of berry juices. Fresh juices from
strawberry, blueberry, and raspberry showed significant inhibition of mutagenesis caused by methyl
methanesulfonate and B[α]P [36]. Berries contain several phytochemicals, such as proanthocyanidins,
anthocyanins, and other flavonoids. Boivin et al. [14] examined the anticancer effects of 13 different
berry juices in a wide range of human cancer cells. The juices of raspberry, black currant, white cur-
rant, gooseberry, velvet leaf blueberry, low-bush blueberry, sea buckthorn, and cranberry juice sig-
nificantly inhibited the growth of various cancer cells, including those of stomach, prostate, intestine,
and breast carcinomas. The antiproliferative effect of berry juices resulted from cell cycle arrest, but
not through caspase-dependent apoptosis, as evidenced by the downregulation of the expression of
cell cycle regulatory proteins, such as cyclin-dependent kinase (Cdk)-4, and Cdk-6 and cyclin-D1 and
cyclin-D3. Of the 13 berries tested, the juice of raspberry, black currant, gooseberry, sea buckthorn,
cranberry, and blackberry significantly inhibited TNFα-induced expression of COX-2 and the activa-
tion of NF-κB [14].
5.3.2.1 Chokeberry
Fruits of Aronia melanocarpa [Michx] Elliot and Aronia arbutifolia [L] Elliot are commonly known as
black choke berry and red chokeberry, respectively [37]. Because of the high content of anthocyanins,
chokeberries have long been used as a food colorant. The astringent taste of chokeberry limits the con-
sumption of this fruit, but recent advances in fruit juice blending technology improve the taste of the
juice by mixing with other fruit juices, such as that of apple, pear, or black currant [37]. Phytochemicals
present in chokeberry include anthocyanins (cyanidin-glycosides), phenolic acids (chlorogenic acid,
cryptochlorogenic acid, and neochlorogenic acid), and carotenoids (β-carotene, β-cryptoxanthin, and
violaxanthin) [37]. Anthocyanins, which represent about 25% of total polyphenols in chokeberry juice
[38], showed inhibitory effects on the mutagenicity of B[α]P and 2-aminoflourene [39]. Administration
of chokeberry juice inhibited the endogenous formation of N-nitrosamine in rats challenged with ami-
nopyrene and sodium nitrite and protected against liver damage [40]. Since N-nitrosamine is a potent
hepatocellular carcinogen, this study suggests the potential of chokeberry juice in preventing liver can-
cer. Treatment with chokeberry juice inhibited the proliferation of colon cancer (Caco2) cells in culture
by inducing G2/M phase cell cycle arrest and restoring the expression of a tumor suppressor protein
carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1), which is diminished in early
adenomas and carcinomas [41]. Moreover, the anthocyanin-rich extract of chokeberry induced apoptosis
in human colon cancer (HT29 cells) without affecting the growth of normal colon epithelial (NCM460)
cells [42]. The colon cancer-preventive effect of chokeberry juice extract can be attributed to its antioxi-
dant effect [43]. Moreover, the chokeberry juice constituents, such as β-cryptoxanthin [44], chlorogenic
acid [45], and cyanidin-3-glucosides [46], exhibited colon cancer chemopreventive effects.
5.3.2.2 Cranberry
Cranberry juice is well known for its beneficial effects in urinary tract infections [47]. The a ntiproliferative
activity of cranberry juice in various human cancer cells has been reported [14]. Prasain et al. [48] demon-
strated that intervention with cranberry juice concentrate prevented chemically induced urinary bladder
carcinogenesis in rats. According to this study, rats were first treated with N-butyl-N-(4-hydroxybutyl)-
nitrosamine (OH-BBN) for 8 weeks and feeding with cranberry juice concentrate was started 1 week
after the final dose of OH-BBN and continued for 6 months. Consumption of cranberry juice inhibited
OH-BBN-induced urinary bladder papilloma and carcinoma formation in rats by 51% and 38%, respec-
tively. The study also identified the major chemopreventive constituent of cranberry juice as quercetin
and its methylated derivatives [48]. These findings were supported by a previous study demonstrating
that rats receiving a 5% quercetin diet were protected against chemically induced rat urinary bladder
carcinomas [49]. In another study, rats receiving 20% cranberry juice decreased the total number of
AOM-induced ACF formation and showed enhanced GST activity in the colon [28]. One of the major
causes of gastric cancer is Helicobacter pylori infection. In a prospective, randomized, double-blind, and
placebo-controlled human intervention trial, consumption of cranberry juice (500 mL/day) for 90 days
inhibited the H. pylori infection [50], suggesting the potential of cranberry juice in gastric cancer pre-
vention. A nondialyzable fraction derived from cranberry juice inhibited the growth of cultured murine
lymphoma (Rev-2-T-6) cells and attenuated the invasion of these cells through the extracellular matrix
[51]. Moreover, intraperitoneal administration of this fraction diminished the growth of Rev-2-T-6 cell
xenograft tumors in mice and enhanced the generation of antilymphoma antibodies [51]. Cranberry
juice was reported to be one of the effective fruit juices in inducing cell cycle arrest in various cancer
cells through the downregulation of the expression of D series cyclins and Cdk-4 and Cdk-6. In addi-
tion, cranberry juice inhibited TNFα-induced NF-κB activation and COX-2 expression in different
cancer cells [14].
5.3.2.3 Other Berry Juices

Although the extract and powder of different varieties of raspberries and their phenolic constituents have
been shown to possess anticarcinogenic effects in vitro and in vivo [52,53], it is beyond the scope of this
chapter to elaborate discussion of organic extracts of raspberries. However, many of the chemopreventive
phytochemicals, such as anthocyanins, are common in organic extracts and juices of raspberries. Liu
et al. [54] examined the phenolic content and the beneficial health effects of juice extracts from different
varieties of raspberry, such as Heritage, Kiwigold, Goldie, and Anne. The total phenolic, flavonoid, and
anthocyanin content were highest in the Heritage variety that appeared with the darkest color and highest
antioxidant activity, whereas the Anne variety with a pale color appearance showed the lowest phyto-
chemical content and minimum antioxidant property. The juice extract of all four varieties of raspberry
exhibited excellent antiproliferative effects in human hepatoma (HepG2) cells [54]. The fruit juice of
black currant was found to contain a polysaccharide-rich substance, designated as cassis polysaccharide
(CAPS), which was separated from the fruit juice of black currant. Oral administration of black currant
juice and CAPS to Ehrlich carcinoma-bearing mice retarded the growth of the solid tumor by 45% and
51%, respectively [55].
5.3.3 Cherry Juice

Epidemiological studies have shown that consumption of cherries lowers the risk of various chronic
diseases including cancer. Damar and Ekşi [56] analyzed the antioxidant activity, total polyphenolics,
and monomeric anthocyanin content of 11 varieties of sour cherry juices. The antioxidant capacity of
sour cherry juices was well correlated with the total polyphenolic content but not with the monomeric
anthocyanin content. Cyanidin-3-glucosylrutinoside, cyanidin-3-rutinoside, cyanidin-3-sophoroside, and
cyanidin-3-glucoside have been identified as the major polyphenols in sour cherry juice [56]. Incubation
of human breast cancer (MCF-7) cells with tart cherry juice significantly reduced bromodeoxyuri-
dine incorporation, suggesting the antiproliferative potential of this fruit juice. Whereas, a 3% cherry
juice enriched with monomeric anthocyanin-induced apoptosis, a 10% juice caused necrosis of MCF-7
cells [57]. In a randomized and double-blind study, consumption of tart cherry juice concentrate for
7 days elevated the serum melatonin level as compared to a placebo group and increased the sleep dura-
tion and quality. Since melatonin exerts antioxidant, anti-inflammatory [58], apoptosis-inducing [59],
and antiangiogenic [60] properties, the elevation of melatonin by cherry juice signifies the potential of
cherry juice in cancer prevention.
5.3.4 Pomegranate Juice

Pomegranate juice is a rich source of polyphenolic anticancer principles. Major polyphenolics present
in pomegranate juice include ellagitannins such as punicalins and punicalagin A, punicalagin B, and
ellagic acid. Consumption of pomegranate fruit juice for 4 weeks decreased the total hepatic CYP content
as well as the expression of CYP1A2 and CYP3A, suggesting that this fruit juice may inhibit tumor initia-
tion by blocking the activation of procarcinogens [61]. The administration of pomegranate juice instead of
drinking water for 6 weeks to rats challenged with AOM showed a decreased number of ACF and inhib-
ited the expression of cell proliferation markers Ki67 and proliferating cell nuclear antigens (PCNAs) in
the colonic epithelium. Moreover, treatment with pomegranate juice attenuated the expression of COX-2,
inducible nitric oxide synthase (iNOS), and vascular cell adhesion molecule-1 (VCAM-1) at both mRNA
and protein levels by blocking the phosphatidyl inositol-3 kinase (PI3K)/Akt signaling via upregulation of
microRNA (miR)-126 in the colon mucosa of AOM-treated rats [12]. Likewise, the incubation of human
colon cancer (HT-29) cells with pomegranate juice extract containing polyphenols decreased the cell
viability, which was associated with elevated expression of miR-126, activation of casapse-3 and cleav-
age of poly (ADP-ribose) polymerase (PARP), and diminished mRNA and protein expression of vari-
ous inflammatory markers, such as NF-κB p65, VCAM-1, intercellular adhesion molecule-1 (ICAM-1),
COX-2, and phosphorylated-Akt [12]. Transfecting cells with antagomiR-126 abrogated the antiprolif-
erative and anti-inflammatory effects of pomegranate juice in HT-29 cells. This study also revealed that
incubation with pomegranate juice polyphenols reduced the expression of vascular endothelial growth
factor (VEGF) in HT-29 cells, indicating the antiangiogenic potential of this fruit juice [12]. Adams et al.
[62] reported that treatment of HT-29 cells with pomegranate juice or its total tannin content attenuated
TNFα-induced expression of COX-2, phosphorylation p65, and DNA binding of NF-κB. In another study,
rats treated with pomegranate juice instead of drinking water ameliorated dextran sulfate sodium (DSS)-
induced colitis by blocking the activation of p70S6K, mitogen-activated protein kinase kinase (MEK),
and extracellular signal-regulated kinase (ERK1/2) and inducing the expression of miR145 in colonic
epithelial tissue [63].
5.3.5 Mango Juice

The mango fruit (Mangifera indica L.) contains high levels of carotenoids (all-trans-violaxanthin and
all-trans-β-carotene) and phenolic compounds (xanthone glycosides and hydrolyzable tannins) [64–67].
Botting et al. [68] have reported the antimutagenic activity of mango fruit. Analysis of the chemical com-
position of different forms of mango fruit showed that the juice and fresh fruit slice had high content of
α- and β-carotenes and a minor amount of cryptoxanthin and zeaxanthin, whereas the dry fruit slice had
a comparatively low amount of β-carotene with a trace amount of other carotenoids [69]. Administration
of mango fruits in three different forms, such as juice, fresh slice, or dried slice to healthy volunteers for
2 weeks showed that serum retinol level was highest in groups receiving juice and fresh fruit as com-
pared to dry mango fruits [69]. Because of the presence of polyphenolics, carotenoids, and antimutagens,
mango possesses significant anticancer activity. Analysis of different varieties of mango fruit pulp has
revealed that its polyphenolic fraction contains mangiferin, gallic acid, and gallotannins [65]. Treatment
with the polyphenolic fraction of mango fruit induced G2/M phase cell cycle arrest and apoptosis in
human colon cancer SW480 cells through increased gene expression of Bax, Bim, caspase-8, and p21
without affecting that of normal colonic myofibroblasts [65]. Mango juice, prepared from pulp after
removal of seeds and peel, exhibited in vitro antioxidant activity as evidenced by DPPH, oxygen radical
absorbance capacity (ORAC), and Folin–Ciocalteu assay and inhibited B[a]P-induced transformation
of BALB/c 3T3 cells [70]. This study also demonstrated that whole mango juice induced S- and G2/M
phase cell cycle arrest in human leukemia (HL-60) cells [70]. Administration of mango juice ame-
liorated DSS-induced colitis in Sprague-Dawley rats by blocking insulin-like growth factor-1 receptor
(IGF1-R)-Akt/mTOR-mediated signaling via upregulation of miR-126 [63]. These authors also dem-
onstrated that mango juice decreased the serum concentration of proinflammatory cytokines, such as
IL-1β and granulocyte-macrophage colony-stimulating factor (GM-CSF), and increased the level of anti-
inflammatory cytokine IL-10 in DSS-challenged rats. Moreover, feeding of mango juice increased fecal
short-chain free fatty acids without altering the composition of intestinal microbiota [71]. These findings
suggest the potential of colon cancer prevention with mango fruit juice.
5.3.6 Noni Juice

Fruits of Morinda citrifolia, known as noni, are widely consumed in the Pacific Islands and India. Noni,
which possesses a range of beneficial health effects, has been reported that its juice exerts antioxidant
[72], antimutagenic [15], anti-inflammatory, and anticancer activities [73]. Rats treated with deacetylas-
parulosidic acid, a constituent of noni juice, reduced serum malondialdehyde (MDA) level and induced
SOD activity [74]. A polysaccharide-rich fraction of the fruit juice of M. citrifolia significantly reduced
the growth of sarcoma 180 ascites tumor [75] and intraperitoneally administered Lewis lung carcinoma
(LLC) [76] in allogeneic mice. The antitumor effect of noni juice was potentiated by combined treatment
with several other anticancer drugs including cisplatin, adriamycin, bleomycin, etoposide, camptothecin,
and 5-fluorouracil [75,76]. The latter study further demonstrated that noni juice induced cytotoxicity in
LLC cells when co-cultured with murine peritoneal cells exudate which increased the release of cytokines
and NO upon treatment with noni juice [76]. These findings suggest that noni juice stimulates host immune
responses and holds the cancer immunotherapeutic potential. Other glycosides present in noni juice
diminished 12-O-tetradecanoyl phorbol-13-acetate (TPA)- or extracellular growth factor (EGF)-induced
transformation of mouse epidermal JB6 cells by blocking the activation of activator protein-1 (AP-1) [77].
Hornick et al. [78] suggested that noni juice disrupted newly formed human vascular networks, indicating
its antiangiogenic activity. A recent study reported that treatment of human lung adenocarcinoma (A549)
cells with noni juice inhibited manganese-induced expression of hypoxia-inducible factor-1α (HIF-1α), an
angiogenic switch in cancer, by blocking the activation of ERK1/2, c-Jun-N-terminal kinase-1 (JNK1),
protein kinase B (PKB), and ribosomal protein S6 kinase. This study also suggested that noni juice attenu-
ated HIF-1α protein expression in A549 cells stimulated with IL-1β [79].
5.3.7 Tomato Juice

Tomato juice is commercially available in markets and is widely consumed among many fruit juices.
Analysis of tomato juice revealed the presence of carotenoids, such as lycopene, β-carotene, and phy-
toene, as the major constituents, which possess anticancer effects [80]. Several animal model studies
have shown that tomato juice intake can prevent experimentally induced carcinogenesis of the skin,
colon, and urinary bladder. De and Das [81] demonstrated that oral feeding of tomato juice daily
for 12 weeks significantly decreased the incidence of skin papillomas in mice topically treated with
7,12-dimethylabenz[a]anthracene (DMBA) and croton oil as well as induced the activities of hepatic
antioxidant and detoxification enzymes, such as GST, GPx, and SOD, which enhance the metabolic
detoxification of chemical carcinogens and protect against oxidative and/or electrophilic DNA damage.
Drinking of diluted tomato juice for 12 weeks reduced the number, but not the incidence, of urinary blad-
der transitional cell carcinomas in rats challenged with N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN)
[82]. In another study, rats allowed to drink diluted tomato juice containing 17 ppm of lycopene for 35
weeks were protected against N-methylnitrosourea (MNU)-induced colon cancer, whereas treatment
with only lycopene-water was ineffective in reducing the colon tumor burden. Thus, tomato juice as
a whole, but not its isolated constituents alone, can prevent rat colon carcinogenesis [83]. In contrast,
administration of tomato juice (330 mL/day) for 2 weeks increased the concentration of lycopene in fecal
water, which induced apoptosis in HT-29 cells, without altering bile acid concentrations or the bacterial
enzymes and fecal water pH, suggesting that consumption of tomato juice had only minor effects on the
luminal biomarkers relevant to colon carcinogenesis [84]. However, a subsequent study reported that the
anticancer effects of tomato juice may be attributed to its antioxidant effects [85]. Thus, daily intake of
tomato juice equivalent to 15 mg lycopene per day for 5 weeks significantly reduced the serum level of
8-oxodG, a marker of oxidative DNA damage, in healthy volunteers undertaking strenuous exercise [85].
Bohn et al. [86] studied the compliance and bioavailability of a soy-fortified lycopene-rich tomato juice
and reported that the juice constituents were well absorbed and showed no signs of toxicity.
5.3.8 Citrus Juices

Orange juice is the most consumed among different types of citrus juices. Major antioxidant and chemo-
preventive constituents present in orange juice include ascorbic acid, flavonoid glycosides (hesperidin),
carotenoids (xanthophylls and cryptoxanthins), terpenoids (α-terpineol and limonene), and folic acid [87,88].
Administration of orange juice components, such as flavanone glycosides, carotenoids, ascorbic acid,
and folate, to healthy volunteers showed the remarkable plasma concentrations of these juice constitu-
ents and markedly reduced the level of an oxidative DNA damage marker 8-hydroxydeoxyguanosine
in white blood cells, indicating the antioxidant potential of orange juice [87]. Analysis of polyphenol
content and antioxidant potential of different citrus juices revealed that mandarin and lemon juices
had the highest total flavonoids and antioxidant activity as determined by the β-carotene bleach-
ing assay (26.67% and 22.67%, respectively), while bitter orange juice exhibiting the highest content
of total polyphenols (784.67 mg gallic acid equivalents [GAE]/L) exhibiting free radical scavenging
activity [89]. Drinking of freshly prepared orange juice (20 fluid ounce [equivalent to 592 mL] daily) for
90 days significantly increased the total plasma antioxidant capacity and inhibited lipid peroxidation in
subjects with hyperlipidemia [90]. Juices prepared by hand-squeezing peeled Moro fruits (Citrus sinen-
sis L. Osbeck) contained a variety of C- and O-glycosylated flavonoids, such as lucenin-2, vicenin-2,
stellarin-2, lucenin-2 4′-methyl ether, scoparin, chrysoeriol 7-O-neohesperidoside, narirutin, and hes-
peridin [21]. Stella et al. [91] demonstrated that total polyphenol content in commercially available
ready-to-drink orange juice and nectar ranged from 18.7 to 54.2 mg GAE/100 mL and the total antioxi-
dant activity varied between 57.88 and 349.32 μmol trolox equivalents (TE)/100 mL. The antioxidant
capacity was more strongly correlated with the total polyphenol rather than vitamin C alone. Intake of
orange juice with a diet rich in high fat–high carbohydrate (HFHC) reduced the oxidative and inflamma-
tory markers, such as the gene expression of Toll-like receptor (TLR)-2, TLR4, nicotinamide-adenine
dinucleotide phosphate oxidase (NOX), and matrix metalloproteinase (MMP)-9 in blood mononuclear
cells of healthy volunteers [92]. Moreover, orange juice component α-terpineol reduced IL-6 production
in buccal epithelial cells [88].
Because of its antioxidant and anti-inflammatory properties, orange juice holds the promise of can-
cer chemopreventive activity. Consumption of orange juice has been shown to prevent experimentally
induced colon and mammary carcinogenesis [93,94]. Administration of orange juice delayed the devel-
opment of mammary tumors in rats challenged with DMBA or high-fat diet [94]. Orange juice supple-
mentation for 28 weeks after AOM administration also reduced the incidence of colon tumors in rats.
Hesperidin flavonoid and limonoid glucosides were identified as the major bioactive principles in orange
juice that could contribute to the chemopreventive effect of orange juice [93]. In fact, the proliferation of
human mammary carcinoma (MDA-MB-435) cells was markedly inhibited by treatment with hesperitin
in combination with grape fruit juice component quercetin or naringenin [94].
Drinking of juice prepared from Satsuma mandarin or its juice enriched with β-cryptoxanthin or
hesperidin for 36 weeks starting 1 week after AOM administration significantly decreased the incidence
and multiplicity of colonic adenocarcinomas in rats [95]. Mandarin juice rich in hesperidin showed the
strongest inhibitory effects. Moreover, mandarin juice consumption increased the apoptotic index and
inhibited the expression of cell proliferation markers PCNA and cyclin D1 in colonic adenocarcinomas
[95]. Likewise, administration of mandarin juices in drinking water starting at 1 week after challenge
with 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) attenuated the incidence and multiplicity
of lung tumorigenesis in male A/J mice [96]. Treatment with juice prepared by squeezing fresh bergamot
fruit (Citrus bergamia) inhibited the proliferation and invasion of neuroblastoma (SK-N-SH and LAN1)
cells in vitro without altering the weight of LAN1 cell xenograft tumors in vivo. However, bergamot juice
inhibited lung metastasis of LAN1 neuroblastoma cells in severe combined immunodeficiency (SCID)
mice [97]. Zhu et al. [98] isolated 7-O-neohesperidoside of isosakuranetin (poncirin) from different parts
including the juice sac of Ougan fruit (Citrus reticulata cv. Suavissima) and reported the antiproliferative
effect of the compound in human gastric cancer (SGC-7901) cells.
5.3.9 Miscellaneous Fruit Juices

Food products made from Japanese apricot (Prunus mume Sieb. et Zucc) are traditionally known for
their diverse beneficial health effects. Administration of 1% or 3% of the fruit juice concentrate of
Japanese apricot in drinking water for 10 weeks reduced gastritis and gastric mucosal hyperplasia in
H. pylori–infected Mongolian gerbils. Moreover, the average relative urease-A gene dosage in the glan-
dular stomach was significantly reduced upon intervention with apricot juice concentrate as compared to
animals inoculated with H. pylori alone [99]. These findings suggest that apricot juice may be effective
in preventing gastric carcinogenesis, which merits further investigation. Another commonly consumed
fruit juice is mangosteen juice, which contains major xanthone derivatives, such as α-mangostin, gar-
cinones, γ-mangostin, and gartanin [100]. The well-documented anticancer activities of α-mangostin
[101], garcinones [102], and gartanin [103] suggest that mangosteen juice as a whole might have cancer
chemopreventive potential.
Fruits of the plants belonging to the genus Opuntia spp. are known as cactus pear or prickly pear.
Fruit juices of three different varieties of prickly pears, such as red-purple pear, white-green pear, or
yellow-orange pear, exhibited DPPH radical scavenging activity with red-purple pear juices being the
most active. When the juice of red-purple pear was given by gavage to mice treated with a mutagen
methyl methanesulfonate, the number of micronucleated polychromatic erythrocytes was significantly
decreased, suggesting the antioxidant and antimutagenic effects of red-purple pear juice [104]. In another
study, juices of nine different varieties of prickly pears were analyzed for their phenolic content, anti-
oxidant potential, and anticancer activity. Flavonoids, betaxanthins, and betacyanins were identified as
major constituents of these pear juices, which exhibited strong antioxidant activity and inhibited the
proliferation of human cancer cells. Among the nine varieties, juice of the Rastrero pear exhibited anti-
proliferative effects against prostate, colon, mammary, and liver cancer cells in vitro, while Moradillo
pear juice had the highest flavonoid content and diminished the growth of only prostate and colon cancer
cells. Interestingly, none of the pear juices reduced the viability of normal human fibroblasts, suggesting
that the prickly pear juices induce cytotoxicity selectively in cancer cells [105].
5.4 Conclusion
Over the last several decades, dietary cancer chemoprevention has received immense interest as a ratio-
nal strategy to reduce the incidence of and mortality from cancer. Numerous dietary phytochemicals,
especially those present in edible fruits, vegetables, and spices, have been identified as promising candi-
dates for cancer chemoprevention. Accumulating evidence from epidemiological and laboratory-based
studies has shown that regular intake of fruits can reduce the risk of various cancers. In addition to their
nutritional value, fruits are rich sources of a large number of chemical compounds, which have been
reported to possess anticancer activity. There is now increasing trend in consuming a wide variety of
ready-made fruit juices available in supermarkets. Since fruit juices contain a whole bunch of chemo-
preventive phytochemicals, it is considered that consumption of fruit juices may prevent carcinogenesis.
Since cancer is a heterogeneous disease, the combination of chemopreventive agents present in fruit
juices makes them a good choice for achieving multitargeted cancer chemoprevention. Although the
experimental findings accumulated to date strongly suggest the chemopreventive potential of fruit juices,
additional studies are warranted to evaluate the anticancer efficacy of fruit juices in an expanded area
covering many different types of cancer. Moreover, further studies are necessary to elucidate the phyto-
chemical and pharmacokinetic profiling of fruit juices and understanding their molecular mechanisms of
anticancer activity. Whereas the majority of chemoprevention research with fruit juices was performed
with freshly prepared juices, practically commercial fruit juices are consumed by humans. Thus, focus
should be given on assessing the chemopreventive potential and safety of commercially available fruit
juices. Nonetheless, fruit juices as a natural blend of anticancer principles are promising dietary measure
for cancer chemoprevention and a large variety of functional foods can be developed to help prevent
carcinogenesis.
Acknowledgments
This work has been supported by the Settlement Research Grant #2012-0195 of Keimyung University
allocated to Joydeb Kumar Kundu.
REFERENCES
1. Moolgavkar, S.H., The multistage theory of carcinogenesis and the age distribution of cancer in man.
J. Natl. Cancer Inst., 61, 49–52, 1978.
2. Surh, Y.J., Cancer chemoprevention with dietary phytochemicals. Nat. Rev. Cancer, 3, 768–780, 2003.
3. Sporn, M.B., Dunlop, N.M., Newton, D.L., and Smith, J.M., Prevention of chemical carcinogenesis by
vitamin A and its synthetic analogs (retinoids). Fed. Proc., 35, 1332–1338, 1976.
4. Kundu, J.K. and Surh, Y.J., Breaking the relay in deregulated cellular signal transduction as a rationale
for chemoprevention with anti-inflammatory phytochemicals. Mutat. Res., 591, 123–146, 2005.
5. Surh, Y.J., Chun, K.S., Cha, H.H., Han, S.S., Keum, Y.S., Park, K.K., and Lee, S.S., Molecular mecha-
nisms underlying chemopreventive activities of anti-inflammatory phytochemicals: Down-regulation of
COX-2 and iNOS through suppression of NF-kappa B activation. Mutat. Res., 480–481, 243–268, 2001.
6. Temple, N.J. and Gladwin, K.K., Fruit, vegetables, and the prevention of cancer: Research challenges.
Nutrition, 19, 467–470, 2003.
7. Terry, P., Terry, J.B., and Wolk, A., Fruit and vegetable consumption in the prevention of cancer:
An update. J. Intern. Med., 250, 280–290, 2001.
8. Miller, A.B., Altenburg, H.P., Bueno-de-Mesquita, B., Boshuizen, H.C., Agudo, A., Berrino, F., Gram,
I.T. et al., Fruits and vegetables and lung cancer: Findings from the European Prospective Investigation
into Cancer and Nutrition. Int. J. Cancer, 108, 269–276, 2004.
9. van Duijnhoven, F.J., Bueno-De-Mesquita, H.B., Ferrari, P., Jenab, M., Boshuizen, H.C., Ros, M.M.,
Casagrande, C. et al., Fruit, vegetables, and colorectal cancer risk: The European Prospective
Investigation into Cancer and Nutrition. Am. J. Clin. Nutr., 89, 1440–1452, 2009.
10. Smith-Warner, S.A., Spiegelman, D., Yaun, S.S., Albanes, D., Beeson, W.L., van den Brandt, P.A.,
Feskanich, D. et al., Fruits, vegetables and lung cancer: A pooled analysis of cohort studies. Int. J.
Cancer, 107, 1001–1011, 2003.
11. Kundu, J.K. and Surh, Y.J., Cancer chemopreventive effects of selected dried fruits, in Dired Fruits:
Phytochemicals and Health Effects, Alasalvar, C. and Shahidi, F., Eds., Wiley-Blackwell, Oxford, UK,
2013.
12. Banerjee, N., Kim, H., Talcott, S., and Mertens-Talcott, S., Pomegranate polyphenolics suppressed
azoxymethane-induced colorectal aberrant crypt foci and inflammation: Possible role of miR-126/
VCAM-1 and miR-126/PI3K/AKT/mTOR. Carcinogenesis, 34, 2814–2822, 2013.
13. Barth, S.W., Fahndrich, C., Bub, A., Dietrich, H., Watzl, B., Will, F., Briviba, K., and Rechkemmer, G.,
Cloudy apple juice decreases DNA damage, hyperproliferation and aberrant crypt foci development in
the distal colon of DMH-initiated rats. Carcinogenesis, 26, 1414–1421, 2005.
14. Boivin, D., Blanchette, M., Barrette, S., Moghrabi, A., and Beliveau, R., Inhibition of cancer cell prolif-
eration and suppression of TNF-induced activation of NFkappaB by edible berry juice. Anticancer Res.,
27, 937–948, 2007.
15. Franchi, L.P., Guimaraes, N.N., De Andrade, L.R., De Andrade, H.H., Lehmann, M., Dihl, R.R., and
Cunha, K.S., Antimutagenic and antirecombinagenic activities of noni fruit juice in somatic cells of
Drosophila melanogaster. An. Acad. Bras. Cienc., 85, 585–594, 2013.
16. Gerhauser, C., Cancer chemopreventive potential of apples, apple juice, and apple components. Planta
Med., 74, 1608–1624, 2008.
17. Seeram, N.P., Adams, L.S., Henning, S.M., Niu, Y., Zhang, Y., Nair, M.G., and Heber, D., In vitro
antiproliferative, apoptotic and antioxidant activities of punicalagin, ellagic acid and a total pomegran-
ate tannin extract are enhanced in combination with other polyphenols as found in pomegranate juice.
J. Nutr. Biochem., 16, 360–367, 2005.
18. Rocha, A., Wang, L., Penichet, M., and Martins-Green, M., Pomegranate juice and specific components
inhibit cell and molecular processes critical for metastasis of breast cancer. Breast Cancer Res. Treat.,
136, 647–658, 2012.
19. Vicinanza, R., Zhang, Y., Henning, S.M., and Heber, D., Pomegranate juice metabolites, ellagic acid
and urolithin A, synergistically inhibit androgen-independent prostate cancer cell growth via distinct
effects on cell cycle control and apoptosis. Evid. Based Complement. Alternat. Med., 2013, 247504,
2013.
20. Schaefer, S., Baum, M., Eisenbrand, G., and Janzowski, C., Modulation of oxidative cell damage by
reconstituted mixtures of phenolic apple juice extracts in human colon cell lines. Mol. Nutr. Food Res.,
50, 413–417, 2006.
21. Barreca, D., Bellocco, E., Leuzzi, U., and Gattuso, G., First evidence of C- and O-glycosyl flavone in
blood orange (Citrus sinensis (L.) Osbeck) juice and their influence on antioxidant properties. Food
Chem., 149, 224–252, 2014.
22. Barth, S.W., Faehndrich, C., Bub, A., Watzl, B., Will, F., Dietrich, H., Rechkemmer, G., and Briviba, K.,
Cloudy apple juice is more effective than apple polyphenols and an apple juice derived cloud fraction in
a rat model of colon carcinogenesis. J. Agric. Food Chem., 55, 1181–1187, 2007.
23. Sporn, M.B., Combination chemoprevention of cancer. Nature, 287, 107–108, 1980.
24. George, J., Singh, M., Srivastava, A.K., Bhui, K., and Shukla, Y., Synergistic growth inhibition of
mouse skin tumors by pomegranate fruit extract and diallyl sulfide: Evidence for inhibition of activated
MAPKs/NF-kappaB and reduced cell proliferation. Food Chem. Toxicol., 49, 1511–1520, 2011.
25. Kowalczyk, M.C., Walaszek, Z., Kowalczyk, P., Kinjo, T., Hanausek, M., and Slaga, T.J., Differential
effects of several phytochemicals and their derivatives on murine keratinocytes in vitro and in vivo:
Implications for skin cancer prevention. Carcinogenesis, 30, 1008–1015, 2009.
26. Zessner, H., Pan, L., Will, F., Klimo, K., Knauft, J., Niewohner, R., Hummer, W. et al., Fractionation of
polyphenol-enriched apple juice extracts to identify constituents with cancer chemopreventive potential.
Mol. Nutr. Food Res., 52(Suppl. 1), S28–S44, 2008.
27. Platt, K.L., Edenharder, R., Aderhold, S., Muckel, E., and Glatt, H., Fruits and vegetables protect against
the genotoxicity of heterocyclic aromatic amines activated by human xenobiotic-metabolizing enzymes
expressed in immortal mammalian cells. Mutat. Res., 703, 90–98, 2010.
28. Boateng, J., Verghese, M., Shackelford, L., Walker, L.T., Khatiwada, J., Ogutu, S., Williams, D.S. et al.,
Selected fruits reduce azoxymethane (AOM)-induced aberrant crypt foci (ACF) in Fisher 344 male rats.
Food Chem. Toxicol., 45, 725–732, 2007.
29. Soyalan, B., Minn, J., Schmitz, H.J., Schrenk, D., Will, F., Dietrich, H., Baum, M., Eisenbrand, G., and
Janzowski, C., Apple juice intervention modulates expression of ARE-dependent genes in rat colon and
liver. Eur. J. Nutr., 50, 135–143, 2011.
30. Shi, D. and Jiang, B.H., Antioxidant properties of apple juice and its protection against Cr(VI)-induced
cellular injury. J. Environ. Pathol. Toxicol. Oncol., 21, 233–242, 2002.
31. Jung, M., Triebel, S., Anke, T., Richling, E., and Erkel, G., Influence of apple polyphenols on inflamma-
tory gene expression. Mol. Nutr. Food. Res., 53, 1263–1280, 2009.
32. Wilms, L.C., Hollman, P.C., Boots, A.W., and Kleinjans, J.C., Protection by quercetin and quercetin-rich
fruit juice against induction of oxidative DNA damage and formation of BPDE-DNA adducts in human
lymphocytes. Mutat. Res., 582, 155–162, 2005.
33. Gosse, F., Guyot, S., Roussi, S., Lobstein, A., Fischer, B., Seiler, N., and Raul, F., Chemopreventive
properties of apple procyanidins on human colon cancer-derived metastatic SW620 cells and in a rat
model of colon carcinogenesis. Carcinogenesis, 26, 1291–1295, 2005.
34. Kern, M., Pahlke, G., Balavenkatraman, K.K., Bohmer, F.D., and Marko, D., Apple polyphenols affect
protein kinase C activity and the onset of apoptosis in human colon carcinoma cells. J. Agric. Food
Chem., 55, 4999–5006, 2007.
35. Teller, N., Roth, M., Esselen, M., Fridrich, D., Boettler, U., Blust, V., Will, F. et al., Apple procyanidins
affect several members of the ErbB receptor tyrosine kinase family in vitro. Food Funct., 4, 689–697,
2013.
36. Smith, S.H., Tate, P.L., Huang, G., Magee, J.B., Meepagala, K.M., Wedge, D.E., and Larcom, L.L.,
Antimutagenic activity of berry extracts. J. Med. Food, 7, 450–455, 2004.
37. Kulling, S.E. and Rawel, H.M., Chokeberry (Aronia melanocarpa)—A review on the characteristic
components and potential health effects. Planta Med., 74, 1625–1634, 2008.
38. Oszmianski, J. and Wojdylo, A., Aronia melanocarpa phenolics and their antioxidant activity. Eur.
Food Res. Technol., 221, 809–813, 2005.
39. Gasiorowski, K., Szyba, K., Brokos, B., Kolaczynska, B., Jankowiak-Wlodarczyk, M., and Oszmianski,
J., Antimutagenic activity of anthocyanins isolated from Aronia melanocarpa fruits. Cancer Lett., 119,
37–46, 1997.
40. Atanasova-Goranova, V.K., Dimova, P.I., and Pevicharova, G.T., Effect of food products on endogenous
generation of N-nitrosamines in rats. Br. J. Nutr., 78, 335–345, 1997.
41. Bermudez-Soto, M.J., Larrosa, M., Garcia-Cantalejo, J.M., Espin, J.C., Tomas-Barberan, F.A., and
Garcia-Conesa, M.T., Up-regulation of tumor suppressor carcinoembryonic antigen-related cell adhe-
sion molecule 1 in human colon cancer Caco-2 cells following repetitive exposure to dietary levels of a
polyphenol-rich chokeberry juice. J. Nutr. Biochem., 18, 259–271, 2007.
42. Malik, M., Zhao, C., Schoene, N., Guisti, M.M., Moyer, M.P., and Magnuson, B.A., Anthocyanin-rich
extract from Aronia meloncarpa E induces a cell cycle block in colon cancer but not normal colonic
cells. Nutr. Cancer, 46, 186–196, 2003.
43. Zheng, W. and Wang S.Y., Oxygen radical absorbing capacity of phenolics in blueberries, cranberries,
chokeberries, and lingonberries. J. Agric. Food Chem., 51, 502–509, 2003.
44. Narisawa, T., Fukaura, Y., Oshima, S., Inakuma, T., Yano, M., and Nishino, H., Chemoprevention by
the oxygenated carotenoid beta-cryptoxanthin of N-methylnitrosourea-induced colon carcinogenesis in
F344 rats. Jpn. J. Cancer Res., 90, 1061–1065, 1999.
45. Park, H.J., Davis, S.R., Liang, H.Y., Rosenberg, D.W., and Bruno, R.S., Chlorogenic acid differentially
alters hepatic and small intestinal thiol redox status without protecting against azoxymethane-induced
colon carcinogenesis in mice. Nutr. Cancer, 62, 362–370, 2010.
46. Lea, M.A., Ibeh, C., desBordes, C., Vizzotto, M., Cisneros-Zevallos, L., Byrne, D.H., Okie, W.R., and
Moyer, M.P., Inhibition of growth and induction of differentiation of colon cancer cells by peach and
plum phenolic compounds. Anticancer Res., 28, 2067–2076, 2008.
47. Howell, A.B. and Foxman, B., Cranberry juice and adhesion of antibiotic-resistant uropathogens. JAMA,
287, 3082–3083, 2002.
48. Prasain, J.K., Jones, K., Moore, R., Barnes, S., Leahy, M., Roderick, R., Juliana, M.M., and Grubbs,
C.J., Effect of cranberry juice concentrate on chemically-induced urinary bladder cancers. Oncol. Rep.,
19, 1565–1570, 2008.
49. Hirose, M., Fukushima, S., Sakata, T., Inui, M., and Ito, N., Effect of quercetin on two-stage carcinogen-
esis of the rat urinary bladder. Cancer Lett., 21, 23–27, 1983.
50. Zhang, L., Ma, J., Pan, K., Go, V.L., Chen, J., and You, W.C., Efficacy of cranberry juice on Helicobacter
pylori infection: A double-blind, randomized placebo-controlled trial. Helicobacter, 10, 139–145, 2005.
51. Hochman, N., Houri-Haddad, Y., Koblinski, J., Wahl, L., Roniger, M., Bar-Sinai, A., Weiss, E.I., and
Hochman, J., Cranberry juice constituents impair lymphoma growth and augment the generation of
antilymphoma antibodies in syngeneic mice. Nutr. Cancer, 60, 511–517, 2008.
52. Wang, L.S., Kuo, C.T., Cho, S.J., Seguin, C., Siddiqui, J., Stoner, K., Weng, Y.I. et al., Black
raspberry-derived anthocyanins demethylate tumor suppressor genes through the inhibition of DNMT1
and DNMT3B in colon cancer cells. Nutr. Cancer, 65, 118–125, 2013.
53. Zhang, Z., Knobloch, T.J., Seamon, L.G., Stoner, G.D., Cohn, D.E., Paskett, E.D., Fowler, J.M., and
Weghorst, C.M., A black raspberry extract inhibits proliferation and regulates apoptosis in cervical
cancer cells. Gynecol. Oncol., 123, 401–406, 2011.
54. Liu, M., Li, X.Q., Weber, C., Lee, C.Y., Brown, J., and Liu, R.H., Antioxidant and antiproliferative
activities of raspberries. J. Agric. Food Chem., 50, 2926–2930, 2002.
55. Takata, R., Yamamoto, R., Yanai, T., Konno, T., and Okubo, T., Immunostimulatory effects of a
polysaccharide-rich substance with antitumor activity isolated from black currant (Ribes nigrum L.).
Biosci. Biotechnol. Biochem., 69, 2042–2050, 2005.
56. Damar, I. and Eksi, A., Antioxidant capacity and anthocyanin profile of sour cherry (Prunus cerasus L.)
juice. Food Chem., 135, 2910–2914, 2012.
57. Martin, K.R. and Wooden, A., Tart cherry juice induces differential dose-dependent effects on apop-
tosis, but not cellular proliferation, in MCF-7 human breast cancer cells. J. Med. Food., 15, 945–954,
2012.
58. Aparicio-Soto, M., Alarcon-de-la-Lastra, C., Cardeno, A., Sanchez-Fidalgo, S., and Sanchez-Hidalgo,
M., Melatonin modulates microsomal PGE synthase 1 and NF-E2-related factor-2-regulated antioxidant
enzyme expression in LPS-induced murine peritoneal macrophages. Br. J. Pharmacol., 171, 134–144, 2014.
59. Bizzarri, M., Proietti, S., Cucina, A., and Reiter, R.J., Molecular mechanisms of the pro-apoptotic
actions of melatonin in cancer: A review. Expert Opin. Ther. Targets, 17, 1483–1496, 2013.
60. Carbajo-Pescador, S., Ordonez, R., Benet, M., Jover, R., Garcia-Palomo, A., Mauriz, J.L., and Gonzalez-
Gallego, J., Inhibition of VEGF expression through blockade of Hif1alpha and STAT3 signalling medi-
ates the anti-angiogenic effect of melatonin in HepG2 liver cancer cells. Br. J. Cancer, 109, 83–91, 2013.
61. Faria, A., Monteiro, R., Azevedo, I., and Calhau, C., Pomegranate juice effects on cytochrome P450S
expression: In vivo studies. J. Med. Food, 10, 643–649, 2007.
62. Adams, L.S., Seeram, N.P., Aggarwal, B.B., Takada, Y., Sand, D., and Heber, D., Pomegranate juice,
total pomegranate ellagitannins, and punicalagin suppress inflammatory cell signaling in colon cancer
cells. J. Agric. Food Chem., 54, 980–985, 2006.
63. Kim, H., Banerjee, N., Ivanov, I., Talcott, S., and Mertens-Talcott, S., Comparison of anti-inflammatory
mechanisms of mango (Mangifera indica L.) and pomegranate (Punica granatum L.) in DSS-induced
colitis in rats (372.8). FASEB J., 28, 372–374, 2014.
64. Mercadante, A.Z. and Rodriguez-Amaya, D.B., Effects of ripening, cultivar differences, and processing
on the carotenoid composition of mango. J. Agric. Food Chem., 46, 128–130, 1998.
65. Noratto, G.D., Bertoldi, M.C., Krenek, K., Talcott, S.T., Stringheta, P.C., and Mertens-Talcott, S.U.,
Anticarcinogenic effects of polyphenolics from mango (Mangifera indica) varieties. J. Agric. Food
Chem., 58, 4104–4112, 2010.
66. Schieber, A., Berardini, N., and Carle, R., Identification of flavonol and xanthone glycosides from
mango (Mangifera indica L. Cv. “Tommy Atkins”) peels by high-performance liquid chromatography-
electrospray ionization mass spectrometry. J. Agric. Food Chem., 51, 5006–5011, 2003.
67. Schieber, A., Ullrich, W., and Carle, R., Characterization of polyphenols in mango puree concentrate
by HPLC with diode array and mass spectrometric detection. Innov. Food Sci. Emerg. Technol., 1,
161–166, 2000.
68. Botting, K.J., Young, M.M., Pearson, A.E., Harris, P.J., and Ferguson L.R., Antimutagens in food plants
eaten by Polynesians: Micronutrients, phytochemicals and protection against bacterial mutagenicity of
the heterocyclic amine 2-amino-3-methylimidazo[4,5-f]quinoline. Food Chem. Toxicol., 37, 95–103,
1999.
69. Gouado, I., Schweigert, F.J., Ejoh, R.A., Tchouanguep, M.F., and Camp, J.V., Systemic levels of carot-
enoids from mangoes and papaya consumed in three forms (juice, fresh and dry slice). Eur. J. Clin.
Nutr., 61, 1180–1188, 2007.
70. Percival, S.S., Talcott, S.T., Chin, S.T., Mallak, A.C., Lounds-Singleton, A., and Pettit-Moore, J.,
Neoplastic transformation of BALB/3T3 cells and cell cycle of HL-60 cells are inhibited by mango
(Mangifera indica L.) juice and mango juice extracts. J. Nutr., 136, 1300–1304, 2006.
71. Kim, H., Minamoto, Y., Markel, M., Suchodolski, J., Talcott, S., and Mertens-Talcott, S., Mango and
pomegranate polyphenolics in the modification of microbiota and short chain fatty acids in DSS-induced
colitis (1045.6). FASEB J., 28, 1045–1046, 2014.
72. Lin, Y.L., Chou, C.H., Yang, D.J., Chen, J.W., Tzang, B.S., and Chen, Y.C., Hypolipidemic and anti-
oxidative effects of noni (Morinda citrifolia L.) juice on high- fat/cholesterol-dietary hamsters. Plant
Foods Hum. Nutr., 67, 294–302, 2012.
73. Dussossoy, E., Brat, P., Bony, E., Boudard, F., Poucheret, P., Mertz, C., Giaimis, J., and Michel, A.,
Characterization, anti-oxidative and anti-inflammatory effects of Costa Rican noni juice (Morinda
citrifolia L.). J. Ethnopharmacol., 133, 108–115, 2011.
74. Ma, D.L., Chen, M., Su, C.X., and West, B.J., In vivo antioxidant activity of deacetylasperulosidic acid
in noni. J. Anal. Methods Chem., 2013, 804504, 2013.
75. Furusawa, E., Hirazumi, A., Story, S., and Jensen, J., Antitumour potential of a polysaccharide-rich
substance from the fruit juice of Morinda citrifolia (noni) on sarcoma 180 ascites tumour in mice.
Phytother. Res., 17, 1158–1164, 2003.
76. Hirazumi, A. and Furusawa, E., An immunomodulatory polysaccharide-rich substance from the fruit
juice of Morinda citrifolia (noni) with antitumour activity. Phytother. Res., 13, 380–387, 1999.
77. Liu, G., Bode, A.M., Ma, W.Y., Sang, S., Ho, C.-T., and Dong, Z., Two novel glycosides from the fruits
of Morinda citrifolia (noni) inhibit AP-1 transactivation and cell transformation in the mouse epidermal
JB6 cell line. Cancer Res., 61, 5749–5756, 2001.
78. Hornick, C.A., Myers, A., Sadowska-Krowicka, H., Anthony, C.T., and Woltering, E.A., Inhibition
of angiogenic initiation and disruption of newly established human vascular networks by juice from
Morinda citrifolia (noni)i. Angiogenesis, 6, 143–149, 2003.
79. Jang, B.C., The fruit juice of Morinda citrifolia (noni) downregulates HIF-1alpha protein expression
through inhibition of PKB, ERK-1/2, JNK-1 and S6 in manganese-stimulated A549 human lung cancer
cells. Int. J. Mol. Med., 29, 499–504, 2012.
80. Tiziani, S., Schwartz, S.J., and Vodovotz, Y., Profiling of carotenoids in tomato juice by one- and two-
dimensional NMR. J. Agric. Food Chem., 54, 6094–6100, 2006.
81. De, S. and Das, S., Protective effects of tomato juice on mouse skin carcinogenesis, Asian Pac. J.
Cancer Prev., 2, 43–47, 2001.
82. Okajima, E., Tsutsumi, M., Ozono, S., Akai, H., Denda, A., Nishino, H., Oshima, S., Sakamoto, H., and
Konishi, Y., Inhibitory effect of tomato juice on rat urinary bladder carcinogenesis after N-butyl-N-(4-
hydroxybutyl)nitrosamine initiation. Jpn. J. Cancer Res., 89, 22–26, 1998.
83. Narisawa, T., Fukaura, Y., Hasebe, M., Nomura, S., Oshima, S., Sakamoto, H., Inakuma, T., Ishiguro,
Y., Takayasu, J., and Nishino, H., Prevention of N-methylnitrosourea-induced colon carcinogenesis
in F344 rats by lycopene and tomato juice rich in lycopene. Jpn. J. Cancer Res., 89, 1003–1008,
1998.
84. Schnabele, K., Briviba, K., Bub, A., Roser, S., Pool-Zobel, B.L., and Rechkemmer, G., Effects of car-
rot and tomato juice consumption on faecal markers relevant to colon carcinogenesis in humans. Br. J.
Nutr., 99, 606–613, 2008.
85. Harms-Ringdahl, M., Jenssen, D., and Haghdoost, S., Tomato juice intake suppressed serum concentra-
tion of 8-oxodG after extensive physical activity. Nutr. J., 11, 29, 2012.
86. Bohn, T., Blackwood, M., Francis, D., Tian, Q., Schwartz, S.J., and Clinton, S.K., Bioavailability of
phytochemical constituents from a novel soy fortified lycopene rich tomato juice developed for targeted
cancer prevention trials. Nutr. Cancer, 65, 919–929, 2013.
87. Franke, A.A., Cooney, R.V., Henning, S.M., and Custer, L.J., Bioavailability and antioxidant effects of
orange juice components in humans. J. Agric. Food Chem., 53, 5170–5178, 2005.
88. Held, S., Schieberle, P., and Somoza, V., Characterization of alpha-terpineol as an anti-inflamma-
tory component of orange juice by in vitro studies using oral buccal cells. J. Agric. Food Chem., 55,
8040–8046, 2007.
89. Tounsi, M.S., Wannes, W.A., Ouerghemmi, I., Jegham, S., Ben-Njima, Y., Hamdaoui, G., Zemni, H.,
and Marzouk, B., Juice components and antioxidant capacity of four Tunisian citrus varieties. J. Sci.
Food Agric., 91, 142–151, 2011.
90. Foroudi, S., Potter, A.S., Stamatikos, A., Patil, B.S., and Deyhim, F., Drinking orange juice increases
total antioxidant status and decreases lipid peroxidation in adults. J. Med. Food., 17, 612–617, 2014.
91. Stella, S.P., Ferrarezi, A.C., dos-Santos, K.O., and Monteiro, M., Antioxidant activity of commercial
ready-to-drink orange juice and nectar. J. Food Sci., 76, C392–C397, 2011.
92. Ghanim, H., Sia, C.L., Upadhyay, M., Korzeniewski, K., Viswanathan, P., Abuaysheh, S., Mohanty, P.,
and Dandona, P., Orange juice neutralizes the proinflammatory effect of a high-fat, high-carbohydrate
meal and prevents endotoxin increase and Toll-like receptor expression. Am. J. Clin. Nutr., 91, 940–949,
2010.
93. Miyagi, Y., Om, A.S., Chee, K.M., and Bennink, M.R., Inhibition of azoxymethane-induced colon can-
cer by orange juice. Nutr. Cancer, 36, 224–229, 2000.
94. So, F.V., Guthrie, N., Chambers, A.F., Moussa, M., and Carroll, K.K., Inhibition of human breast cancer
cell proliferation and delay of mammary tumorigenesis by flavonoids and citrus juices. Nutr. Cancer,
26, 167–181, 1996.
95. Tanaka, T., Kohno, H., Murakami, M., Shimada, R., Kagami, S., Sumida, T., Azuma, Y., and Ogawa,
H., Suppression of azoxymethane-induced colon carcinogenesis in male F344 rats by mandarin juices
rich in beta-cryptoxanthin and hesperidin. Int. J. Cancer, 88, 146–150, 2000.
96. Kohno, H., Taima, M., Sumida, T., Azuma, Y., Ogawa, H., and Tanaka, T., Inhibitory effect of m andarin
juice rich in beta-cryptoxanthin and hesperidin on 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone-
induced pulmonary tumorigenesis in mice. Cancer Lett., 174, 141–150, 2001.
97. Navarra, M., Ursino, M.R., Ferlazzo, N., Russo, M., Schumacher, U., and Valentiner, U., Effect of Citrus
bergamia juice on human neuroblastoma cells in vitro and in metastatic xenograft models. Fitoterapia,
95, 83–92, 2014.
98. Zhu, X., Luo, F., Zheng, Y., Zhang, J., Huang, J., Sun, C., Li, X., and Chen, K., Characterization, purifi-
cation of poncirin from edible citrus ougan (Citrus reticulate cv. Suavissima) and its growth inhibitory
effect on human gastric cancer cells SGC-7901. Int. J. Mol. Sci., 14, 8684–8697, 2013.
99. Otsuka, T., Tsukamoto, T., Tanaka, H., Inada, K., Utsunomiya, H., Mizoshita, T., Kumagai, T.,
Katsuyama, T., Miki, K., and Tatematsu, M., Suppressive effects of fruit-juice concentrate of Prunus
mume Sieb. et Zucc. (Japanese apricot, Ume) on Helicobacter pylori-induced glandular stomach lesions
in Mongolian gerbils. Asian Pac. J. Cancer Prev., 6, 337–341, 2005.
100. Chitchumroonchokchai, C., Riedl, K.M., Suksumrarn, S., Clinton, S.K., Kinghorn, A.D., and Failla,
M.L., Xanthones in mangosteen juice are absorbed and partially conjugated by healthy adults. J. Nutr.,
142, 675–680, 2012.
101. Won, Y.S., Lee, J.H., Kwon, S.J., Kim, J.Y., Park, K.H., Lee, M.K., and Seo, K.I., Alpha-mangostin-
induced apoptosis is mediated by estrogen receptor alpha in human breast cancer cells. Food Chem.
Toxicol., 66, 158–165, 2014.
102. Ho, C.K., Huang, Y.L., and Chen, C.C., Garcinone E, a xanthone derivative, has potent cytotoxic effect
against hepatocellular carcinoma cell lines. Planta Med., 68, 975–979, 2002.
103. Liu, Z., Antalek, M., Nguyen, L., Li, X., Tian, X., Le, A., and Zi, X., The effect of gartanin, a naturally
occurring xanthone in mangosteen juice, on the mTOR pathway, autophagy, apoptosis, and the growth
of human urinary bladder cancer cell lines. Nutr. Cancer, 65(Suppl. 1), 68–77, 2013.
104. Madrigal-Santillan, E., Garcia-Melo, F., Morales-Gonzalez, J.A., Vazquez-Alvarado, P., Munoz-Juarez,
S., Zuniga-Perez, C., Sumaya-Martinez, M.T., Madrigal-Bujaidar, E., and Hernandez-Ceruelos, A.,
Antioxidant and anticlastogenic capacity of prickly pear juice. Nutrients, 5, 4145–4158, 2013.
105. Chavez-Santoscoy, R.A., Gutierrez-Uribe, J.A., and Serna-Saldivar, S.O., Phenolic composition, anti-
oxidant capacity and in vitro cancer cell cytotoxicity of nine prickly pear (Opuntia spp.) juices. Plant
Foods Hum. Nutr., 64, 146–152, 2009.
6
Fruit Juices and the Prevention of
Postprandial Metabolic Stress in Humans
Giuseppa Morabito and Mauro Serafini
CONTENTS
6.1 Introduction..................................................................................................................................... 69
6.2 Postprandial Metabolic Stress......................................................................................................... 69
6.3 Effects of Fruit Juices on Postprandial Stress: Evidence in Humans............................................. 70
6.4 Bioactives: Fruit and Fruit Juice Polyphenols................................................................................. 77
6.5 Conclusion....................................................................................................................................... 78
References................................................................................................................................................. 79
6.1 Introduction
A large body of epidemiological evidence strongly suggests a primary role for plant-based dietary
patterns in reducing the risk of diseases, such as cardiovascular disease (CVD) and cancer, impacting
overall mortality [1,2]. However, identification of the molecules involved in the protective effect of plant
foods and their mechanism of action is ongoing [3]. Dietary polyphenols contained in plant food–derived
products have been suggested to be involved in the prevention of oxidative stress, reducing free-radical-
related cellular damage, potentiating redox defense of the body, and contributing to the reduction of the
risk of developing free-radical-related diseases [4].
The consumption of unbalanced meals, consisting of foods rich in lipids and/or carbohydrates and
calories, typical of a Western diet style, has been shown to increase the susceptibility of the organism
toward oxidative damage; metabolic and transcriptional pathways are activated leading to a massive
increase in the production of free radicals and proinflammatory markers. Growing evidence suggests
that association of fruit-derived products, such as fruit juices, to a calorie-dense meal, may help attenuate
the onset of postprandial metabolic and inflammatory stress. The available evidence investigating the
effects of polyphenol-rich fruit juices on the modulation of postprandial-induced oxidative and inflam-
matory stress in humans are reviewed and critically discussed.
6.2 Postprandial Metabolic Stress

The postprandial state is characterized by a dynamic metabolic response following meals. During this
period, the organism must undergo an intense metabolic activity aimed to metabolize the absorbed
substrates, including lipids, carbohydrates, proteins, and other dietary constituents. In this phase of
short-term disturbance, major biological systems are engaged in the activation of compensatory mecha-
nisms to restore body homoeostasis. Most of the time, the stress is modest, and physiological balance
is rapidly restored; however, daily stress might give raise to clusters of different metabolic stress fac-
tors. Postprandial stress has received great attention in the last decades due to the fact that, in Western
societies, most people consume three or more meals a day, spending a considerable amount of time in
a nonfasting postprandial state. Moreover, it should take into account that the Western diet is mainly
69
characterized by the consumption of refined foods and unbalanced meals consisting of foods rich in
lipids and carbohydrates. Indeed, after the consumption of unbalanced meals, the susceptibility of the
organism toward oxidative damage is increased, and metabolic and transcriptional pathways are acti-
vated leading to a massive increase in the production of free radicals and pro-inflammatory cytokines
[5–7]. In the long term, consistent acute postprandial stress arising from repeated dietary stressors
may turn into a chronic state of oxidative stress and inflammation associated with an increased risk for
chronic and metabolic diseases [8–12].
Recent in vivo studies have demonstrated a significant increase of a wide range of pro-oxidative vari-
ables such as glucose, insulin, triacylglycerols (TAG), and consequently reactive oxygen species (ROS)
and inflammatory variables such as plasma interleukin (IL)-6, IL-17, IL-8, tumor necrosis factor-α (TNF-
α), neutrophils count, soluble intercellular adhesion molecule-1, and vascular adhesion molecule-1, and
concomitantly a decrease in antioxidant variables following a dietary stressor [11,13–16]. All the afore-
mentioned factors, indicative of chronic low-grade systemic inflammation, are believed to be involved
in the pathogenesis of CVD, insulin resistance, and type-2 diabetes [17,18]. Chronic low-grade systemic
inflammation generated by consecutive intake of hypercaloric meals is also a hallmark of obesity [19].
6.3 Effects of Fruit Juices on Postprandial Stress: Evidence in Humans

In the last decade, it has been hypothesized that postprandial metabolic stress induced by hypercaloric
meals can be attenuated by the concomitant ingestion of antioxidant-rich foods and beverages. Fruit-
derived products, such as fruit juices, contain bioactive molecules that have long been recognized to play
a role in decreasing the risk of development of degenerative diseases. A description of the characteristics,
classification, and distribution of a group of bioactive molecules, and polyphenols, will subsequently be
provided.
In vitro studies widely show the antioxidant potential of fruit polyphenols; however, the high in vitro
antioxidant capacity of fruit juices may not be a real indicator of their antioxidant potential in vivo due to
their low bioavailability and to the extensive metabolism in the human body. In humans, only 15 dietary
intervention trials investigating the effects of fruit juices on the modulation of postprandial metabolic
stress are available in the literature. Except for two interventions in which a 6-week polyphenols treat-
ment was provided to subjects and then the effect of acute test meal ingestion was evaluated [20,21], all
the others followed an acute (1 day) study design with concomitant consumption of polyphenol-rich fruit
juices to a test meal. The test meals were designed to provide a dietary stress to volunteers with an energy
intake comprised between 200 and 1344 kcal and consisting of food rich in carbohydrates (either high-
carbohydrate meal [HCM] or high fat meal [HFM] or both high-carbohydrate, high-fat meal [HCHFM]).
In the analyzed studies, the number of volunteers enrolled was comprised between 6 and 24, with 53%
(8/15) of the interventions involving healthy subjects and the remaining 47% (7/15) overweight subjects.
Within the latter four trials, they involved dyslipidemic (1/8) or hyperlipidemic (2/8) or are character-
ized by atherosclerosis-prone phenotype (1/8) volunteers. Given that, overweight subjects have a strong
tendency to transition to obesity, identifying whether inflammation perturbations occur during the over-
weight period in the postprandial state that may be clinically relevant and is currently not well known.
For convenience of analysis, the selected studies were split in two different tables. Table 6.1 summa-
rizes all the interventions (n = 12) in which clinical metabolic parameters such as total cholesterol (TC),
TAG, insulin, and glucose have been measured. Table 6.2 reviews intervention studies (n = 11) focused
on markers of oxidative stress such as nonenzymatic antioxidant capacity (NEAC) and ROS and markers
of inflammation (cytokines, and so on) after intake of a test meal together with fruit juices.
In the postprandial phase, following the ingestion of the test meals, an expected increase in plasma
TAG was observed in all the reviewed studies. Five trials out of eight reported no significant changes in
plasma TAG levels when consuming the meal together with orange juice and apple [22], black currant–
based juice [23], blueberry drink [24], or a mixed fruit juice [16]. A chronic consumption of a milk-based
strawberry beverage for 6 weeks, as part of the usual diet of the subjects, was ineffective in lowering the
postprandial TAG levels [21]. On the contrary, Burton-Freeman [20] showed that acute strawberry bever-
age consumption (single intake) significantly reduced postprandial TAG levels compared to a placebo in
TABLE 6.1
Human Intervention Studies Reporting Circulating Levels of Triacylglycerol, Total Cholesterol, Glucose, and Insulin Measurements Following Ingestion
of a Stressor Meal Associated with Fruit Juices
Number and Health Total
Study Design Status of Subjects Treatment Triacylglycerols Cholesterol Glucose Insulin References
Acute parallel placebo 20 healthy (M, F) HFM (802 kcal) (59.9% fat + 190 mL orange ↔ ↔ ↑ nd [22]
controlled juice and 400 g apple)
Acute crossover 24 overweight HFM + strawberry beverage (containing 10 g ↓ ↔ nd nd [20]
placebo controlled hyperlipidemic (10M, 14F) freeze-dried fruit/serving = 110 g/fresh weight)
Long-term crossover 24 overweight HFM (6 weeks of strawberry beverage containing ↓ ↓ nd nd [20]
placebo controlled hyperlipidemic (10M, 14F) 10 g freeze-dried fruit/serving = 110 g/fresh
weight)
Acute crossover 24 overweight dyslipidemic HCHFM + milk-based strawberry beverage nd nd ↔ ↓ [26]
placebo controlled (10M, 14F)
Long-term parallel 24 overweight (10M, 14F) HCF meal (960 kcal) (6 weeks of strawberry ↔ ↔ nd nd [21]
placebo controlled beverage; TP, 64.7 mg/305 mL)
Acute crossover 10 healthy (M, F) HCHFM (900 kcal) + orange juice (300 kcal) nd nd ↔ ↔ [27]
placebo controlled
Acute crossover 11 healthy (4M, 7F) HCM (70 g bread) + bilberry drink (bilberry 10% nd nd ↓ ↓ [28]
placebo controlled and oatmeal 5%), fermented with Lactobacillus
plantarum 299v (BFOMD) (Pro-Viva®)
Acute crossover 11 healthy (4M, 7F) HCM (70 g bread) + bilberry drink BBFOMD nd nd ↓ ↓ [28]
placebo controlled (bilberries enriched fermented oat meal drink),
BFOMD (bilberries fermented oat meal drink)
added with frozen, thawed, and homogenized
Fruit Juices and the Prevention of Postprandial Metabolic Stress in Humans
bilberries
Acute crossover 11 overweight HFM (30% fat) + 250 g black currant–based ↔ ↔ ↔ ↔ [23]
placebo controlled atherosclerosis-prone juice
phenotype (M)
Acute crossover 8 healthy (M) HFM (853 kcal) (49.3% fat, 35.4% carbohydrates, ↔ ↔ ↑ nd [24]
placebo controlled 15.3% protein + 100 g freeze-dried wild
blueberry powder in 500 mL water)
(Continued)
71
72

Human Intervention Studies Reporting Circulating Levels of Triacylglycerol, Total Cholesterol, Glucose, and Insulin Measurements Following Ingestion
of a Stressor Meal Associated with Fruit Juices
Number and Health Total
Study Design Status of Subjects Treatment Triacylglycerols Cholesterol Glucose Insulin References
Acute crossover 14 overweight (12M, 2F) HFM (1344 kcal) (55% fat, 30% carbohydrates, ↔ ↔ ↔ ↔ [16]
placebo controlled and 15% protein) + 500 mL fruit juice (40%
pineapple, 18% black currant, and 5% plum)
Acute crossover 14 overweight (12M, 2F) HFM (1344 kcal) (55% fat, 30% carbohydrates, ↓ ↓ ↔ ↔ [25]
placebo controlled and15% protein) + 500 mL fruit-based juice
drink (mix of apple, red grape, raspberry,
pomegranate juice, grape skin, grape seed,
green tea extracts, ascorbic acid, potassium
citrate, apple polyphenols, lemon flavonoids,
sucralose, and acesulfame K)
Abbreviations:
↑, increase; ↓, decrease; ↔, no changes; HCF, high carbohydrate/fat; HCHFM, high-carbohydrate, high-fat meal; HCM, high-carbohydrate meal; HFM, high-fat meal; nd,
not determined; TAG, triacylglycerols; TC, total cholesterol; TP, total polyphenols.
TABLE 6.2
Human Intervention Studies Reporting Circulating Levels of Nonenzymatic Antioxidant Capacity, Reactive Oxygen Species, and Inflammatory Marker
Measurements Following Ingestion of a Test Meal Associated with Fruit Juices
Nonenzymatic Reactive
Number and Health Antioxidant Oxygen Inflammatory
Study Design Status of Subjects Treatment Capacity Polyphenols Species Markers References
Acute parallel 20 healthy (M, F) HCM + 190 mL orange juice and 400 g nd nd ↔ nd [22]
placebo apple (802 kcal)
controlled
Acute 24 overweight HCHFM (960 kcal) + milk-based nd ↑ pelargonidin sulfate, nd ↓ hs-CRP and [26]
crossover dyslipidemic strawberry beverage pelargonidin-3-O- IL-6
placebo (10M, 14F) glucoside (plasma) ↔ IL-1β, PAI-1,
controlled and TNF-α
Long-term 24 overweight HCF meal (960 kcal) (6 weeks of nd nd nd ↓ IL-1β and [21]
parallel (10M, 14F) milk-based strawberry beverage and PAI-1
placebo TP 64.7 mg/305 mL) ↔ IL -6, hs-CRP,
controlled and TNF-α
Acute 10 healthy (M, F) HCHFM (900 kcal) + orange juice nd nd ↓ ROS by ↔ p38 MAPK [27]
crossover (300 kcal) PMNs protein
placebo ↔ p47phox
controlled ↔ MMP-9,
↓TLR-2 and
TLR-4
Acute 11 overweight HFM (30% fat) + 250 g black currant– ↑ ORAC ↑ 1,3,5-trihydroxybenzene nd ↑ TNF-α, ex vivo [23]
crossover atherosclerosis- based juice (plasma) ↔IL-1β ex vivo
placebo prone phenotype (M) ↔IL-6 plasma
controlled
Fruit Juices and the Prevention of Postprandial Metabolic Stress in Humans
Acute 8 healthy (M) HFM (853 kcal) (49.3% fat, 35.4% ↔ORAC nd nd nd [24]
crossover carbohydrates, and 15.3% protein + 100 g ↑ ORAC PCA
placebo freeze-dried wild blueberry powder in ↑ TEAC
controlled 500 mL water)
Acute 14 overweight HFM (1344 kcal) (55% fat, 30% ↑ TRAP nd nd nd [30]
crossover (12M, 2F) carbohydrates, and 15% protein) + 500 mL ↔ FRAP
placebo fruit juice (86% of a mix of apple, grape, ↑ FRAP (urine)
controlled blueberry, pomegranate juices, grape skin,
grape seed, and green tea extracts)
(Continued)
73
74

Human Intervention Studies Reporting Circulating Levels of Nonenzymatic Antioxidant Capacity, Reactive Oxygen Species, and Inflammatory Marker
Measurements Following Ingestion of a Test Meal Associated with Fruit Juices
Nonenzymatic Reactive
Number and Health Antioxidant Oxygen Inflammatory
Study Design Status of Subjects Treatment Capacity Polyphenols Species Markers References
Acute 14 overweight HFM (1344 kcal) (55% fat, 30% ↔ TRAP nd nd nd [30]
crossover (12M, 2F) carbohydrates, and 15% protein) + ↔ FRAP
placebo 500 mL fruit juice (63% of a mix of ↑ UA
controlled pineapple, black currant, and plum juices)
Acute 16 young healthy HCM 28 g of Kellogg’s Corn Flakes and ↑ ORAC ↔ TP nd nd [29]
crossover (M, F) 118 mL of 2% milk + in 591 mL of navel
placebo orange juice
controlled
Acute 14 overweight HFM (1344 kcal) (55% fat, 30% nd nd nd ↓ IL-17 [16]
crossover (12M, 2F) carbohydrates, and 15% protein) + ↓ IL-6
placebo 500 mL fruit juice (40% pineapple, ↓ TNF-α
controlled 18% black currant, and 5% plum)
Acute 14 overweight HFM (1344 kcal) (55% fat, 30% nd nd nd ↓ IL-6 [25]
crossover (12M, 2F) carbohydrates, and 15% protein) + ↓ TNF-α
placebo 500 mL fruit-based juice drink (mix of
controlled apple, red grape, raspberry, pomegranate
juices, grape skin, grape seed, green tea
extracts, ascorbic acid, potassium citrate,
apple polyphenols, lemon flavonoids,
sucralose, and acesulfame K)
Abbreviations:
↑, increase; ↓, decrease; ↔, no changes; FRAP, ferric-reducing antioxidant potential; HCF, high carbohydrate/fat; HCHFM, high-carbohydrate, high-fat meal; HCM, high-
carbohydrate meal; HFM, high-fat meal; hs-CRP, high-sensitivity C-reactive protein; IL, interleukin; MAPK, mitogen-activated protein kinase; MMP, metalloproteinase; nd,
not determined; NEAC, nonenzymatic antioxidant capacity; ORAC, oxygen radical absorbance capacity; PAI-1, plasminogen activator inhibitor-1; PCA, perchloric acid;
PMN, polymorphonuclear leukocytes; ROS, reactive oxygen species; TEAC, trolox equivalent antioxidant capacity; TNF-α, tumor necrosis factor-α; TP, total polyphenols;
TRAP, total radical-trapping antioxidant potential; TRL, toll-like receptor; UA, uric acid.
Fruit Juices and the Prevention of Postprandial Metabolic Stress in Humans 75
overweight hyperlipidemic subjects; moreover, they confirmed this result after chronic consumption of
the same beverage for 6 weeks as part of the usual diet when significantly lower TAG (P < 0.0001) levels
were recorded in response to the HFM compared to a placebo.
A significant decrease in postprandial TAG was also observed between 4 and 8 h (P < 0.05) compared
to a placebo following the intake of a fruit-based juice drink containing apple, red grape, raspberry,
pomegranate, grape skin, seed, and green tea extracts in overweight volunteers [25] (Table 6.1).
The intake of the test meal induced no changes in plasma TC levels in all the reviewed trials except for
two studies [16,25], in which a slight but significant increase was recorded 8 h after HFM consumption.
Postprandial TC concentration did not change significantly when orange juice and apple [22], strawberry
beverage [20], milk-based strawberry beverage [21], black currant–based juice [23], blueberry-based
juice [24], and a mixed fruit juice [16] were given to volunteers with the test meal. The ingestion of a
fruit-based juice drink [25] concomitantly to the HFM significantly reduced HFM-induced increases in
plasma TC levels (P < 0.05) at 6–8 h. An interesting result was obtained by Burton-Freeman [20] who
provided a 6-week treatment with a strawberry beverage to overweight hyperlipidemic subjects, who sub-
sequently showed a significant decrease in TC levels (P < 0.0001) after consuming an HFM (Table 6.1).
As expected, glucose and insulin plasma concentrations also increased when consuming a HFM,
HCM, or HCHFM with fruit juices. Available data showed that postprandial glucose levels did not
change significantly following the consumption of milk-based strawberry beverages [26], black currant–
based juices [23], or mixed fruit juices [16,25] in overweight subjects and orange juice [27] in healthy
volunteers. Conversely, changes in postprandial glucose levels were observed in four trials conducted in
healthy subjects, half of which reported an increase after orange juice and apple [22], or blueberry drink
(P < 0.05) [24], while the other half showed a significant reduction (P < 0.05) following the intake of a
fermented oatmeal drink added with bilberries, and this effect was concomitant to an insulin reduction
[28]. Postprandial insulin concentration was also significantly reduced (P < 0.01) after consumption of
a milk-based strawberry beverage as compared to the placebo beverage in overweight subjects [26];
however, no effect was reported in postprandial glucose level, as described earlier. In the remaining four
interventions, in which insulin measurements were recorded following the ingestion of orange juice [27],
black currant–based juice [23], or mixed fruit juices [16,25] with a test meal, no changes in postprandial
insulin concentration were noted (Table 6.1).
Plasma NEAC was measured in 5 out of 11 reviewed trials. Among these, different analytical meth-
odologies, including oxygen radical antioxidant capacity (ORAC), ferric-reducing antioxidant power
(FRAP), trolox equivalent antioxidant capacity (TEAC), and total radical-trapping antioxidant param-
eter (TRAP), have been used for plasma NEAC evaluation as shown in Table 6.2.
Plasma ORAC was shown to significantly increase (P < 0.05) at 1.5–2 h after HFM and black currant
juice intake compared to a placebo in overweight subjects [23]. A similar increase (P < 0.05) in serum
ORAC, determined in samples without proteins, treated with perchloric acid (PCA) and TEAC was
observed by Kay and Holub [24] at 1 h (P = 0.02) after intake of an HFM in association with blueberry
drink, compared to the control group in healthy individuals. Increased postprandial ORAC was also
measured 2 h following navel orange juice consumption in association with a HCM in young healthy
volunteers, but the effect disappeared by 3 h [29].
Recently, Miglio et al. [30] investigated the effects of antioxidant-rich fruit juice drinks on the endog-
enous antioxidant response induced by HFM. The two tested beverages had different composition
resulting in different antioxidant properties: AB1 contained 86% of a mix of apple, grape, blueberry,
and pomegranate juices and grape skin, grape seed, and green tea extracts. It contained high levels
of flavan-3-ols (407 mg/L), along with lower concentrations of hydroxycinnamic acids, flavonols, and
anthocyanins, while AB2 contained 63% of a mix of pineapple, black currant, and plum juices and
mainly anthocyanins (32 mg/L). AB1 displayed a higher in vitro NEAC values than AB2. In agreement
with in vitro findings, the previously mentioned study showed that AB1 ingestion, but not AB2 drink,
resulted in a higher antioxidant effect in vivo through an increase in plasma TRAP and urinary FRAP
concentrations. A significant increase in plasma uric acid (UA) concentration was observed following
HFM (placebo beverage) consumption. This finding is important in view of the fact that UA increase
following dietary stressors might represent a mechanism of endogenous antioxidant protection, pro-
viding an explanation for the association of high UA level with development of inflammatory-related
diseases [31]. Additionally, we showed that AB2 ingestion led to similar urinary excretion of UA recorded
after p lacebo consumption but higher than that observed for AB1. This result suggests that AB2 drink,
providing less amount of exogenous antioxidants, do not provide the same antioxidant efficiency than
AB1 in reducing endogenous antioxidant response to dietary stressors carried out by UA [30].
Circulating levels of polyphenols or their metabolites have been evaluated only in 3 out of 10 studies
[23,26,29]. Analyzing the single trials, the intake of an HCHFM associated with a milk-based straw-
berry beverage containing 28.11 mg of pelargonidin-3-O-glucoside significantly increased plasma post-
prandial pelargonidin sulfate and pelargonidin-3-O-glucoside (P < 0.001) concentration in overweight
dyslipidemic volunteers [26]. Blood samples collected after the consumption of HFM and black currant–
based juice showed increased anthocyanin metabolites concentration such as 1,3,5-trihydroxybenzene
at 130 min that decreased to basal level only after 20 min [23]. In contrast, plasma total polyphenols
(TPs), assessed using the Folin–Ciocalteu assay, resulted to be unaffected after intake of navel orange
juice and HCM [29]. In only one of these three studies in which circulating polyphenol levels were mea-
sured, a parallel increase in plasma polyphenol metabolites and NEAC was observed [23]. In another
trial, although an increase in plasma polyphenols has been observed, no effects on plasma NEAC were
detected [26], whereas in the third study, despite an increase in plasma NEAC, no effects on plasma
polyphenols were noticed [29].
The consumption of an HCM was associated with an increase in oxidative stress characterized by
increased production of ROS occurring during the metabolism of carbohydrates and the utilization of
oxygen. As displayed in Table 6.2, the generation of ROS by polymorphonuclear cells (PBMCs) was
measured in only two trials [22,27]. Concomitant consumption of orange juice and HCHFM led to
a lower ROS generation when compared with water [27], while no changes were observed following
orange juice and apple intake in association with HCM [22].
Six trials reported measurements of inflammatory markers including high-sensitivity C-reactive pro-
tein (hs-CRP), plasminogen activator inhibitor-1 (PAI-1), and cytokines such as TNF-α, IL-6, IL-1β, and
IL-17. For what concerns plasma IL-6 levels, a postprandial reduction was observed after consumption
of HCHFM and a strawberry beverage [26] or a fruit juice containing pineapple, black currant, and plum
[16] or containing apple, grape, blueberry, pomegranate, grape skin and seed, and green tea extracts [25],
whereas no effect on meal-induced IL-6 production was reported either after black currant juice intake
in healthy individuals [23] or after consumption of a strawberry beverage for 6 weeks as part of the daily
diet in overweight subjects [21].
Postprandial plasma TNF-α concentration was significantly reduced when a mixed fruit juices was
associated with the HFM [16,25], while no effects were reported in plasma TNF-α after HFM consump-
tion together with a milk-based strawberry beverage [26], and only one study reported increased ex vivo
TNF-α concentration following intake of HFM and black currant–based juice [23]. For the first time, we
have shown that IL-17 production is also increased following ingestion of a stressor meal, but the pres-
ence of a juice drink rich in bioactive molecules is enough to significantly inhibit IL-17 as well as TNF-α
and IL-6 production already described earlier. Thus, an active role of the diet in modulating IL-17 and,
in general, the inflammatory response is suggested [16].
In overweight subjects, the intake of a strawberry beverage with an HCHFM [26] or a black currant
juice with a HFM had no effect on IL-1β, differently from chronic consumption of a strawberry bever-
age for 6 weeks that leads to decreased plasma IL-1β concentration following HCHFM loading [21].
Meanwhile, hs-CRP was reduced following consumption of HCHFM with a strawberry beverage in
overweight subjects [26]. In contrast, no effects on postprandial hs-CRP were noted following chronic
consumption of a strawberry beverage, whereas the treatment provided reduced PAI-1 concentrations
following a high-carbohydrate/fat (HCF) loading [21].
The postprandial levels of toll-like receptor (TLR)-2 and TLR-4 as well as other inflammatory mark-
ers such as metalloproteinase (MMP)-9, plasma lipopolysaccharide (LPS), p38 mitogen-activated pro-
tein kinase (MAPK) protein, 7-kilodalton cytosolic subunit of NADPH oxidase (47phox), and nuclear
factor erythroid 2–related factor 2 binding activity have been measured by Ghanim et al. [27], showing
that only HFM intake together with orange juice induces a decrease in plasma TLR-2 and TLR-4, but no
changes in MMP-9, p38MAPK protein, or p47phox.
6.4 Bioactives: Fruit and Fruit Juice Polyphenols

Other than vegetables, fruit is the most abundant source of polyphenols in the human diet. Polyphenols
are essential for the growth and reproduction of plants and are produced as a response for defending
injured plants against pathogens. They have received increasing attention due to their antioxidant poten-
tial and for their possible use in processed foods as natural antioxidants.
The potential health benefits of polyphenols are getting more and more recognition, as reports indicate that
these compounds inhibit the harmful effects of ROS, which act as oxidants, thus protecting macromolecules
such as proteins, lipids, and DNA from oxidative degradation [32]. Their functions have long been recog-
nized, at epidemiological level, to display a role in reducing the development of degenerative diseases [33].
According to the number of phenol rings contained and to the structural elements that bind these
rings to each other, polyphenols may be divided into several classes such as flavonoids, phenolic acids,
stilbenes, and lignans [34].
The most abundant class of polyphenols in fruits is flavonoids, which have a C6–C3–C6 structure with dif-
ferent substitution patterns. The majority of flavonoids occur naturally as glycosides rather than aglycones.
The main subclasses of these compounds are flavonols, flavones, flavanones, flavanols, and anthocyanins.
Within flavonoids, flavonols are the most ubiquitous flavonoids in foods, the most common of them are
kaempferol, quercetin, isorhamnetin, and myricetin, as glycosides with conjugation at the 5, 7, 3′, 4′, and 5′
positions. They are present in many fruits, such as berry (20–40 mg/100 g), grape, plum, strawberry, and
apple (1–5 mg/100 g), and fruit beverage, such as apple juice (1–5 mg/100 mL) [35]. Fruits often contain
between 5 and 10 different flavonol glycosides. Marked differences exist in concentration between fruits on
the same tree and even between different sides of a single piece of fruit, depending on exposure to sunlight.
Flavones are much less common than flavonols in fruit. They consist chiefly of glycosides of luteo-
lin and apigenin. Polymethoxylated flavones are contained in large quantities in the skin of fruits, for
example, in the skin of mandarin at up to 6.5 g/L of mandarin essential oil [34].
Flavanones are characterized by the presence of a saturated three-carbon chain and an oxygen atom
in the C4 position. They are generally glycosylated with a disaccharide in the C7 position. Flavanones
are present in high concentrations only in citrus fruits. The main aglycones are naringenin in grapefruit,
hesperetin in orange, and eriodictyol in lemon. Orange juice contains 200–600 mg hesperidin/L and
15–85 mg narirutin/L, and a single glass of orange juice may contain between 40 and 140 mg flavanone
glycosides [36]. The solid parts of citrus fruit, in particular the white spongy portion (albedo) and the
membranes separating the segments, have very high flavanone content. This is the reason why the whole
fruit may contain flavanone at up to fivefold higher amounts than a glass of orange juice.
Flavanols is a class of flavonoids containing a saturated three-carbon chain with a hydroxyl group in
the C3 position. They exist in both the monomeric and polymeric forms (catechin and proanthocyani-
dins, respectively). Different from other flavonoid classes, flavanols are not glycosylated in fruits. The
main representative flavanols in fruit are catechin and epicatechin. Catechins are found in many fruits
such as apricot (250 mg/kg), cherry (250 mg/kg), plum (60 mg/100 g), and berry (100 mg/kg) [34].
Anthocyanins are water-soluble pigments, responsible for the red, blue, and purple colors of fruits.
Although they are highly unstable in the aglycone form (anthocyanidins), in plants, they are resistant
to light, pH, and oxidation conditions that are likely to degrade them. Degradation is prevented by gly-
cosylation, generally with a glucose unit at position 3, and esterification with various organic acids
(citric and malic acids) and phenolic acids. In addition, anthocyanins are stabilized by the formation of
complexes with other flavonoids (copigmentation). Cyanidin is the most common anthocyanin in fruits.
In human diet, anthocyanins are abundant in many fruits such as black currant, black chokeberry, and
black elderberry (100 mg/100 g); lingonberry (40–80 mg/100 g); grape, plum, raspberry, strawberry, and
blueberry (10–40 mg/100 g); cherry, cranberry, and red currant (1–20 mg/100 g); and pomegranate and
grape juices (1–10 mg/100 g) [34]. Anthocyanins are found mainly in the skin, except for certain types
of red fruit, in which they also occur in the flesh (cherry and strawberry). The content of anthocyanins is
generally proportional to color intensity, reaching values of up to 2–4 g/kg in black currant or blackberry,
and may increase as the fruit ripens [37].
With regard to phenolic acids, they could be divided in two classes of benzoic and cinnamic acid deriv-
atives. Hydroxybenzoic acids, such as gallic acid, are found in very few fruits eaten by humans, except
for certain red fruits, for example, blackberry containing up to 270 mg/kg and raspberry that contains
up to 100 mg/kg of protocatechuic acid. The hydroxycinnamic acids consist chiefly of coumaric, caffeic,
and ferulic acids that are rarely found in the free form. The bound forms are glycosylated derivatives or
ester of quinic, shikimic, or tartaric acid. Caffeic and quinic acids combine to form chlorogenic acid,
which is found in many types of fruits. Blueberry contains 2 g hydroxycinnamic acids/kg. Caffeic acid is
the most abundant phenolic acid, representing 75%–100% of the total hydroxycinnamic acids content in
most fruits. Kiwi contains up to 1 g caffeic acid/kg [34]. Hydroxycinnamic acids are present in all part of
the fruit, although the highest concentrations are seen in the outer part of the ripe fruit.
Stilbenes (1,2-diphenylethylene) are phytoalexins with a C6–C2–C6 carbon skeleton and are produced
by plants in response to disease, injury, or stressors. Stilbenes are present in fruits such as grape and
berry as cis and trans isomeric forms of resveratrol, mostly glycosylated. The fresh skin of red grapes is
particularly rich in resveratrol (50–100 g/kg) that contributes to a relatively high concentration of resve-
ratrol in grape juice (up to 7 mg aglycones/L) [38].
Lignans are rarely present in fruits and are produced by oxidative dimerization of two phenylpropane
units and are mostly present in nature in the free form, with their glycoside derivatives being only a
minor form. Fruits like kiwi, apricot, strawberry, and peach are packed with lignans. Pear, nectarine,
pink grapefruit, and cherry are also good sources; however, the richest dietary source is linseed, which
contains secoisolariciresinol (up to 3.7 g/kg) and low quantities of matairesinol [39]. Lignans are metabo-
lized to enterodiol and enterolactone by the intestinal microflora. The low quantities of secoisolarici-
resinol and matairesinol that are ingested as part of our normal diet do not account for the concentrations
of the metabolites enterodiol and enterolactone.
The health effect of polyphenols strongly depends on both their intake and bioavailability. Flavonoids,
and polyphenols in general, once absorbed are targeted as xenobiotics and metabolized, resulting in a
scarce bioavailability and low circulating concentrations postingestion. Generally, the aglycones can be
absorbed from the small intestine [40]. However, most polyphenols are present in food in the form of
ester, glycosides, or polymers that cannot be absorbed in the native form. Prior to their absorption, these
compounds must be hydrolyzed by intestinal enzymes or by the colonic microflora. During the course of
the absorption, polyphenols undergo extensive modification; in fact they are conjugated in the intestinal
cells and later in the liver by methylation, sulfation, and/or glucuronidation. As a consequence, the forms
reaching the blood and tissues are often different from those present in food, and it is very difficult to
identify all the metabolites and to evaluate their biological activity.
Most of the absorbed metabolites are excreted via urine, but it is possible that part of them is recycled
back into the small intestine through the enterohepatic transport [41]. It is believed that extensively con-
jugated metabolites are eliminated through the bile, while the smaller ones are found mainly in the urine
[34]. Although data on bioavailability indicate a low absorption of flavonoids, great metabolic variability
among different compounds has been observed, depending upon the chemical structure [42], type of gly-
coside [43], and food matrix [44]. Interindividual variations have also been observed, probably due to dif-
ferences in the colonic microflora [45], which can differently affect flavonoid metabolism. It is important
to notice that the most common polyphenols in the human diet are not necessarily the most active ones
within the body, because either they have a lower intrinsic activity or they are poorly absorbed from the
intestine, highly metabolized, or rapidly eliminated. In addition, the metabolites that are found in blood
and target organs and that result from digestive or hepatic activity may differ from the native substances
in terms of biological activity.
6.5 Conclusion
The experimental evidence from studies conducted in humans over the last decades, despite few con-
trasting results, establishes that a single meal rich in fat or carbohydrate increases blood TAG, insu-
lin, and glucose, which are considered as risk factors for CVD. Moreover, it has been shown that such
a meal induces the release of inflammatory cytokines, such as IL-6, TNF-α, IL-1β, and IL-17, into
the bloodstream. Scientific evidence suggests a direct role for plant foods and their juices, in modulating
endogenous mechanisms of defenses (e.g., antioxidants, inflammation, and immunity). However, avail-
able data on the effects of polyphenols on the modulation of postprandial oxidative and inflammatory
stress are still scarce and controversial. Most of the intervention studies lack assessment of polyphenol
absorption or fail to associate the antioxidant effect following ingestion of fruit juices with changes in
circulating levels of flavonoids or their metabolites. Thus, the effect of fruit juice and plant foods has
been attributed to their bioactives; however, the molecules responsible for this effect have not yet been
clearly identified.
Postprandial circulating pro-inflammatory cytokines have been investigated only in few studies and
results are contrasting. Intake of fruit polyphenols resulted in an inhibition of the meal-induced TNF-α
and IL-6 production in two and three out of six interventions, respectively. The slenderness of the results
reported makes it difficult to draw any firm conclusion and warrant further investigations.
Although overall data suggest that the association of polyphenol-rich fruit juices to a calorie-dense meal
may help to attenuate the postprandial-induced oxidative and inflammatory stress, we have to consider
in the first instance the small number of volunteers enrolled in the studies. Second, in Western societies
where people eat four to five times a day, meal consumption is not an isolated phenomenon throughout
the day; thus investigating only the acute ingestion of a test meal associated with fruit juices may be
misleading, as it is impossible to separate out the potential effects of the background diet. Moreover, if
acute ingestion of a calorie-dense meal is enough to trigger metabolic and inflammatory cascades, in
the long term, repeated dietary stressors may turn into a low-grade inflammatory and chronic oxidative
stress state increasing the chance to develop obesity, diabetes, atherosclerosis, and CVD.
Based on these observations, long-term trials investigating the effects of polyphenol-rich fruit juices on
the modulation of metabolic, oxidative, and inflammatory profiles following calorie-dense meal consump-
tion are urgently needed. It is crucial to establish if daily consumption of fruit-derived products, rich in
polyphenols, may play a role in preventing metabolic stress associated to the body’s postprandial response.
REFERENCES
1. Serafini, M., Jakszyn, P., Luján-Barroso, L., Agudo, A., Bas Bueno-de-Mesquita, H., van Duijnhoven,
F.J., Jenab, M. et al., Dietary total antioxidant capacity and gastric cancer risk in the European prospec-
tive investigation into cancer and nutrition study. Int. J. Cancer, 131, E544–E554, 2012.
2. Estruch, R., Ros, E., Salas-Salvadó, J., Covas, M.I., Corella, D., Arós, F., Gómez-Gracia, E. et al.,
PREDIMED Study Investigators. Primary prevention of cardiovascular disease with a Mediterranean
diet. N. Engl. J. Med., 368, 1279–1290, 2013.
3. Key, T.J., Fruit and vegetables and cancer risk. Br. J. Cancer, 104, 6–11, 2011.
4. Khan, N., Afaq, F., and Mukhtar, H., Cancer chemoprevention through dietary antioxidants: Progress
and promise. Antioxid. Redox Signal, 10, 475–510, 2008.
5. Alipour, A., Elte, J.W.F., van Zaanen, H.C.T., Rietveld, A.P., and Cabezas, M.C., Postprandial inflamma-
tion and endothelial dysfunction. Biochem. Soc. Trans, 35, 464–469, 2007.
6. van Oostrom, A.J., van Wijk, J., and Cabezas, M.C., Lipaemia, inflammation and atherosclerosis: Novel
opportunities in the understanding and treatment of atherosclerosis. Drugs, 64(Suppl. 2), 19S–41S,
2004.
7. Sies, H., Stahl, W., and Sevanian, A., Nutritional, dietary and postprandial oxidative stress. Am. Soc.
Nutr. Sci., 135, 969–972, 2005.
8. Wallace, J.P., Johnson, B., Padilla, J., and Mather, K., Postprandial lipaemia, oxidative stress and endo-
thelial function: A review. Int. J. Clin. Pract., 64, 3, 389–403, 2010.
9. Klop, B., Proctor, S.D., Mamo, J.C., Botham, K.M., and Castro Cabezas, M., Understanding postpran-
dial inflammation and its relationship to lifestyle behaviour and metabolic diseases. Int. J. Vasc. Med.,
2012, 947417, 2012.
10. Ceriello, A., Postprandial hyperglycaemia and diabetes complications: Is it time to treat? Diabetes, 54,
1e7, 2005.
11. Nappo, F., Esposito, K., Coiffi, M., Giugliano, G., Molinari, A.M., Paolisso, G., Marfella, R., and
Giugliano, D., Postprandial endothelial activation in healthy subjects and in type 2 diabetic patients:
Role of fat and carbohydrate meals. J. Am. Coll. Cardiol., 39, 1145–1150, 2002.
12. Ong, P.J.L., Dean, T.S., Hayward, C.S., Della Monica, P.L., Sanders, T.A.B., and Collins, P., Effect of
fat and carbohydrate consumption on endothelial function. Lancet, 354(9196), 2134, 1999.
13. van Oostrom, A.J., Sijmonsma, T.P., Verseyden, C., Jansen, E.H., de Koning, E.J., Rabelink, T.J., and
Castro Cabezas, M., Postprandial recruitment of neutrophils may contribute to endothelial dysfunction.
J. Lipid Res., 44, 576–583, 2003.
14. Gregersen, S., Samocha-Bonet, D., Heilbronn, L.K., and Campbell, L.V., Inflammatory and oxidative
stress responses to high-carbohydrate and high-fat meals in healthy humans. J. Nutr. Metab., 2012,
238056, 2012.
15. Esposito, K., Nappo, F., Marfella, R., Giugliano, G., Giugliano, F., Ciotola, M., Quagliaro, L., Ceriello,
A., and Giugliano, D., Inflammatory cytokine concentrations are acutely increased by hyperglycemia in
humans: Role of oxidative stress. Circulation, 106, 2067–2072, 2002.
16. Peluso, I., Raguzzini, A., Villano, D.V., Cesqui, E., Toti, E., Catasta, G., and Serafini, M., High fat
meal increase of IL-17 is prevented by ingestion of fruit juice drink in healthy overweight subjects.
Curr. Pharm. Des., 18, 85–90, 2012.
17. Pai, J.K., Pischon, T., Ma, J., Manson, J.E., Hankinson, S.E., Joshipura, K., Curhan, G.C., et al.,
Inflammatory markers and the risk of coronary heart disease in men and women. N. Engl. J. Med., 351,
2599–2610, 2004.
18. Festa, A., D’Agostino, R. Jr., Howard, G., Mykkanen, L., Tracy, R.P., and Haffner, S.M., Chronic sub-
clinical inflammation as part of the insulin resistance syndrome: The Insulin Resistance Atherosclerosis
Study (IRAS). Circulation, 102, 42–47, 2000.
19. Gregor, M.F. and Hotamisligil, G.S., Inflammatory mechanisms in obesity. Ann. Rev. Immunol., 29,
415–445, 2011.
20. Burton-Freeman, B., Postprandial metabolic events and fruit-derived phenolics: A review of the science.
Br. J. Nutr., 104(Suppl. 3), S1–S14, 2010.
21. Ellis, C.L., Edirisinghe, I., Kappagoda, T., and Burton-Freeman, B., Attenuation of meal-induced
inflammatory and thrombotic responses in overweight men and women after 6-week daily strawberry
(Fragaria) intake. A randomized placebo-controlled trial. J. Atheroscler. Thromb., 18, 318–327, 2011.
22. Bae, J.H., Bassenge, E., Kim, K.B., Kim, Y.N., Kim, K.S., Lee, H.J., Moon, K.C., Lee, M.S., Park,
K.Y., and Schwemmer M., Postprandial hypertriglyceridemia impairs endothelial function by enhanced
oxidant stress. Atherosclerosis, 155, 517–523, 2001.
23. Huebbe, P., Giller, K., de Pascual-Teresa, S., Arkenau, A., Adolphi, B., Portius, S., Arkenau, C.N., and
Rimbach, G., Effects of blackcurrant-based juice on atherosclerosis-related biomarkers in cultured mac-
rophages and in human subjects after consumption of a high-energy meal. Br. J. Nutr., 108, 234–244,
2012.
24. Kay, C.D. and Holub, B.J., The effect of wild blueberry (Vaccinium angustifolium) consumption post-
prandial serum antioxidant status in human subjects. Br. J. Nutr., 88, 389–397, 2002.
25. Peluso, I., Villano, D.V., Roberts, S.A., Cesqui, E., Raguzzini, A., Borges, G., Crozier, A., Catasta, G.,
Toti, E., and Serafini, M., Consumption of mixed fruit-juice drink and vitamin C reduces postprandial
stress induced by a high fat meal in healthy overweight subjects. Curr. Pharm. Des., 20, 1020–1024,
2014.
26. Edirisinghe, I., Banaszewski, K., Cappozzo, J., Sandhya, K., Ellis, C.L., Tadapaneni, R., Kappagoda,
C.T., and Burton-Freeman, B.M., Strawberry anthocyanin and its association with postprandial inflam-
mation and insulin. Br. J. Nutr., 106, 913–922, 2011.
27. Ghanim, H., Sia, C.L., Upadhyay, M., Korzeniewski, K., Viswanathan, P., Abuaysheh, S., Mohanty, P.,
and Dandona, P., Orange juice neutralizes the proinflammatory effect of a high-fat, high-carbohydrate
meal and prevents endotoxin increase and Toll-like receptor expression. Am. J. Clin. Nutr., 91, 940–949,
2010.
28. Granfeldt, Y.E. and Björck, I.M., A bilberry drink with fermented oatmeal decreases postprandial insu-
lin demand in young healthy adults. Nutr. J., 10, 57 2011.
29. Snyder, S.M., Reber, J.D., Freeman, B.L., Orgad, K., Eggett, D.L., and Parker, T.L., Controlling for
sugar and ascorbic acid, a mixture of flavonoids matching navel oranges significantly increases human
postprandial serum antioxidant capacity. Nutr. Res., 31, 519–526, 2011.
30. Miglio, C., Peluso, I., Raguzzini, A., Villaño, D.V., Cesqui, E., Catasta, G., Toti, E., and Serafini, M.,
Fruit juice drinks prevent endogenous antioxidant response to high-fat meal ingestion. Br. J. Nutr., 12,
1–7, 2013.
31. So, A. and Thorens, B., Uric acid transport and disease. J. Clin. Invest., 120, 1791–1799, 2010.
32. Halliwell, B., How to characterize an antioxidant: An update. Biochem. Soc. Symp., 61, 73–101, 1995.
33. Lampe, J.W., Health effects of vegetables and fruit: Assessing mechanisms of action in human experi-
mental studies. Am. J. Clin. Nutr., 70(Suppl. 3), 475S–490S, 1999.
34. D’Archivio, M., Filesi, C., Di Benedetto, R., Gargiulo, R., Giovannini, C., and Masella, R., Polyphenols,
dietary sources and bioavailability. Annali dellIstituto Superiore di Sanità, 43, 348–361, 2007.
35. Neveu, V., Perez-Jimeńez, J., Vos, F., Crespy, V., du Chaffaut, L., Mennen, L., Knox, C. et al., Phenol-
explorer: An online comprehensive database on polyphenol contents in foods 2010. Published online at:
http://dx. doi.org/10.1093/database/bap024 (accessed July 26, 2014).
36. Tomas-Barberan, F.A. and Clifford, M.N., Flavanones, chalcones and dihydrochalcones-nature, occur-
rence and dietary burden. J. Sci. Food Agric., 80, 1073–1080, 2000.
37. Clifford, M.N., Anthocyanins-nature, occurrence and dietary burden. J. Food Sci. Agric., 80, 1063–1072,
2000.
38. Bertelli, A., Bertelli, A.A., Gozzini, A., and Giovannini, L., Plasma and tissue resveratrol concentra-
tions and pharmacological activity. Drugs Exp. Clin. Res., 24, 133–138, 1998.
39. Milder, I.E.J., Arts, I.C., van de Putte, B., Venema, D.P., and Hollman, P.C., Lignan contents of Dutch
plant foods: A database including lariciresinol, pinoresinol, secoisolariciresinol and matairesinol. Br. J.
Nutr., 93, 393–402, 2005.
40. Crespy, V., Morand, C., Besson, C., Manach, C., Demigne, C., and Remesy, C., Quercetin, but not its
glycosides, is absorbed from the rat stomach. J. Agric. Food Chem., 50, 618–621, 2002.
41. Donovan, J.L., Manach, C., Faulks, R.M., and Kroon, P., Absorption and metabolism of dietary plant
secondary metabolites, in Plant Secondary Metabolites: Occurrence, Structure and Role in the Human
Diet, Crozier, A., Clifford, M.N., and Ashihara, H., Eds., Blackwell Publishing, Oxford, UK, 2006,
pp. 303–351.
42. Simons, A.L., Renouf, M., Murphy, P.A., and Hendrich, S., Greater apparent absorption of flavonoids
is associated with lesser human fecal flavonoid disappearance rates. J. Agric. Food Chem., 58, 141–147,
2010.
43. Hollman, P.C.H., de Vries, J.H.M., van Leeuwen, S.D., Mengelers, M.J., and Katan, M.B., Absorption
of dietary quercetin glycosides and quercetin in healthy ileostomy volunteers. Am. J. Clin. Nutr., 62,
1276–1282, 1995.
44. Serafini, M. and Crozier, A., Milk and absorption of dietary flavonols. Nature, 426, 788, 2003.
45. Setchell, K.D., Brown, N.M., and Lydeking-Olsen, E., The clinical importance of the metabolite equol-a
clue to the effectiveness of soy and its isoflavones. J. Nutr., 132, 3577–3584, 2002.
Section II
Fruit Juices
7
Acerola Juice
Delia B. Rodriguez-Amaya
CONTENTS
7.1 Introduction..................................................................................................................................... 85
7.2 Nutritional Characteristics.............................................................................................................. 85
7.3 Bioactives and Antioxidant Efficacy............................................................................................... 86
7.3.1 Bioactive Compounds......................................................................................................... 86
7.3.2 Antioxidant Capacity.......................................................................................................... 87
7.4 Health Effects.................................................................................................................................. 87
7.5 Novel Products/Formulations and Future Trends........................................................................... 88
7.6 Conclusion....................................................................................................................................... 89
References................................................................................................................................................. 90
7.1 Introduction
Tropical fruits are gaining wide attention because of their high content of nutrient and bioactive com-
pounds, aside from their unique flavors. One of these fruits is acerola (Malpighia emarginata DC.). It is
claimed to have originated in northern South America or Central America or in the West Indies. Thus, it
is called the Antilles cherry, Barbados cherry, or West Indian cherry.
The cherry-sized fruit has bright red skin and an orange-red, soft, and succulent pulp, with a pleas-
ant tart flavor. It is one of the richest natural sources of vitamin C with levels varying from 695 to
4827 mg/100 g ripe fruit [1]. It is also a good source of bioactive compounds such as anthocyanins [2–6],
other phenolic compounds [2,4,7,8], and carotenoids [9–11].
In Brazil, acerola is generally consumed as a fresh or processed juice. It is used to be a garden fruit,
but is now commercially produced and processed. The main marketed products are the fruit itself, the
single-strength pasteurized juice, and frozen pulp used for preparing the juice in restaurants and fruit
juice stands. The processed juice is either aseptically packaged in Tetra Pak cartons or bottled (hot
filled). This chapter highlights the composition of acerola juice in terms of its nutrients, bioactive com-
pounds, antioxidant activity as well as potential health effects, and development of new products.
7.2 Nutritional Characteristics

Acerola juice is a good to excellent source of minerals and vitamins. Except for the marked reduction in
potassium, sodium, vitamin A, vitamin B6, and vitamin C, the levels of these micronutrients are essentially
maintained in the juice as compared to the fresh fruit (Table 7.1) [12]. A 100 g acerola juice contains an
average of 94.30 g water, 0.40 g protein, 0.30 lipid, 4.80 g carbohydrate, 0.3 g total dietary fiber, and 4.50 g
total sugars. Compared with the widely consumed vitamin C–rich orange juice, acerola juice contains
considerably higher vitamin C (1600 vs. 50 mg/100 g), vitamin A (25 vs. 10 µg retinol activity equivalents
[RAE]/100 g), sodium (3 vs. 1 mg/100 g), and zinc (0.10 vs. 0.05 mg/100 g) [12]. On the other hand, acerola
85
TABLE 7.1
Compositional and Nutritional Characteristics of Acerola Fruit and Juice (per 100 g)
Nutrient Unit Fruit Juice
Proximate Composition
Water g 91.41 94.30
Energy kcal 32 23
Protein g 0.40 0.40
Lipid (fat) g 0.30 0.30
Carbohydrate g 7.69 4.80
Total sugars g na 4.50
Total dietary fiber g 1.1 0.3
Minerals
Calcium mg 12 10
Iron mg 0.20 0.50
Magnesium mg 18 12
Phosphorus mg 11 9
Potassium mg 146 97
Sodium mg 7 3
Zinc mg 0.10 0.10
Vitamins
Folate (DFE) μg 14 14
Niacin mg 0.400 0.400
Riboflavin mg 0.060 0.060
Thiamin mg 0.020 0.020
Vitamin A (RAE) μg 38 25
Vitamin B6 mg 0.009 0.004
Vitamin C mg 1678 1600
Vitamin E mg na 0.18
Vitamin K μg na 1.4
Source: Adapted from the U.S. Department of Agriculture (USDA), USDA National Nutrient Database for Standard
Reference, Release 26, 2013, Published online at: http://www.ars.usda.gov/ba/bhnrc/ndl (accessed June 1, 2014).
Abbreviations: DFE, dietary folate equivalents; RAE, retinol activity equivalents; na, not available.
juice is lower in potassium (97 vs. 200 mg/100 g), phosphorus (9 vs. 17 mg/100 g), folate (14 vs. 30 µg
dietary folate equivalents [DFE]/100 g), and total sugars (4.5 vs. 8.4 g/100 g) than orange juice.
7.3 Bioactives and Antioxidant Efficacy

7.3.1 Bioactive Compounds
Concentrated acerola juice has an average of 1.4 mg/100 g of quercetin and 0.4 mg/100 g of kaemp-
ferol [8]. The commercial cultivar “Longa Vida” fruit has 4.1 and 0.9 mg/100 g of quercetin and
kaempferol, respectively. The corresponding values for the commercial cultivar “Olivier” fruit are 5.3
and 1.0 mg/100 g. These results indicate loss of flavonol during processing.
Concentrated and ready-to-drink juices have much lower carotenoid concentrations than the fresh ripe
fruit [9]. Increases in cis-isomers, epoxidation to form the 5,6-epoxides, and rearrangement of the 5,6-
to the 5,8-epoxides have been reported. Three brands of processed juices were found to have variable
carotenoid levels, these concentrations being markedly lower than those of the ripe fruit (Table 7.2) [11].
Brand A consistently had lower values.
The lower carotenoid and flavonoid content in the processed juices indicate that optimization of
processing is warranted. Moreover, influencing preprocessing factors, such as cultivar and maturity
Acerola Juice 87
TABLE 7.2
Carotenoid Content (µg/g) of Acerola Fruit and Juice
Food Description β-Carotene β-Cryptoxanthin Lutein α-Carotene Violaxanthin References
Ripe fruit 5.4 4.2 1.0 nd 4.0 [9]
Concentrated juice 6.3 0.9 2.1 nd nd [9]
Ready-to-drink juice 2.2 0.5 0.1 nd nd [9]
Ripe fruit 38 1.2 1.1 0.7 3.1 [11]
Juice, Brand A 2.7 0.4 0.2 0.3 nd [11]
Juice, Brand B 8.0 1.0 0.6 0.7 0.04 [11]
Juice, Brand C 10 1.0 0.5 0.2 0.1 [11]
Abbreviation: nd, not detected.
at harvest, should be considered. Acerola cultivars have been developed for commercial production.
Among these cultivars, acerola “Olivier” appears to be the raw material of choice because it has a higher
anthocyanin [6], flavonol [8], and carotenoid [9–11] content than others. These bioactive compounds are
higher in the ripe fruit than immature fruit [4,9,11]. On the other hand, vitamin C is higher in the imma-
ture fruit than that of its ripe counterpart [13].
7.3.2 Antioxidant Capacity

Although numerous publications reporting the antioxidant activity/capacity of foods measured in vitro
have been published, the voluminous data obtained have been criticized as being inconsistent, difficult to
compare and interpret, and, above all, lacking biological relevance. Antioxidant capacity data on acerola
have been obtained by these chemical assays and can only be considered as indicative of their potential
antioxidant activity.
The antioxidant capacity of hydrophilic extracts of acerola pulps and juices were evaluated by the
2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonate (ABTS), oxygen radical absorbance capacity (ORAC),
and 2,2-diphenyl-1-picrylhydrazyl (DPPH) methods [7]. The antioxidant activity values obtained for
acerola juice were higher than those reported for other fruit juices particularly rich in polyphenols, such
as strawberry, grape, and apple juices. Five polyphenolic compounds were identified in the samples,
namely, chlorogenic acid, (−)-epigallocatechin gallate, (−)-epicatechin, procyanidin B1, and rutin, the
last two predominating. Three polyphenolic fractions (phenolic acids, anthocyanins, and flavonoids)
were separated and their respective antioxidant activities calculated. Phenolic acids were found to be the
main contributors to the antioxidant activity.
In a linoleic acid model system, acerola juice exhibited high antioxidant capacity [13]. Copper-mediated
low-density lipoprotein oxidation measured in vitro and in the presence of cultured rabbit aortic endothe-
lial cells was inhibited in the presence of commercial soy and alfalfa extracts, and this effect was further
enhanced in the presence of acerola extract [14].
7.4 Health Effects

Studies on the effects of acerola on human health are still limited. Supplementation with acerola juice
was shown to successfully improve vitamin C serum levels in vitamin C–deficient preschool children
[15] and institutionalized elderly [16] in Brazil.
The effects of unripe, ripe, and industrial acerola juice on relevant inflammatory and lipolysis proteins
in the adipose tissue of mice with cafeteria diet–induced obesity were examined [17]. Acerola juice
prevented weight gain (measured in terms of body weight and the adiposity index) and dyslipidemia
(measured by the triacylglycerol levels) and increased the interleukin 10 and-tumor-necrosis-factor-alpha
ratio in the adipose tissue. In addition, acerola juice intake led to reductions in the level of phosphory-
lated jun N-terminal kinase and to increases in the phosphorylation of inhibitor of kappa B alpha and
OH H
OH OH
HO O+ HO O+
O OH O OH
O O
CH3 CH3
OH OH
OHOH OH OH
Cyanidin-3-O-rhamnoside Pelargonidin-3-O-rhamnoside
OH
OH
HO O
O OH
O
CH3
OH O
OH OH
Quercetin-3-O-rhamnoside
FIGURE 7.1 Chemical structures of major polyphenols found in acerola juice.
hormone-sensitive lipase at serine 660. Taken together, the results suggest that acerola juice reduces low-
grade inflammation and ameliorates obesity-associated defects in the lipolytic processes.
In mice, acerola fruit inhibited increases in the levels of proliferating nuclear cell antigen and orni-
thine decarboxylase and also suppressed the activation of Ras signal pathway at the promotion stage [18].
These results suggest that acerola regulates abnormal cell growth at the promotion stage of lung tumori-
genesis in mice treated with 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (a potent carcinogen)
through suppression of the initiation stage.
Acerola had higher cytotoxic activity against tumor cell lines such as human oral squamous cell
carcinoma and human submandibular gland carcinoma than against normal cells such as human peri-
odontal ligament fibroblasts and human gingival fibroblasts [19]. Electron spin resonance spectroscopy
showed that radical-mediated oxidation was not involved in the induction of the tumor-specific cyto-
toxic activity.
Cyanidin-3-O-rhamnoside, pelargonidin-3-O-rhamnoside, and quercetin-3-O-rhamnoside (Figure 7.1)
isolated from acerola juice were found to have O2-scavenging activity and inhibitory effects on both
α-glucosidase and advanced glycation end product formation in vitro [3]. Based on these results,
these polyphenols could be beneficial in the prevention of diabetes mellitus and its complications.
Subsequently, Hanamura et al. [20] showed that a crude acerola polyphenol fraction, prepared by
extraction with ethanol and elution from a C18 cartridge with ethanol containing 10% acetic acid, sig-
nificantly suppressed the plasma glucose level in mice after the administration of both glucose and
maltose. This suggested that the extract could have a preventive effect on hyperglycemia in the post-
prandial state, the suggested mechanism being both suppression of the intestinal glucose transport and
inhibition of α-glucosidase.
7.5 Novel Products/Formulations and Future Trends

Acerola is gaining wide acceptance in the world market, especially in Japan and the United States, where
it is used as a source of natural vitamin C in a variety of products. Product development with this fruit
has been pursued in recent years. There is conscientious effort to ensure maximum retention during
processing and stability during storage of the constituent nutrients and bioactive compounds.
Acerola Juice 89
Aside from being time- and energy-consuming, pressing and enzymatic maceration, the classical tech-
niques for juice extraction, have low extraction efficiency. After optimizing the conditions of pectolysis
for maximum juice yield, application of ultrasound and pectinase preparation on the extraction yield of
acerola juice was investigated [21]. The maximum yield of simultaneous treatment of acerola mash by
ultrasound and pectinase preparation was 3.2% and 15.5% higher than the ultrasonic treatment and the
enzymatic treatment, respectively. In another study of the same group, ultrasound-assisted extraction
took only 6 min to achieve the highest level of vitamin C and phenolic compounds, as well as antioxidant
activity determined by the DPPH and ABTS methods, in acerola juice, while enzyme-assisted extraction
took up to 120 min to obtain the maximal values [22].
Extraction and concentration of acerola juice using commercial pectinolytic enzymes and hydro-
gel and silica sol as clarifying aids was efficient in removing substances that cause turbidity, result-
ing in a clear and concentrated juice adequate for consumption [23]. The ascorbic acid content was
3.4-fold higher than that of the original juice. Enzymatic hydrolysis, microfiltration, and reverse
osmosis were used for the same purpose [24]. Microfiltration reduced mold, yeast, and bacteria,
and the clarified and concentrated (from 7 to 29.2 oBrix) product had the required microbiological
counts. Vitamin C increased from 1234 mg/100 g in the original juice to 5229 mg/100 g in the
concentrated juice.
Fruit juice blends continue to attract attention because these products combine the sensory and nutri-
tional properties of different fruits. A blend containing acerola juice and green coconut water with added
caffeine was microbiologically stable and acceptable according to sensory analyses during 6 months of
storage at room temperature (27°C) [25]. Although still relatively high, vitamin C decreased significantly
throughout storage and anthocyanin was completely lost.
Mixed tropical fruit nectars consisting of acerola, cashew apple, guava, papaya, and passion fruit, with
added caffeine, were well accepted by consumers and were microbiologically stable during 6 months of
storage at room temperature [26]. However, their vitamin C content decreased significantly throughout
the storage time, although it still remained relatively high.
Degradation of vitamin C of green acerola juice and synthetic ascorbic acid, encapsulated with malto-
dextrin DE20 and a mixture of this with gum Arabic, was studied at 15°C, 25°C, 35°C, and 45°C [27].
The juice was obtained from the immature fruit because of its higher content of vitamin C compared to the
ripe fruit. Vitamin C degradation followed first-order kinetics. Neither encapsulating materials showed a
prominent protection against degradation at the lower storage temperatures. At the higher temperatures,
the formulation containing a mixture of maltodextrin and gum Arabic (3:1) was the most effective in
protecting vitamin C. Higher temperature (35°C or 45°C) had a greater impact on the degradation of
synthetic vitamin C than that of green acerola, the difference in effect attributed, by the authors, possibly
to the presence of phenolic compounds with antioxidant activity.
Acerola nectar with added microencapsulated probiotic [28] has also been investigated in line with a
growing interest in developing nondairy probiotic foods. This type of product could be useful for people
with lactose intolerance [29].
The production of a beverage containing both whey butter cheese and acerola juice was considered to
be a good commercialization potential, since it aggregated the benefits provided by both, including the
ingestion of essential amino acids and increasing the vitamin C content [30]. The beverage produced had
good sensory performance and low caloric value.
7.6 Conclusion
Because of its excellent composition of nutrients and bioactive compounds, acerola and its prod-
ucts will continue to be investigated. Commercial processing of this fruits is expected to continue
to expand, but optimization of the processing conditions or applications of new technologies (e.g.,
nonthermal processing) are warranted to ensure the retention of the health-promoting constituents
during processing and storage. Health effects should be investigated through well-designed human
intervention studies.
REFERENCES
1. Mezadri, T., Fernández-Pachón, M.S., Villano, D., Gracía-Parilla, M.C., and Troncoso, A.M., El fruto
de la acerola: composición y posibles usos alimenticios. Arch. Latinoam. Nutr., 56, 101–109, 2006.
2. Vendramini, A.L.A. and Trugo, L.C., Phenolic compounds in acerola fruit (Malpighia punicifolia, L.).
J. Braz. Chem. Soc., 15, 664–668, 2004.
3. Hanamura, T., Hagiwara, T., and Kawagishi, H., Structural and functional characterization of poly-
phenols isolated from acerola (Malpighia emarginata DC.) fruit. Biosci. Biotechnol. Biochem.,
69, 280–286, 2005.
4. Hanamura, T., Uchida, E., and Aoki, H., Changes of the composition in acerola (Malpighia emarginata
DC.) fruit in relation to cultivar, growing region and maturity. J. Sci. Food Agric., 88, 1813–1820, 2008.
5. De Brito, E.S., de Araújo, M.C.P., Alves, R.E., Carkeet, C., Clevidence, B.A., and Novotny, J.A.,
Anthocyanins present in selected tropical fruits: Acerola, jambolão, jussara, and guajiru. J. Agric. Food
Chem., 55, 9389–9394, 2007.
6. De Rosso, V.V., Hillebrand, S., Montilla, E.C., Bobbio, F.O., Winterhalter, P., and Mercadante, A.Z.,
Determination of anthocyanins from acerola (Malpighia emarginata DC.) and açai (Euterpe oleracea
Mart.) by HPLC-PDA-MS/MS. J. Food Comp. Anal., 21, 291–299, 2008.
7. Mezadri, T., Villano, D., Fernández-Pachón, M., García-Parilla, M., and Troncoso, A.M., Antioxidant
compounds and antioxidant activity in acerola (Malpighia emarginata DC.) fruits and derivatives.
J. Food Comp. Anal., 21, 282–290, 2008.
8. Hoffmann-Ribani, R., Huber, L.S., and Rodriguez-Amaya, D.B., Flavonols in fresh and processed
Brazilian fruits. J. Food Comp. Anal., 22, 263–268, 2009.
9. Mezadri, T., Pérez-Gálvez, A., and Hornero-Méndez, D., Carotenoid pigments in acerola fruits
(Malpighia emarginata DC.) and derived products. Eur. Food Res. Technol., 220, 63–69, 2005.
10. De Rosso, V.V. and Mercadante, A.Z., Carotenoid composition of two Brazilian genotypes of acerola
(Malpighia punicifolia L.) from two harvests. Food Res. Int., 38, 1073–1077, 2005.
11. Porcu, O.M. and Rodriguez-Amaya, D.B., Variation in the carotenoid composition of acerola and its
processed products. J. Sci. Food Agric., 86, 1916–1920, 2006.
12. U.S. Department of Agriculture (USDA), USDA National Nutrient Database for Standard Reference,
Release 26, 2013. Published online at: http://www.ars.usda.gov/ba/bhnrc/ndl (accessed June 1, 2014).
13. Rhighetto, A.M., Netto, F.M., and Carraro, F., Chemical composition and antioxidant activity of juices
from mature and immature acerola (Malpighia emarginata DC.). Food Sci. Technol. Int., 11, 315–321,
2005.
14. Hwang, J., Hodis, H.N., and Sevanian, A., Soy and alfalfa phytoestrogen extracts become potent low-
density lipoprotein antioxidants in the presence of acerola cherry extract. J. Agric. Food Chem., 49,
308–314, 2001.
15. Costa, M.J.C., Terto, A.L.Q., Santos, L.M.P., Rivera, M.A.A., and Moura, L.S.A., Supplementation with
West Indian cherry and its effects on the blood levels of vitamin C and hemoglobin in preschool chil-
dren. Rev. Nutr. Campinas, 14, 13–20, 2001.
16. Aranha, F.Q., Moura, L.S.A., Simoes, M.O.S., Barros, Z.F., Quirino, I.V.L, Metri, J.C., and Barros,
J.C., Normalization of the ascorbic acid serum levels by supplementation with acerola juice (Malpighia
glabra L.) and pills in institutionalized elderly. Rev. Nutr. Campinas, 17, 309–317, 2004.
17. Dias, F.M., Leffa, D.D., Daumann, F., Marques, S.O., Luciano, T.F., Possato, J.C., de Santana, A.A.
et al., Acerola (Malpighia emarginata DC.) juice intake protects against alterations to proteins involved
in inflammatory and lipolysis pathways in the adipose tissue of obese mice fed a cafeteria diet. Lipids
Health Dis., 13, 24–32.
18. Nagamine, I., Akiyama, T., Kainuma, M., Kumagai, H., Satoh, H., Yamada, K., Yano, T., and Sakurai,
H., Effect of acerola cherry extract on cell proliferation and activation of Ras signal pathway at the pro-
motion stage of tumorigenesis in mice. J. Nutr. Sci. Vitaminol., 48, 69–72, 2002.
19. Motohashi, N., Wakabayashi, H., Kurihara, T., Fukushima, H., Yamada, T., Kawase, M., Sohara, Y.
et al., Biological activity of Barbados cherry (acerola fruits, fruit of Malpighia emarginata DC.) extracts
and fractions. Phytother. Res., 18, 212–223, 2004.
20. Hanamura, T., Mayama, C., Aoki, H., Hirayama, Y., and Shimizu, M., Antihyperglycemic effect of poly-
phenols from acerola (Malpighia emarginata DC.) fruit. Biosci. Biotechnol. Biochem., 70, 1813–1820,
2006.
Acerola Juice 91
21. Dang, B.K., Huynh, T.V., and Lee, V.V.M., Simultaneous treatment of acerola mash by ultrasound and
pectinase preparation in acerola juice processing: Optimization of the pectinase concentration and pec-
tolytic time by response surface methodology. Int. Food Res. J., 19, 509–513, 2012.
22. Le, H.V. and Le, V.V.M., Comparison of enzyme-assisted and ultrasound-assisted extraction of vitamin
C and phenolic compounds from acerola (Malpighia emarginata DC.) fruit. Int. J. Food Sci. Technol.,
47, 1206–1214.
23. Montenegro, I., Teles, K.H., de Oliveira, G.S.F., Maia, G.A., and de Figueiredo, R.W., Physicochemical
changes during extraction and concentration of acerola (Malpighia emarginata DC.) using pectinases
and clarifying agents. Braz. J. Food Technol., 10, 266–270, 2007.
24. Matta, V.M., Moretti, R.H., and Cabral, L.M.C. Microfiltration and reverse osmosis for clarification and
concentration of acerola juice. J. Food Eng., 61, 477–482.
25. Lima, A.S., Maia, G.A., de Sousa, P.H.M., do Prado, G.M., and Rodrigues, S., Storage stability of a
stimulant coconut water-acerola fruit juice beverage. Int. J. Food Sci. Technol., 44, 1445–1451, 2009.
26. De Sousa, P.H.M., Maia, G.A., de Azeredo, H.M.C., de Souza Filho, M.S.M., Garruti, D.S., and de
Freitas, C.A.S., Mixed tropical fruit nectars with added energy components. Int. J. Food Sci. Technol.,
42, 1290–1296, 2007.
27. Rhighetto, A.M. and Netto, F.M., Vitamin C stability in encapsulated green West Indian cherry juice
and in encapsulated synthetic ascorbic acid. J. Sci. Food Agric., 86, 1202–1208, 2006.
28. Antunes, A.E.C., Liserre, A.M., Coelho, A.L.A., Menezes, C.R., Moreno, I., Yotsuyanagi, K., and
Azambuja, N.C., Acerola nectar with added microencapsulated probiotic. LWT—Food Sci. Technol.,
54, 125–131, 2013.
29. Granato, D., Branco, G.F., Nazzaro, F., Cruz, A.G., and Faria, J.A.F., Functional foods and nondairy
probiotic food development: Trends, concepts, and products. Comp. Rev. Food Sci. Food Safety,
9, 292–302, 2010.
30. Cruz, A.G., Sant’anna, A.S., Macchione, M.M., Teixeira, A.M., and Schmidt, F.L., Milk drink using whey
butter cheese (Queijo manteiga) and acerola juice as a potential source of vitamin C. Food Bioprocess
Technol., 2, 368–373, 2009.
8
Apple Juice
H.P. Vasantha Rupasinghe and Surangi Thilakarathna
CONTENTS
8.1 Introduction..................................................................................................................................... 93
8.2 Nutritional Characteristics.............................................................................................................. 94
8.3 Bioactives and Antioxidant Efficacy............................................................................................... 96
8.4 Health Effects................................................................................................................................ 100
8.5 Novel Products/Formulation and Future Trends........................................................................... 102
8.6 Conclusion..................................................................................................................................... 103
References............................................................................................................................................... 104
8.1 Introduction
Apple juice is the second most widely consumed fruit juice in the world and is popular among adults and
children owing to its authentic taste. Compared to whole fruits, it is convenient to consume fruit and veg-
etable juices to satisfy “5 A Day” dietary requirement of fruits and vegetables [1]. According to Health
Canada [2], “a functional food is similar in appearance to, or may be a conventional food, is consumed as
part of a usual diet, and is demonstrated to have physiological benefits and/or reduce the risk of chronic
disease beyond basic nutritional functions’’. Rather than apple juice is considered as a functional food
due to its health-promoting properties apart from the basic nutritional characteristics. It is a good source
of polyphenolic compounds and has shown antiatherosclerotic, anti-inflammatory, and neuroprotective
effects, which are briefly discussed in this chapter.
In Canada, per capita annual consumption of apple juice in 2009 was 7.23 L [3], which was relatively
unchanged from 2005 to 2009 [4]. China is the world’s largest apple producer and thus has a greater
influence on the world apple juice production [5]. The United States is the top apple juice importer in the
world and, in year 2006, imported around 35% of the total apple juice traded [5]. In addition to China,
countries such as Argentina, Chile, New Zealand, and South Africa produce and supply apple juice to
the world market.
In Canada, apple juice production declined from 320 million liters per year in 1986 to 243 million
liters per year in 2008 due to the decline in apple juice consumption [3]. Several reasons for this declined
consumption could be its higher acidity, sugar content, calories, and attractive alternative products on
the market. Regarding the higher acidity, numerous attempts such as deacidification using novel methods
(electrodialysis, malolactic conversion, etc.), blending, and formulation, among others [6], have been
investigated to reduce the acidity of apple juice.
There are different types of apple juice on the market; pure single strength juice (not from concentrates,
100% juice), juice from concentrate, cloudy juice, clear juice, juice blends, and many more. On the other
hand, there are juice drinks, cocktails, and beverages where the juice content is less (10%–20%). Figure 8.1
shows a simplified process flowchart for clear and cloudy apple juice production. Despite other alterna-
tives on the market, apple juice is still popular among consumers due to its health benefits especially those
associated with antioxidant compounds, vitamins, and minerals [6]. Therefore, countless efforts are being
made to further improve the quality and consumer acceptance of apple juice. This chapter highlights the
93
Receiving apples
Sanitizing
Grinding/crushing
Enzyme treatment
Pressing and juice

Coarse filtration Apple pomace
extraction
Pasteurization Clarification
Cloudy apple juice Clear apple juice Pasteurization
Concentration
Clear concentrate
FIGURE 8.1 Simplified process flowchart for cloudy and clear apple juice production.
composition of apple juice in relation to nutrients, antioxidant activity, bioactive phytochemicals, poten-
tial health benefits of apple juice consumption, and future perspective of apple juice industry.

Numerous factors contribute to the nutrient composition of apple juice. Different cultivars of apples con-
tain varying amounts of sugar, protein, ascorbic acid, minerals, as well as bioactives such as polyphenols.
Processing methods can also play a vital role in the nutrient composition of the resultant juice. Basic
nutrients in apple juice are discussed under this section.
As mentioned before, the basic nutritive value of apple juice can vary depending on the variety of
apple used for processing (Table 8.1) [55]. Among 10 different apple cultivars tested, sugar, vitamin
C, total nitrogen, phosphorus, potassium, and calcium content ranged between 9.53%–12.34%, 25.75–
77.00 mg/100 g, 0.67%–0.11%, 0.15%–0.24%, 0.40%–0.75%, and 2.50–7.80 mg/100 g, respectively
[7]. Compositional data obtained for apple juice prepared from 175 noncommercial varieties of apples
reported the following ranges for the sugars analyzed: sucrose 0.38–5.65 (g/100 mL), glucose 1.05–
3.23 (g/100 mL), fructose 3.84–8.01 (g/100 mL), and sorbitol 0.17–1.40 (g/100 mL) [8]. The reported
ranges for sodium, potassium, magnesium, calcium, iron, chloride, and phosphate (ppm) were 0.5–73.4,
766–2712, 35.2–101, 18.7–80.3, 0.7, 18, and 86–459, respectively. As reported by Health Canada [9],
ready-to-drink vitamin C–added apple juice (125 mL) may contain traces of protein and lipid and 15 g
of carbohydrate, 14 g of total sugars and 0.1 g of total dietary fiber, 9 mg calcium, 0.5 mg iron, 4 mg
sodium, 150 mg potassium, 4 mg magnesium, 9 mg phosphorous, 52 mg of vitamin C, and traces of
folate, while no vitamin A or vitamin B12.
Apple Juice 95
TABLE 8.1
Compositional and Nutritional Characteristics of Apple Juices (per 100 g)
Nutrient Unit Apple Juice 1 Apple Juice 2
Water g 88.24 88.24
Energy kcal 46 46
Protein g 0.10 0.10
Lipid (fat) g 0.13 0.13
Total sugars g 9.62 9.62
Minerals
Calcium mg 8 8
Iron mg 0.12 0.12
Magnesium mg 5 5
Phosphorus mg 7 7
Potassium mg 101 101
Sodium mg 4 4
Zinc mg 0.02 0.02
Vitamins
Folate (DFE) µg 0 0
Niacin mg 0.073 0.073
Vitamin A (RAE) µg 0 0
Vitamin C mg 0.9 38.5
Vitamin E (ATE) mg 0.01 0.01
Reference, Release 27, National Technical Information Service, USDA, Springfield, VA, 2014.
Notes: Apple juice 1 (canned or bottled, unsweetened, and without added ascorbic acid) and apple juice 2 (canned or bottled,
unsweetened, and with added ascorbic acid).
Abbreviations: DFE, dietary folate equivalents; RAE, retinol activity equivalents; ATE, alpha-tocopherol equivalents.
Not only nutrients but also allergenic compounds, agrochemicals, and toxins in foods are a great
concern to health-cautious consumers as their presence can cause deleterious effects to health. Apple
is known to contain two main allergenic compounds, namely, Mal d 3 and Mal d 1. Apple Mal d 3 is a
member of the prolamine family and is a nonspecific lipid transfer protein where Mal d 1 is associated
with pollen-related fruit allergy. Numerous attempts have been made to reduce the allergenicity of apple
allergens, but results showed that high-pressure/-temperature treatments or pulsed electric field treat-
ments had minor effects on the purified allergen structure [10]. It was reported that treating apple juice
and homogenates at 450–550 MPa for 3–10 min at 30°C did not change the allergenicity or the structure
of recombinant Mal d 1 [11].
Patulin is a mycotoxin found in apples and is produced by certain species of Aspergillus, Penicillium,
and Byssochlamys. In rodents, patulin showed neurotoxic, immunotoxic, genotoxic, and gastrointestinal
effects [12], and therefore, a maximum tolerable daily intake of 0.4 µg/kg body weight/day had been
established [13]. Because of the human health concerns and possibility of using patulin as a quality
indicator in food products, World Health Organization (WHO) has set 50 µg/L as the maximum recom-
mended concentration of patulin in apple juice [13]. Apple juice is known as the most important dietary
source of patulin for humans [13]. A survey was carried out in Spain to determine the dietary intake of
patulin from apple juice, which the authors tested 100 samples of apple juice marketed [14]. Among the
tested samples, 66% contained patulin at above 0.7 µg/L, while 11% contained more than 50 µg/L.
During processing of apple juice concentrates, it is possible to reduce patulin levels at all stages of
production, and thus patulin level is being used as a quality indicator of apple juice. It was previously
reported that patulin level may be reduced by 75% when apple juice concentrates are produced from
whole apples [15]. Pasteurization, enzymatic treatment, microfiltration, and evaporation process as well
as dilution of juice to get the desired Brix value reduced the level of patulin in the concentrated apple
juice [15]. A seven-pass UV treatment (99.4 mJ/cm2) reduced an initial patulin level of 1000 µg/L in
reconstituted apple juice to below the maximum recommended concentration of 50 µg/L [16]. The reduc-
tion of patulin level after the first pass (14.2 mJ/cm2) and after the seven passes (99.4 mJ/cm2) was from
72.5% to 5.14%, respectively. Although UV treatment resulted in loss of organoleptic properties, it has
the potential to be developed as a novel approach to reduce patulin levels in apple juice.
Organophosphorus pesticides are commonly used to control infestations in agricultural crops. It is
known that these pesticides show cholinesterase inhibitory activity, reproductive toxicity, immunotoxic-
ity, genotoxicity, and bioaccumulation, all of which pose a threat to human health. It was reported that
not only conventional apple juice but also organic apple juice contained dialkylphosphates, a metabolic
product of organophosphorus pesticides [17].

Apples are a significant source of fruit phenolics as they are the most widely consumed fruit by Western
populations [18]. There are five major phenolic groups in apples. These include flavonols, flavanols,
anthocyanins, hydroxycinnamic acids, and dihydrochalcones, but hydroxycinnamic acids and flavonoids
are among the most abundant [19]. A serving of one medium apple (200 g) provides around 400 mg of
total polyphenols expressed as gallic acid equivalents (GAE) [20]. The apples produced in the world are
mostly used for commercial juice making, while a smaller amount is used for fresh eating [21]. Juice pro-
duction usually involves clarification, which aims at removing certain flavonoids, and thus, polyphenol
content in apple juice can be considerably less [22] compared to whole apples (Table 8.2). Structures of
some common phenolic compounds found in apple juice are shown in Figure 8.2.
According to research findings, cloudy apple juice is better in terms of total phenolics and thereby
antioxidant activity compared to clear apple juice [23]. According to Mullen et al. [23], cloudy apple
juice had greater amounts of hydroxycinnamates, flavanols, and flavonols compared to clear apple juice,
but clear apple juice had more hydroxychalcones compared to cloudy apple juice. Among 24 commer-
cial juice products tested, cloudy apple juice contained a total polyphenol content ranging from 152 to
459 mg/L, while the total polyphenols in clear apple juice ranged from 110 to 173 mg/L [21]. The same
study reported that fresh juice produced from cider apples (seven cultivars) had considerably more poly-
phenols compared to the tested dessert apple cultivars (four cultivars). In general, fresh juices prepared
from either dessert or cider cultivars contained more polyphenols compared to commercial cloudy or
clear apple juices [24]. Polyphenol-rich apple varieties are known to produce juices with greater poly-
phenol content (clear and cloudy); hydroxycinnamic acids were the most abundant group of polyphenolic
compounds followed by flavanols including monomers and procyanidins [24,25]. The U.S. Department
of Agriculture (USDA) released a database of flavonoid contents in 500 selected foods in 2011 [26]. The
data reported on bottled or canned unsweetened apple juice without/with the addition of ascorbic acid
are listed in Table 8.2. Unsweetened, canned or bottled apple juice can be a good source of epicatechin,
catechin, and quercetin as the data implied. According to the database released by USDA on the proan-
thocyanidin contents of selected foods [27], unsweetened, bottled, or canned apple juice without ascorbic
acid addition contained proanthocyanidins in the order of monomers > dimmers > trimers > 4–6mers
> 7–10mers, and contained no polymers. When comparing the proanthocyanidin contents in raw apples
with skin with the mentioned type of apple juice, this order was in the opposite trend where proanthocy-
anidin polymers were the most abundant and monomers were the least (Table 8.3).
Countless reports can be found as evidence of the high antioxidant activity of apples and apple prod-
ucts. The antioxidant activity of apples and apple products has also been associated with numerous health
benefits linked to cardiovascular disease (CVD), cancers, and cognitive health, among others. According
to Karaman et al. [28], the contribution of ascorbic acid to the total antioxidant capacity of fresh apple
Apple Juice 97
TABLE 8.2
Polyphenolic Composition of Apple Juice
Concentration
Food Description Class Flavonoid (mg/100 g) References
Apple cider (European) Flavan-3-ols (−)-Epicatechin 0.32 [26]
(+)-Catechin 1.95 [26]
Flavonols Quercetin 0.48 [26]
Apple juice, canned or Anthocyanidins Cyanidin 0.01 [26]
bottled, unsweetened, Flavan-3-ols (−)-Epicatechin 4.71 [26]
without added ascorbic acid Catechin 1.25 [26]
Flavonols Myricetin 0.01 [26]
Quercetin 0.58 [26]
Apples, red, delicious, raw, Anthocyanidins Cyanidin 3.74 [26]
with skin Delphinidin 0.01 [26]
Pelargonidin 0.01 [26]
Peonidin 0.05 [26]
Flavan-3-ols (−)-Epicatechin 9.83 [26]
(−)-Epigallocatechin 0.37 [26]
(−)-Epigallocatechin-3-gallate 0.13 [26]
(+)-Catechin 2.00 [26]
Flavonols Myricetin 0.01 [26]
Quercetin 3.86 [26]
A commercial clear apple Hydroxycinnamic Chlorogenic acid 95.6 [21]
juice (units: mg/L) acids 4-p-Cumaroylquinic acid 21.8 [21]
Caffeic acid 4.4 [21]
Dihydrochalcone Phloretin-2′-O-xyloglucoside 23.0 [21]
derivatives Phloridzin 9.7 [21]
Phloretin nd [21]
Flavan-3-ols Procyanidin B2 <DL [21]
(+)-Catechin <DL [21]
(−)-Epicatechin 14 [21]
Flavonols Quercetin-3-O-glucoside nd [21]
Quercetin-3-O-galactoside 1.6 [21]
Quercetin-3-O-rhamnoside 1.7 [21]
Quercetin nd [21]
Total polyphenols 172.7 [21]
A commercial cloudy apple Hydroxycinnamic Chlorogenic acid 181.5 [21]
juice (units: mg/L) acids 4-p-Cumaroylquinic acid 37.2 [21]
Dihydrochalcone Phloretin-2′-O-xyloglucoside 45.4 [21]
derivatives Phloridzin 23.2 [21]
Phloretin nd [21]
Flavan-3-ols Procyanidin B2 19.7 [21]
(+)-Catechin 2.8 [21]
(−)-Epicatechin 48.6 [21]
(Continued)

Polyphenolic Composition of Apple Juice
Concentration
Food Description Class Flavonoid (mg/100 g) References
Flavonols Quercetin-3-O-glucoside 1.8 [21]
Quercetin-3-O-galactoside 2.0 [21]
Quercetin-3-O-xyloside 5.0 [21]
Quercetin-3-O-rhamnoside 3.2 [21]
Quercetin nd [21]
Total polyphenols 380.6 [21]
Abbreviations: nd, not detected; DL, detection limit.
OH
OH
O OH
HO O
OH
HO HO OH OH
Phloretin (–)-Epicatechin
H
O
O-
O O
H
O
O
O OH
O H O O
H O O
OH OH
O HO
H O OH
H HO OH
Quercetin-3-O-glucoside Chlorogenic acid
FIGURE 8.2 Common polyphenolic compounds found in apple juice.
juice was 1.9%–13%, depending on the apple variety tested. Therefore, phytochemicals such as polyphe-
nolic compounds present in apple juice can mainly contribute to its antioxidant activity. Procyanidins
can contribute to more than 80% of the antioxidant capacity in apples and apple products like juices due
to their abundance [19]. A procyanidin-rich apple juice extract recorded the highest antioxidant capacity
followed by a hydroxycinnamic acid–rich apple juice extract as measured by trolox equivalent antioxi-
dant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) assays [29]. Hydroxycinnamic
acid was abundant in flesh compared to peel and can contribute to the antioxidant capacity of apple juices
[19]. Not only fresh juice but also six commercial apple juices tested at a level of 5 µmol GAE/L inhibited
oxidation of human low-density lipoprotein (LDL) at a range of 9%–34% [30]. Human LDL oxidation
inhibition is widely used as a method to assess the antioxidant activity of test compounds or extracts.
Processing methods greatly impact the antioxidant activity of apple juice. Raw apple juice obtained
by pulping and straight pressing or pulp enzyme treatment of apples from cultivar “Jonagold” showed
antioxidant activities of 10% and 3%, respectively, compared to the antioxidant activity of fresh apples
[31]. Rather than transfer to juice, most of the antioxidant compounds retained in the apple pomace as
those were absorbed to the solid matter. The levels of catechin and chlorogenic acid were reduced to 3%
and 50%, respectively. Total quercetin glycosides and cyanidin galactoside were not affected by extended
Apple Juice 99
TABLE 8.3
Proanthocyanidin Content of Apple Juice
Food Description Proanthocyanidin Mean (mg/100 g)
Apple juice, canned or bottled, unsweetened, without added Monomers 4.96
ascorbic acid Dimmers 4.04
Trimers 2.74
4–6mers 0.38
7–10mers 0.10
Polymers 0.00
Apples, red, delicious, raw, with skin Monomers 8.31
Dimmers 15.12
Trimers 10.11
4–6mers 28.59
7–10mers 25.12
Polymers 40.54
Source: Adapted from the U.S. Department of Agriculture (USDA), Database for the proanthocyanidin content of selected
foods, August 2004, Published online at: http://www.nal.usda.gov/fnic/foodcomp (accessed April 20, 2013).
oxidation periods during pulp enzyme treatment as these compounds were not substrates for polyphenol
oxidase [31]. It was suggested that extracting antioxidant compounds from the apple pomace and later
adding them back to apple juice can be a possible way of enhancing the antioxidant activity of apple juice
[31]. A recent study tested this idea and evaluated the pomace maceration by pectinase enzyme treatment
in producing puree-enriched cloudy apple juice [32]. The resultant mixed juice contained more polyphe-
nols, namely, phloretin 2′-O-xyloglucose and phloretin 2′-O-glucose, flavonol glucosides, and polymeric
procyanidins compared to fresh apple juice and did not affect the other polyphenolic compounds. This
technique was impressive as it increased the flavonol glucoside content in the juice, which was quite low
in apple juice.
Comparing three apple juices, namely, a commercial apple juice, an apple juice made by straight
pressing of apples from cultivar “Jonagold,” and juice prepared by pulp enzyme treatment of “Jonagold”
apples showed that all three juices had lower quercetin glycoside content compared to whole apples
[33]. The commercial apple juice and that prepared by straight pressing had a higher antioxidant activ-
ity compared to the juice prepared by pulp enzyme treatment. It was suggested that stirring during pulp
enzyme treatment could be the possible cause for losing the antioxidant (except quercetin glycosides) due
to enzymatic oxidation and loss of antioxidant activity [33].
Enzymatic mash treatment with pectinase in producing cloudy apple juice increased the polyphenolic
content and juice yield [34]. The procyanidins in the juice after the treatment were more stable compared
to the control sample where 50% of the polymeric procyanidins were degraded after 6 months of storage
at 4°C. The radical scavenging activity of the cloudy apple juice resulted from the pectinase treatment
was increased 4%–19% compared to the untreated apple juice samples. However, the antioxidant activi-
ties of the treated and untreated juice were reduced during storage [34]. Apple juice production using
pulsed electric field as a treatment improved the juice extraction and considered as a novel food pro-
duction method. This method was tested as an alternative for enzymatic mash maceration. Although
the juice yield increased significantly with increasing field intensities, it did not change the polyphenol
content or antioxidant capacity of the resultant cloudy apple juice compared to the controlled samples
[35]. In contrast, a study carried out in 2010 revealed that treating whole apples with pulsed electric field
increased the antioxidant capacity of the resultant juice [36]. It not only increased the juice yield but also
increased its total polyphenol content.
Antioxidant activity of apple juice has previously been tested in humans, and promising results were
reported. Acute intake of apple juice by human subjects contributed to improved antioxidant activity
and reduced lipid peroxidation biomarkers in the serum [37]. Apples of two Brazilian cultivars, “Golden
Delicious” and “Catarina,” were used in the study; juice of both cultivars showed similar effects.
Consumption of unsupplemented apple juice (375 mL) for 6 weeks increased ex vivo copper-mediated
LDL oxidation lag time by 20% compared with base line and reduced conjugated diene formation [38].
Contribution of polyphenols to the antioxidant activity of apple juice had been a topic of debate. A study
reported that rapid consumption of 1 L of apple juice increased the plasma antioxidant status of healthy
subjects [39]. The authors concluded that the effect was due to the fructose-induced increase of uric acid
levels but not due to the presence of antioxidant polyphenols in the juice. They further stated that rapid
acute intake of apple juice might not be the most effective way to increase the plasma antioxidant activity.
8.4 Health Effects

There is evidence that consumption of apple juice can promote numerous health effects. Cardiovascular
health-promoting effects, cancer chemoprevention, and neurodegenerative disease preventive effects are
briefly discussed under this section and summarized in Table 8.4.
Consumption of apple juice by hamsters fed with an atherogenic diet for 12 weeks reduced the plasma
total cholesterol level by 24%, high-density lipoprotein (HDL) cholesterol level by 55%, and the a ortic
fatty streak lesion area by 60% and improved the plasma antioxidant capacity by 27% [40]. The study
further revealed that hepatic superoxide dismutase activity, glutathione peroxidase activity, and thiobar-
bituric acid–reactive substances (TBARS) were reduced by 25%, 85%, and 52%, respectively, in hyper-
cholesterolemic hamsters that received the apple juice treatment. Interestingly, the efficiency of fruit was
less compared to the juice. Not only the risk of atherosclerosis but also its progression was reduced by
apple juice consumption. Apple juice reduced the progression of atherosclerosis in hypercholesterolemic
rabbits [41]. The antiatherosclerotic effect was exerted through the reduction of fibrinogen, factor VII
(factor of coagulation systems), C-reactive protein (CRP), total cholesterol, triacylglycerols (TAG), and
LDL cholesterol and increased level of nitrite, nitrate, and HDL cholesterol [41].
The genotoxic effect exerted by an indirect carcinogen 1,2-dimethylhydrazine (DMH) in rat colon
mucosal cells was significantly reduced by a cloudy apple juice treatment but not by a clear apple juice [42].
The total number of aberrant crypt foci per colon was reduced in rats receiving cloudy apple juice com-
pared to the control receiving DMH but not the treatment. Although both apple juice samples contained
similar amounts of dihydrochalcones, hydroxycinnamates, and quercetin glycosides, cloudy apple juice
contained around 30% more procyanidin B1 and B2, and the pectin concentration was fourfold more
compared to clear apple juice. Further studies were carried out to identify the effective bioactive classes
in the cloudy apple juice, which exerted the antiproliferative effects. The results revealed that cloudy
apple juice significantly reduced DNA damage induced by DMH in colon mucosal cells isolated from
rats where none of its fractions were effective [43]. The fractions studied were a polyphenol fraction and
a cloud fraction (consisting of proteins, fatty acids, polyphenols, and cell wall polysaccharides), and rats
received either cloudy apple juice, its fractions separately, or as a combination. Apart from genotoxicity,
cloudy apple juice and its fractions separately and combined significantly reduced the hyperproliferative
effect of DMH in colon mucosal cells. It was suggested that polyphenol fraction alone was not respon-
sible for the exerted effects as the cloud fraction itself showed certain antiproliferative effects.
Constituents of apple juice that contributed to the chemopreventive activity have previously been stud-
ied by fractionating apple juice extracts and testing those constituents using in vitro bioassays [44].
The authors revealed that procyanidins alone or in combination with low-molecular-weight polyphenolic
compounds contributed to the chemopreventive activity measured. Quercetin aglycone was found to be a
potent cytochrome P450 1A inhibitor, while phloretin and (−)-epicatechin were potent cyclooxygenase-1
inhibitors. Aromatase and cytochrome P450 1A inhibitory activity and cytotoxicity toward a human
colon cancer cell line (HCT116) increased toward increasing levels of procyanidins [44].
Consumption of an apple juice concentrate showed promising results in preventing the decline in
cognitive performance due to dietary and genetic deficiencies and aging [45–48]. Accumulation of
amyloid-β is neurotoxic and can be involved in oxidative damage to neuron cells that ultimately cause
neuronal death, which is a characteristic of Alzheimer’s disease. It is believed that at least part of the
neurotoxicity of amyloid-β is due to oxidative stress. Acetylcholine is a neurotransmitter that is known to
decline with normal aging process and with Alzheimer’s disease.
Apple Juice 101
TABLE 8.4
Summary of the Health Effects of Apple Juice
Health Effects Experimental Model Possible Mode of Action References
Apple juice Prevent Hypercholesterolemic Reduced plasma total cholesterol, [40]
diet-induced hamsters non-HDL-C, aortic fatty streak
atherosclerosis lesion area, and improved plasma
antioxidant capacity. Further, the
treatment reduced the hepatic
superoxide dismutase and
glutathione peroxidase activities
and TBARS.
Apple juice Reduce Hypercholesterolemic Reduced factor of coagulation [41]
progression of rabbits systems (fibrinogen, factor VII),
atherosclerosis CRP, total cholesterol, TAG, and
LDL-C; and increasing the levels
of nitrite, nitrate, and HDL-C.
Cloudy and Antiproliferative Rat colon mucosal Cloudy apple juice but not clear [42,43]
clear apple effect cells apple juice significantly reduced
juice the genotoxic effect of DMH, total
number of aberrant crypt foci per
colon, and DNA damage in colon
mucosal cells.
Fractions of Chemopreventive In vitro bioassays Procyanidins alone or in [44]
apple juice activity combination with low-molecular-
weight polyphenolic compounds
contributed to the measured
chemopreventive activity.
Quercetin aglycone: potent
cytochrome P450 1A inhibitory
activity, phloretin, and
(−)-epicatechin: potent COX-1
inhibitory activity. Increasing
levels of procyanidins increased
aromatase and cytochrome P450
1A inhibitory activity and
cytotoxicity toward a human colon
cancer cell line.
Apple juice Neuroprotective Cultured human Reduced generation of ROS, [45]
concentrate in effects neuroblastoma cells calcium influx, and apoptosis
drinking water induced normally by amyloid-β.
Normal aged mice Counteracted the neurodegenerative [46]
effects of a vitamin-deficient,
prooxidant diet.
Normal adult mice and Alleviated the increase of [47]
transgenic Apo amyloid-β.
E−/− mice Alleviated the decrease of [48]
acetylcholine.
Normal aged mice, Attenuated the increased expression [46]
juvenile and adult of presinil-1.
normal C57B1/6J and
Apo E−/− mice
HDL-C, high-density lipoprotein cholesterol; TBARS, thiobarbituric acid–reactive substances; CRP,
Abbreviations:
C-reactive protein; TAG, triacylglycerols; LDL-C, low-density lipoprotein cholesterol; DMH,
1,2-dimethylhydrazine; DNA, deoxyribonucleic acid; COX-1, cyclooxygenase-1; ROS, reactive oxygen
species.
An apple juice concentrate reduced the amyloid-β-induced oxidative damage in cultured SH-SY5Y
human neuroblastoma cells [45]. This particular apple juice concentrate reduced the generation of reac-
tive oxygen species (ROS) and calcium influx and apoptosis induced by amyloid-β. Supplementation of a
vitamin-deficient (folate and vitamin E), oxidative stress–provoking diet with an apple juice concentrate
in drinking water improved neuroprotection in normal, aged mice [46]. The normal, 2.0–2.5-year-aged
mice consumed the aforementioned diet for 1 month and showed a significantly higher oxidative dam-
age in the brain and poorer performance in a standard Y-maze test. These neurodegenerative effects
were counteracted by supplementation with the apple juice concentrate. The mice aging 9–10 months
were not affected by this deficient diet. This vitamin-deficient diet along with dietary iron addition as
a prooxidant has shown to increase amyloid-β levels and decrease acetylcholine levels in normal adult
mice and potentiated in transgenic mice homozygously lacking Apo E (Apo E−/−). Supplementation with
apple juice concentrate in drinking water alleviated the increase of amyloid-β [47] and the decrease of
acetylcholine [48] in both mouse models.
Overexpression of the gene presenilin-1 is associated with the increased production of the neurotoxic
peptide amyloid-β, which in turn increases the generation of ROS. A folate-deficient diet was involved
in the overexpression of presenilin-1. A study showed that folate- and vitamin E–deficient diet increased
the expression of presenilin-1 in aged normal mice and juvenile and adult normal C57B1/6J and Apo
E−/− mice [49]. Interestingly, supplementation with an apple juice concentrate introduced in the drinking
water prevented or attenuated the increased expression of presenilin-1 [49].
8.5 Novel Products/Formulation and Future Trends

As mentioned previously in this chapter, apple juice production and consumption had declined, and
many reasons have been predicted as causative factors for the decline. There have been numerous stud-
ies and surveys carried out to understand the consumer’s perception of apple juice and how to overcome
hurdles to produce the ultimate product that will attract consumers. To satisfy today’s health-cautious
consumers, improvements on the functionality of apple juice are of a greater concern to the apple juice
production industry. Introducing new cultivars, investigating alternative processing methods that pre-
serve bioactive compounds, and blending and formulation of apple juice to further improve the health
benefits and functionality are being studied in this regard. Some of the studies briefly discussed in this
section aimed to improve the final product, and some studies have attempted to address the problems
associated with a specific processing operation.
There had been interest for improving the polyphenol content in apple juice through novel process-
ing techniques. One approach would be to introduce new improved cultivars or assess existing cultivars
that have a greater polyphenol content that is not currently considered as a variety for juice making.
Cyanidin-3-O-galactoside is common in the skin of apples, but certain crab apple varieties contain this
pigment in their flesh as well. Juice made from 14 crab apple genotypes had cyanidin-3-O-galactoside in
the juice, and the genotype “Roberts crab” contained the highest level of 39 mg/L [50]. The red-fleshed
genotypes “Babine” and Malus pumila Niedzwetzkyana showed the greatest antioxidant capacity where
the measured antioxidant capacity was over fivefold of those from the commercial apple cultivars and
commercial juices analyzed. Interestingly, there was no significant difference (P > 0.05) among the com-
mercial apple cultivars, commercial apple juices, and the red-fleshed apple genotypes tested, for the juice
quality measurements such as pH, total soluble solids, or titratable acidity. Therefore, these crab apple
varieties have a greater potential for producing apple juice with higher bioactives.
Among novel processing techniques, enzymatic pomace treatment with pectolytic enzymes improved
polyphenolic content and juice yield in puree-enriched cloudy apple juice [34]. This novel apple juice
was rich in procyanidins, and the use of apple pomace as a source of bioactive compounds can serve as
a good approach to add value to the final product as well as to better handle the waste and underutilized
products of the apple processing industry.
Blending different types of juices together not only enhances the sensory properties of the resultant
juice but also greatly improves its nutritional quality and functionality. A study tested the possibility of
Apple Juice 103
adding apple juice to carrot juice as a method of acidification in order to preserve the carrot juice [51].
Blending apple juice with carrot juice at a ratio of 90:10 significantly (P < 0.05) reduced the total aero-
bic viable count of the juice blend. The same study tested ultrasound treatment as an alternative to the
conventional thermal pasteurization as heat treatments destroy heat-liable nutrients as well as bioactive
compounds in apple juice. The juice quality parameters such as pH, total soluble solids, titratable acid-
ity, color, antioxidant capacity, and β-carotene content were not significantly different (P > 0.05) among
ultrasound and pasteurization treatments, while turbidity and total aerobic viable count showed similar
trends for both treatments. The authors suggested ultrasound treatment as a potential method that can be
used as an alternative to thermal pasteurization.
Another recent study confirmed the finding of Gao and Rupasinghe [51] who evaluated a number of
juice quality parameters as well as antioxidant capacity and nutritional characteristics of ultrasound-
treated apple juice [52]. Apart from the similar findings on quality parameters of the juices to the previ-
ous study, Abid et al. [52] further showed that ultrasound treatment significantly improved ascorbic acid
content, cloud value, phenolic compounds, antioxidant capacity, and differences in Hunter color values.
These results confirmed that ultrasound treatment can be used as an alternative to the conventional ther-
mal pasteurization as these thermal treatments cause loss of nutritional and physicochemical parameters.
Osmotic evaporation was tested as a method to produce concentrated apple juice products with high
quality and low volume and as a substitution for thermal evaporation techniques [53]. The use of mem-
brane processes can be beneficial as concentration was carried out at room temperature. During the
process, some of the phenolics (overall loss of 18%) and volatile compounds were lost, while less volatile
compounds were concentrated. However, the reconstituted juices were accepted by consumers.
Another potential substitution for thermal pasteurization was ultra-high-pressure homogenization
treatment, which did not disturb the original ascorbic acid and dehydroascorbic acid content of the apple
juice compared to the raw juice [54]. Further, the antioxidant capacity and phenolic compounds in apple
juice was preserved when ultra-high-pressure homogenization was applied at 300 MPa. The weakness of
this method was the oxidation of some bioactives under these particular processing conditions.
Production of apple juice involves pressing combined with enzyme pretreatment of the apple mash.
These conventional methods involve higher degradation of nutrients, energy consumption, and loss of
juice quality. A combination of pressing and pulsed electric fields has been tested in this regard as an
alternative to the traditional mechanical thermal treatments and showed promising results [36]. Pulsed
electric field treatment applied to whole apples or cut apple slices before pressing improved juice yield
and clarity without affecting pH and conductivity. This treatment increased the polyphenol content of the
juice and improved its antioxidant capacity. The drawback of this treatment was the accelerated brown-
ing process of the juice. However, it was suggested that identifying the optimum time for pulsed electric
field application to whole apples and combine it along with pressing can cut down the energy consump-
tion and improve juice quality and quantity. A similar study had previously been carried out where apple
mash was treated with pulsed electric field before pressing [35]. The juice yield increased with increasing
field intensities, but contrastingly, overall composition, nutritional content, and antioxidant capacity were
not affected.
8.6 Conclusion
Apple juice is popular among adults and children due to its desirable flavor. Health-conscious shoppers
consume apple juice for its nutritive value, health-promoting constituents, and a convenient substitute for
the whole fruit. Apple juice has shown numerous health benefits including antiatherosclerotic, cardiovas-
cular health effects, cancer chemopreventive effects, and neurodegenerative disease preventative effects,
studied under various experimental model systems. The bioactives present in apple juice are responsible
for health-promoting effects, and apart from their antioxidant activity, many other cellular mecha-
nisms have been suggested for the bioactivity of these compounds. To make apple juice the u ltimate
health-promoting convenient drink for consumers, research for improving its biological functionality is
required, which would greatly benefit the consumers as well as the producers.
REFERENCES
1. Food and Agriculture Organization (FAO), Principles and Practices of Small-and Medium-Scale Fruit
Juice Processing, Agricultural Services Bulletin 146, FAO, Rome, Italy, 2001.
2. Health Canada (HC), Nutraceuticals/Functional Foods and Health Claims on Foods, Policy Paper,
Therapeutic Products Programme and the Food Directorate from the Health Protection Branch, Health
Canada, Ottawa, Ontario, Canada, 1998.
3. Statistics Canada (SC), Food statistics, 2009, May 2010. Published online at: http://www.statcan.gc.ca/
pub/21–020-x/21–020-x2009001-eng.htm (accessed April 12, 2013).
4. Agriculture and Agri-Food of Canada (AAFC), A snapshot of the Canadian apple industry, 2010,
2012. Published online at: http://www4.agr.gc.ca/AAFC-AAC/display-afficher.do?id=1334147419910
5. U.S. Department of Agriculture (USDA), World Markets and Trade: Apple Juice, Foreign Agricultural
Services, USDA, Springfield, VA, 2007.
6. Gao, J. and Rupasinghe, H.P.V., Characterization of malolactic conversion by Oenococcus oeni to reduce
the acidity of apple juice. Int. J. Food Sci. Technol., 48, 1018–1027, 2013.
7. Campeanu, G., Neata, G., and Darjanschi, G., Chemical composition of the fruits of several apple culti-
vars growth as biological crop. Not. Bot. Horti. Agrobot., 37, 161–164, 2009.
8. Eisele, T.A. and Drake, S.R., The partial compositional characteristics of apple juice from 175 apple
varieties. J. Food Comp. Anal., 18, 213–221, 2005.
9. Health Canada (HC), Nutrient values of some common foods, 2008. Published online at: http://www.
hc-sc.gc.ca/fn-an/nutrition/fiche-nutri-data/nutrient_value-valeurs_nutritives-tc-tm-eng.php (accessed
April 15, 2013).
10. Johnson, P.E., van der Plancken, I., Balasa, A., Husband, F.A., Grauwet, T., Hendrickx, M., Knorr,
D., Mills, E.N., and Mackie, A.R., High pressure, thermal and pulsed electric-field-induced structural
changes in selected food allergens. Mol. Nutr. Food Res., 54, 1701–1710, 2010.
11. Houska, M., Heroldova, M., Vavrova, H., Kucera, P., Setinova, I., Havranova, M., Honzova, S. et al.
Is high-pressure treatment able to modify the allergenicity of the main apple juice allergen, Mal d1?
High Pres. Res., 29, 14–22, 2009.
12. Hopkins, J., The toxicological hazards of patulin. Food Chem. Toxicol., 31, 455–456, 1993.
13. World Health Organization (WHO), WHO 44th Report of the Joint FAO/WHO Expert Committee on
Food Additives, Technical Report Series, 859, 36, Geneva, Switzerland, 1995.
14. Murillo-Arbizu, M., Amézqueta, S., González-Peñas, E., and de Cerain, A.L., Occurrence of patulin
and its dietary intake through apple juice consumption by the Spanish population. Food Chem., 113,
420–423, 2009.
15. Welke, J.E., Hoeltz, M., Dottori, H.A., and Noll, I.B. Effect of processing stages of apple juice concen-
trate on patulin levels. Food Control, 20, 48–52, 2009.
16. Assatarakul, K., Churey, J.J., Manns, D.C., and Worobo, R.W., Patulin reduction in apple juice from
concentrate by UV radiation and comparison of kinetic degradation models between apple juice and
apple cider. J. Food Prot., 75, 717–724, 2012.
17. Lu, C., Bravo, R., Caltabiano, L.M., Irish, R.M., Weerasekera, G., and Barr, D.B., The presence of
dialkylphosphates in fresh fruit juices: Implication for organophosphorus pesticide exposure and risk
assessments. J. Toxicol. Environ. Health A, 68, 209–227, 2005.
18. Boyer, J. and Liu, R.H., Apple phytochemicals and their health benefits. Nutr. J., 3, 5–11, 2004.
19. Rupasinghe, H.P.V., Thilakarathna, S.H., and Nair, S., Polyphenols of apples and their potential health
benefits, in Polyphenols: Chemistry, Dietary Sources and Health Benefits, Sun, J., Prasad, K.N., Ismail,
A., Yang, B., You, X., and Li, L., Eds., Nova Science Publishers Inc., Hauppauge, NY, 2013, pp. 333–368.
20. Thilakarathna, S.H. and Rupasinghe H.P.V., Anti-atherosclerotic effects of fruit bioactive compounds:
A review of current scientific evidence. Can. J. Plant Sci., 92, 407–419, 2012.
21. Kahle, K., Kraus, M., and Richling, E., Polyphenol profiles of apple juices. Mol. Nutr. Food Res., 49,
797–806, 2005.
22. Manach, C., Scalbert, A., Morand, C., Remesy, C., and Jimenez, L., Polyphenols: Food sources and
bioavailability. Am. J. Clin. Nutr., 79, 727–747, 2004.
23. Mullen, W., Marks, S.C., and Crozier, A., Evaluation of phenolic compounds in commercial fruit juices
and fruit drinks. J. Agric. Food Chem., 55, 3148–3157, 2007.
Apple Juice 105
24. Hyson, D.A., A comprehensive review of apples and apple components and their relationship to human
health. Adv. Nutr., 2, 408–420, 2011.
25. Guyot, S., Marnet, N., Sanoner, P., and Drilleau, J.F., Variability of the polyphenolic composition of
cider apple (Malus domestica) fruits and juices. J. Agric. Food Chem., 51, 6240–6247, 2003.
26. U.S. Department of Agriculture (USDA), Database for the flavonoid content of selected foods, Release 3,
September 2011. Published online at: http://www.ars.usda.gov/Services/docs.htm?docid=6231 (accessed
April 20, 2013).
27. U.S. Department of Agriculture (USDA), Database for the proanthocyanidin content of selected foods,
August 2004. Published online at: http://www.nal.usda.gov/fnic/foodcomp (accessed April 20, 2013).
28. Karaman, Ş., Tütem, E., Sözgen Başkan, K., and Apak, R., Comparison of total antioxidant capacity
and phenolic composition of some apple juices with combined HPLC–CUPRAC assay. Food Chem.,
120, 1201–1209, 2010.
29. Bellion, P., Digles, J., Will, F., Dietrich, H., Baum, M., Eisenbrand, G., and Janzowski, C., Polyphenolic
apple extracts: Effects of raw material and production method on antioxidant effectiveness and reduc-
tion of DNA damage in Caco-2 cells. J. Agric. Food Chem., 58, 6636–6642, 2010.
30. Pearson, D.A., Tan, C.H., German, J.B., Davis, P.A., and Gershwin, M.E., Apple juice inhibits human
low density lipoprotein oxidation. Life Sci., 64, 1913–1920, 1999.
31. van Der Sluis, A.A., Dekker, M., Skrede, G., and Jongen, W.M., Activity and concentration of poly-
phenolic antioxidants in apple juice. 1. Effect of existing production methods. J. Agric. Food Chem.,
50, 7211–7219, 2002.
32. Oszmianski, J., Wojdylo, A., and Kolniak, J., Effect of pectinase treatment on extraction of antioxi-
dant phenols from pomace, for the production of puree-enriched cloudy apple juices. Food Chem.,
127, 623–631, 2011.
33. van der Sluis, A.A., Dekker, M., Verkerk, R., and Jongen, W.M., An improved, rapid in vitro method to
measure antioxidant activity: Application on selected flavonoids and apple juice. J. Agric. Food Chem.,
48, 4116–4122, 2000.
34. Oszmianski, J., Wojdylo, A., and Kolniak, J., Effect of enzymatic mash treatment and storage on phe-
nolic composition, antioxidant activity, and turbidity of cloudy apple juice. J. Agric. Food Chem., 57,
7078–7085, 2009.
35. Schilling, S., Alber, T., Toepfl, S., Neidhart, S., Knorr, D., Schieber, A., and Carle, R., Effects of pulsed
electric field treatment of apple mash on juice yield and quality attributes of apple juices. Innov. Food
Sci. Emerg., 8, 127–134, 2007.
36. Grimi, N., Mamouni, F., Lebovka, N., Vorobiev, E., and Vaxelaire, J., Impact of apple processing modes
on extracted juice quality: Pressing assisted by pulsed electric fields. J. Food Eng., 103, 52–61, 2011.
37. Vieira, F.G.K., di Pietro, P.F., da Silva, E.L., Borges, G.S.C., Nunes, E.C., and Fett, R., Improvement
of serum antioxidant status in humans after the acute intake of apple juices. Nutr. Res., 32, 229–232,
2012.
38. Hyson, D., Studebaker-Hallman, D., Davis, P.A., and Gershwin, M.E., Apple juice consumption reduces
plasma low-density lipoprotein oxidation in healthy men and women. J. Med. Food, 3, 159–166, 2000.
39. Godycki-Cwirko, M., Krol, M., Krol, B., Zwolinska, A., Kolodziejczyk, K., Kasielski, M., Padula, G.
et al., Uric acid but not apple polyphenols is responsible for the rise of plasma antioxidant activity after
apple juice consumption in healthy subjects. J. Am. Coll. Nutr., 29, 397–406, 2010.
40. Décordé, K., Teissèdre, P.L., Auger, C., Cristol, J.P., and Rouanet, J.M., Phenolics from purple grape,
apple, purple grape juice and apple juice prevent early atherosclerosis induced by an atherogenic diet in
hamsters. Mol. Nutr. Food Res., 52, 400–407, 2008.
41. Setorki, M., Asgary, S., Eidi, A., Rohani, A.H., and Esmaeil, N., Effects of apple juice on risk factors
of lipid profile, inflammation and coagulation, endothelial markers and atherosclerotic lesions in high
cholesterolemic rabbits. Lipids Health Dis., 8, 39–48, 2009.
42. Barth, S.W., Fahndrich, C., Bub, A., Dietrich, H., Watzl, B., Will, F., Briviba, K., and Rechkemmer, G.,
Cloudy apple juice decreases DNA damage, hyperproliferation and aberrant crypt foci development in
the distal colon of DMH-initiated rats. Carcinogenesis, 26, 1414–1421, 2005.
43. Barth, S.W., Faehndrich, C., Bub, A., Watzl, B., Will, F., Dietrich, H., Rechkemmer, G., and Briviba, K.,
Cloudy apple juice is more effective than apple polyphenols and an apple juice derived cloud fraction in
a rat model of colon carcinogenesis. J. Agric. Food Chem., 55, 1181–1187, 2007.
44. Zessner, H., Pan, L., Will, F., Klimo, K., Knauft, J., Niewohner, R., Hummer, W. et al., Fractionation of
polyphenol-enriched apple juice extracts to identify constituents with cancer chemopreventive potential.
Mol. Nutr. Food Res., 52(Suppl. 1), 28S–44S, 2008.
45. Ortiz, D. and Shea, T., Apple juice prevents oxidative stress induced by amyloid-beta in culture.
J. Alzheimers Dis., 6, 27–30, 2004.
46. Tchantchou, F., Chan, A., Kifle, L., Ortiz, D., and Shea, T., Apple juice concentrate prevents oxidative
damage and impaired maze performance in aged mice. J. Alzheimers Dis., 8, 283–287, 2005.
47. Chan, A. and Shea, T.B., Dietary supplementation with apple juice decreases endogenous amyloid-beta
levels in murine brain. J. Alzheimers Dis., 16, 167–171, 2009.
48. Chan, A., Graves, V., and Shea, T.B., Apple juice concentrate maintains acetylcholine levels following
dietary compromise. J. Alzheimers Dis., 9, 287–291, 2006.
49. Chan, A. and Shea, T.B., Supplementation with apple juice attenuates presenilin-1 overexpression
during dietary and genetically-induced oxidative stress. J. Alzheimers Dis., 10, 353–358, 2006.
50. Rupasinghe, H.P.V., Huber, G.M., Embree, C., and Forsline P.L., Red-fleshed apple as a source for
functional beverage., Can. J. Plant Sci., 90, 95–100, 2010.
51. Gao, J. and Rupasinghe, H.P.V., Characterization of ‘Honeycrisp’ and ‘McIntosh’ apple juice quality in
relation to delayed cooling treatments. Open Food Sci. J., 6, 12–15, 2012.
52. Abid, M., Jabbar, S., Wu, T., Hashim, M.M., Hu, B., Lei, S., Zhang, X., and Zeng, X., Effect of ultra-
sound on different quality parameters of apple juice. Ultrason. Sonochem., 20, 1182–1187, 2013.
53. Aguiar, I.B., Miranda, N.G.M., Gomes, F.S., Santos, M.C.S., Freitas, D.d.G.C., Tonon, R.V., and Cabral,
L.M.C., Physicochemical and sensory properties of apple juice concentrated by reverse osmosis and
osmotic evaporation. Innov. Food Sci. Emerg., 16, 137–142, 2012.
54. Suarez-Jacobo, A., Rufer, C.E., Gervilla, R., Guamis, B., Roig-Sagues, A.X., and Saldo, J., Influence
of ultra-high pressure homogenisation on antioxidant capacity, polyphenol and vitamin content of clear
apple juice. Food Chem., 127, 447–454, 2011.
Release 27. National Technical Information Service, USDA, Springfield, VA, 2014.
9
Apricot Juice/Nectar
Emine Aytunga Arık Kibar and Hatice İmge Oktay Başeğmez
CONTENTS
9.1 Introduction................................................................................................................................... 107
9.2 Nutritional Characteristics............................................................................................................ 108
9.3 Bioactives and Antioxidant Efficacy............................................................................................. 109
9.4 Health Effects.................................................................................................................................112
9.5 Novel Products/Formulations and Future Trends..........................................................................114
9.5.1 Novel Processing Technologies.........................................................................................114
9.5.2 Apricot Juice Blends and Fortification..............................................................................115
9.5.3 Apricot Juice Processing By-Products..............................................................................115
9.6 Conclusion......................................................................................................................................116
References................................................................................................................................................116
9.1 Introduction
Apricot (Prunus armeniaca L.) is classified under the Prunus species of Prunoideae subfamily of the
Rosaceae family of the Rosales group and categorized under “stone fruits,” due to its seed being enclosed
in a hard, “stone-like” endocarp. It is produced by cultivation of a wild apricot, called “Zerdali,” and has
an important place in human nutrition [1]. Turkey is the leading apricot producer followed by Iran. These
two countries accounted for about 32% of the share of the total world production in 2012 [2].
Apricot is a seasonal fruit and must either be consumed rapidly or treated in some manner to retard
spoilage. About 15%–20% of apricots produced are consumed as fresh and the remainder are processed
as juice, nectar, dried, jam, and canned, among others [3]. The Turkish Juice Industry Association
(MEYED) reported that 36,500 tons of apricot was processed to pulp in Turkey in 2010, which is 8% of
the apricot production and ranks as the second largest production of pulp after peaches [4].
The processing of apricots to apricot pure includes pretreatment steps such as selecting, washing,
cleaning, destoning, and mashing, which are similar to those for other fruits. The following two crucial
unit operations are applied, namely, (1) heat processing ranging from 85°C to 145°C for microbial and
enzyme inactivation and (2) enzyme treatment aids in the extraction of juice, higher yields, and higher
soluble solid contents. Other steps such as filtration and clarification are similar to those for other juices.
Apricot juice and nectar contain several bioactive class of compounds such as polyphenols, carot-
enoids, polysaccharides, minerals, sugars, and vitamins [5,6], which contribute to their characteristic fla-
vor (taste and aroma) and nutritive value as well as attracting considerable interest recently. This chapter
highlights the nutritional and phytochemical content of apricot juice and apricot nectar, their effect on
human health with regard to clinical and in vitro studies, and the novel processing methods and future
trends in the apricot juice industry.
107

A comparative nutritional data of fresh apricot, juice, and nectar are provided in Table 9.1. Nutritional
characteristics of apricots may vary among cultivars and different processing steps. Apricot is not very
suitable to be produced and consumed as 100% fruit juice individually due to its natural dense structure.
Therefore, it is commonly processed as pulpy nectar by adding water and sugar.
Despite the processing conditions, several nutritional constituents of fresh apricot remain unchanged
in the juice and nectar. However, energy and total sugars content are slightly higher in nectar than fresh
apricots and juice, probably due to the added sugar during processing. Fresh apricot, its juice and nec-
tar are good sources of soluble fiber (Table 9.1), which is important for a healthy diet [3]. Dietary fiber
is the edible part of plants or analogous carbohydrates that are resistant to digestion and absorption in
the human small intestine, with complete or partial fermentation in the large intestine [7]. The British
Nutrition Foundation (BNF) [8] has recommended a minimum fiber intake of 18 g/day for healthy adults.
Soluble fiber can help to regulate blood sugar level by binding water and forming a viscous gel dur-
ing digestion, slowing the emptying of the stomach and intestinal transit, shielding carbohydrates from
TABLE 9.1
Compositional and Nutritional Characteristics of Apricot, Juice, and Nectar (per 100 g)
Nutrient Unit Apricot Juice Nectar
Water g 86.35 86.62 84.87
Energy kcal 48 48 56
Protein g 1.40 0.63 0.37
Fat (lipid) g 0.39 0.04 0.09
Carbohydrate g 11.12 12.34 14.39
Total sugars g 9.24 10.74 13.79
Total dietary fiber g 2.0 1.6 0.6
Minerals
Calcium mg 13 12 7
Iron mg 0.39 0.30 0.38
Magnesium mg 10 10 5
Phosphorus mg 23 20 9
Potassium mg 259 165 114
Sodium mg 1 4 3
Zinc mg 0.20 0.11 0.09
Vitamins
Niacin mg 0.60 0.344 0.260
Riboflavin mg 0.040 0.019 0.014
Thiamin mg 0.030 0.018 0.009
Folate (DFE) μg 9 2 1
Vitamin A (RAE) μg 96 85 66
Vitamin B6 mg 0.054 0.054 0.022
Vitamin B12 μg 0.00 0.00 0.00
Vitamin C mg 10 4.9 0.6
Vitamin E (ATE) mg 0.89 0.60 0.31
Vitamin K μg 3.3 2.2 1.2
Source: Adapted from the U.S. Department of Agriculture (USDA), USDA National Nutrient Database for
Standard Reference, Release 27, National Technical Information Service, USDA, Springfield, VA, 2014.
Abbreviations: DFE, dietary folate equivalents; RAE, retinol activity equivalents; ATE, alpha-tocopherol
equivalents.
Apricot Juice/Nectar 109
enzymes, and delaying absorption of glucose [9]. Moreover, it is reported that soluble fiber lowers total
and low-density lipoprotein (LDL) cholesterol, which may reduce the risk of cardiovascular disease
(CVD) [10].
Fresh apricot and apricot juice/nectar are considered nutritious due to their mineral content. Potassium
is the most abundant mineral in apricot, followed by calcium. Other important minerals found in apricot
and its products include iron and magnesium. One serving of apricot juice, nectar (200 mL) or fresh
apricot contains ~7%–8%, 3%–5%, or 3% of Recommended Dietary Allowances (RDA) for potassium,
iron, and magnesium, respectively [11].
Apricot is a good source of vitamins, including vitamins A and C. Niacin, riboflavin, thiamin, folate,
vitamin B6, vitamin E, and vitamin K are other vitamins that are found in fresh apricot, juice, and nectar
in low concentrations. The most abundant vitamins in fresh apricot is vitamin C, followed by niacin
and vitamin A, but these are in considerably lower amounts in apricot nectar compared to fresh apricot
probably due to their high sensitivity to oxidation during processing. However, apricot juice/nectar is
still a good source of vitamin A. Indeed, drinking one glass (200 mL) of apricot juice/nectar provides
approximately 15%–24% of the RDA for vitamin A [11].

Apricot is a good source of various phytochemicals mainly based on polyphenols and carotenoids
(Table 9.2), but it is not suitable to be processed as 100% fruit juice due to its natural dense structure. Hence,
apricot is commonly processed as pulpy nectar by adding water and sugar. In the available literature on
TABLE 9.2
Bioactive Compositions of Apricot, Juice, and Nectar (per 100 mL or 100 g)
Class Unit Compound Apricot Juice Nectar References
Phenolic acids mg p-Coumaric acid 0.02–8.29 na 0.007–0.594 [12,13,15]
mg Chlorogenic acid 1.58–5.67 2.0–3.1 0.06–2.59 [12,13,15,18,19]
mg Neochlorogenic acid 0.12–0.15 na 2.44 [12,14]
mg Ferulic acid 0.30–0.70 na 0.005–0.644 [12,13,15]
mg Caffeic acid 0.81–2.72 na 0.009–0.396 [12,13,15]
mg p-Hydroxybenzoic acid na na 0.006–0.039 [15]
mg Vanillic aldehyde na na 0.001–0.016 [15]
mg Syringic aldehyde na na 0.001–0.007 [15]
mg 3,4-Dihydroxybenzoic na na 0.004–0.008 [15]
aldehyde
Flavan-3-ols mg (+)-Catechin 0.31–7.34 na 0.18–2.65 [12,13,16]
mg (−)-Epicatechin 4.19–8.29 na 0.35–2.36 [12,13]
Flavonols mg Quercetin 0.38–2.90 na 0.004–0.016 [15,16]
mg Isoquercetin 0.65–2.85 na 0.002–0.128 [14,15]
mg Rutin 0.46–2.27 1.55–2.83 0.037–0.813 [15,18,19]
mg Kaempferol 0.00–1.32 na na [16]
mg Kaempferol-3-rutinoside 0.01–2.81 0.12–0.19 0.11–0.21 [13,18]
mg Astragalin na na 0.008–0.075 [15]
Hydoxycoumarins mg Scopoletin na na 0.003–0.007 [15]
mg Aesculetin na na 0.002–0.005 [15]
Carotenoids µg β-Carotene 163–2554 na 1625 [12,14,16]
µg α-Carotene nd-44 na 38.3 [12,14]
µg Lutein 3–188 na 0.62 [12,14]
µg Zeaxantin nd-38.96 na 1.44 [12]
µg β-Crytoxanthin na na 4.90 [12]
Abbreviations: nd, not detected; na, not available.
O O
OH OH
HO HO
O OH O OH
HO O O OH HO O O OH
H H
Chlorogenic acid Neochlorogenic acid
OH OH
OH OH
HO O HO O
OH
OH
OH OH
(+)-Catechin (–)-Epicatechin
OH
HO O
OH
OH
O
HO OH O OH
OH O
HO
O O CH3
OH
O O OH OH2 O O OH
O
HO OH HO OH
O OH
OH OH
Kaempferol 3-Ο-rutinoside Rutin
β-Carotene
FIGURE 9.1 Chemical structures of major phytochemicals found in apricot nectar.
the bioactive content of apricot products, only a few studies are related to apricot juice and the rest are
mainly related to phytochemicals of apricot nectar. Therefore, the bioactive composition and antioxidant
activity of apricot nectar are discussed in detail in this section rather than apricot juice. The most abundant
phenolic acids, flavan-3-ols, flavonols, and carotenoids found in apricot nectar are shown in Figure 9.1.
When the bioactive compositions of apricot nectar are considered, chlorogenic acid is the most abun-
dant phenolic acid found in apricot nectar, followed by neochlorogenic acid, which is another hydroxy-
cinnamic acid derivative [12–14]. Caffeic, p-coumaric, and ferulic acids are other phenolic acids, which
are found in fresh apricot and apricot nectar in low amounts [12,14,15]. Furthermore, one benzoic acid
(p-hydroxybenzoic acid) and three benzoic aldehydes (3,4-dihydroxybenzoic, vanillic, and syringic) are
also detected in apricot nectar in ample amounts (Table 9.2).
The presence of two well-known flavan-3-ol monomers, (+)-catechin and (−)-epicatechin, in fresh
apricot and apricot nectar is documented [12,13,16]. However, a great variation in the reported concen-
trations of those flavanols in apricot nectar is noteworthy. This can probably be attributed not only to
cultivar differences [14] but also to the differences in processing. A comprehensive study was carried
out for detecting the phenolic profiles of 21 apricot genotypes. It was presented that (+)-catechin and
(−)-epicatechin concentrations showed 15- and 50-fold variations, respectively, in different genotypes
[17]. Processing conditions such as the degree of pressing during leaching and whether the peeled fruit or
whole fruit is used in the process are also important factors that affect the flavanol concentration in apri-
cot products. Flavanols are mainly located in the skin of apricot; therefore some of these compounds may
remain in the peel during processing, and this may lead to a decrease in their concentrations in nectar
[18]. In addition, flavanols are natural substrates for polyphenol oxidase, and they are involved in brown-
ing, polymerization, and haze formation phenomena leading to losses during storage or processing [15].
Rutin, a flavonoid glycoside, is reported as the most dominant flavonol in apricot nectar [15,18,19].
Quercetin, isoquercetin, kaempferol, kaempferol-3-rutinoside, and astragalin are the other flavonols
detected in apricot nectar [13–16,18,19]. Moreover, Fernandez de Simon et al. [15] detected small quan-
tities of two hydroxycoumarins (scopoletin and aesculetin) in apricot nectar. In addition, β-carotene is
the main carotenoid found in apricot nectar, accounting for approximately 97% of total carotenoids, fol-
lowed by α-carotene, β-cryptoxanthin, zeaxanthin, and lutein (Table 9.2).
Polyphenol and carotenoid content of fresh apricot and apricot nectar vary considerably. This varia-
tion can be attributed to differences in three important factors: (1) cultivars, (2) cultivation practices, and
(3) the processing methods. A study on phenolic profiles of 11 apricot cultivars revealed that individual
phenolic compounds showed up to 12-fold variation according to the cultivar. Among the cultivation
parameters, season, weather condition, ripeness, and cultivation techniques played significant roles in the
formation of colorants of apricots, particularly carotenoids [17,20,21]. In apricot fruits, the predominant
pigment, β-carotene, increased rapidly during ripening, because biosynthesis of carotenoids is improved
significantly during ripening [14]. Interestingly, farming practices may impact the content of phytochem-
icals present in fresh apricots and apricot juice. A comprehensive study was carried out among 26 Italian
commercial apricot nectars obtained from apricots produced with organic, integrated, and conventional
agriculture practices [19]. It was determined that chlorogenic acid and rutin concentrations were higher
in nectars produced from apricots obtained by organic farming than those of integrated and conventional
agriculture practices.
As it can be seen from Table 9.2, similar bioactive profiles are present in fresh apricot and apricot
nectar. However, the amounts of phenolics and carotenoids are considerably lower in nectar than in fresh
apricot. This result may indicate that bioactives of fresh apricot are decreased during processing to nec-
tar [13,16]. Dragovic-Uzelac et al. [13] revealed that during processing of apricot nectar, (+)-catechin and
(−)-epicatechin concentrations in fresh apricot decreased by ~65% and ~40%, respectively. β-Carotene
content of fresh apricot and canned apricot nectar were 1094 µg/100 g and 786 µg/100 mL, respectively
[16], reflecting ~28% decrease in β-carotene concentration. Therefore, alternative processing technolo-
gies are needed to be evaluated in order to conserve functional properties of apricot products.
Recently, Huang et al. [12] conducted a comparative study of phenolics and carotenoids of apricot
nectar produced by alternative processing technologies, such as high hydrostatic pressure (HHP) and
high-temperature short-time (HTST) methods. They measured total phenolics of untreated, HHP-, and
HTSH-treated apricot nectars as 366, 403, and 721 mg gallic acid equivalents (GAE)/L, respectively.
All treated samples presented a significant increase in total phenolics as compared to untreated apricot
nectar. This increase was attributed to cell disruption entailed by HHP, which would release substrates
and promote changes in total phenolic content (TPC). It was also noted that easier extraction of phenolic
compounds in the pulps and complete inactivation of polyphenol oxidase enzyme during HTST also
made contribution to phenolics retention [12]. Moreover, as compared to unprocessed nectar, HHP and
HTST treatments induced a significant increase (P < 0.05) in neochlorogenic acid, (+)-catechin, chloro-
genic acid, and caffeic acid concentrations. HTST and HHP exhibited no effect (P > 0.05) on total carot-
enoids in apricot nectar except that the HHP treatment at 500 MPa/20 min increased total carotenoids
and β-carotene [12].
Table 9.3 shows the TPC, total carotenoid content (TCC), and total antioxidant activities (TAA) of
fresh apricot, juice, and nectar. TPC of fresh apricot, juice, and nectar were in the range of 208–2451 mg
GAE/kg, 186–626 mg GAE/L, and 366–457 mg GAE/L, respectively[12,17,20–26]. Similarly, TCC of
apricot, juice, and nectar were 148–919, 10.1, and 3.3–16.7 mg β-carotene equivalents (βCE)/kg, respec-
tively (Table 9.3). The significant variations in TPC and TCC of fresh apricot can be attributed to the
TABLE 9.3
Total Phenolics, Carotenoids, and Antioxidant Activities of Apricot, Juice, and Nectar (per L or kg)
Assay Unit Apricot Juice Nectar References
Total phenolics mg GAE 208–2451 186–626 366–457 [12,17,21–26]
Total carotenoids mg βCE 148–919 10.1 3.3–16.7 [5,12,26,43]
Antioxidant activity TEAC mmol TE 1.76–12.73 1.25–2.48 na [21,25]
FRAP mmol Fe+2 0.16–4.02 2.2–7.15 5.68 [22,23,26]
TRAP mmol TE 2.29 2.19 na [22]
ORAC mmol TE 13.41 na na [52]
DPPH mmol TE 0.11–7.4 na na [17]
Abbreviations: na, not available; GAE, gallic acid equivalents; βCE, β carotene equivalents; TE, trolox equivalents; TEAC,
trolox equivalent antioxidant capacity; FRAP, ferric reducing antioxidant power; TRAP, total radical-
trapping antioxidant parameter; ORAC, oxygen radical absorbance capacity; DPPH, 1,1-diphenyl-2
picrylhydrazyl.
aforementioned cultivar and cultivation differences. In addition, both the TPC and TCC of juices and
nectars are lower than fresh apricots, probably due to unfavorable processing conditions.
Several methods have been developed to assess TAA of food and beverages such as trolox equivalent
antioxidant capacity (TEAC), ferric reducing antioxidant power (FRAP), and total radical-trapping anti-
oxidant parameter (TRAP) in which chemical principals behind the assays are different. Various anti-
oxidant compounds may act in vivo through different mechanisms. Therefore, no single method can fully
evaluate the TAA of foods [22]. For example, Herken and Guzel [23] determined TPC and in vitro TAA
of seven commercial fruit juices including apricot juice. They measured TAA using different assays,
the first one was using Fe+2-o-dianisidine complex, the second one was based on the decolorization of
(2,2-azino-bis [3-methybenzothiazoline-6-sulfonate]) (ABTS) radical, and the last one was the com-
monly used FRAP method. The TAA values were 26.5, 3.0, and 2.2 mmol trolox equivalents (TE)/L
using three methods, respectively. Therefore, it is emphasized that the measurement of antioxidant activ-
ity of fruit juice, thus largely depended upon the free radical or the oxidants used in the assays and the
amount and type of antioxidants.
It is generally accepted that there is a high correlation between phenolic content and antioxidant
activity of fruits [24]. For instance, Herken and Guzel [23] showed that a significant correlation existed
between TPC and TAA of apricot juice (r 2 = 0.832). In another study, the correlation coefficient between
TPC and TAA of fresh apricot juices was calculated as r 2 = 0.922, which indicated that phenolics were
major contributors to the antioxidant activity of apricots juice [25].
When fresh apricot and apricot nectars were compared with respect to TAA, significant differences
might be detected due to vitamin C addition to commercial apricot nectars. In a comparative study,
TEAC, FRAP, and TRAP values of fresh apricot were measured as 1.44 mmol TE/kg, 4.02 mmol
Fe2+/kg, and 2.29 mmol TE/kg, respectively, while values for apricot nectar were 2.48 mmol TE/kg,
7.15 mmol Fe2+/kg, and 2.19 mmol TE/kg, respectively [22]. According to these values, antioxidant
activity of apricot nectar was slightly higher than that of fresh apricot due to vitamin C addition dur-
ing processing. However, in another study, it was shown that antioxidant activity of commercial apricot
nectar was threefold lower than that of laboratory-scale produced apricot juice [25]. Similarly, Mahdavi
et al. [24] reported that fresh apricot juice had higher TPC (237.5 mg GAE/L) than commercial apricot
nectar (185.7 mg GAE/L). As a result, it might be stated that processing techniques, clarification, and
pasteurization might reduce the antioxidant capacity of apricot nectars.
9.4 Health Effects

Although epidemiological studies have documented that diets rich in fruits and vegetables can reduce
the risk of cancer and other chronic diseases with their high antioxidant content, the health benefits of
apricot juice/nectar consumption are poorly investigated.
Phenolic compounds present in fruits and vegetables act as metal chelators and can have antimicrobial
properties. However, inappropriate processing and storage conditions may cause contamination of fresh
cut fruits and unpasteurized fruit juices, with pathogenic and/or deteriorative microorganisms, and thus
increase the risk of microbial diseases and spoilage [27]. Parallel to the growing interest in food industry
for natural preservatives against pathogenic microorganisms, Krisch et al. [28] studied in vitro effect of
fruit juices including apricot juice on the growth of Gram-positive (Bacillus subtilis and B. cereus) and
Gram-negative (Escherichia coli and Serratia marcescens) bacteria. Apricot juice showed poor growth-
reducing effects, compared with black currant, cornelian cherry, and European rowan juice, probably
due to the higher anthocyanin content of these dark-colored fruits. This assumption was supported by
many studies demonstrating that juices from purple and red fruits with high content of anthocyanins had
strong antimicrobial effects [29,30].
Several studies show that dietary antioxidants can scavenge reactive oxygen species (ROS) in the
body and hence favor lowering the oxidative stress involved in tissue damage, accelerated aging and
chronic degenerative diseases, such as cancer and CVD. Furthermore, the protective effects of naturally
occurring antioxidants against oxidative stress using human colon cancer cell line (Caco-2 cells) have
been investigated by different researchers. Thus, monitoring the antioxidant effects of carotenoids [31]
and flavonoids [32] on the accumulation of ROS was comprehensively reported. There are only a few
studies addressing solely the health benefits of apricot juice/nectar [33,34]. Cilla et al. [35] investigated
in vitro effect of fruit juice mixture including apricot juice on the inhibition of oxidative stress induced
by hydrogen peroxide (H2O2) on Caco-2 cells. Fruit juice mixtures from grape concentrate, orange
concentrate, and apricot puree with concentrations of 7.2, 4.2, and 24.5 g/100 g were used, respectively.
After the digestion of fruit juice mixture using simulation of human gastrointestinal digestive process,
Caco-2 cell cultures were incubated with the bioaccessible fractions of digested samples. Despite fruit
juice mixture failed to prevent intracellular ROS accumulation, cytoprotective effect was achieved by
increased mitochondrial integrity and unaltered mitochondrial enzyme function. The maintenance of
mitochondrial integrity is essential for metabolizing intracellular ROS accumulation and for preserv-
ing the activity of the antioxidant enzyme systems. Due to the high mitochondrial integrity of prein-
cubated cultures, the cytoptotective effect was attributed to the bioaccessible fractions of fruit juice
mixture [35]. As a result, the cytoprotective effect of fruit beverages was revealed which was based
on an intestinal epithelial model and could simulate the in vivo situation. After the demonstration of
cytoprotective effect on Caco-2 cells against oxidative stress, Cilla et al. [36] aimed to answer questions
of antiproliferative activity of the mixture of grape, orange, and apricot juices against Caco-2 cells and
the potential mechanism behind the possible antiproliferative activity if it was either apoptosis and/or
cell cycle arrest. Caco-2 cells were incubated with 2%, 5%, and 7.5% of fruit juice digest for repetitive
exposure (4 h for 4 days) and continuously for 24 h after in vitro gastric and intestinal digestion of the
fruit juice mixture. The highest antiproliferative effect (53% inhibition) was observed after continuous
incubation with 7.5% digest (~50 µM total phenolics) for 24 h. However, it was not clear whether the
antiproliferative effect was dose dependent or not. Moreover, the mechanism behind the antiprolifera-
tive effect was associated with the arresting in the S-phase of the cell cycle by decreasing cyclins B1
and D1 levels.
On the other hand, the single human intervention study involved apricot juice consumption as a mix-
ture of beverages, pointed out the effect of daily consumption of fruit juice (mixture of apple, mango,
orange, lime, apricot, and green tea) on DNA damage, antioxidant, and immune system status [34].
Twenty-seven nonsmoking, healthy men with normal body weight consumed 330 mL/day juice mixture
with the main dish avoiding consuming polyphenol rich foods for 2 weeks. The total polyphenols in fruit
juice mixture determined as 684 mg/L that mainly comprised of flavanols and phenolic acids. The most
abundant polyphenols identified in the juice mixture was epigallocatechin gallate (155 mg/L) and gentisic
acid (278 mg/L). Consumption of fruit juices provided 226 mg polyphenols with epigallocatechin gallate
as the major polyphenolic compound. The study revealed a reduction of thiobarbituric acid reactive sub-
stances (TBARS) in human plasma, despite the fact that LDL oxidation did not change after consumption
of polyphenol-rich fruit juice. Furthermore, antioxidant status was improved after consumption of juice
mixture measured by FRAP. The results of this study showed that juice consumption reduced the num-
ber of oxidized DNA bases in peripheral blood mononuclear cells reaching highest effect after 5 weeks
of consumption. Researchers concluded that juice consumption was related to the reduction in DNA
damage through not only the results of direct ROS scavenging by polyphenols but also rather the result
of induced protective enzymes. Finally, fruit juice mixture can specifically change functions of immune
cells without interfering with immune cells [34].
These studies could be inadequate in order to evaluate health benefits of apricot juice/nectar. Because
research samples contain a mixture of various juices, positive effects of bioaccessible fraction cannot
be dedicated to only apricot juice/nectar. Nevertheless, it can be concluded that apricot juice has also a
positive impact on human health with its polyphenol, carotenoid, and nutrient constituents.

9.5.1 Novel Processing Technologies
The processing of apricot juice/nectar includes thermal processing ranging from 85°C to 145°C for
microbial and enzyme inactivation and for improving pressing yield via softening of fruit tissue.
However, thermal processing may cause undesired detrimental effects such as degradation of heat-
sensitive phytochemicals, off-flavor formation, and darkening of the product [12,37]. Today, consum-
ers look for foods that fulfill not only safety requirements but also high-quality standards. In order to
satisfy these demands, nonthermal technologies such as pulsed electric fields (PEF), HHP, and novel
heat treatments such as HTST application may be considered as alternative technologies for apricot
nectar production.
HHP is a promising alternative processing technique to pasteurization due to its limited effects on
covalent bonds resulting in minimal modifications in nutritional and sensory quality [38]. It is applied
to process many fruit and vegetable products, and some reports are available on its application to apricot
nectar [39,40]. Huang et al. [12] showed that the effect of HHP treatment on the total and individual
phenolics, TCC, individual carotenes, and color of apricot nectar was closely related to the pressure
levels and treatment times. Moreover, they reported that treatment at 500 MPa for 20 min increased
total carotenoids and β-carotene in apricot nectar. PEF is another processing technology that can be
applied for apricot nectar processing. Evrendilek et al. [37] showed that processing of apricot nectar
via PEF did not cause a significant difference in the concentration of mineral ions, ascorbic acid, and
β-carotene, besides inactivation of all microorganisms was significantly increased with increased elec-
tric field strength and treatment time (P < 0.05). HTST process, which is known to result in a greater
retention of quality factors than pasteurization, has been used for apricot nectar processing [12]. It was
reported that HTST resulted in a significant increase in TPC, exhibited no effect on the TCC and indi-
vidual carotenes except α-carotene, and produced major color difference with increased lightness and
higher color intensity.
The production of apricot juice concentrates is mainly based on heat evaporation, which is one of the
most used traditional technologies in food industry. The quality of the end product mainly depends on
the temperature gradient between the product and the heat exchanger and high gradient causes local
degradation of the product. Membrane technology is an effective and environmentally friendly alterna-
tive to evaporation. There are some patented applications of membrane technologies such as microfiltra-
tion, ultrafiltration, and reverse osmosis for the production of apricot juice concentrate [41]. Membrane
technologies offer several advantages such as low production cost and operation at ambient tempera-
ture giving the possibility for maximum conservation of the aromatic and nutritional potential of the
juice. However, membrane technologies present limits such as fouling of the membranes and tear due
to high pressure. Therefore, periodic cleaning and replacement of the membranes are inevitable [42].
Cryoconcentration is also a promising alternative technology due to its environmentally friendly, effec-
tive, and low energy consumption nature as well as its ability to preserve sensory and nutritional char-
acteristics. The dry matter content of apricot juice increased up to 35 g/100 g by applying two freezing
temperatures (−10°C and −20°C) and three cryoconcentration stages [43]. In addition, vitamin C content
and aroma number increased by increasing cryoconcentration stage and release of membrane bound
phytochemicals was enhanced during freezing [44].
9.5.2 Apricot Juice Blends and Fortification

During the last decade, ongoing consumer health and wellness trend and the aging population in many
countries offer strong potential for increasing market segmentation for enhanced juices. On the producer
site, the innovation is focused on juice blends due not only to the opportunity for managing production
costs, but also to the intrinsic functional benefits that flavor mixes can offer.
Apricot juice can be enriched by mixing with some additives and other juices. For example, black
carrot is an excellent source of anthocyanins, which are the best-known natural red colorant used in
foods. In a recent study, apricot juice was colored with black carrot juice concentrate and stability of
anthocyanins was studied during heating and storage at 4°C−37°C [45]. It was reported that black car-
rot anthocyanins had good stability in apricot nectar during both heating and storage at refrigerator
temperature.
In another study, bergamot juice, which was a by-product of bergamot oil production, was added to
apricot juice [46]. They reported that apricot juice fortified with bergamot juice concentrate showed a
significant increase in its antioxidant properties and ascorbic acid conservation during typical production
steps. Ascorbic acid content of thermally treated apricot juice was increased from 0.036 to 0.124 mg/
mL by the addition of 20% bergamot juice. Similarly, antioxidant activities of apricot juices with and
without bergamot addition after thermal treatment were evaluated by N,N-dimethyl-p-phenylenediamine
(DMPD) method and measured as 4.85 and 22.76 mmol ascorbic acid equivalents (AAE)/mL, respec-
tively. A consumer test was then carried out which encouraged the production of bergamot fortified
apricot juice. Moreover, the phenolic pattern of bergamot juice was determined in the study, and it was
shown that bergamot juice was rich in phenolic compounds such as narirutin, naringin, isorhoifolin,
rhoifolin, rutin, eriocitrin, neoponcirin, hesperidin, and neodiosmin [46], which would contribute sig-
nificantly to apricot juice’s phenolic content.
Fruit juice–milk mixtures are recently new commercial beverages on market shelves. Zulueta et al.
[47] studied vitamin C, vitamin A, phenolic compounds, and TAA of 17 different commercial juice–
milk beverages. The values of various phytochemical compounds of apricot juice–skim–milk mixtures
were measured as vitamin C: 39.3 mg/100 mL, TPC: 99.9 mg GAE/100 mL, TEAC: 3.31 mmol TE/L,
β-carotene: 75.6 μg/100 mL, α-carotene: 6.24 μg/100 mL, β-cryptoxanthin: 146 μg/100 mL, and retinol:
235 μg/100 mL. When those reported values were compared with the literature data given in Tables 9.2
and 9.3, it was seen that TAA, phenolic content, and vitamin C content of juice–milk beverage were
significantly higher than that of apricot nectar. The authors reported that the main contribution to the
TAA of beverage was provided by vitamin C and percentage of juice did not interact significantly with
the parameters analyzed [47].
Apricot is occasionally considered as the cause of some allergic responses, mainly of the oral allergy
syndrome type, in sensitive subjects. This is because of the presence of a protein in the peel of apricot
named as Pru p3 and Pru ar3, respectively [48]. Brenna et al. [48] showed that it might be possible to
eliminate allergenic proteins from apricot pulp by chemical peeling and enzymatic treatment with pec-
tinases and cellulases followed by ultrafiltration using 10 kDa cutoff membranes. In this way, hypoal-
lergenic apricot pulp and anallergenic apricot juice can be produced and used as ingredients in the
production of other foods such as toffees, jellies, yogurts, and fruit-stuffed snacks, among others.
9.5.3 Apricot Juice Processing By-Products

There are two main by-products of apricot juice processing: apricot pomace (press cake) and kernel.
In a typical processing scheme, 10% (w/w) of apricot flesh is wasted as press cake. Today, it does not
have added value and is generally used as feed or bioenergy processing. However, it can be a good
source of phytochemicals such as β-carotene. Sanal et al. [49] accomplished to extract 88 µg/g dry
pomace β-carotene by supercritical CO2 extraction at 40.5 MPa and 328 K, and they reported that
extraction yield might be improved by adding entrained or applying ultrasonic pretreatment. Apricot
kernel is another by-product that is left after processing and comprises 12.7%–22.2% (w/w) of apri-
cot weight. The apricot kernel contains 45%–50% oil, 23.6%–26.2% protein, 4.2% ash, 5.42% crude
fiber, and 8.2% carbohydrate. The apricot kernel oil is rich in oleic and linoleic acid and a good source
of vitamin E (72–107 mg/100 g) [50]. Moreover, Yiğit et al. [51] showed in vitro antimicrobial and
antioxidant activity of methanol and water extracts of apricot kernel. They determined TPC of sweet
kernels as 5.7–7.9 GAE μg/mL.
9.6 Conclusion
Due to the apricot’s dense structure with high soluble fiber content, it is commonly processed as a pulpy
nectar by adding water and sugar. The soluble fiber content remains at a high level during apricot juice/
nectar processing, although enzymatic extraction is not applied. In addition to the soluble fiber content,
apricot juice/nectar is also rich in vitamin A, potassium, β-carotene, and polyphenols such as chlorogenic
acid. Even though there is a lack of studies related to the health effects of apricot juice/nectar as a whole
food, it can be concluded that apricot juice/nectar has beneficial effects on human health with its content
of carotenoids, polyphenols, and soluble fiber. Generally, juice/nectar is a suitable food product in terms
of digestion of phytochemicals. The bioactive components may be better absorbed from the juice than
the fruit. The consumption of fruit juice contributes to fulfilling the recommended fruit servings.
REFERENCES
1. Haciseferogullari, H., Gezer, I., Ozcan, M., and Murat, A.B., Postharvest chemical and physical-
mechanical properties of some apricot varieties cultivated in Turkey. J. Food Eng., 79, 364–373, 2007.
2. FAO, Word primary crops data, Food and Agriculture Organization of the United Nations Statistics
Division, August 2014. Published online at: http://faostat.fao.org/site/567/default.aspx#ancor (accessed
October 10, 2014).
3. Siddiq, M., Apricots, in Handbook of Fruits and Fruit Processing, 2nd edn., Hui, Y.H., Ed., Wiley-
Blackwell, Oxford, UK, 2006, pp. 279–291.
4. Turkish Juice Industry Association (MEYED), Türkiye Meyve Suyu vb Ürünler Sanayi Raporu,
MEYED, Istanbul, Turkey, 2011 (in Turkish).
5. Akin, E., Karabulut, I., and Topcu, A., Some compositional properties of main Malatya apricot (Prunus
armeniaca L.) varieties. Food Chem., 107, 939–948, 2008.
6. Ali, S., Masud, T., and Abbasi, K., Physico-chemical characteristics of apricot (Prunus armeniaca L.)
grown in northern areas of Pakistan. Sci. Hortic., 130, 386–392, 2011.
7. American Association of Cereal Chemists (AACC), The definition of dietary fiber: Report of the dietary
fiber definition committee to the board of directors of the American Association of Cereal Chemists.
Cereals Food World, 46, 112–126, 2001.
8. British Nutrition Foundation, Dietary fiber definition. Published online at: http://www.nutrition.org.uk/
nutritionscience/nutrients/dietary-fibre.html (accessed December 15, 2014).
9. Sareen, S.G., Smith, J.L., and Groff, J.L., Advanced Nutrition and Human Metabolism, 5th edn.,
Wadsworth, Cengage Learning, Belmont, CA, 2008.
10. Food and Nutrition Board Institute of Medicine of the National Academies, Dietary Reference
Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and Amino Acids
(Macronutrients), The National Academies Press, Washington, DC, 2005.
11. Food and Nutrition Board Institute of Medicine of the National Academies, Dietary Reference Intakes
for Vitamin A, Vitamin K, Arsenic, Boron, Chromium, Copper, Iodine, Iron, Manganese, Molybdenum,
Nickel, Silicon, Vanadium, and Zinc, The National Academies Press, Washington, DC, 2001.
12. Huang, W., Bi, X., Zhang, X., Liao, X., Hu, X., and Wu, J., Comparative study of enzymes, phenolics,
carotenoids and color of apricot nectars treated by high hydrostatic pressure and high temperature short
time. Innov. Food Sci. Emerg. Technol., 18, 74–82, 2013.
13. Dragovic-Uzelac, V., Pospisil, J., Levaj, B., and Delonga, K., The study of phenolic profiles of raw apri-
cots and apples and their purees by HPLC for the evaluation of apricot nectars and jams authenticity.
Food Chem., 91, 373–383, 2005.
14. Dragovic-Uzelac, V., Levaj, B., Mrkic, V., Bursac, D., and Boras, M., The content of polyphenols and
carotenoids in three apricot cultivars depending on stage of maturity and geographical region. Food
Chem., 102, 966–975, 2007.
15. Fernandez de Simon, B., Perezilzarbe, J., Hernandez, T., Gomezcordoves, C., and Estrella, I., Importance
of phenolic-compounds for the characterization of fruit juices. J. Agric. Food Chem., 40, 1531–1535,
1992.
17. Sochor, J., Zitka, O., Skutkova, H., Pavlik, D., Babula, P., and Krska, B., Content of phenolic compounds
and antioxidant capacity in fruits of apricot genotypes. Molecules, 15, 6285–6305, 2010.
18. Garcia-Viguera, C., Bridle, P., Ferreres, F., and Tomasbarberan, F., Influence of variety, maturity
and processing on phenolic compounds of apricot juices and jams. Z. Lebensm.Unters. Forsch. 199,
433–436, 1994.
19. Versari, A., Parpinello, G., Mattioli, A., and Galassi, S., Characterisation of Italian commercial apricot
juices by high-performance liquid chromatography analysis and multivariate analysis. Food Chem.,
108, 334–340, 2008.
20. Sass-Kiss, A., Kiss, J., Milotay, P., Kerek, M., and Toth-Markus, M., Differences in anthocyanin and
carotenoid content of fruits and vegetables. Food Res. Int., 38, 1023–1029, 2005.
21. Leccese, A., Bartolini, S., and Viti R., Total antioxidant capacity and phenolics content in fresh apricots.
Acta Alimentaria, 37, 65–76, 2008.
22. Pellegrini, N., Serafini, M., Colombi, B., Del Rio, D., Salvatore, S., and Bianchi, M., Total antioxidant
capacity of plant foods, beverages and oils consumed in Italy assessed by three different in vitro assays.
J. Nutr., 133, 2812–2819, 2003.
23. Herken, E. and Guzel, S., Total antioxidant capacity and total phenol contents of selected commercial
fruit juices in Turkey. Int. J. Food Prop., 13, 1373–1379, 2010.
24. Mahdavi, R., Nikniaz, Z., Rafraf, M., and Jouyban A., Determination and comparison of the total
polyphenol contents of fresh and commercial fruit juices. Br. Food J., 113, 744–752, 2011.
25. Sercan, K. and Aziz, E., Antioxidant capacity and total phenolic contents of peach and apricot cultivars
harvested from different regions of Turkey. Int. J. Food Nutr. Sci., 1, 13–17, 2012.
26. Tosun, I. and Ustun, N.S., An investigation about antioxidant capacity of fruit nectars. Pak. J. Nutr., 2,
167–169, 2003.
27. Beuchat, L., Pathogenic microorganisms associated with fresh produce. J. Food Protect., 59, 204–216,
1996.
28. Krisch, J., Galgoczy, L., Tölgyesi, M., Papp, T., and Vagvölgyi, J., Effect of fruit juices and pomace
extracts on the growth of Gram-positive and Gram-negative bacteria. Acta Biol. Szeged., 52, 267–270,
2008.
29. Cavanagh, H.M.A., Hipwell, M., and Wilkinson, J.M., Antibacterial activity of berry fruits used for
culinary purposes. J. Med. Food, 6, 57–61, 2003.
30. Lee, Y., Cesario, T., Wang, Y., Shanbrom, E., and Thrupp, L., Antibacterial activity of vegetables and
juices. Nutrition, 19, 994–996, 2003.
31. Bestwick, C. and Milne, L., Effects of beta-carotene on antioxidant enzyme activity, intracellular reac-
tive oxygen and membrane integrity within post confluent Caco-2 intestinal cells. Biochim. Biophys.
Acta, 1474, 47–55, 2000.
32. Yokomizo, A. and Moriwaki, M., Effects of uptake of flavonoids on oxidative stress induced by hydro-
gen peroxide in human intestinal Caco-2 cells. Biosci. Biotechnol. Biochem., 70, 1317–1324, 2006.
33. Bermudez-Soto, M.J., Larrosa, M., Garcia-Cantalejo, J.M., Espin, J.C., Tomas-Barberan, F.A., and
Garcia-Conesa, M.T., Up-regulation of tumor suppressor carcinoembryonic antigen-related cell adhe-
sion molecule 1 in human colon cancer caco-2 cells following repetitive exposure to dietary levels of a
polyphenol-rich chokeberry juice. J. Nutr. Biochem., 18, 259–271, 2007.
34. Bub, A., Watzl, B., Blockhaus, M., Briviba, K., Liegibel, U., and Muller, H., Fruit juice consumption
modulates antioxidative status, immune status and DNA damage. J. Nutr. Biochem., 14, 90–98, 2003.
35. Cilla, A., Laparra, J.M., Alegria, A., Barbera, R., and Farre, R., Antioxidant effect derived from bioac-
cessible fractions of fruit beverages against H2O2-induced oxidative stress in Caco-2 cells. Food Chem.,
106, 1180–1187, 2008.
36. Cilla, A., González-Sarrías, A., Tomás-Barberán, F.A., Espín, J.C., and Barberá, R., Availability of
polyphenols in fruit beverages subjected to in vitro gastrointestinal digestion and their effects on pro-
liferation, cell-cycle and apoptosis in human colon cancer Caco-2 cells. Food Chem., 114, 813–820,
2009.
37. Evrendilek, G.A., Altuntas, J., Sangun, M.K., and Zhang, H.Q., Apricot nectar processing by pulsed
electric fields. Int. J. Food Prop., 16, 216–227, 2011.
38. Oey, I., Lille, M., Van Loey, A., and Hendrickx, M., Effect of high-pressure processing on colour,
texture and flavour of fruit- and vegetable-based food products: A review. Trends Food Sci. Technol.,
19, 320–328, 2008.
39. Bayındırlı, A., Alpas, H., Bozoğlu, F., and Hızal, M., Efficiency of high pressure treatment on inactiva-
tion of pathogenic microorganisms and enzymes in apple, orange, apricot and sour cherry juices. Food
Control., 17, 52–58, 2006.
40. Patrignani, F., Vannini, L., Kamdem, S.L.S., Lanciotti, R., and Guerzoni, M.E., Effect of high pressure
homogenization on Saccharomyces cerevisiae inactivation and physico-chemical features in apricot and
carrot juices. Int. J. Food Microbiol., 136, 26–31, 2009.
41. Patent, Preparation of Unsuspended Property Japanese Apricot-Fruit Juice, Involves Filtering Japanese
Apricot-Fruit Juice with Ultrafiltration Film/Membrane. FUJI SHOKKEN KK (FUJI-Non-standard),
Patent no. JP2003334017, 2002.
42. Cassano, A., Conidi, C., Timpone, R., D’Avella, M., and Drioli, E., A membrane-based process for the
clarification and the concentration of the cactus pear juice. J. Food Eng., 80, 914–921, 2007.
43. Aider, M. and de Halleux, D., Production of concentrated cherry and apricot juices by cryoconcentration
technology. LWT—Food Sci. Technol., 41, 1768–1775, 2008.
44. Leong, S. and Oey, I., Effects of processing on anthocyanins, carotenoids and vitamin C in summer
fruits and vegetables. Food Chem., 133, 1577–1587, 2012.
45. Kirca, A., Ozkan, M., and Cemeroglu, B., Stability of black carrot anthocyanins in various fruit juices
and nectars. Food Chem., 97, 598–605, 2006.
46. Pernice, R., Borriello, G., Ferracane, R., Borrelli, R., Cennamo, F., and Ritieni, A., Bergamot: A source
of natural antioxidants for functionalized fruit juices. Food Chem., 112, 545–550, 2009.
47. Zulueta, A., Esteve, M., Frasquet, I., and Frigola, A., Vitamin C, vitamin A, phenolic compounds and
total antioxidant capacity of new fruit juice and skim milk mixture beverages marketed in Spain. Food
Chem., 103, 1365–1374, 2007.
48. Brenna, O., Pompei, C., Pravettoni, V., Farioli, L., and Pastorello, E., Production of hypoallergenic foods
from apricots. J. Food Sci., 70, 38–41, 2005.
49. Sanal, I., Guvenc, A., Salgin, U., Mehmetoglu, U., and Calimli, A., Recycling of apricot pomace by
supercritical CO2 extraction. J. Sup. Fluids, 32, 221–230, 2004.
50. Gupta, A., Sharma, P.C., Tilakratne, B.M.K.S., and Verma Anil, K., Studies on physico-chemical
charactersitics and fatty acid composition of wild apricot (Prunus armenaica Linn.) kernel oil. Indian
J. Nat. Prod. Resour, 3, 366–370, 2012.
51. Yiğit, D., Yigit, N., and Mavi, A., Antioxidant and antimicrobial activities of bitter and sweet apricot
(Prunus armeniaca L.) kernels. Braz. J. Med. Biol. Res., 42, 346–352, 2009.
52. Wu, X., Beecher, G., Holden, J., Haytowitz, D., Gebhardt, S., and Prior, R., Lipophilic and hydrophilic
antioxidant capacities of common foods in the United States. J. Agric. Food Chem., 52, 4026–4037,
2004.
10
Aronia Juice
Maria Glibetić and Aleksandra Konić-Ristić
CONTENTS
10.1 Introduction....................................................................................................................................119
10.3 Bioactives and Antioxidant Efficacy..............................................................................................121
10.4 Health Effects................................................................................................................................ 124
10.4.1 In Vitro Studies................................................................................................................. 125
10.4.2 Animal Studies................................................................................................................. 125
10.4.3 Human Intervention Studies............................................................................................. 126
10.5 Novel Products/Formulations and Future Trends......................................................................... 128
10.6 Conclusion..................................................................................................................................... 130
References............................................................................................................................................... 130
10.1 Introduction
Black aronia (Aronia melanocarpa L.), also known as black chokeberry, chokeberry, or just aronia, is a
shrub with purple-black pomes that originates from the eastern parts of North America. At the beginning
of twentieth century, it was transferred to the southeast and central European countries where it became
very popular. It was cultivated as a crop and its fruits were used in the production of juices, jams, and
wines, as well as natural food colorants. Today, this fruit is widely grown all over the world and used as
raw material for foods, dietary supplements, or herbal remedies [1]. Black aronia is the most extensively
investigated among the species of the Aronia genus (Rosaceae family, Maloideae subfamily), and its ber-
ries are almost exclusively used in the production of aronia juices. As a rich source of polyphenols and
based on their profile and antioxidant activity, the berries of black aronia are often considered superior to
other berries [2], including the berries of two other species of the Aronia genus, red chokeberry (Aronia
arbutifolia L.) and purple chokeberry (Aronia prunifolia L.) [3].
Accordingly, juice from the aronia berry, due to the high polyphenol content, atypical composition
of individual polyphenols, specific composition of nutrients, and exceptional antioxidant potential, has
attracted the attention of both consumers and field experts and initiated numerous research activities
aimed at elucidating its health benefits.
A large body of scientific evidence supports the opinion that polyphenol-rich aronia juice can be
considered a functional food that “beneficially affects one or more target functions in the body, beyond
adequate nutritional effects in a way that is relevant to either an improved state of health and well-being
and/or reduction of risk of disease” [4]. However, many questions still remain unanswered, while novel
targets for the beneficial effects of this unique juice are continuously being discovered. In parallel to the
evaluation of its health properties, scientific efforts have been made toward the optimization of its com-
position, stability of its compounds, sensory characteristics, factors influencing its biological activity,
and potential synergy with other healthy foods.
This chapter highlights the most relevant facts and scientific results on aronia juice’s nutritional com-
position and its biological effects. It also aims to support further research to elucidate if the juice obtained
from this small berry fruit can provide health benefits to consumers worldwide.
119

Pure aronia juice is generally considered as a low-energy beverage, rich in vitamins and minerals. The
nutrient composition of aronia juice is given in Table 10.1. It comprises literature data on fresh and
pasteurized aronia juice compositions [2] and original data on pasteurized juices produced from berries
grown in mountain regions of western Serbia, compiled into the Serbian Food Composition Database [5].
Aronia juice is a rich source of various nutrients including sugars, vitamins, and minerals. Sugars are
the major nutrients in aronia juice, including glucose, fructose, and sorbitol as routinely analyzed com-
pounds. The content of both glucose and fructose content is approximately 40 g/L, while sorbitol content
in aronia juice is found to be 50–80 g/L [2,5]. Sorbitol has a refreshing effect in the mouth and has a
laxative and cariostatic effect, and hence may be considered as a multipurpose food additive [6]. Aronia
juice is one of the richest sources of sorbitol among fruit juices. Thus, sorbitol content is routinely used
as a marker of authenticity of aronia juice or adulteration of juices from other berries with aronia juice.
With regard to the mineral content, aronia juice is a rich source of potassium, compared to other berry
juices, with the average content of 2.85 g/L that might contribute to the observed vasodilatory effects of
aronia juice. Freshly pressed aronia juice contains a high level of vitamin C (200 mg/L) [2], but a large
TABLE 10.1
Compositional and Nutritional Characteristics of Aronia Juice (per L)
Nutrient Unit Fresh Aronia Juice [2] Pasteurized Aronia Juice [2,5]
Water g 80.9 84.5–86.7
Energy kcal 545 469–540
Protein g na na
Lipid (fat) g na na
Total dietary fiber g trace na
Fructose g 38 37–40
Glucose g 41 40–51
Sorbitol g 80 50–55.6
Minerals
Sodium mg 5 4.7–5.7
Potassium mg 2850 1969–2200
Calcium mg 150 152–185
Magnesium mg 140 139–160
Iron mg 2–8 0.4–4
Zinc mg 1.3 0.58–0.6
Iodine µg na <5
Vitamins
Vitamin C mg 200 29.2
Thiamin µg 500 381
Riboflavin µg 600 708
Niacin µg 3400 2500
Pantothenic acid µg 2200 1500
Pyridoxine µg 550 442
Folate (DFE) µg na 35
Organic Acids
Malic acid mg 9.0 11.1
Citric acid mg na na
Succinic acid mg 1.5 0.16
Abbreviations: na, not analyzed; DFE, dietary folate equivalents.
Aronia Juice 121
amount is degraded during processing and storage (Table 10.1). The supplementation of pasteurized aronia
juice with vitamin C to the levels found in fresh juice is a promising approach in the improvement of nutri-
tional properties of aronia juice. Other vitamins present belong mostly to the vitamin B group, including
thiamin, riboflavin, niacin, pantothenic acid, and pyridoxine. However, the intake of vitamins by a single
portion of the juice (100–200 mL) does not reach 10% of Recommended Dietary Allowances (RDA).
In general, aronia juice lacks most of the dietary fibers present in aronia berries (3.4–5.8 g/kg) [2].
Pectins, present in fresh pressed juice (3.7 g/kg) [2] are destroyed by the action of pectinase during aronia
juice processing [7].
Organic acid, present in substantial quantities, contribute to the characteristic flavor of aronia juice,
mostly defined as astringent and almond-like, due to the high content of procyanidins and the presence
of amygdalin. The genus name, Aronia, has now replaced the former common name of the plant, choke-
berry, which indicated the astringency of both berries and the juice.
An important approach in the functional drinks industry includes the addition of functional ingredients
and fortification with micronutrients. Data on the exact nutrient composition that vary due to environ-
mental conditions, varietal changes, and processing, are crucial in designing healthy and safe products,
taking into account the general intake of particular micronutrients. Based on literature data, aronia juice
is a good source of certain minerals and vitamins and has a relatively low energy content. Accordingly,
as a regular component of a healthy and balanced diet, aronia juice intake can contribute to the daily
intake of macro- and micronutrients. However, open-access food composition databases often lack data
on nutrient composition of aronia juice and aronia berries [8–10]. Analytical and published data on the
nutritional composition of both aronia berry and aronia juice, locally produced and imported, fresh, and
pasteurized, have been included in the Serbian Food Composition Database [5], harmonized with the
European Food Information Resource (EuroFIR) criteria [11,12].

Polyphenols are the most abundant bioactive compounds present in aronia juice (Table 10.2 and
Figure 10.1) and the major contributor to its antioxidant activity [13]. Based on available data in the litera-
ture [2,14–17], the content of total phenolics and total anthocyanins of aronia juice varies within the range
of 3.9–9.1 g gallic acid equivalents (GAE)/L and 0.15–3.04 g cyanidin-3-glucoside equivalents (C3GE)/L,
respectively. Suggested determinants of variation include the origin of aronia berries (e.g., environmen-
tal conditions), processing method, maturation of the juice, and differences in analytical methodology.
Taheri et al. [18] have shown that polyphenol content of aronia juice depends on the harvesting time, and
it was 20% higher at week 1 than week 7 of ripening. However, maximum juice anthocyanins occurred at
week 5. Processing and storage have a major influence on the content of bioactive compounds, inducing
mainly the loss of monomeric anthocyanidins and a raise in the degree of polymerization [7].
Based on available data, the major subclasses of polyphenolic compounds found are polymeric pro-
anthocyanidins, representing up to 40% of total phenolics, and contribute mostly to the astringent taste
of the juice [19–22]. Proanthocyanidin profile of the juice is complex, characterized by the presence of
monomers to decamers and exceptionally high levels of polymers [20]. Polymeric flavan-3-ols are com-
posed predominantly of (−)-epicatechin with a degree of polymerization of up to 40 [20]. Anthocyanins
come in second at approximately 25% of the total phenol composition. Both the aronia berry and the
juice are exceptionally rich in anthocyanins and considered to be one of the best dietary sources of
anthocyanin. Aronia juice anthocyanins consist mainly of cyanidin glycosides (glucoside, xyloside,
galactoside, and arabinoside), but the galactoside and arabinoside generally account for 60%–75% of
the total anthocyanins [19,21–24]. The juice also contains high levels of hydroxycinnamic acids (chlo-
rogenic acid and neochlorogenic acid) and considerable levels of flavonols, mostly quercetin derivatives
(quercetin 3-rutinoside, quercetin 3-galactoside, and quercetin 3-glucoside) (Table 10.2 and Figure 10.1).
Aronia polyphenols are susceptible to the treatments applied during juice processing and the envi-
ronmental factors they are exposed to, during the storage, resulting in substantial decline of bioactives.
Wilkes et al. [25] investigated the stability of anthocyanins, total proanthocyanidins, hydroxycinnamic
acids, and flavonols at different stages of aronia juice processing and over 6 months of storage at 25°C.
TABLE 10.2
Phytochemicals in Aronia Juice
Phytochemicals Unit Content References
Phenolics
Total phenolics g GAE/L 3.9–9.1 [2,5,14–17]
Total flavonoids g CE/L 1.9 [15]
Anthocyanins
Total anthocyanins g C3GE/L 0.15–3.04 [14–17]
Cyanidin 3-galactoside mg/L 125–210 [22,23]
Cyanidin 3-glucoside mg/L 5.12 [22]
Cyanidin 3-arabinoside mg/L 0.71–1.00 [22,23]
Cyanidin 3-xyloside mg/L 0.59 [22]
Cyanidin glycosides mg/L 357 [24]
(−)-Epicatechin mg/L 1.48 [22]
Polymeric procyanidins mg/L 280–293 [22,23]
Flavonols
Quercetin glycosides mg/L 7.85 [20]
Quercetin 3-rutinoside mg/L 1.68 [22]
Quercetin 3-galactoside mg/L 2.68 [22]
Quercetin 3-glucoside mg/L 2.25 [22]
Quercetin 3-O-β-arabinosyl-β-glucoside mg/L 1.15 [22]
Quercetin 3-O- R-rhamnosyl-β-galactoside mg/L 1.17 [22]
Phenolic Acids
Chlorogenic acid mg/L 45.50–80 [22,23]
Neochlorogenic acid mg/L 49.21–120 [22,23]
Total hydroxycinnamic acids mg/L 202 [24]
Carotenoids µg/L 70 [2]
Amigdalin mg/kg 57.5 [2]
Abbreviations: GAE, gallic acid equivalents; CE, catechin equivalents; C3GE, cyanidin-3-glucoside equivalents.
Processing stages, including blanching (95°C, 3 min), treatment with pectinase (45°C, 1 h), mash pressing,
and pasteurization (90°C, 10 min), had different effects on different types of polyphenols. Anthocyanins
have been shown to be highly sensitive to thermal treatments, as bleaching reduced the content of indi-
vidual anthocyanins up to 63% compared to the content in frozen fruits, with further decrease observed
during the pasteurization, resulting in their retention of 30%–52% in pasteurized compared to unpasteur-
ized juice. Thermal treatment had no effect on hydroxycinnamic acids, total proanthocyanidins, and
flavonols, while enzyme treatment even induced an increase in their content without effects on antho-
cyanins. Pressing caused losses in the content of polyphenolic constituents mostly due to the removal
of seeds and skins, shown by substantial levels of individual polyphenols in the press cake. Finally,
anthocyanin, proanthocyanidin, hydroxycinnamic acid, and flavonol contents in fresh, pasteurized aronia
juice were 7%, 55%, 63%, and 52%, respectively, compared to the contents in frozen berries [25]. Storage
over 6 months at 25°C caused further linear decrease in anthocyanin content (up to 83% of the content in
fresh pasteurized juice) and to a much lesser extent, in the content of other polyphenolics. Anthocyanins
remaining in aronia juice were in large part in the polymeric form. The main degradation products of
anthocyanins detected in aronia juice after thermal treatment were protocatechuic acid and phlorogluci-
naldehyde, although in concentrations equivalent to only 3% of the starting anthocyanin level [25].
Nonnutritive bioactive compounds other than polyphenols present in aronia juice are carotenoids and
amygdalin with the content of 70 µg/L and 57.5 mg/kg, respectively (Table 10.2), responsible for the
almond aroma of fresh juice [2].
Polyphenols are considered as major dietary antioxidants. Based on exceptional polyphenolic content,
compared to other polyphenol-rich foods and other berry juices, aronia juice attracted the attention as
Aronia Juice 123
OH OH
OH OH
HO O+ HO O +
O O
OH OH OH OH
O O
HO HO
OH OH
OH OH
Cyanidin 3-O-β-D-galactopyranoside Cyanidin 3-O-β-D-glucopyranoside
OH OH
HO O+ HO O+
OH
O O
OH OH OH OH
O O
OH OH
OH OH
Cyanidin 3-O-α-D-arabinopyranoside Cyanidin 3-O-β-D-xylopyranoside
OH OH
HO HO
O O
HO OH HO OH
O O
OH OH
O O
HO HO
Chlorogenic acid Neochlorogenic acid
OH
OH OH
HO O HO O
OH
O O
OH O O OH OH OO OH
HO HO
OH OH
OH OH
Quercetin 3-O-β-D-galactopyranoside Quercetin 3-O-β-D-glucopyranoside
OH
HO O
OH
O
OH O O OH
H3C O O
OH
HO OH OH
OH
Quercetin 3-O-rutinoside
FIGURE 10.1 Chemical structures of major polyphenols present in aronia juice.

TABLE 10.3
Antioxidant Activity of Aronia Juice
Method Unit Content References
DPPH mg TE/mL 18.1 [17]
FRAP mmol Fe2+/100 g 9.8 [13]
ABTS mmol TE/100 g 9.7 [13]
ORAC mmol TE/100 g 11.5 [13]
Abbreviations: DPPH, 2,2-diphenyl-1-picryl-hydrazyl-hydrate free radical method; FRAP, ferric ion reduc-
ing antioxidant power; ABTS, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) free
radical method; ORAC, oxygen radical absorbance capacity; TE, trolox equivalents.
a powerful dietary antioxidant source, which is confirmed from published data on antioxidant activ-
ity of aronia juice in chemically based assays and cellular models. Reported antioxidant activity of
aronia juice, as measured by scavenging 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis
(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radicals, or assessed as ferric ion reducing antioxi-
dant power (FRAP), and oxygen radical absorbance capacity (ORAC), is given in Table 10.3. By using
electron spin resonance spectroscopy, Valcheva-Kuzmanova et al. [26] also demonstrated antiradical
activity of aronia juice toward galvinoxyl free radicals. However, Müller et al. [13] compared the
antioxidant activity of reconstituted aronia juice concentrate, rich in vitamin C (200 mg/100 g) and
total polyphenols (766 mg/100 g), with other juices and purees with different contents of vitamin C and
polyphenols, including their ratio. The results so obtained indicated that both vitamin C and polyphenols
contributed to the antioxidant activity of aronia juice. Contribution of the vitamin C content to the
antioxidant activity determined by FRAP and ABTS assays was higher compared to the contribution to
the activity obtained by ORAC assay. The effects of polyphenols were opposite as they mainly contribute
to the antioxidant activity assessed by the ORAC assay [13].
Although antioxidant activity of the juice, assessed by chemical assays, do not guarantee its effects
in vivo, it represents an important characteristics for the functionality of aronia juice bioactives within a
food matrix or as a part of a meal, with potential to enhance the stability of other compounds and prevent
the load of oxidized products and their deleterious effects on human organism.
In addition to the use of chemically based assays, antioxidant activity of aronia juice was assessed in
cellular models as well. Slatnar et al. [24] investigated the cellular antioxidant effects of different berry
juices including aronia juice, by using Saccharomyces cerevisiae as in vitro cellular model. They reported
that results obtained in the yeast cells were markedly different from the data obtained by DPPH assay and
concluded that a major factor found to influence in vivo antioxidant activity was cellular availability of
polyphenols present in juices, and also the ratio of individual polyphenols consumed by the cell, with the
favorable effects of high anthocyanin content and low content of hydroxycinnamic acids. They also found
that when polyphenols’ uptake by the yeast cells was low, antioxidant activity increased. High hydroxycin-
namic acid uptake with low anthocyanin intake induced higher intracellular oxidation. Aronia juice was
characterized by relatively high uptake of anthocyanins (84%) and the lowest uptake of hydroxycinnamates
[24]. Along with the effects of anthocyanin-rich fractions of aronia berries, Konić-Ristić et al. [14] investi-
gated the cellular antioxidant activity of protocatechuic acid that was found to be one of the major metabo-
lite of cyanidin glucosides in humans [27]. The decrease in intracellular reactive oxygen species (ROS)
obtained indicated antioxidant activity of protocatechuic acid at low doses. The presence of an inverse dose
response correlation, with the lowest dose inducing the highest reduction in intracellular ROS, suggests
that antioxidant activity of protocatechuic acid is not mediated by its direct antioxidant action.
10.4 Health Effects

Beneficial health effects of aronia juice, shown in experimental models and clinical trials, indicate that
mechanisms of action of its bioactive constituents and/or metabolites are not limited to their antioxidant
activity. Interactions of aronia juice bioactive constituents with molecules, cells and tissues of animals,
Aronia Juice 125
and humans include both kinetic and dynamic interactions. Most of the studies performed up to date are
targeted to the effects on biomarkers and risk factors for cardiovascular disease (CVD) and cancer, and
interactions with the relevant cells and molecules, although the scientific evidence on its effects in the
prevention of other types of the diseases are emerging.
10.4.1 In Vitro Studies

Among numerous studies investigating effects of aronia juice and aronia berry extracts in different
cellular models in vitro, the most relevant to the putative effects in humans are studies performed in cul-
tures of endothelial cells of the gastrointestinal tract (GIT) that are exposed to the bioactive components
of the juice after the consumption. In the study performed in CaCo-2 cells, as human model of colon
cancer, recurrent treatment with nontoxic doses of predigested aronia juice (80 µM of phenolics; 2 h/day
for 4 days) induced inhibition of both cell proliferation (30%–40%) and viability (20%) in comparison
to untreated cells [28]. Observed effects were accompanied by a modulation in transcription of several
genes including upregulation of tumor suppressor genes and downregulation of genes involved in tumor
invasion and metastasis [28].
The treatment of other type of cells with aronia juice, however, indicated potential of bioactives pres-
ent in the juice in interacting with numerous cellular and molecular targets. It was recently shown that
aronia juice exhibits strong anticancer activity through a redox-sensitive mechanism. Consecutive expo-
sure of p53-deficient Jurkat cells to aronia juice, with the total polyphenols content of 7.15 g GAE/L,
induced inhibition of cell proliferation, associated with cell cycle arrest in G(2)/M phase and induction
of apoptosis [29]. Mechanisms of antitumor action included upregulation of the expression of tumor
suppressor p73 and active caspase 3, downregulation of the expression of cyclin B1 and the epigenetic
integrator UHRF1, increase in ROS formation, decrease in mitochondrial membrane potential, and the
release of cytochrome c into the cytoplasm. Pretreatment with intracellular ROS scavengers prevented
the aronia juice-induced apoptosis and upregulation of the expression of p73 and active caspase 3. The
fractionation of the juice and the use of identified isolated compounds indicated that the anticancer
activity was associated predominantly with chlorogenic acids, some cyanidin glycosides, and derivatives
of quercetin. Aronia juice treatment also induced apoptosis of different human lymphoblastic leukemia
cells (HSB-2, Molt-4, and CCRF-CEM). In addition, aronia juice exerted a strong proapoptotic effect
in human primary lymphoblastic leukemia cells, but not in human normal primary T-lymphocytes [29].
Further studies are needed to investigate if aronia juice, the bioactives present or their metabolites, exerts
some of the activities shown previously for other types of polyphenols [30–33].
The bioactives of aronia juice have also been shown to exert inhibitory effects on the activity of
cytochrome P450 3A4 activity, an important enzyme of xenobiotic metabolism in humans. The most
effective was procyanidin B5, while the inhibitory effect of anthocyanins depended on sugar moiety,
with the stronger inhibitory effects of cyanidin 3-arabinoside compared to cyanidin 3-galactoside and
cyanidin 3-glucoside [34]. This indicates putative synergistic effects with therapeutics and favors further
research in the area of drug–food compound interactions, determining interindividual variations.
10.4.2 Animal Studies

Antioxidant and anti-inflammatory effects of aronia juice was also confirmed in animal models.
Components of aronia juice also exerted anti-inflammatory effects, as shown in animal model of paw
edema induced by histamin and serotonin. The observed effects were more pronounced than the effects
of rutin used as a positive control [35]. In other experiments performed on rats, aronia juice consumption
prevented indomethacin-induced damage of gastric mucosa [36], beneficially affected disturbed levels
of glucose and lipids in streptozocin-induced diabetes, but without any effects on these parameters in
normal rats [37], and prevented harmful effects of carbon tetrachloride on the activities of liver enzymes,
lipid peroxidation, and glutathione depletion in the liver of rats [38]. In rats fed with high c holesterol
diet, aronia juice supplementation prevented elevation of plasma cholesterol, low-density lipoprotein
(LDL)-cholesterol, and triacylglycerols (TAG), but no significant effects were observed in animals fed
a standard diet [15]. The observed effect of aronia juice consumption on deleterious effects of foods,
and other stress factors, rather than on fasting, normal or even disturbed, metabolic and p hysiological
biomarkers and functions, were similar to the action of other polyphenols [39].
Detoxifying effects of aronia juice, mediated by its inhibition of phase I and stimulation phase II
enzymes, were shown in an animal study by Krajka-Kuzniak et al. [22]. However, simultaneous admin-
istration of procarcinogen and forced feeding with aronia juice augmented DNA damage and aggravated
the effects of procarcinogens [22].
A current trend in the area of functional beverages is the development of new products guided by their
bioactivity in addition to the composition and sensory characteristics. An example is work performed
by Auger et al. [40] who investigated the effects of five different juice blends on endothelium-dependent
relaxations in porcine arteries. The formulation of juice blends are performed based on the effects of 13
different fruit juices and purees. Aronia juice has been found to be one of the tested products with the
most pronounced effects, and in blends with other less active fruit juices it significantly contributes to
their in vitro vasodilatory effects [40].
10.4.3 Human Intervention Studies

Similar to the studies performed in vitro, a majority of studies that investigated the effects of aronia con-
sumption on human health in vivo were performed by using extracts of aronia berries. However, several
studies specifically investigated the effects of aronia juice consumption, showing numerous effects on
antioxidant/prooxidant status, biomarkers and risk factors for CVD, prevention of urinary tract infec-
tions, or the beneficial effects on the skin.
Regarding the effect of aronia juice as in vivo antioxidant, it is a general opinion that the effects
of polyphenol-rich foods or native polyphenols shown in chemically based assays and cellular mod-
els do not guarantee their potential in vivo. The major factor precluding extrapolation between in vitro
and in vivo systems is extensive metabolism of polyphenols, especially anthocyanins, and consecutively
“poor” bioavailability in the native form. Bioactive compounds of aronia juice are absorbed in dif-
ferent parts of the GIT and subsequently undergo excessive transformations in different tissues of the
human body providing different anthocyanin metabolites but also, simple phenolics, aromatic acids,
and other products, with many of them are still unidentified, mainly as a result of a transformation by
colon microflora [41]. In the bioavailability study performed in healthy subjects, Wiczkowski et al. [42]
determined native anthocyanins and corresponding metabolites in plasma and urine after the inges-
tion of a physiological dose of aronia juice (250 mL/60 kg), providing 0.8 mg of anthocyanins/kg body
weight (0.67 mmoL cyanidin/L of juice). Three native anthocyanins (cyanidin-3-galactoside, cyanidin-
3-arabinoside, and cyanidin-3-glucoside) and four anthocyanin metabolites (cyanidin monoglucuro-
nide, peonidin-3-galactoside, peonidin monoglucuronide, and peonidin-3-arabinoside) were detected in
human plasma and urine, as a result of both methylation and glucuronidation. Metabolites appeared in
plasma after 30 min with a maximum concentration of 32.7 nmol/L at 1.3 h. A cumulative excretion of
anthocyanins in urine was 0.25% of the total amount of anthocyanins ingested [42]. Small catabolic by-
products, produced in the small and large intestine, were not under the scope of the study.
Concentrations of anthocyanins’ metabolites in circulation are very low, compared with the constitu-
tive antioxidants of human body [43]. Thus, it is postulated that the effects of aronia juice consumption
on antioxidant status, observed in human intervention studies, in vivo, are mainly the result of their
indirect effect, mediated through various signaling pathways and the effects on enzymes, cytokines,
and receptors. It should be noted that berry phytochemicals may also influence cellular processes that
are completely independent of antioxidant mechanisms [44]. Although direct correlation of disturbed
antioxidant/prooxidant status and the risk of chronic diseases has not been established, several studies
demonstrated the beneficial effects of aronia juice consumption on various parameters as markers of the
redox status. In healthy women, long-term consumption of aronia juice as a part of regular diet induced
beneficial changes in several markers of antioxidant/prooxidant status, including the decrease in bio-
markers of lipid peroxidation and prooxidant–antioxidant balance and the increase in paroxonase-1
activity [45]. Anthropometric and biochemical parameters, blood pressure values, and other markers of
oxidative status in this population were not significantly influenced by the intervention [45]. Pilaczynska-
Szczesniak et al. [46] investigated the effects of a 4-week long consumption of aronia juice (150 mL/day)
Aronia Juice 127
in rowers, by assessing thiobarbituric acid reactive substances (TBARS) levels, glutathione-peroxidase

(GPx), and superoxide-dismutase (SOD) activity in erythrocytes and creatine kinase (CK) in plasma
at rest, immediately after exercise, and 24 h following the exercise. Results have shown significant
decrease in TBARS levels after an exercise test (baseline and 24 h) and increase in GPx activity and
CK levels only immediately after the exercise. SOD was not influenced by the aronia juice consumption
[46]. Antioxidant enzyme activity in erythrocytes influenced by long-term aronia juice consumption
(100 mL/day for 12 weeks) was investigated in healthy population by Kardum et al. [16], in addition
to the effects on membrane lipid status. Compared to the baseline, aronia juice intervention induced
significant increase in polyunsaturated fatty acids (PUFA), total PUFA, and unsaturation index as well
as significant decrease in monounsaturated fatty acids (MUFA) and n-6/n-3 ratio. Activities of both
enzymes assessed (SOD and GPX) were significantly higher at the end of the study, while TBARS level
was significantly reduced [16].
CVD and cancer, being the major burden on health worldwide, are in the epicenter of nutritional
research. Among the first studies that initiated further investigation of berry consumption benefits for
cardiovascular health, was a study by Knekt et al. [47] showing that high intake of berries, alongside
other factors, was associated with a 60% decline in coronary heart disease (CHD) and stroke. Taking
epidemiological studies as a starting point, and based on numerous in vitro data, beneficial effects of
berries on cardiovascular health are investigated in numerous human intervention studies using “tradi-
tional” risk factors (e.g., dyslipidemia, hyperglycemia, obesity, and blood pressure) and other biomarkers
(e.g., platelet function, chronic inflammation, endothelial function, and oxidative stress) as “nontradi-
tional” factors of risk.
Several studies have investigated the effects of aronia juice consumption on lipid status. Skoczyñska
et al. [48] demonstrated that the consumption of 250 mL of aronia juice during the course of 6 weeks
beneficially affected lipid status of men with mild hypercholesterolemia by significant reduction in total
cholesterol, LDL cholesterol, and TAG and increased the high-density lipoprotein (HDL) cholesterol.
Significant decreases in the serum glucose, homocysteine, and fibrinogen concentrations were also
observed [48]. In an uncontrolled study, Simeonov et al. [49] showed that long-term consumption of
200 mL of pure aronia juice induced a significant decrease in total cholesterol in patients with non-
insulin-dependent diabetes mellitus.
The potential of aronia juice and the present bioactives on blood pressure was evaluated in several
human dietary intervention studies. Five studies (three with an aronia extract and two with aronia juice)
reported a marked decrease in systolic and diastolic blood pressure, including a study in patients with myo-
cardial infarction with regular consumption of aronia extract (255 mg/day) during 6 weeks [50]; in subjects
with mild hypercholesterolemia who consumed aronia juice (250 mL/day) during 6 weeks [48]; and in
patients with non-insulin-dependent diabetes mellitus during a 12-week long consumption of aronia juice
(200 mL/day) [49]. Aronia extract consumption in combination with statins significantly decreased the
serum levels of interleukine-6 (IL-6), C-reactive protein (CRP), intercellular adhesion molecule (ICAM),
vascular cell adhesion molecule (VCAM), monocyte chemotactic protein-1 (MCP-1), and adiponectin,
compared to the control statin-only-treated group [50]. In subjects with mild hypercholesterolemia, con-
sumption of 250 mL of aronia juice during 6 weeks improved serum nitric oxide (NO) level, flow-mediated
dilatation (FMD), and brachial artery diameter (BAD) [51]. In addition, consumption of different berries
or berry juice blends that included aronia juice during 8 weeks was also effective in systolic blood pressure
reduction compared to the control in patients with cardiovascular risk factors [52].
Consumption of aronia juice was also shown to significantly reduce fasting glucose levels in subjects
with risk factors compared to the control group [48] and fasting glucose levels and the levels of oxidized
hemoglobin (HbA1C) in patients with non-insulin-dependent diabetes mellitus, compared to the baseline
values [49].
One of the novel targets of bioactive compounds in the scope of their beneficial effects on cardiovascu-
lar health are platelets, based on their role in the processes of thrombosis and atherosclerosis, mediated
through their interactions with other platelets, monocytes, and neutrophils. Disturbed platelet function
correlates with other CVD risk factors and the polyphenol’s effects on platelets is generally accepted as
beneficial for cardiovascular health as defined by the European Food Safety Association (EFSA) [53].
A preliminary study in healthy subjects showed that the acute intake of aronia juice (150 mL) induced
significant decrease in the expression of P-selectin, as a marker of platelet activation, as well as a decrease
in platelet–monocyte and platelet–neutrophil aggregation, both in vivo and ex vivo, after the action of
platelet agonists (adenosine-diphosphate and arachidonic acid) [54]. The observed results support further
studies, aiming to elucidate both acute and chronic effects of aronia juice as antiplatelet agent.
The enhancement of aronia juice effects on health by its combination with other bioactive compounds
represents a promising strategy in the functional foods industry. Aiming to respond to the consumers
demand for enhanced functional products with beneficial effects on various biomarkers and risk fac-
tors (serum glucose, lipids, body weight, and antioxitative/prooxidative status), our team designed a
novel aronia juice–based functional beverage by combining pure aronia juice and commercial Luralean®
fibers. This product was a result of collaboration with the producer of aronia juice from Serbia (Nutrika
d.o.o., Irig, Serbia) as a joint effort of researchers from our center and specialists from the industry.
Luralean fibers (Shimitzu, Japan) are stable, enzyme-free glucomannans from Amorphilus konjac, for-
mulated to be used as a supplement to liquid products. It has been shown that glucomannan fibers influ-
ence appetite and satiety by increasing the volume of gastrointestinal content, without adverse effects
due to their extremely high viscosity. As a barrier between fat and sugars and the endothelium of the
GIT, they impair their digestion and absorption and regulate postprandial insulin levels. All these are
suggested mechanisms of their beneficial effects in obesity, dyslipidemia, and hyperglycemia, although
clinical trials do not provide sufficient evidence of the proposed effect [55].
The synergistic effects of aronia juice and glucomannan were evaluated within a clinical trial in 30
obese postmenopausal women at high risk of CVD. Dietary intervention included daily consumption of
100 mL of aronia juice supplemented with 2 g of Luralean during 4 weeks, as part of a regular dietary
regiment monitored by a food frequency questionnaire. Consumption of Luralean-supplemented aronia
juice induced significant reduction in total body weight, fat free mass, waist and hip circumference, and
blood pressure, accompanied by significant enhancement in the activity of antioxidant enzymes and
subjective improvement in well-being [56].
Recent results providing evidence of the deleterious postprandial effects of high-sugar and high-fat
meals, especially in subjects with the disturbed metabolism of fat and sugars [57], support the observed
in vivo effects of various polyphenols on postprandial state [39]. Novel research on diets rich in bioac-
tives induced favorable changes in fatty acid profiles in healthy, as well as in many, diseased patients
[58]. The effects of a diet rich in plant and fruit bioactives on the metabolism of sugars and dietary fats,
rationalize future work toward the formulation of meals supplemented with bioactives from aronia juice
as a functional ingredient.
In vitro data and the results of dietary interventions in animal models suggested the potential of aronia
juice as a functional food demonstrating ability “to affect beneficially one or more target functions in the
body, beyond adequate nutritional effects in a way that is relevant to either an improved state of health
and well-being and/or reduction of risk of disease” [4]. However, according to the European legislative,
the substantiation of claims indicating beneficial health effects can be exclusively based on the results
from human trials, in addition to the in vitro and data from animal models. The number of human stud-
ies aiming to evaluate the effects of aronia juice (Table 10.4) is far less than the number of studies that
involved aronia extracts, and most of them are not designed as randomized, controlled studies, consid-
ered as the “gold” standard of clinical trials. However, their results, mostly showing the effects of aronia
juice consumption on different risk factors and biomarkers of cardiovascular risk in humans, are promis-
ing and rationalize further research in this area toward final confirmation of its functional properties.

The rising awareness of numerous health benefits of aronia juice initiates the need for further research
toward the development of novel aronia juice–based beverages and functional foods. The development
of original products in addition to the traditional fruit juice is targeted to the improvement of the stabil-
ity of phytochemicals, enhancement of its nutritional composition, and evaluation of novel composite
products providing synergistic effects with other bioactive compounds and foods with beneficial effects
on health.
Aronia Juice 129
TABLE 10.4
List of Human Intervention Studies with Aronia Juice and Their Key Results
Outcomes Key Results References
Markers of Consumption of 100 mL of AJ/day during 12 weeks (uncontrolled study in [45]
antioxidant/ healthy females) induced a significant decrease in TBARS level, pro-oxidant-
pro-oxidant status antioxidant balance, and paroxonase-1 activity.
Markers of Consumption of 50 mL AJ/day during 4 weeks (randomized controlled trial in [46]
antioxidant/ rowers) induced a significant decrease in TBARS, creatine kinase, and lactate
pro-oxidant status levels and significant increase in the postexercise status of antioxidant enzymes
activity.
Markers of Consumption of 100 mL AJ/day during 12 weeks (uncontrolled study in healthy [16]
antioxidant/ females) induced a significant increase in PUFA, total PUFA, unsaturation
pro-oxidant status index, and the activities of antioxidant enzymes. A significant decrease in
MUFA status, n-6/n-3 ratio, and TBARS level.
Biochemical Consumption of 250 mL AJ/day during 6 weeks (uncontrolled study in males [48]
parameters, vitamin with hypercholesterolemia) induced a significant decrease in glucose, total
status, and blood cholesterol, LDL, TAG, vitamin A, and fibrinogen levels as well as diastolic and
pressure systolic blood pressure (confirmed with intervention repeated after wash out).
Glycemic status and Consumption of 200 mL AJ/day acutely in insulin-dependent and nondependent [49]
lipid levels diabetics (comparative uncontrolled study) resulted in an acute decrease in
postprandial glucose levels. The 12-week-long consumption induced a
significant decrease in glucose levels, HbA1c, TC, and lipid levels.
Biomarkers of Consumption of 250 mL AJ/day during 6 weeks resulted in a significant increase [51]
vascular endothelial in brachial artery diameter, flow-mediated vasodilatation, and serum nitric oxide
function levels.
Abbreviations: AJ, aronia juice; TBARS, thiobarbituric acid-reactive substances; PUFA, polyunsaturated fatty acids;
MUFA, monounsaturated fatty acids; LDL, low-density lipoprotein; TAG, triacylglycerols; HbA1c, oxi-
dized hemoglobin; TC, total cholesterol.
Different strategies have been applied in attempting to enhance the stability of polyphenols, mainly
anthocyanins as the most vulnerable molecules during the processing and storage of aronia juice. Based
on the fact that anthocyanins form inclusion complexes with cyclodextrin that may protect anthocy-
anins from hydration and polymerization reactions [59], Howard et al. [60] evaluated the effects of
β-cyclodextrin addition, in various ratios of up to 3%, and in combination with different pH conditions,
on the stability of anthocyanins in aronia juice during pasteurization and storage. Optimal results were
obtained after the addition of 3% of β-cyclodextrin to aronia juice at the natural pH of 3.6, with the
retention of 81% and 95% of individual anthocyanins (at 25°C and 4°C, respectively) after 8 months of
storage. However, lowering the pH from 3.6 to 2.8 had no significant effects on anthocyanin losses dur-
ing processing and storage [60]. In addition to its contribution to the stability of anthocyanins in juices
and other liquid forms, cyclodextrin alone or in combination with other matrices, such as soy protein,
maltodextrin or gum Arabic, can be used during spray drying of aronia juice, based on the data for other
polyphenols and sugar-rich juices [61]. Spray drying is reported to be the optimal drying method of aro-
nia juice in relation to anthocyanins’ stability and is recommended for obtaining powder forms of aronia
juice. It preserves the presence of bioactives and has a minimum influence on their antioxidant activity.
At the same time, spray drying is the most common and cheap technique to be used in the industrial
production of food material [62].
Gonzalez-Molina et al. [63] evaluated the stability of bioactives and antioxidant activity of functional
beverage designed by combining lemon juice with 2.5%, or 5% of aronia juice concentrate, providing
13.75 and 27.21 mg of total anthocyanins per 100 mL of the beverage, respectively. Results have shown
that the addition of aronia juice concentrate (5%, w/w) to lemon juice provides significant reduction in
anthocyanin loss, the highest in vitro antioxidant activity and best sensory properties during 60 days of
storage at room temperature in the dark, compared to the control aronia and lemon juices, and lemon
juice with lower content of aronia juice concentrate.
The bioactives of aronia juice can also be used to improve both quality and health-promoting char-
acteristics of dairy products. The supplementation of goat’s yogurt with aronia juice (5 g/kg) prior to
the addition of starter cultures induced coagulation at a lower acidity and in a shorter time, enhanc-
ing the number of lactic bacteria for up to 80% with the prolonged effects during the storage, and also
increased the content of unsaturated fatty acids compared to the control yogurt [64]. Anthocyanins are
shown to possess prebiotic characteristics, which indicate aronia juice as a suitable functional ingredient
in probiotic dairy products [64], and also a possibility for designing encapsulated powdered forms of
aronia juice by using milk or fermented milk proteins as carriers. Almost parallel to the polyphenol con-
taining foods, functional foods with probiotics show increasing trend worldwide, providing numerous
health benefits to the humans as the host organisms. Although the main probiotic bacteria (Lactobacillus
and Bifidobacterium) are traditionally used in the production of fermented dairy products, they can
also be used to formulate non-diary functional foods [65]. With other sources of prebiotics, including
polyphenol-rich juices, and advanced techniques for the protection of probiotic cells, these products can
fulfill all defined criteria and can be used as the convenient way to improve and maintain health [65].
In addition to novel trends in the formulation of functional products based on aronia juice, novel
concepts also emerge in the scope of evaluation of different modes of action of bioactive compounds.
Putative synergy of aronia juice beneficial effects and the effects of various therapeutics, based on phar-
macokinetic interactions (e.g., inhibition of xenobiotic-metabolizing enzymes by present bioactives),
pharmacodynamic interactions, or putative palliative effects are promising fields to be explored in the
future.
10.6 Conclusion
Aronia juice may be considered as a valuable component of a healthy and balanced diet, providing
significant amounts of vitamins, minerals, carbohydrates, and phytochemicals. Regular consumption
of aronia juice is a promising approach in health promotion and in the reduction of risk of suffering
from a variety of chronic diseases, including CVD and cancer. Numerous effects on health are the
result of high content of polyphenols as major dietary antioxidants. Further research is needed to
elucidate the complete portfolio of aronia juice effects on health compared to other polyphenol-rich
foods, including factors influencing interindividual variation, and with special focus on the mecha-
nisms of action. In addition to traditional fruit juice, novel products in liquid or solid forms with
functional properties based on aronia juice bioactives are emerging, providing enhanced nutritional
and functional benefits.
REFERENCES
1. Kokotkiewicz, A., Jaremicz, Z., and Luczkiewicz, M., Aronia plants: A review of traditional use,
biological activities, and perspectives for modern medicine. J. Med. Food, 13, 255–269, 2010.
2. Kulling, S.E. and Rawel, H.M., Chokeberry (Aronia melanocarpa)–A review on the characteristic com-
ponents and potential health effects. Planta Med., 74, 1625–1634, 2008.
3. Taheri, R., Connolly, B.A., Brand, M.H., and Bolling, B.W., Underutilized chokeberry (Aronia
melanocarpa, Aronia arbutifolia, and Aronia prunifolia) accessions are rich sources of anthocyanins,
flavonoids, hydroxycinnamic acids, and proanthocyanidins. J. Agric. Food Chem., 61, 8581–8588, 2013.
4. Roberfroid, M.B., A European consensus of scientific concepts of functional foods. Nutrition, 16,
689–691, 2000.
5. Institute for Medical Research (IMR), Serbian Food Composition Database, Centre of Research
Excellence in Nutrition and Metabolism Research, IMR, Belgrade, Serbia, 2009. Published online at:
http://www.serbianfood.info/ (accessed June 14, 2014).
6. Wolever, T.M. and Miller, J.B., Sugars and blood glucose control. Am. J. Clin. Nutr., 62(Suppl. 1),
212S–221S, 1995.
7. Howard, L.R., Prior, R.L., Liyanage, R., and Lay, J.O., Processing and storage effect on berry polyphe-
nols: Challenges and implications for bioactive properties. J. Agric. Food. Chem., 60, 6678–6693, 2012.
Aronia Juice 131
8. Finnish National Institute for Health and Welfare, Nutrition Unit, Fineli Food Composition Database
Release 16, December 2013. Published online at: http://www.nal.usda.gov/fnic/foodcomp (accessed
June 14, 2014).
9. National Food Institute, Technical University of Denmark (DTU), Danish Food Composition Database,
March 2009. Published online at: http://www.foodcomp.dk/v7/fvdb_search.asp (accessed June 14,
2014).
10. U.S. Department of Agriculture (USDA), USDA National Nutrient Database for Standard Reference
Release 27, 2014. Published online at: http://www.nal.usda.gov/fnic/foodcomp (accessed September 16,
2014).
11. Westenbrink, S., Oseredczuk, M., Castanheira, I., and Roe, M., Food composition databases: The
EuroFIR approach to develop tools to assure the quality of the data compilation process. Food Chem.,
113, 759–767, 2009.
12. Glibetić, M., Kadvan, A., Tepsić, J., Martacić, J.D., Djekić-Ivanković, M., and Gurinović, M.,
Management of food composition database harmonized with EuroFIR criteria using a web application.
J. Food Comp. Anal., 24, 741–743, 2011.
13. Müller, L., Gnoyke, S., Popken, A.M., and Böhma, V., Antioxidant capacity and related parameters of
different fruit formulations. LWT—Food Sci. Technol., 43, 992–999, 2010.
14. Konić-Ristić, A., Srdić-Rajić, T., Kardum, N., and Glibetić M., Biological activity of Aronia melano-
carpa antioxidants pre-screening in an intervention study design. J. Serb. Chem. Soc., 78, 429–443,
2013.
15. Valcheva-Kuzmanova, S., Kuzmanov, K., Mihova, V., Krasnaliev, I., Borisova, P., and Belcheva, A.,
Antihyperlipidemic effect of Aronia melanocarpa fruit juice in rats fed a high-cholesterol diet. Plant
Foods Hum. Nutr., 62, 19–24, 2007.
16. Kardum, N., Takić, M., Šavikin, K., Zec, M., Zdunić, G., Spasić, S., and Konić-Ristić, A., Effects of
polyphenol-rich chokeberry juice on cellular antioxidant enzymes and membrane lipid status in healthy
women. J. Funct. Foods, 9, 89–97, 2014.
17. Jakobek, L., Seruga, M., Medvidović-Kosanović, M., and Novak, L., Anthocyanin content and antioxi-
dant activity of various red fruit juices. Deut. Lebensm.-Rund., 103, 57–64, 2007.
18. Taheri, R., Durocher, S., Brand, M., and Bolling, B.W., Impact of harvest timing on the polyphenol
content of black chokeberry (Aronia melanocarpa, cv ‘Viking’) juice. FASEB J., 27, lb272, 2013.
19. Wu, X., Gu, L., Prior, R.L., and McKay, S., Characterization of anthocyanins and proanthocyanidins in
some cultivars of Ribes, Aronia, and Sambucus and their antioxidant capacity. J. Agric. Food Chem., 52,
7846–7856, 2004.
20. Handeland, M., Grude, N., Torp, T., and Slimestad, R., Black chokeberry juice (Aronia melanocarpa)
reduces incidences of UTI among nursing home residents in the long term—A pilot study. Nutr. Res.,
DOI: 10.1016/j.nutres.2014.05.005.
21. Oszmiański, J. and Wojdylo, A., Aronia melanocarpa phenolics and their antioxidant activity. Eur. Food
Res. Technol., 221, 809–813, 2005.
22. Krajka-Kuzniak, V., Szaefer, H., Ignatowicz, E., Adamska, T., Oszmianski, J., and Baer-Dubowska, W.,
Effect of chokeberry (Aronia melanocarpa) juice on the metabolic activation and detoxication of carci-
nogenic N-nitrosodiethylamine in rat liver. J. Agric. Food Chem., 57, 5071–5077, 2009.
23. Hellstrom, J.K., Shikov, A.N., Makarova, M.N., Pihlanto, A.M., Pozharitskaya, O.N., Ryhanen, E.-L.,
Kivijarvi, P., Makarov, V.G., and Mattil, P.H., Blood pressure-lowering properties of chokeberry (Aronia
mitchurinii, var. Viking). J. Funct. Foods, 2, 163–169, 2010.
24. Slatnar, A., Jakopic, J., Stampar, F., Veberic, R., and Jamnik, P., The effect of bioactive compounds on
in vitro and in vivo antioxidant activity of different berry juices. PLoS One, 7, e47880, 2012.
25. Wilkes, K., Howard, L.R., Brownmiller, C., and Prior, R.L., Changes in chokeberry (Aronia melano-
carpa L.) polyphenols during juice processing and storage. J. Agric. Food Chem., 62, 4018–4025, 2014.
26. Valcheva-Kuzmanova, S., Blagović, B., and Valić, S., Electron spin resonance measurement of radical
scavenging activity of Aronia melanocarpa fruit juice. Pharmacogn. Mag., 8, 171–174, 2012.
27. Vitaglione, P., Donnarumma, G., Napolitano, A., Galvano, F., Gallo, A., Scalfi, L., and Fogliano, V.,
Protocatechuic acid is the major human metabolite of cyanidin-glucosides. J. Nutr., 137, 2043–2048, 2007.
28. Bermudez-Soto, M.J., Larrosa, M., Garcia-Cantalejo, J., Espin, J.C., Tomas-Barberan, F.A., and Garcia-
Conesa, M.T., Transcriptional changes in human Caco-2 colon cancer cells following exposure to a
recurrent non-toxic dose of polyphenol-rich chokeberry juice. Genes Nutr., 2, 111–113, 2007.
29. Sharif, T., Alhosin, M., Auger, C., Minker, C., Kim, J.H., Etienne-Selloum, N., Bories, P. et al., Aronia
melanocarpa juice induces a redox-sensitive p73-related caspase 3-dependent apoptosis in human leu-
kemia cells. PLoS One, 7, e32526, 2012.
30. Konić-Ristić, A., Srdić-Rajić, T., Kardum, N., Aleksić-Velicković, V., Kroon, P.A., Hollands, W.J.,
Needs, P.W. et al., Effects of bioactive-rich extracts of pomegranate, persimmon, nettle, dill, kale, and
Sideritis and isolated bioactives on arachidonic acid induced markers of platelet activation and aggrega-
tion. J. Agric. Food Chem., 93, 3581–3587, 2013.
31. Hollands, W.J., Saha, S., Hayran, O., Boyko, N., Glibetić, M., Konić-Ristić, A., Jorjadze, M., and Kroon,
P.A., Lack of effect of bioactive-rich extracts of pomegranate, persimmon, nettle, dill, kale, and Sideritis
and isolated bioactives on platelet function. J. Sci. Food. Agric., 93, 3588–3594, 2013.
32. Danesi, F., Saha, S., Kroon, P.A., Glibetić, M., Konić-Ristić A., D’Antuono, L.F., and Bordoni, A.,
Bioactive-rich Sideritis scardica tea (mountain tea) is as potent as Camellia sinensis tea at inducing
cellular antioxidant defences and preventing oxidative stress. J. Sci. Food. Agric., 93, 3558–3564, 2013.
33. Woodcock, M.E., Hollands, W.J., Konić-Ristić, A., Glibetić, M., Boyko, N., Kocaoglu, B., and Kroon,
P.A., Bioactive-rich extracts of persimmon, but not nettle, Sideritis, dill or kale, increase eNOS activa-
tion and NO bioavailability and decrease endothelin-1 secretion by human vascular endothelial cells.
J. Sci. Food. Agric., 93, 3574–3580, 2013.
34. Braunlich, M., Christensen, H., Johannesen, S., Slimestad, R., Wangensteen, H., Malterud, K.E., and
Barsett, H., In vitro inhibition of cytochrome P450 3A4 by Aronia melanocarpa constituents. Planta
Med., 79, 137–141, 2013.
35. Borissova, P., Valcheva, S., and Belcheva, A., Anti-inflammatory effect of flavonoids in the natural juice
from Aronia melanocarpa, rutin and rutin-magnesium complex on an experimental model of inflamma-
tion induced by histamine and serotonin. Acta Physiol. Pharmacol. Bulg., 20, 25–30, 1994.
36. Valcheva-Kuzmanova, S., Marazova, K., Krasnaliev, I., Galunska, B., Borisova, P., and Belcheva, A.,
Effect of Aronia melanocarpa fruit juice on indomethacin-induced gastric mucosal damage and oxida-
tive stress in rats. Exp. Toxicol. Pathol., 56, 385–392, 2005.
37. Valcheva-Kuzmanova, S., Kuzmanov, K., Tancheva, S., and Belcheva, A., Hypoglycemic and hypolip-
idemic effects of Aronia melanocarpa fruit juice in streptozotocin-induced diabetic rats. Methods Find.
Exp. Clin. Pharmacol., 29, 101–105, 2007.
38. Valcheva-Kuzmanova, S., Borisova, P., Galunska, B., Krasnaliev, I., and Belcheva, A., Hepatoprotective
effect of the natural fruit juice from Aronia melanocarpa on carbon tetrachloride-induced acute liver
damage in rats. Exp. Toxicol. Pathol., 56, 195–201, 2004.
39. Burton-Freeman, B., Postprandial metabolic events and fruit-derived phenolics: A review of the science.
Br. J. Nutr., 104(Suppl. 3), 1S–14S, 2010.
40. Auger, C., Kim, J.H., Trinh, S., Chataigneau, T., Popken, A.M., and Schini-Kerth, V.B., Fruit juice-
induced endothelium-dependent relaxations in isolated porcine coronary arteries: Evaluation of differ-
ent fruit juices and purees and optimization of a red fruit juice blend. Food Funct., 2, 245–250, 2011.
41. Williamson, G. and Clifford, M.N., Colonic metabolites of berry polyphenols: The missing link to
biological activity? Br. J. Nutr., 104(Suppl. 3), 48S–66S, 2010.
42. Wiczkowski, W., Romaszko, E., and Piskula, M.K., Bioavailability of cyanidin glycosides from natural
chokeberry (Aronia melanocarpa) juice with dietary-relevant dose of anthocyanins in humans. J. Agric.
Food Chem., 58, 12130–12136, 2010.
43. Hollman, P.C., Cassidy, A., Comte, B., Heinonen, M., Richelle, M., Richling, E., Serafini, M., Scalbert,
A., Sies, H., and Vidry, S., The biological relevance of direct antioxidant effects of polyphenols for
cardiovascular health in humans is not established. J. Nutr., 141(Suppl.), 989S–1009S, 2011.
44. Prior, R.L., Gu, L., Wu, X., Jacob, R.A., Sotoudeh, G., Kader, A.A., and Cook, R.A., Plasma antioxidant
capacity changes following a meal as a measure of the ability of a food to alter in vivo antioxidant status.
J. Am. Coll. Nutr., 26, 170–181, 2007.
45. Kardum, N., Konić-Ristić, A., Savikin, K., Spasić, S., Stefanović, A., Ivanisević, J., and Miljkovic,
M., Effects of polyphenol-rich chokeberry juice on antioxidant/pro-oxidant status in healthy subjects.
J. Med. Food, 17, 869–874, 2014.
46. Pilaczynska-Szczesniak, L., Skarpanska-Steinborn, A., Deskur, E., Basta, P., and Horoszkiewicz-
Hassan, M., The influence of chokeberry juice supplementation on the reduction of oxidative stress
resulting from an incremental rowing ergometer exercise. Int. J. Sport Nutr. Exerc. Metab., 15, 48–58,
2005.
Aronia Juice 133
47. Knekt, P., Jarvinen, R., Reunanen, A., and Maatela, J., Flavonoid intake and coronary mortality in
Finland: A Cohort Study. Br. Med. J., 312, 478–481, 1996.
48. Skoczyñska, A., Jêdrychowska, I., Porêba, R., Affelska-Jercha, A., Turczyn, B., Wojakowska, A., and
Andrzejak, R., Influence of chokeberry juice on arterial blood pressure and lipid parameters in men with
mild hypercholesterolemia. Pharm. Rep., 59(Suppl. 1), 177S–182S, 2007.
49. Simeonov, S.B., Botushanov, N.P., Karahanian, E.B., Pavlova, M.B., Husianitis, H.K., and Troev, D.M.,
Effects of Aronia melanocarpa juice as part of the dietary regimen in patients with diabetes mellitus.
Folia Med. (Plovdiv), 44, 20–23, 2002.
50. Naruszewicz, M., Łaniewska, I., Millo, B., and Dłużniewski, M., Combination therapy of statin with
flavonoids rich extract from chokeberry fruits enhanced reduction in cardiovascular risk markers in
patients after myocardial infraction (MI). Atherosclerosis, 194, 179–184, 2007.
51. Poreba, R., Skoczynska, A., Gac, P., Poreba, M., Jedrychowska, I., Affelska-Jercha, A., Turczyn, B.,
Wojakowska, A., Oszmianski, J., and Andrzejak, R., Drinking of chokeberry juice from the ecological
farm Dzieciolowo and distensibility of brachial artery in men with mild hypercholesterolemia. Ann.
Agric. Environ. Med., 16, 305–308, 2009.
52. Erlund, I., Koli, R., Alfthan, G., Marniemi, J., Puukka, P., Mustonen, P., Mattila, P., and Jula, A.,
Favorable effects of berry consumption on platelet function, blood pressure, and HDL cholesterol.
J. Am. Clin. Nutr., 87, 323–331, 2008.
53. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA), Guidance on the scientific require-
ments for health claims related to antioxidants, oxidative damage and cardiovascular health. EFSA J.,
9, 2474–2786, 2011.
54. Konić-Ristić, A., The impact of plant polyphenols on platelet activation and their aggregation with endo-
thelial, blood and malignant cells (in Serbian), PhD thesis, University of Belgrade, Belgrade, Serbia,
2013.
55. Onakpoya, I., Posadzki, P., and Ernst, E., The efficacy of glucomannan supplementation in overweight
and obesity: A systematic review and meta-analysis of randomized clinical trials. J. Am. Coll. Nutr., 33,
70–78, 2014.
56. Kardum, N., Petrović-Oggiano, G., Takić, M., Glibetić, N., Zec, M., Debeljak-Martacić, J., and Konić-
Ristić, A., Effects of glucomannan-enriched, aronia juice-based supplement on cellular antioxidant
enzymes and membrane lipid status in subjects with abdominal obesity. Sci. World J., 2014, 869250,
2014.
57. Lefebvre, P.J. and Scheen, A.J., The postprandial state and risk of cardiovascular disease. Diabetic
Med., 15(Suppl. 4), 63S–68S, 1998.
58. Ristić-Medić, D., Suzić, S., Vucić, V., Takić, M., Tepsić, J., and Glibetić, M., Serum and erythrocyte
membrane phospholipids fatty acid composition in hyperlipidemia: Effects of dietary intervention and
combined diet and fibrate therapy. Gen. Physiol. Biophys., 28(Suppl.), 190S–199S, 2009.
59. Mourtzinos, I., Makris, D.P., Yannakopoulou, K., Kalogeropoulos, N., Michali, I., and Karathanos, V.T.,
Thermal stability of anthocyanin extract of Hibiscus sabdariffa L. in the presence of beta-cyclodextrin.
J. Agric. Food Chem., 56, 10303–10310, 2008.
60. Howard, L.R., Brownmiller, C., Prior, R.L., and Mauromoustakos, A., Improved stability of chokeberry
juice anthocyanins by beta-cyclodextrin addition and refrigeration. J. Agric. Food Chem., 61, 693–699,
2013.
61. Yousefi, S., Emam-Djomeh, Z., and Mousavi, S.M., Effect of carrier type and spray drying on the physi-
cochemical properties of powdered and reconstituted pomegranate juice (Punica granatum L.). J. Food
Sci. Technol., 48, 677–684, 2011.
62. Horszwald, A., Julien, H., and Andlauer, W., Characterisation of aronia powders obtained by different
drying processes. Food Chem., 141, 2858–2863, 2013.
63. Gonzalez-Molina, E., Moreno, D.A., and Garcia-Viguera, C., Aronia-enriched lemon juice: A new
highly antioxidant beverage. J. Agric. Food Chem., 56, 11327–11333, 2008.
64. Boycheva, S., Dimitrov, T., Naydenova, N., and Mihaylova, G., Quality characteristics of yogurt from
goat’s milk, supplemented with fruit juice. Czech J. Food Sci., 29, 24–30, 2011.
65. Gawkowski, D. and Chikindas, M.L., Non-dairy probiotic beverages: The next step into human health.
Benef. Microbes, 4, 127–142, 2013.
11
Blackberry Juice
Mirela Kopjar and Vlasta Piližota
CONTENTS
11.1 Introduction....................................................................................................................................135
11.3 Bioactives and Antioxidant Efficiency.......................................................................................... 137
11.4 Health Effects................................................................................................................................ 140
11.6 Conclusion......................................................................................................................................143
References............................................................................................................................................... 144
11.1 Introduction
The blackberry belongs to the genus Rubus and has a complex genetic background, growth characteristics,
and a number of species. It is a specific fruit that consists of aggregates of drupelets [1]. Blackberries
contain a high content of bioactive compounds, especially anthocyanins and ellagitannins, possessing
high antioxidant activity [2–4]. Blackberry juice can be extracted either from fresh or frozen fruits.
Frozen fruit may be cold pressed. The benefits of this method are a higher yield and that the plant may
operate outside the harvest season. The final product has a better color than that of fresh cold press juice
and nearly as good as that from heated fresh berries and seems to have a fresh fruit flavor not present
in the heated juice. In hot processing method, clean, ripe, and wholesome fruit is heated and agitated in
a steam-jacketed kettle between 60°C and 82°C. When this temperature range is reached, the crushed
berries can be pressed in a hydraulic rack and cloth press. Usually, with thermal treatment, pretreatment
of fruit with pectolytic enzymes is used causing the release of more juice due to cell wall breakdown.
The resultant juice contains high amounts of pectin making the juice viscous and an enzymatic depec-
tinization is necessary in the production of clear juices and concentrates. The addition of pectinase
improves juice and color of the extract while retaining the organoleptic properties of the fruit [5]. Detailed
blackberry juice production can be found elsewhere [6–8].
Due to high nutritional value, it is important that bioactive compounds are transferred from raw mate-
rial to the product. During processing of fruit into juice, chemical and biochemical changes cannot be
avoided, causing degradation of bioactive compounds, thus decreasing antioxidant activity of blackberry
juice as well as reducing beneficial effects on human health. For the production of highly nutritional
valuable juice, it is necessary to slow down and/or reduce those degradation reactions as much as pos-
sible and to enhance bioactive compounds in blackberry juice. Some efforts have already been made,
such as application of nonthermal processing, instead of conventional heat processing and addition of
some ingredients that could enhance antioxidant capacity of the juice and/or minimize degradation reac-
tion of bioactive compounds. This chapter highlights the nutritional and phytochemical content of black-
berry juice, its antioxidant capacity, human health, as well as novel formulations that may improve this
functionality.
135

Blackberry varieties vary in their antimutagenic potency, chemical, and genetic bases. For these differ-
ences, they should be established and defined to produce fruit with enhanced anticancer properties or
to isolate compounds from those verities [9]. Blackberry juice contains 90.90 g/100 g of water and has
an energy value of 38 kcal. Carbohydrate is most abundant (7.80 g/100 g), followed by fat (0.60 g/100 g)
and protein (0.30 g/100 g). Sugars are the major part of carbohydrates in blackberry juice, consisting
of glucose, fructose, and, to a lesser extent, sucrose. Blackberry juice is also a source of some vitamins
(choline, folate, niacin, riboflavin, thiamin, vitamins A, B6, C, E, and K), minerals (calcium, copper,
iron, magnesium, phosphorus, potassium, selenium, sodium and zinc), and carotenoids (Table 11.1) [10].
TABLE 11.1
Compositional and Nutritional Characteristcis of Blackberry Juice (per 100 g)
Nutrient Unit Value
Water g 90.90
Energy kcal 38
Protein g 0.30
Lipid (fat) g 0.60
Carbohydrate g 7.80
Total sugars g 7.70
Total dietary fiber g 0.10
Ash g 0.40
Minerals
Calcium mg 12
Copper mg 0.114
Iron mg 0.48
Magnesium mg 21
Phosphorus mg 12
Potassium mg 135
Selenium μg 0.3
Sodium mg 1
Zinc mg 0.41
Vitamins
Choline mg 6.6
Folate (DFE) μg 10
Niacin mg 0.446
Riboflavin mg 0.018
Thiamin mg 0.012
Vitamin A (REA) μg 6
Vitamin B6 mg 0.021
Vitamin C mg 11.3
Vitamin E (ATE) mg 0.90
Vitamin K μg 15.2
Carotenoids
β-Carotene μg 74
Lutein + zeaxanthin μg 38
Source: Adapted from the U.S. Department of Agriculture (USDA), USDA National
Nutrient Database for Standard Reference, Release 26, 2013, Published online
at: http://www.nal.usda.gov/fnic/foodcomp/search/ (accessed May 26, 2014).
Abbreviations: DFE, dietary folate equivalents; RAE, retinol activity equivalents; ATE,
alpha-tocopherol equivalents.
Blackberry Juice 137
11.3 Bioactives and Antioxidant Efficiency

There have been an increasing number of studies relating to diet rich in fruits and vegetables with
a decreasing risk of cardiovascular disease (CVD) and some types of cancer. There are more and
more nutritional guidelines emphasizing the importance of fruits and fruit products in everyday life.
Generally, berries are widely recognized for their high content of bioactive compounds, including
flavonoids, phenolic acids, and tannins, having high antioxidant activity. Bioactive compounds, both
individually and/or synergistically, may help protect against CVD, cancer, inflammation, obesity, dia-
betes, and other chronic diseases.
Total phenolics, anthocyanins, ellagitannins, and antioxidant activities of blackberry juices are given in
Tables 11.2 and 11.3. As can be seen from these tables, large variations exist among samples. These varia-
tions are a consequence of using different blackberry varieties in juice production. Total phenol content
ranged from 0.31 to 3.66 g gallic acid equivalents (GAE) and anthocyanin content from 76.9 to 1100 mg
cyanidin-3-glucoside equivalents (C3GE). Nonclarified and clarified blackberry juices had almost the same
anthocyanin content, while ellagitannins were higher by 38.6% in nonclarified juice. Ultrafiltrated juice
had around 44% higher anthocyanin and ellagitannin content than microfiltrated juice. Five anthocyanins
were detected in blackberry fruit, and they were identified as cyanidin aglycones [6,7,14,15]. Blackberry
juice contains higher percentage of cyanidin-3-O-glucoside (80%–91% of the total anthocyanins)
than cyanidin-3-O-malonyl glucoside, cyanidin-3-O-xyloside, and cyanidin-3-O-rutinoside [6,7,16–19]
(Figure 11.1). Cyanidin-3-O-glucoside, as the most abundant cyanidin in blackberry juice, was determined
at 90 mg/kg fresh weight (FW) [7], 743.3 mg/L [16], and 410 mg/kg [17], while other cyanidins were
TABLE 11.2
Total Phenolics, Anthocyanins, and Ellagitannins in Blackberry Juice
Total Total
Sample Unit Phenolics Unit Anthocyanins Unit Ellagitannins References
Juice g GAE/L 2.49 mg C3GE/L 516.4 — [39]
g GAE/L 2.44 mg C3GE/L 480.3 — [40]
g GAE /L 3.66 — — [11]
g GAE/kg FW 0.48 — — [7]
g GAE/L 1.83 mg C3GE/L 740 — [16]
g GAE/L 1.15 mg C3GE/L 173 — [12]
g GAE/L 0.52 mg C3GE/L 150 — [42]
g GAE/L 1.541 mg C3GE/L 401 — [23]
— mg C3GE/L 203 — [24]
g GAE/L 2.94 mg C3GE/L 554 — [25]
— mg C3GE/L 906 — [44]
— mg C3GE/L 1100 — [45]
g GAE/L 0.31 mg C3GE/L 76.9 — [11]
g GAE/kg DW 45.0 g C3GE/kg DW 11.5 — [28]
Juice g GAE/kg DW 24.9 g C3GE/kg DW 5.6 — [28]
(microfiltrated)
— mg C3GE/L 569 mg EAE/L 1419 [33]
Juice — mg C3GE/L 1017 mg EAE/L 2545 [33]
(ultrafiltrated)
Juice — mg C3GE/kg 836 mg EAE/kg 104.1 [6]
(nonclarified)
Juice (clarified) — mg C3GE/kg 820 mg EAE/kg 63.9 [6]
Concentrate g GAE/L 1.55 mg C3GE/L 418 — [23]
Abbreviations: GAE, gallic acid equivalents; C3GE, cyanidin-3-glucoside equivalents; EAE, ellagic acid equivalents;
FW, fresh weight; DW, dry weight.
TABLE 11.3
Antioxidant Activities of Blackberry Juice Using Different Methods
Sample Method Unit Concentration References
Juice ORAC µmol TE/g FW 4.6 [7]
Juice ORAC μmol TE/g DW 306 [25]
Juice ORAC μmol TE/g DW 547 [28]
Juice ORAC µmol TE/mL 180 [30]
Juice ORAC µmol TE/g FW 20.3–24.6 [13]
Juice H-ORAC µmol TE/g 43 [17]
Juice H-ORAC µmol TE/mL 48 [45]
Juice DPPH µmol TE/g FW 3.3 [7]
Juice DPPH µmol TE/mL 8.75 [16]
Juice DPPH mg GAE/mL 0.747 [12]
Juice ABTS mg GAE/mL 0.522 [12]
Juice ABTS µmol TE/mL 4.2 [11]
Juice ABTS µmol TE/mL 5.2 [42]
Juice (microfiltrated) ORAC μmol TE/g DW 417 [28]
Juice (nonclarified) ORAC µmol TE/g 42.9 [6]
Juice (nonclarified) PCL µmol TE/g 9.4 [6]
Juice (clarified) ORAC µmol TE/g 43.8 [6]
Juice (clarified) PCL µmol TE/g 10.1 [6]
Abbreviations: ORAC, oxygen radical absorbance capacity; H-ORAC, hydrophilic-ORAC; DPPH, 2,2-diphenyl-
1-p icrylhydrazyl; ABTS, 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid); PCL, photo-
chemiluminescent; TE, trolox equivalents; GAE, gallic acid equivalents; FW, fresh weight; DW,
dry weight.
present in much lower amounts. Cyanidin-3-O-malonyl glucoside was present at 6.3 mg/kg FW [7] and
30 mg/kg [17]. In addition, cyanidin-3-O-xyloside was present at 68.1 mg/kg and cyanidin-3-O-rutinoside
was 10.8 mg/kg [16]. Blackberry juice is also a good source of ellagitannins, especially lambertianin C and
sanguiin H-6 [7,8,17,18] (Figure 11.1). Lambertianin C was found at a higher amount than sanguiin H-6.
Gancel et al. [7] determined lambertianin C and sanguiin H-6 at 31 and 3 mg/kg FW, respectively, while
Acosta et al. [17] found higher values, 380 and 230 mg/kg, respectively.
Investigation of the influence of processing steps on the bioactive compounds in blackberry juice in
comparison to the fruit revealed that thermal treatment is the main cause of degradation of bioactives
during processing [6–8]. Juice production resulted in degradation of total phenols (around 45%), ellagi-
tannins (70%–82%), and anthocyanins (around 67%) with increase of polymeric color compounds [6–8].
Monitoring degradation of the most important bioactives in blackberry juice through the whole pro-
duction process, the content of cyanidin-3-O-glucoside and cyanidin-3-O-malonyl glucoside decreased
by 52% and 64%, respectively, and lambertianin C and sanguiin H-6 were reduced by 80% and 50%,
respectively [7].
Several mechanisms are involved in the degradation of anthocyanins, such as hydrolysis, oxidation,
and condensation with other polyphenols. All of these reactions occur during juice production and stor-
age, and their intensity depends on processing and other conditions in the system. For example, the
degradation of cyanidin-3-O-glucoside (the main anthocyanin in blackberry juice) in aqueous solution
begins with hydrolysis of glucosidic bonds and the opening of the pyrylium ring upon heating [20] or
by anthocyanase [21]. Further degradation includes the formation of protocatechuic acid and phloroglu-
cinaldehyde. Cyanidin-3-O-glucoside can also be cooxidized with other phenolics such as chlorogenic
acid by polyphenol oxidase with the formation of O-quinones that generate polymerized products by
quinine–phenol reactions [22]. Since thermal treatment is one of the steps during juice production and
it is one of the most important ones through which degradation of anthocyanins occurs, achievement of
R1
OH
HO O+
O
OH R2
Anthocyanin R1 R2
Cyanidin-3-O-glucoside OH Glucose
Cyanidin-3-O-malonyl glucoside OH Malonyl-glucose
Cyanidin-3-O-xyloside OH Xylose
Cyanidin-3-O-rutinoside OH Rutinose
HO
O
HO
HO O OH
HO O O O
HO
O HO OH
HO O O
HO O HO O O O O OH
O
HO HO O O O
O HO OH
HO O O
HO OH HO OH OH
HO O O O O O HO
O OH OH
HO O O O
HO OH
O O
HO OH OHHO OH OH
O OO O HO
OH OH
HO OH OH
HO
OH OH
(a) Lambertianin C
HO
O
HO
HO O
OH
HO HO O O O
O HO OH
O O
HO
HO O HO O OH
O O O O
HO O O O
HO OH
O O
HO OH HO OH
O O OH OH
O O HO
OH OH
HO OH OH
HO
OH OH
(b) Sanguiin H-6
FIGURE 11.1 The most abundant anthocyanins and ellagitannins (a: lambertianin C; b: sanguiin H-6) found in
blackberry juice.
thermal stability of anthocyanins is very important. An investigation of thermal stability of blackberry

juice revealed that the thermal degradation of anthocyanins follows a first-order reaction kinetics with
activation energy values of 24–92 kJ/mol [23–27].
Analysis of fresh and microfiltrated blackberry juice showed less polyphenols in the microfiltrated
juice. However, values for biological activity, such as protection of lipid peroxidation, was not significantly
different between juices. These results suggest that compounds responsible for antioxidant activity are
maintained even after microfiltration and the free radical scavenging capacity of these compounds could
protect against the initiation of lipid peroxidation. Microfiltration could be used as an industrial technique
to produce blackberry juice that maintains the biological activity of its polyphenols [28]. Blackberry juice
can be applied in food products for achieving oxidative stability of oil-in-water emulsions with differ-
ent concentrations of whey protein and rapeseed oil due to the presence of antioxidants [29]. On the one
hand, the oxidative stability of emulsions can be achieved, but absorption of phenols after consumption of
such foods is questionable. Hassimotto et al. [18] observed that a food matrix had a great influence on the
absorption of anthocyanins and ellagitannins. The rate of absorption of anthocyanins slowed down when
blackberry juice was taken with defatted milk, without any change in total amount of anthocyanins that
were absorbed. For ellagitannins, a total inhibition of their absorption was observed when juice was pre-
pared by using defatted milk. It was necessary to determine if blackberry juice and blackberry juice with
defatted milk had any effect on the antioxidant capacity of human plasma and urine. The consumption of
blackberry juice caused an increase in the plasma antioxidant capacity, which could be associated more to
ascorbic acid content than polyphenols present in blackberry juice [18]. In the urine, antioxidant activity
was associated with anthocyanin and urate content, but further studies are necessary to determine exactly
which compounds are responsible for the observed effects [18].
11.4 Health Effects

Beneficial health effects of blackberry juice are summarized in Table 11.4. Antioxidant activity of black-
berry juice and cyanidin-3-O-glucoside on endothelial dysfunction in cells and vascular rings after
exposition to peroxynitrite was investigated [19]. Peroxynitrite caused a significant suppression of mito-
chondrial respiration, DNA strand breakage, and activation of poly-adenosine diphosphate-ribosyl syn-
thetase in human umbilical vein endothelial cells. Peroxynitrite-induced damage of vascular rings could
also be suppressed by application of blackberry juice and cyanidin-3-O-glucoside. Blackberry juice con-
tains a high amount of cyanidin-3-O-glucoside, thus a scavenger of peroxynitrite and has a protective
effect against endothelial dysfunction and vascular failure induced by peroxynitrite [19]. According to
Boivin et al. [30], blackberry juice slightly inhibited cancer cell lines (stomach, prostate, intestine, and
breast) but had a high anti-inflammatory potential. The development of metastatic cancer is a complex
process [9]. Tate et al. [9] also investigated the influence of eight varieties of blackberry on the sup-
pression of mutagenesis induced by metabolically activated carcinogen 2-aminoanthracene or methyl
methanesulfonate. Blackberry juice was found to suppress mutagenesis by the metabolically activated
carcinogen 2-aminoanthracene, but the degree of suppression of mutagenesis depended on the variety
examined. However, blackberry juice did not have any significant influence on mutagenesis induced by
direct-acting alkylating agent methyl methanesulfonate. Since no effect on mutagenesis was found, the
juice was treated with hydrochloric acid to pH 1 overnight and then neutralized in order to simulate the
influence of stomach acid on blackberry composition. This treatment did not also suppress mutagenesis
induced by the direct-acting carcinogen methyl methanesulfonate. It reacts directly with DNA in vitro,
while 2-aminoanthracene is activated by the cytochrome P450 system. It is most likely that suppression
of mutagenesis induced by 2-aminoanthracene by blackberry is a consequence of their inhibition of
cytochrome P450 activation [9].
Liver diseases are one of the most serious health problems, and hepatocellular carcinoma is one of
the world’s deadliest cancers in the world. Agha et al. [31] studied the influence of blackberry juice on
the hepatotoxicity and oxidative stress induced by carbon tetrachloride in male rats. Blackberry juice
had a significant hepatoprotective effect, as indicated by a significant inhibition of the activity of serum
enzymes, bilirubin, and lipid peroxidation levels accompanied by significant elevation in the activity of
TABLE 11.4
Positive Actions of Blackberry Juice
Sample Action References
Juice and Reduction of peroxynitrite-induced mitochondrial respiration, DNA strand [19]
cyanidin-3-O- breakage, and activation of poly-ADP-ribosyl synthetase in human umbilical
glucoside vein endothelial cells.
Reduction of peroxynitrite-induced damage of vascular rings. [19]
Scavenger of peroxynitrite and protective effect against endothelial [19]
dysfunction and vascular failure that were induced by peroxynitrite.
Juice Slight inhibition effect on cancer cell line (stomach, prostate, intestine, and [30]
breast).
High anti-inflammatory potential. [30]
No effect on mutagenesis induced by the direct-acting carcinogen methyl [9]
methanesulfonate.
Suppression of mutagenesis induced by 2-aminoanthracene is a consequence [9]
of the inhibition of cytochrome P450 activation.
Significant hepatoprotective effect as indicated by a significant inhibition of [31]
the activity of serum enzymes, bilirubin, and lipid peroxidation levels.
Significant elevation in the activity of hepatic antioxidants and serum [31]
glutathione-S transferase activity.
Reduction of the hepatic lesions induced by carbon tetrachloride. [31]
Protective effect against sodium fluoride–induced hepatotoxicity by [32]
antagonizing the free radicals generation and enhancement of the antioxidant
defense mechanisms.
Facilitate UVB-induced effects by reducing DNA damage and increasing [33]
apoptosis of damaged cells by the following:
• Reduction of the formation of UVB (25 mj/cm2)-mediated cyclobutane
pyrimidine dimers and 8-oxo-7,8-dihydro-2′-deoxyguanosine in normal
human epidermal keratinocytes.
• Increased UVB-mediated poly(ADP-ribose) polymerase cleavage and
activation of caspases 3, 8, and 9 in normal human epidermal keratinocytes.
• Decreased formation of cyclobutane pyrimidine dimmers 9 in normal
human epidermal keratinocytes.
Preservative in food processing and a preventive in food-borne infections as a [37]
natural antimicrobial by the following:
• Reduction of the growth of Listeria monocytogenes, Salmonella
typhimurium, and Escherichia coli O157:H7.
• Stimulation of the growth of Lactobacillus strains.
Increase in the plasma antioxidant capacity. [18]
Achieving oxidative stability of oil-in-water emulsions. [29]
Berry juice Decrease of lipid peroxidation, which is important for cardiovascular disease. [34]
containing Protection of the serum lipids against oxidation and therefore natural [34]
blackberry juice anti-atherosclerotic.
Increase of plasma antioxidant activity. [35]
Decrease of plasma malondialdehyde. [35]
Prevention of chronic diseases such as cancer and cardiovascular disease in [36]
hemodialysis patients exposed to enhanced oxidative stress by the following:
• Decrease of DNA oxidation damage, protein and lipid peroxidation, and
nuclear factor-κB binding activity.
• Increase of level and status of glutathione.
Abbreviations: ADP, adenosine diphosphate; UVB, ultraviolet B.
hepatic antioxidants and serum glutathione-S transferase activity. The hepatic lesions induced by carbon
tetrachloride were significantly reduced by blackberry juice pretreatment. These results suggest that
blackberry juice, due to its antioxidant properties, could prevent liver dysfunction induced by carbon
tetrachloride and prevent hepatic disorders.
Antioxidants can be used for the management of fluorosis and as antidotes for fluoride toxicity. When
blackberry juice was applied together with sodium fluoride, harmful effects of sodium fluoride on
antioxidant parameters were decreased, thus blackberry juice has a protective effect against sodium
fluoride–induced hepatotoxicity by antagonizing free radical generation and enhancing the antioxidant
defense mechanisms [32].
Solar ultraviolet radiation, especially ultraviolet B spectrum (UVB, radiation from 280 to 320 nm), is
the primary environmental stimulus leading to skin carcinogenesis. Several botanical species, includ-
ing blackberry, which are exhibiting antioxidant properties, have shown photochemopreventive effects
against UVB damage. Calvo-Castro et al. [33] investigated the photochemopreventive effect of black-
berry juice on UVB-mediated responses in human epidermal keratinocytes and in a 3D reconstituted
normal human skin equivalent. They determined that pretreatment (for 2 h) and posttreatment (for 24 h)
of normal human epidermal keratinocytes with blackberry juice reduced UVB (25 mJ/cm2)-mediated
cyclobutane pyrimidine dimers and 8-oxo-7,8-dihydro-2′-deoxyguanosine formation. In addition,
treatment of normal human epidermal keratinocytes with blackberry juice increased UVB-mediated
poly(ADP-ribose) polymerase cleavage and activation of caspases 3, 8, and 9. Immunochemistry studies
on reconstituted normal human skin equivalent also showed decreased formation of cyclobutane pyrimi-
dine dimmers. Based on the preceding text, blackberry juice seems to facilitate UVB-induced effects by
reducing DNA damage and increasing apoptosis of damaged cells.
Some investigations with mixed fruit juices have revealed their potential benefits. García-Alonso et al.
[34] investigated the short-term influence of the intake of a phenolic-rich juice (26% grape, 2% cherry,
0.6% blackberry, 0.6% blackcurrant, and 1% raspberry) on the oxidative stress status on healthy subjects.
They suggested that increase of total antioxidant capacity after ingestion of the juice might be respon-
sible for an increase in serum uric acid due to rapid metabolism of fructose and/or other sugars from the
juice, but not from the serum phenols content. Phenols were probably absorbed in sufficient amounts to
decrease lipid peroxidation, which is important for CVD, meaning that phenols from the juice could pro-
tect serum lipids against oxidation and therefore be natural anti-atherosclerotic compounds in the diet.
However, increase of protein oxidation associated with juice intake was observed, probably due to the
naturally present soluble sugars that are capable of mediating oxidative stress in vivo, by increasing car-
bonyl proteins after intake [34]. Consumption of berry juice (30% white grape, 25% blackcurrant, 15%
elderberry, 10% sour cherry, 10% blackberry, and 10% aronia) caused increase of plasma antioxidant
activity and decrease of plasma malondialdehyde. It was also demonstrated that after juice consumption
valuable ingredients such as anthocyanins and ascorbic acid are available for humans and are active as
antioxidants in vivo [35]. Red fruit juice (40% red grape juice, 20% blackberry juice, 15% sour cherry
juice, 15% black currant juice, and 10% elderberry juice) with high anthocyanin/polyphenol content was
shown to reduce oxidative damage, thus this fruit juice was tested on hemodialysis patients to evaluate
its preventive effect on the patients. Those patients are facing an elevated risk of cancer, arteriosclerosis,
and other diseases, which are related to increase in oxidative stress [36]. Results of this study showed
that juice uptake significantly decreased DNA oxidation damage, protein and lipid peroxidation, and
nuclear factor-κB (NF-κB) binding activity, as well as an increase of glutathione level. Such beneficial
effects of the juice were attributed to its high anthocyanin/polyphenol content, and its consumption can
be promising for prevention of chronic diseases such as cancer and CVD in patients exposed to enhanced
oxidative stress.
New potential of blackberry juice application in food products may be by providing oxidative sta-
bility [29], but it can also be used as a preservative material in food processing and an agent against
food-borne infections as a natural antimicrobial agent [37]. The growth of Listeria monocytogenes,
Salmonella typhimurium, and Escherichia coli O157:H7 were significantly reduced, while the growth of
Lactobacillus strains was stimulated in milk and broth by blackberry juice [37].
It is very important to keep in mind that phenolics that are the most common in the human diet are not
necessarily the most active ones within the body, due to several reasons such as having lower intrinsic
activity, poorly absorbed from the intestine, highly metabolized, or rapidly eliminated. The metabolites
that are also found in blood and target organs and result from digestive or hepatic activity may differ in
biological activity from the native substances consumed [38].

Blackberry juices, such as single strength puree (10.5–18°Brix), a puree concentrate (20–40°Brix), a single
strength juice (10°Brix), and a juice concentrate (45–68°Brix), can be found on the markets [5]. Generally,
blackberry juice has a specific flavor that is not always preferred by consumers. Studies have shown that
consumers prefer red juices (such as blueberry or Concord grape juice) rather than blackberry juice.
Consumers also prefer juice blends that contain lower amounts of blackberry juice [39–41]. However,
Lawless et al. [40] found that when consumers were informed about anthocyanin content of the juice,
they were more likely to buy the juice with higher anthocyanin content since they relate it to the health
benefits. In this way, they gave priority to health benefits over the flavor. Flavor is determined by the
content of sugars, acids, and volatiles, all of which vary with cultivar and growing conditions. The ratio
of sugars to acids plays a major part in determining flavor [1]. New product formulations are directed to
overcome obstacles and problems during processing and storage of blackberry juice. Conventional heat
processing of fruit juice remains the most widely used technology for ensuring shelf-life extension and
preservation of fruit juice. Thermal degradation of bioactive compounds, especially anthocyanins, is one
of the main issues related to the stability of bioactive compounds. Prevention of thermal degradation of
anthocyanins in blackberry juice could be achieved through addition of sugars, especially trehalose or
glucose [24] and plant extracts [42]. These compounds were found to have a positive effect on antho-
cyanins during storage [24,43,44]. Conventional thermal treatments are causing significant reduction of
anthocyanins, but in order to avoid their degradation, high pressure treatment [38], sonication [43,44],
or membrane processes (microfiltration and ultrafiltration) [17,21,33] could be used for juice p roduction.
The combined effects of ultrasound and osmosonication can also be applied on blackberry juice to
reduce lactic acid bacterium, yeasts, and molds without changing the antioxidant capacity, anthocyanins,
ellagitannins, and color [45].
Novel formulations of blackberry juice could also be directed to improve the quality of raw material.
Preharvest application of methyl jasmonate [46] and treatment [47] of the blackberry showed that black-
berries had a higher antioxidant capacity and flavonoid content. Extracts of treated fruits showed an
enhanced inhibition of A549 cell and HL-60 cell proliferation and induced apoptosis of HL-60 cells [46].
Frozen storage of blackberry coated with sugars, especially glucose, could be used to increase retention
of anthocyanins, phenols, and antioxidant activity [48].
Future trends should also include characterization and determination of bioavailability of such com-
pounds in the human body, in addition to the retention of bioactive compounds. It is well known that all
bioactive compounds do not have the same effect after digestion, thus their real impact on human health
should be determined. Different patterns of hydroxylation and glycosylation in anthocyanins can also
modulate their antioxidant activity, thus exerting an influence on human health.
11.6 Conclusion
The blackberry is a very perishable fruit, thus it is necessary to process it into different products such as
juice, jam, and jelly. Blackberry juice is a good source of bioactive compounds and has a high antioxidant
capacity. The bioactive compounds, mostly anthocyanins (cyanidin-3-O-glucoside) and ellagitannins,
both individually and/or synergistically may help protect against CVD, cancer, inflammation, and other
chronic diseases. Due to its beneficial effects on human health, it is necessary to protect the bioactive
compounds and prevent their degradation that can decrease their antioxidant activity. Novel product for-
mulations are becoming more important every day. Researchers are directed to reduce the degradation
of bioactive compounds and/or improve their stability during processing through the application of non-
thermal processes or the addition of some stabilizing ingredients. Nevertheless, for the future, attention
should be paid not only to the retention and/or improvement of bioactive compounds in blackberry juice,
but also toward the bioavailability of those compounds and their characterization, and the true beneficial
potential on human health.
REFERENCES
1. Jennings, D.L., Blackberries and related fruits, in Encyclopedia of Food Science and Nutrition,
Caballero, B., Ed., Academic Press, Amsterdam, the Netherlands, 2003, pp. 546–550.
2. Pantelidis, G.E., Vasilakakis, M., Manganaris, G.A., and Diamantidis, G., Antioxidant activity, phenol,
anthocyanin and ascorbic acid contents in raspberries, blackberries, red currents, gooseberries, and
Cornelian cherries. Food Chem., 102, 777–783, 2007.
3. Moyer, R.A., Hummer, K.E., Finn, C.E., Frei, B., and Wrolstad, R.E., Anthocyanins, phenolics, and
antioxidant capacity, in diverse small fruits: Vaccinium, Rubus, and Ribes. J. Agric. Food Chem.,
50, 519–525, 2002.
4. Siriwoharn, T., Wrolstad, R.E., Finn, C.E., and Pereira, C.B., Influence of cultivar, maturity, and
sampling on blackberry (Rubus L. Hybrids) anthocyanins, polyphenolics, and antioxidant properties.
J. Agric. Food Chem., 52, 8021–8030, 2004.
5. FAO Corporate Document Repository, Principles and practices of small- and medium-scale fruit
juice processing. Published online at: http://www.fao.org/docrep/005/y2515e/y2515e16.htm (accessed
April 28, 2014).
6. Hager, T.J., Howard, L.R., and Prior, R.L., Processing and storage effects on monomeric anthocyanins,
percent polymeric color, and antioxidant capacity of processed blackberry products. J. Agric. Food
Chem., 56, 689–695, 2008.
7. Gancel, A.-L., Feneuil, A., Acosta, O., Pérez, A.M., and Vaillant, F., Impact of industrial processing
and storage on major polyphenols and the antioxidant capacity of tropical highland blackberry (Rubus
adenotrichus). Food Res. Int., 44, 2243–2251, 2011.
8. Hager, T.J., Howard, L.R., and Prior, R.L., Processing and storage effects on the ellagitannins composi-
tion of processed blackberry products. J. Agric. Food Chem., 58, 11749–11754, 2010.
9. Tate, P., Kuzmar, A., Smith, S.W., Wedge, D.E., and Larcom, L.L., Comparative effects of eight varieties
of blackberry mutagenesis. Nutr. Res., 23, 971–979, 2003.
Release 26, 2013. Published online at: http://www.nal.usda.gov/fnic/foodcomp/search/ (accessed May
26, 2014).
11. Kopjar, M., Bilić, B., and Piližota, V., Anthocyanins, phenols, and antioxidant activity in blackberry
juice with plant extracts addition during heating. Acta Alim., 43, 333–343, 2014.
12. Kopjar, M., Jakšetić, K., and Piližota, V., Influence of sugars and chlorogenic acid addition on antho-
cyanin content, antioxidant activity and color of blackberry juice during storage. J. Food Proc. Pres.,
36, 545–552, 2012.
13. Wang, S.Y. and Lin, H., Antioxidant activity in fruits and leaves of blackberry, raspberry, and straw-
berry varies with cultivar and developmental stage. J. Agric. Food Chem., 48, 140–146, 2000.
14. Fan-Chiang, H.-J. and Wrolstad, R.E., Anthocyanin pigment composition of blackberries. J. Food Sci.,
70, C198–C202, 2005.
15. Sapers, G.M., Hicks, K.B., Burgher, A.M., Hargrave, D.L., Sondey, S.M., and Bilyk, A., Anthocyanin
patterns in ripening thornless blackberries. J. Am. Soc. Hortic. Sci., 111, 945–950, 1986.
16. Jakobek, L., Seruga, M., Medvidovic-Kosanovic, M., and Novak, I., Anthocyanin content and antioxi-
dant activity of various red fruit juices. Deut. Lebensm. Rundsch., 103, 58–64, 2007.
17. Acosta, O., Vaillant F., Pérez A.M., and Dornier, M., Potential of ultrafiltaration for separation and
purification of ellagitannins in blackberry (Rubus adenotrichus Schitdl.) juice. Sep. Purif. Technol.,
125, 120–125, 2014.
18. Hassimotto, N.M.A., Pinto, M.D.S., and Lajolo, F.M., Antioxidant status in humans after consump-
tion of blackberry (Rubus fruticosus L.) juices with and without defatted milk. J. Agric. Food Chem.,
56, 11727–11733, 2008.
19. Serraino, I., Dugo, L., Dugo, P., Mondello, L., Mazzon, E., Dugo, G., Caputi, A.P., and Cuzzocrea S.,
Protective effects of cyanidin-3-O-glucoside from blackberry extract against peroxynitrite-induced
endothelial dysfunction and vascular failure. Life Sci., 73, 1097–1114, 2003.
20. Sadilova, E., Carle, R., and Stintzing, F.C., Thermal degradation of anthocyanins and its impact on color
and in vitro antioxidant capacity. Mol. Nutr. Food Res., 51, 1461–1471, 2007.
21. Zhang, Z., Pang, X., Ji, Z., and Jiang, Y., Role of anthocyanin degradation in litchi pericarp browning.
Food Chem., 75, 217–221, 2001.
22. Kader, F., Haluk, J.P., Nicolas, J.P., and Metche, M., Degradation of cyanidin 3-glucoside by blueberry
polyphenol oxidase: Kinetic studies and mechanisms. J. Agric. Food Chem., 46, 3060–3065, 1998.
23. Wang, W.-D. and Xu, S.-Y., Degradation kinetics of anthocyanins in blackberry juice and concentrate.
J. Food Eng., 82, 271–275, 2007.
24. Kopjar, M. and Piližota, V., Prevention of thermal degradation of anthocyanins in blackberry juice with
addition of different sugars. CyTA—J. Food, 9, 237–242, 2011.
25. Jiménez, N., Bohuon, P., Lima, J., Dornier, M., Vaillant, F., and Pérez A.M., Kinetics of anthocyanin
degradation and browning in reconstituted blackberry juice treated at high temperatures (100°C–180°C).
J. Agric. Food Chem., 58, 2314–2322, 2010.
26. Cisse, M., Vaillant, F., Dornier, M., Acosta, O., and Dhuique-Mayer, C., Thermal degradation kinetics
of anthocyanins from blood orange, blackberry and roselle, using Arrhenius, Eyring, and Ball models.
J. Agric. Food Chem., 57, 6285–6291, 2009.
27. Debicki-Pospisil, J., Lovric, T., Trinajstic, N., and Sabljic. A., Anthocyanin degradation in the presence
of furfural and 5-hydroxymethylfurfural. J. Food Sci., 48, 411–416, 1983.
28. Azofeifa, G., Quesada, S., and Pérez, A.-M., Effect of the microfiltration process on antioxidant activity
and lipid peroxidation protection capacity of blackberry juice. Rev. Bras. Farmacogn., 21, 829–834,
2011.
29. Viljanen, K., Halmos, A.L., Sinclair, A., and Heinonen, M., Effect of blackberry and raspberry juice on
whey protein emulsion stability. Eur. Food Res. Technol., 221, 602–609, 2005.
30. Boivin, D., Blanchette, M., Barrette, S., Moghrabi, A., and Béliveau, R., Inhibition of cancer cell pro-
liferation and suppression of TNF-induced activation of NFkB by edible berry juice. Antican. Res.,
27, 937–948, 2007.
31. Agha, F.G., Fyiad, A.A., and El Safi, H., Comparative study of the protective role of black berry juice and
silymarin against liver damage induced by carbon tetrachloride in rats. J. Appl. Sci. Res., 8, 727–738,
2012.
32. Hassan, H.A. and Yousef, M.I., Mitigating effects of antioxidant properties of black berry juice on
sodium fluoride induced hepatotoxicity and oxidative stress in rats. Food Chem. Toxicol., 47, 2332–2337,
2009.
33. Calvo-Castro, L., Syed, D.N., Chamcheu, J.C., Vilela, F.M.P., Pérez, A.M., Vaillant, F., Rojas, M., and
Mukhtar, H., Protective effect of tropical highland blackberry juice (Rubus adenotrichos Schltdl.)
against UVB-mediated damage in human epidermal keratinocytes and in a reconstituted skin equiva-
lent model. Photochem. Photobiol., 89, 1199–1207, 2013.
34. García-Alonso, J., Ros, G., Vidal-Guera, M.L., and Periago M.J., Acute intake of phenolic-rich juice
improves antioxidant status in healthy subjects. Nutr. Res., 26, 330–339, 2006.
35. Netzel, M., Strass, G., Kaul, C., Bitch, I., Dietrich, M., and Bitch, R., In vivo antioxidant capacity of a
composite berry juice. Food Res. Int., 35, 213–216, 2002.
36. Spormann, T.M., Albert, F.W., Rath, T., Dietrich, H., Will, F., Stockis, J.-P., Eisenbrand, G., and
Janzowski, C., Anthocyanin/polyphenolic-rich fruit juice reduces oxidative cell damage in an interven-
tion study with patients on hemodialysis. Cancer Epidemiol. Biomarkers Prev., 17, 3372–3380, 2008.
37. Yang, H., Hewes, D., Salaheen, S., Federman, C., and Biswas, D., Effects of blackberry juice on growth
inhibition of foodborne pathogens and growth promotion of Lactobacillus. Food Cont., 37, 15–20, 2014.
38. Patras, A., Brunton, N.P., Da Pieve, S., and Butler, F., Impact of high pressure processing on total anti-
oxidant activity, phenolic, ascorbic acid, anthocyanin content and colour of strawberry and blackberry
purées. Innov. Food Sci. Emerg. Technol., 10, 308–313, 2009.
39. Lawless, L.J.R., Threlfall, R.T., Howard, L.R., and Meullenet, J.-F., Sensory, compositional, and color
properties of nutraceutical-rich juice blends. Am. J. Enol. Vitic., 63, 529–537, 2012.
40. Lawless, L.J.R., Threlfall, R.T., Meullenet, J.-F., and Howard, L.R., Consumer-based optimization of
blackberry, blueberry and concord juice blends. J. Sens. Stud., 27, 439–450, 2012.
41. Vazquez-Araujo, L., Chambers, E., Adhikari, K., and Carbonell-Barrachina, A.A., Sensory and
physicochemical characterization of juices made with pomegranate and blueberries, blackberries, or
raspberries. J. Food Sci., 75, S398–S404, 2010.
42. Kopjar, M., Bilić, B., and Piližota, V., Influence of different extracts addition on total phenols, anthocy-
anin content and antioxidant activity of blackberry juice during storage. Croat. J. Food Sci. Technol.,
3, 9–15, 2011.
43. Tiwari, B.K., O’Donnell, C.P., and Cullen, P.J., Effect of sonication on retention of anthocyanins in
blackberry juice. J. Food Eng., 93, 166–171, 2009.
44. Tiwari, B.K., O’Donnell, C.P., Muthukumarappan, K., and Cullen, P.J., Anthocyanin and colour degra-
dation in ozone treated blackberry juice. Innov. Food Sci. Emerg. Technol., 10, 70–75, 2009.
45. Wong, E., Vaillant, F., and Pérez A., Osmosonication of blackberry juice: Impact on selected pathogens,
spoilage microorganisms, and main quality parameters. J. Food Sci., 75, M468–M474, 2010.
46. Wang, S.Y., Bowman, L., and Ding, M., Methyl jasmonate enhances antioxidant activity and flavonoid
content in blackberries (Rubus sp.) and promotes antiproliferation of human cancer cells. Food Chem.,
107, 1261–1269, 2008.
47. Chanjirakul, K., Wang, S.Y., Wang, C.Y., and Siriphanich, J., Natural volatile treatments increase free-
radical scavenging capacity of strawberries and blackberries. J. Sci. Food Agric., 87, 1463–1472, 2007.
48. Kopjar, M., Nedić Tiban, N., Piližota, V., and Babić, J., Stability of anthocyanins, phenols and free
radical scavenging activity through sugar addition during frozen storage of blackberries. J. Food Proc.
Pres., 33, 1–11, 2009.
12
Black Currant Juice
Bradley W. Bolling, Derek A. Martin, Ruisong Pei, Liyang Xie, and Diana M. DiMarco
CONTENTS
12.1 Introduction....................................................................................................................................147
12.2 Nutritional Characteristics.............................................................................................................148
12.3.1 Polyphenol Content Reported in the Literature................................................................149
12.3.2 Polyphenol Content Reported in Nutrient Databanks.......................................................151
12.3.3 Antioxidant Efficacy of Bioactives in Black Currant Juice..............................................151
12.3.4 Preharvest Effects on Bioactives.......................................................................................152
12.3.5 Postharvest Effects on Bioactives......................................................................................152
12.3.6 Bioavailability and Biotransformation of Polyphenols from Black Currant Juice...........153
12.4 Health Effects.................................................................................................................................153
12.4.1 Preclinical Evidence for Black Currant Juice Functionality.............................................153
12.4.2 Clinical Evidence for Functionality of Black Currant Juice............................................ 154
12.5.1 Black Currant Juice Blends.............................................................................................. 156
12.5.2 Use of By-Products from Juice Making........................................................................... 157
12.5.3 Novel Processing Methods............................................................................................... 157
12.6 Conclusion..................................................................................................................................... 158
References............................................................................................................................................... 158
12.1 Introduction
Black currants (Ribes nigrum) are cultivated primarily in Europe and Russia, but also in New Zealand,
China, Australia, and North America. Most of the black currant crop is processed into juice or concen-
trate and distributed worldwide. Black currant juice is an established product in European markets and
its presence in the United States and elsewhere is growing. It is marketed for its refreshing taste and
potential health benefits. The benefits attributed to black currant juice consumption include supporting
healthy aging, cardiovascular health, and vision. These claims have growing plausibility based on pre-
clinical and clinical studies. Furthermore, black currant juice is nutrient dense and rich in antioxidant
polyphenols.
Data from epidemiological studies support the health benefits of consuming polyphenol-rich foods.
Increased flavonoid consumption is associated with reduced risk of chronic disease including diabetes,
cardiovascular disease (CVD), and some cancers. Berries and berry products are an important component
of polyphenol intake, as they contribute to more than 30% of cyanidin intake in central and northern
European countries [1]. As a subcategory, fruits and fruit juices contributed 1.7%–13.4% of anthocyani-
din consumption in Europe [1]. In the United States, the median anthocyanin consumption of the lowest
population quartile was 4.7 and 22.3 mg/day in the highest quartile [2]. Therefore, even consuming a
modest 237 mL (8 oz) serving of black currant juice, which contains ~200 mg anthocyanins, has potential
to increase an individual’s anthocyanin intake to the highest quartile of intake.
147
In addition to its dietary polyphenols, black currant juice also has vitamins, minerals, and oligosac-
charides that may contribute to its health-promoting potential. This chapter highlights the nutritional
and phytochemical content of black currant juice, its antioxidant capacity, the preclinical and clinical
evidence for supporting human health, as well as novel formulations that may improve this functionality.

Black currant juice is desirable for its high content of both vitamin C and potassium (Table 12.1). In the
United States, black currant juice could be considered a good source of potassium, since juices may pro-
vide 350–665 mg potassium per serving. The vitamin C content of freshly prepared black currant juices
were as high as 403 mg/100 mL juice and 2.0–2.4 mmol/kg in nectar [3,4]. After 6 months of storage,
black currant juice may lose up to 78%–80% of its initial vitamin C content [5]. Therefore, many black
currant juices are fortified with vitamin C. The average concentration of vitamin C in black currant
beverages prepared from concentrates was 125–148 mg/100 mL, whereas commercial European black
currant juices had 17–115 mg/100 mL [6]. Higher vitamin C content in black currant juices was achieved
by fortification [6].
Black currant juice is acidic and astringent. Polyphenols contribute to astringency, while acidity of the
juice is mainly from its citric acid content [7]. Most ready-to-drink black currant juices and concentrates
contain added sugars to improve palatability. Black currant juices or nectars may therefore contain up
to 27 g of carbohydrate per serving (Table 12.1). Black currants naturally contain fructose and glucose,
TABLE 12.1
Nutrient Content of Black Currant and Its Commercial Juice and Nectar
Nutrient Unit Black Currant (per 100 g) [63] Juice or Nectar (per 237 mL, 8 fl oz) [64–67]
Energy kcal 63 11–100
Protein g 1.4 0
Lipid (fat) g 0.41 0
Carbohydrate g 15.38 1.3–27
Total dietary fiber g 4 <1
Minerals
Calcium mg 28–55 20–25
Copper mg nr nr
Iron mg 0.33–1.54 nr
Magnesium mg 13–24 7–10
Manganese mg 0.17 nr
Phosphorus mg 40–59 nr
Potassium mg 240–322 310–500
Sodium mg 2 <20
Zinc mg 0.27 nr
Vitamins
Niacin mg 0.3–0.59 nr
Riboflavin mg 0.03–0.05 nr
Thiamin mg 0.05 nr
Vitamin A IU <10 <60
Vitamin B12 μg 0 nr
Vitamin B6 mg 0.066 nr
Vitamin C mg 181 (mean) 25–80
Vitamin E mg 1 nr
Abbreviation: nr, not reported.
Black Currant Juice 149
ranging from 39% to 54% and 35% to 45% of total sugars, respectively [7]. Black currants contain
4 g fiber/100 g primarily as lignins, hemicelluloses, and other polysaccharides that are not retained in
significant quantities in commercial juices [8,9]. The use of pectinases during processing may contribute
small amounts of pectins into the juice [10].
Much of the vitamin and mineral content in black currant juice does not appear to have been exten-
sively analyzed, but as the berries contain small amounts of B vitamins, iron, magnesium, and phosphorus,
it is likely that many of these are present in the juice as well. The majority of protein, tocopherols, and
fatty acids are retained in the pomace [8,9]. Black currant seed oil is extracted for its high content of
γ-linolenic acid and fatty acid profile, particularly its favorable n-6/n-3 fatty acid ratio of less than 4:1
[11,12]. Other trace nutrients found in the press cake that may enter the juice in small quantities include
protein and phytosterols [11–13].

12.3.1 Polyphenol Content Reported in the Literature
The major polyphenol classes in black currants are anthocyanins, hydroxycinnamic acids, flavonols,
and proanthocyanins. Nonspecific methods such as total phenols, anthocyanins by pH differential, and
specific methods such as high-performance liquid chromatography (HPLC) coupled with ultraviolet–
visible or mass spectroscopy or gas chromatography have been used to characterize black currant poly-
phenols. The polyphenol content of black currant juice reported in the literature varies considerably
(Table 12.2). This variation can be attributed to differences in methodology (e.g., hydrolyzing polyphe-
nols prior to analysis), as well as variability introduced by currant cultivars, cultivation technique, or
processing methods, which are discussed further in the succeeding text. Reports are generally consistent
that in black currant juice, anthocyanins are the most abundant phenolics, followed by proanthocyanins,
phenolic acids, and flavonols.
TABLE 12.2
Polyphenol Content of Black Currant Juices Reported in the Literature (mg/100 mL)
Polyphenol Commercial Samples (<100% Juice) Laboratory Prepared References
Anthocyanins
Delphinidin 3-O-rutinoside 35.0 9.4–210 [18,49]
Cyanidin 3-O-rutinoside 34.9 6.0–100 [18,49]
Delphinidin 3-O-glucoside 11.3 5.5–73 [18,49]
Cyanidin 3-O-glucoside 6.53 0.9–28 [18,49]
Phenolic Acids
p-Coumaric acid 2.5–3.2 0.9 [17,18]
Caffeic acid 2.1–2.6 0.42 [17,18]
Ferulic acid 0.8 1.66 [18,19]
Protocatechuic acid 1.08 nr [18]
Gallic acid 1.39 nr [18]
Flavonols
Myricetin 3.2 0.5–8.7 [18,49,58]
Quercetin 1.6 0.3–2.7 [18,49,58]
Kaempferol 0.4 0.3–0.5 [18,19,61]
Proanthocyanidins 7.3–59 59.2 [6,58]
Total Anthocyanins 17.0–133 23.0–340 [6,29,49]
Total Phenolic Acids 7.9–21 1.4–5.1 [6,49]
Total Flavonols 5.3–30 0.9–11.3 [6,18,49,58]
Total Phenolics 37.5–243 84.5–415.6 [6,18,49,58]
OH OH
OH HO
OH
HO
HO O O O
HO O+ HO O
R1 OH O O
HO
HO OH
R2
OH O-Glucoside O-Rutinoside
Anthocyanins R1 R2
Delphinidin-3-O-glucoside OH O-Glucoside
Delphinidin-3-O-rutinoside OH O-Rutinoside
Cyanidin-3-O-glucoside H O-Glucoside
Cyanidin-3-O-rutinoside H O-Rutinoside
O O
R1 R1
OH OH
HO HO
R2 R2
Hydroxycinnamic acids R1 R2 Hydroxybenzoic acids R1 R2

p-Coumaric acid H H p-Hydroxybenzoic acid H H
Caffeic acid OH H Protocatechuic acid H OH
Ferulic acid OCH3 H Gallic acid OH OH
R1 OH
OH OH
HO O HO O
R2 R1
OH R2
R3
OH O OH
Flavan-3-ols R1 R2 R3
Flavonols R1 R2 (+)-Catechin H H OH
Myricetin OH OH (–)-Epicatechin H OH H
Quercetin OH H (+)-Gallocatechin OH H OH
Kaempferol H H (–)-Epigallocatechin OH OH H
FIGURE 12.1 Major polyphenols identified in black currant juice.
Anthocyanins account for up to 80% of the total polyphenols in black currants [14,15]. The main
anthocyanins in black currants are identified as the glucoside and rutinoside of delphinidin and cyani-
din (Figure 12.1). Other less abundant anthocyanins, such as delphinidin-3-O-galactoside, delphinidin-
3-O-(6″-p-coumaroyl) glucoside, and petunidin, malvidin, and peonidin conjugates are also found in
black currant fruits [14,15]. However, these anthocyanins are not generally reported in juice. The major
phenolic acids in black currant juice include caffeic, p-coumaric, ferulic, gallic, p-hydroxybenzoic, and
protocatechuic acids [16–18]. Flavonols in black currant juice are mainly glucosides and rutinosides of
myricetin, quercetin, and kaempferol [15,18,19]. The proanthocyanins in black currant juice are primar-
ily B-type procyanidins and prodelphinidins, which are oligomeric compounds of both (epi)catechin and
(epi)gallocatechin subunits with average degree of polymerization ranging from 10 to 16 [5,18].
TABLE 12.3
Polyphenol Content of Black Currant Juice and Related Products (mg/100 mL)
Polyphenol Juice (n = 2–4) [68] Concentrate (n = 1) [68] Wine (n = 1) [69]
Cyanidin 29.76 110.4 nr
Delphinidin 45.27 201.28 nr
Kaempferol nr nr 0.03
Myricetin 1.86 20.85 1.39
Proanthocyanidins nr nr nr
Quercetin 1.15 22.85 0.84
12.3.2 Polyphenol Content Reported in Nutrient Databanks

The polyphenol content of many foods is documented in nutrient databanks such as the USDA fl avonoid
and proanthocyanidin databases and Phenol-Explorer. These values are useful references for epidemio-
logical studies that investigate the association between the consumption of foods and health. However,
values for black currant juice presently included in these nutrient databanks are incomplete compared
to literature reports (Table 12.3). For example, the phenolic acids and proanthocyanidin content are
currently not included for these databases. Furthermore, black currant juice is not included in the Phenol-
Explorer database. Thus, studies that have used these data likely underestimate the contribution of black
currant juice to polyphenol intake.
12.3.3 Antioxidant Efficacy of Bioactives in Black Currant Juice

Black currant juice has potent antioxidant properties in vitro, mainly related to its polyphenol content.
The 2,2,-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of fresh, pressed black currant
juice from 13 cultivars was correlated to total phenols values (r 2 = 0.78) [20]. Likewise, trolox equivalent
antioxidant capacity (TEAC) values of black currant juices from 23 cultivars were significantly cor-
related to total anthocyanins (r 2 = 0.56) and total phenols (r 2 = 0.84), but not ascorbic acid content [21].
Black currant juice also inhibits lipid oxidation and inhibited peroxidation of liposomes prepared from
soybean and had additive antioxidant activity with α-tocopherol [22]. Black currant juice also prolonged
copper-catalyzed human low-density lipoprotein (LDL) oxidation lag time by 2.6–3.6 times at a dose
of 2.5 µM total phenols [23]. When black currant juice was incubated with rat intestinal mucosa, LDL
antioxidant capacity was diminished 33% from the juice, attributable to the degradation of anthocyanins
[23]. Therefore, the metabolism and stability of black currant phenols is important to consider when
evaluating its potential antioxidant capacity.
A yeast-based cell system was also used to assess black currant juice antioxidant activity. Black currant
juice at 0.5% of cell suspension media inhibited ~30% intracellular reactive oxygen species (ROS) in
Saccharomyces cerevisiae ZIM 2155 determined by 2,7-dichlorofluorescein acetate reactivity [24].
Cells were found to internalize 29% of flavonols, 53% of hydroxycinnamic acids, and 65% of antho-
cyanidins [24]. Comparing berry juices, the quantity of polyphenols incorporated was not related to
reductions in ROS.
The antioxidant efficacy of black currant polyphenols has been mainly studied in hydroalcoholic or
acetone extracts from whole berries, rather than juices. Results from these studies give further insight
into the antioxidant activity of black currant juice. Acidified aqueous acetone extracts of six black currant cul-
tivars were assessed for proanthocyanidins, anthocyanin, oxygen radical absorbance capacity (ORAC)
values, and total phenols [25]. Black currant proanthocyanidins were highly polymerized and did not
contribute as highly to ORAC values as did anthocyanins [25]. Borges et al. [15] reported the contribu-
tion of individual polyphenols of acidified methanol extracts of black currant by coupling HPLC analysis
to ferric-reducing antioxidant power (FRAP) values. Black currant anthocyanins contributed 78% to
FRAP values, with lesser contributions by ascorbic acid and flavonols [15]. Analysis of a phosphate
buffer extract of black currant polyphenol classes found that black currant anthocyanins contributed to
more than 55% of FRAP value [26].
Taken together, anthocyanins appear to be the main contributor to the antioxidant potential of black
currant juice. Other polyphenols and ascorbic acid contribute less to total antioxidant capacity values.
Further work is needed to investigate the antioxidant interaction between polyphenol components, as
anthocyanins, neutral phenols, and ascorbic acid have synergies and antagonism in other polyphenol-rich
fruit juices [27].
12.3.4 Preharvest Effects on Bioactives

Before juicing, a number of preharvest conditions can impact the content of antioxidants and phyto-
chemicals present in black currant. Cultivar selection, climate (rainfall, sunlight, and temperature), soil
condition, insect damage, irrigation, and pesticide use all have potential to affect the bioactive content
of berries and may affect the polyphenol content of juices. For example, a comprehensive study of juices
prepared from 23 black currant cultivars grown over 3 years in Germany found significant preharvest
differences in bioactive content [20]. Anthocyanin content of black currants increased during ripening,
and higher temperatures during fruit development were associated with increased antioxidant capacity,
total phenols, and anthocyanins in juices [20]. Among different black currant cultivars, juice ascorbic
acid varied up to 4.4-fold and anthocyanin by 3.4-fold. However, cultivar variation had less impact on
antioxidant activity than polyphenol content, with only a 2.1-fold variation of TEAC values.
Planting location and climate can also lead to differences in black currant bioactives. Black currant
anthocyanins, flavonols, and total phenols were greater in berries grown at lower latitudes in Finland
[28]. However, hydroxycinnamic acids were increased in berries at higher latitudes in two currant culti-
vars [28]. The effect of climate on individual phytochemicals ranged from 8.9% to 38% and was related
primarily to temperature and radiation, with a smaller influence of humidity [28]. In a similar study,
berries from north Finland had 13% more ascorbic acid than those from the south [7]. Temperature and
humidity were associated with differences in ascorbic acid in only one of three black currant cultivars
[7]. Within a single growing season in Finland, cultivation site had a greater impact on polyphenol con-
tent than organic or conventional methods of production. Polyphenol differences in berries were mostly
influenced by anthocyanin, flavonol, and phenolic acid content, which could indicate a difference in fruit
maturity at harvest of black currants [14].
On the basis of these studies, it may be concluded that cultivar selection has a larger impact than
planting location, weather, and organic or conventional production on black currant polyphenol content.
The suitability of underutilized currant cultivars with high polyphenol content for juice making requires
further examination.
12.3.5 Postharvest Effects on Bioactives

Juice processing can be used to increase the polyphenol content of black currant juices. Enzymatic
clarification of pectins generally increases polyphenol content, discussed in more depth in the succeed-
ing text. A marketplace analysis of anthocyanins in commercial black currant juices, jams, and cordials
found significantly (P < 0.05) less anthocyanins than in fresh and frozen berries [29]. Excessive heat
treatment during pasteurization and bottling can cause thermal degradation of polyphenols, which has
been well characterized by Woodward et al. [30], and others. However, relatively little anthocyanin
content is lost during the industrial juicing process [30]. The lower content in juice than fresh berries
can be attributed to inefficient recovery of polyphenols from the fruit skin, rather than degradation of
polyphenols.
Long-term storage of fresh black currant juice generally decreases polyphenol content. Total phenols
of black currant juice declined during refrigerated storage but later increased by 29 days to a level similar
to the initial phenolic content [31]. Casati et al. [32] reported that ~40% of phenolics and 56% of TEAC
values were lost during 30 days of storage of black currant juice. Similar to other anthocyanin-rich fruit
juices, TEAC values of black currant juice were half of initial values following 29 days of storage at 4°C
[31]. Likewise, ascorbic acid content was negligible by 17 days of storage in unfortified black currant
juice [31]. In contrast, frozen storage may increase or have minimal impact on DPPH antioxidant activ-
ity. Storage of juice for 1 year at −18°C led to increased DPPH radical scavenging activity in juices from
several cultivars [20].
12.3.6 Bioavailability and Biotransformation of Polyphenols from Black Currant Juice

The polyphenols in black currant juice are bioavailable and detectable in plasma and urine following
consumption. Anthocyanins, the most abundant polyphenols in black currant juice, have low bioavail-
ability. For example, less than 0.1% of anthocyanins were recovered from the urine of adults within 4 h
after intake of 1.2 g of anthocyanins from black currant juice [33]. Similarly, only 0.036% of anthocyanins
were recovered in urine within 48 h of adults consuming 300 g of a black currant drink containing
33.6 mg anthocyanins [29]. Likewise, the bioavailability of flavonols from dietary levels of black currant
juice is low. Flavonols or their metabolites were not detected when adults consumed black currant juice
containing 16 mg of flavonols [18]. In contrast, urinary quercetin excretion was 0.29%–0.47% of consump-
tion within 24 h after intake of 4.8–9.6 mg quercetin in a black currant and apple juice mixture [34].
Black currant polyphenols are metabolized by deconjugation of sugar moieties followed by glucuroni-
dation, sulfation, or methylation. The main anthocyanins from black currant juice are found in plasma
and urine after consumption, with similar rates of absorption and excretion of delphinidins and cyani-
dins [33]. The major metabolites of black currant quercetin are the methoxylated metabolites isorham-
netin and tamarixetin [35]. Following black currant juice consumption, isorhamnetin was about three
times more abundant than quercetin and tamarixetin in plasma [35]. These intact metabolites were only
less than 1% of the total bioactive compounds found in the juice. Other than these intact metabolites,
bioactives may also be metabolized to other phenolic metabolites, although the mechanism is still not
completely clear.
Hippuric acid and its derivatives were identified as the major phenolic metabolites after black c urrant
juice consumption [18,29]. Other phenolic acids, including dihydroxybenzoic acid sulfate, dihydroxy-
phenylacetic acid sulfate, catechol sulfate, ferulic acid sulfate, and caffeic acid sulfate derivatives, were
also reported [18]. These phenolic metabolites may be derived from the B-ring of anthocyanidins, fla-
vonols, or proanthocyanidins following C-ring fission. Cinnamic acid sulfate derivatives, a phenolic
metabolite derived from flavonoid A-rings, were also reported after black currant juice consumption [18].
These phenolic metabolites may account for an additional 4.68% of anthocyanin bioavailability [29].
Thus, the bioavailability of the bioactives may be higher than previously thought.
Black currant polyphenol bioavailability may also be affected by concurrent food consumption,
although this area is underexplored. Several studies have characterized the impact of fiber and sugar on
the absorption of black currant juice anthocyanins and quercetin. Consumption of black currant juice
with a rice cake, which is rich in fiber and sugar, had no effect on the total anthocyanins found in the
plasma, although the tmax was shifted from 45 to 90 min [33]. Similarly, a rice cake consumed with black
currant juice had no impact on the total quercetin content of plasma or the tmax value of quercetin [36].
12.4 Health Effects

12.4.1 Preclinical Evidence for Black Currant Juice Functionality
Black currant juice has demonstrated preclinical functionality, including improving vascular function,
antioxidant function, and anticarcinogenic activity (Table 12.4).
In a rat model of hypertension, drinking water was replaced with black currant juice containing
39.9 mg anthocyanins, 0.1 mg hydroxybenzoic acids, 4.5 mg hydroxycinnamic acids, 6.5 mg flavonols,
and 4.4 mg/proanthocyanidins per 100 mL for 8 weeks [37]. At the end of the intervention period,
plasma levels of angiotensin II were 50% lower in the intervention group than in the control group,
but blood pressure was not different between groups. Asymmetric dimethylarginine, total nitrate/
nitrite, 8-isoprostanes, and C-reactive protein (CRP) were unaffected by black currant juice treatment.
The mRNA expression of cyclooxygenase-2 in animals consuming black currant juice was about half
TABLE 12.4
Selected Preclinical Studies on the Bioactivity of Black Currant Juice
Preclinical Outcomes References
Reduced angiotensin II in hypertensive rats [37]
Modulated cholesterol metabolism and antioxidant activity in rabbits [38]
Reduced PUFA-induced oxidative stress in pigs [39]
Anticarcinogenic activity in vitro [40]
Reduced hepatocarcinomas in rats [41]
Immunostimulatory activity in vitro [42]
Inhibited tumor growth in mice [42]
Abbreviation: PUFA, polyunsaturated fatty acids.
that of control animals, but angiotensin converting enzyme, monocyte chemotactic protein-1, P-selectin,
adiponectin, and vascular cell adhesion molecule-1 (VCAM-1) were unaffected [37].
Male and female Watanabe heritable hyperlipidemic rabbits were fed purified black currant anthocya-
nins (0.1%) for 16 weeks, while another group received black currant juice instead of drinking water,
containing 58 mg anthocyanins/100 mL [38]. The anthocyanin-fed group had increased plasma total
cholesterol and LDL compared to all other groups at 4, 12, and 16 weeks. In contrast, black currant
juice did not change total cholesterol or triacylglycerols (TAG) and lowered very-low-density lipoprotein
(VLDL) cholesterol. Hepatic malondialdehyde (MDA) was unaffected by either dietary treatment, but
superoxide dismutase activity was increased in both currant groups, and glutathione peroxidase activity
was increased in the anthocyanin-supplemented animals [38]. Thus, black currant juice improves hepatic
antioxidant function and lipoprotein profile in Watanabe heritable hyperlipidemic rabbits. Due to the
differential effects of isolated black currant anthocyanins on VLDL metabolism, other juice components
may modulate this function.
Black currant juice intake also improves antioxidant defenses in pigs. Male pigs were overfed and sup-
plemented with linseed oil to induce oxidative stress and were supplemented with 42 mg vitamin E or
300 g black currant juice daily for 2 weeks [39]. Black currant juice was as effective as vitamin E
supplementation in reducing leukocyte DNA damage as assessed by the comet assay, although it did
not significantly lower (P > 0.05) plasma MDA or 24 h urinary MDA excretion compared to the control
group [39].
Black currant juice inhibits growth of stomach, intestine, prostate, and breast cancer cell lines [40].
The anticarcinogenic activities of black currant have been corroborated in various rodent models. In
a rat model of induced hepatocarcinoma, extracts from black currant skins at 100 or 500 mg/kg body
weight were incorporated into the diet [41]. The highest dose of black currant skin extract significantly
reduced total nodule incidence and multiplicity. Both treatments improved hepatocellular architecture,
with the highest dose showing normal hepatocellular characteristics [41]. Additionally, black currant
skin extract‒treated rats showed decreased hepatic cell proliferation and increased apoptosis [41]. Black
currant polysaccharides also have promising immunostimulatory and anticarcinogenic activity in vitro.
Coincubation of a black currant polysaccharide stimulated the production of interleukin (IL)-1β, tumor
necrosis factor-alpha (TNF-α), IL-12 p70, and nitric oxide (NO) from thioglycollate-elicited murine
macrophages [42]. Administration of black currant juice and isolated black currant polysaccharides
inhibited Ehrlich carcinoma tumor growth in mice by 45% and 51%, respectively [42].
12.4.2 Clinical Evidence for Functionality of Black Currant Juice

A number of human intervention studies have reported improved outcomes following the consumption
of black currant juice or extracts (Table 12.5). Positive outcomes include an improvement in plasma
markers of antioxidant and vascular function, intraocular pressure, and reduced biomarkers related to
kidney stone formation.
In a study of six males consuming 250 mL black currant juice every day for 1 week, there were no
changes (P > 0.05) in serum lipid parameters, glucose, or oxidative stress as measured by the thiobarbituric
TABLE 12.5
Clinical Evidence for the Health Effects of Black Currant Juice
Intervention Outcomes References
330 mL black currant juice × 5 days Alkalinized urine, increased citric acid, and oxalic acid excretion. [47]
475–1000 mL black currant juice or Increased plasma vitamin C levels and increased. [44]
black currant anthocyanin drink/ formamidopyrimidine DNA glycosylase–sensitive site DNA
day × 3 weeks damage.
17 mg/kg black currant juice Increased forearm blood flow at rest; during typing work, [45]
extract, acute increased oxygenated hemoglobin and prevented significant
increase in trapezius muscle stiffness seen in a placebo group.
240 mg powdered black currant Reduced postexercise plasma carbonyl levels, plasma ROS [46]
anthocyanins, 120 mg before and generating capacity, and creatine kinase levels; reduced TNF-α
after exercise and IL-6 secretion from LPS-stimulated postexercise blood.
250 mL black currant juice/day × Increased PON-1 activity 20%, increased serum sulfhydryl groups [43]
1 week 11%, no changes in lipids, glucose, or other measures of
oxidative stress.
250 mL 20% black currant juice, No change of vascular reactivity, concentrations of ICAM-1 or [16]
acute VCAM-1, plasma FRAP, or ORAC compared to control beverage.
50 mg/day black currant Decreased intraocular pressure relative to baseline at 2 and [48]
anthocyanins × 4 weeks 4 weeks.
50 mg/day black currant Reduced intraocular pressure and protected visual field [48]
anthocyanins × 24 months deterioration.
Abbreviations: ROS, reactive oxygen species; TNF-α, tumor necrosis factor-alpha; IL-6, interleukin-6; LPS, lipopolysac-
charide; PON-1, paraoxonase-1; ICAM-1, intercellular adhesion molecule-1; VCAM-1, vascular cell adhe-
sion molecule-1; FRAP, ferric-reducing antioxidant power; ORAC, oxygen radical absorbance capacity.
acid–reactive substances (TBARS) assay [43]. However, juice consumption increased serum sulfhydryl
groups by 11% and paraoxonase (PON)-1 activity by 20% (P < 0.01) [43]. In another study, a group of 60
healthy subjects consumed 475–1000 mL (~400 mg anthocyanin/day) black currant juice with vitamin C
or a black currant anthocyanin drink for 3 weeks [44]. Mononuclear blood cell DNA damage was low in
participants, but damage to formamidopyrimidine DNA glycosylase–sensitive sites increased slightly in
the black currant juice group relative to the control beverage, which was accompanied by a greater vita-
min C level [44]. Further studies have not confirmed increased oxidative damage following black currant
juice consumption, or if black currant polyphenols and vitamin C are pro-oxidative in vivo.
Acute vascular reactivity was unchanged in 20 healthy participants after consumption of 250 mL of
20% black currant juice compared to a control beverage [16]. Plasma concentrations of intercellular
adhesion molecule-1 (ICAM-1) and VCAM-1 tended to fall with consumption of the black currant juice
and the control beverage, but there was not a significant difference (P > 0.05) between the two [16].
Additionally, there was no effect on plasma antioxidant capacity as assessed by the FRAP and ORAC
assays [16]. Another of study of vascular function in small numbers of healthy adults (n = 9–11) found
improved vascular function [45]. Consumption of black currant juice extract providing 17 mg anthocy-
anin/kg body mass significantly increased forearm blood flow [45]. After 2 h, forearm blood flow was
increased relative to a placebo. Further, during typing work, oxygenated hemoglobin was significantly
higher after 7.7 mg/kg intake of the extract [45]. Black currant extract consumption also prevented a
significant increase in the trapezius muscle stiffness that was observed in the placebo group after
typing work. Despite these changes, there were no significant differences in blood pressure or heart rate
between consumption of the black currant extract and the placebo [45].
Consumption of a commercial black currant extract consisting of 240 mg anthocyanins in four
capsules consumed around a 30 min acute exercise bout at 80% VO2max reduced postexercise plasma
carbonyl levels, ROS generating capacity, and creatine kinase activity levels [46]. Exercise induced a
reduction in TNF-α and IL-6 secretion from lipopolysaccharide-stimulated peripheral blood, and black
currant supplementation exaggerated these effects [46]. Further study of the exercise-related functional
outcomes is needed to determine if these changes affect performance or recovery.
Black currant juice has also reduced risk factors associated with kidney stone formation in men.
Healthy men (n = 12) that consumed 330 mL black currant juice daily for 5 days resulted in an alkaliza-
tion of urine from pH 6.35–6.56 [47]. Citric acid and oxalic acid excretion were increased by 25% and
22%, respectively, following consumption of black currant juice [47]. Due to its alkalizing effect, black
currant juice may be effective in the treatment of uric acid urolithiasis [47].
Consumption of black currant anthocyanins also decreases intraocular pressure, a risk factor of glau-
coma. In a crossover study, healthy adults (n = 12) consuming 50 mg black currant anthocyanins daily
for 4 weeks had decreased intraocular pressure relative to baseline at 2 and 4 weeks with no changes in
intraocular pressure during a placebo period [48]. In another parallel-design trial, adults with glaucoma
consuming 50 mg black currant anthocyanins daily for 24 months had reduced intraocular pressure at
the end of the study period, whereas the placebo group did not [48]. The placebo group experienced dete-
rioration in the visual field at 12 and 18 months, but there were no changes in the intervention group [48].
To summarize, interventions have mainly tested dietary-relevant doses of black currant juice, typically
providing 50–240 mg anthocyanins or 250 mL or more of juice. Nearly, all studies have been performed
in healthy individuals, so further work is needed to assess the functionality of black currant juice in
populations at risk for chronic disease. A limited number of studies have evaluated the bioavailability of
black currant juice polyphenols. Further work is needed to determine how polyphenol bioavailability
affects treatment efficacy.

Black currants are rarely consumed fresh and are processed to juice, jams, syrups, concentrates, extracts,
wines, and other products. Black currant juice may be blended or consumed alone, but fresh black cur-
rant juice is perceived as sour and astringent [49]. By-products from processing black currants contain
functional components and may be further processed into functional products and ingredients.
12.5.1 Black Currant Juice Blends

Black currant juice is sold as 100% juice, pulp-containing or clarified nectars, or blended with other
juices or carbonated water. Concentration of black currant juice is used to produce a stabilized product
that can be used for further blending. Evaporative concentration, membrane, and freeze concentration
have been used to concentrate black currant juice. Freeze concentration can produce up to 39°Brix con-
centrates while maintaining the original aroma and fruit flavor of black currant juice [50]. Membrane
concentration produced a 58.2°Brix product following the preconcentration of juice by reverse osmosis
[51]. Membrane concentration of black currant juice is potentially economically feasible on an industrial
scale provided a moderate capital investment [52].
Black currant juice and other fruit juices can be used as vehicles to deliver probiotics. Probiotics may
improve intestinal and general health by increasing immune function, lowering cholesterol, and reduc-
ing cancer risk. Several studies have reported that black currant juice is an appropriate vehicle to deliver
probiotics. Bifidobacterium longum NCIMB 8809 was stable in commercial black currant juice, losing
0.7 log at 6 weeks when held at 4°C [53]. Citric acid, protein, and dietary fiber enhanced Bifidobacterium
survival, while fruit phenolics may have inhibited survival [53]. Likewise, Lactobacillus plantarum with
black currant juice powder was more stable through a 12-month storage and reconstitution than straw-
berry, pomegranate, and cranberry powders, losing less than 0.5 log from initial values [54]. Therefore,
both dried and refrigerated black currant juices appear to be appropriate vehicles to deliver probiotics.
Given the commercial availability of acid and heat-stable probiotics, other probiotic strains could also be
used to formulate novel black currant juice products.
Other additives can enhance the functionality of black currant juice. For example, a carbonated black
currant juice drink formulated with 410 mg calcium/L was less erosive to tooth enamel than a conven-
tional carbonated orange control drink [55]. Adding powdered crowberry (Empetrum nigrum) to black
currant juice increased juice polyphenols [18]. Further, the crowberry/black currant blend improved
postprandial glycemic response of ~36 g sugar dose, which corresponded to increased bioavailability of
polyphenols [18]. Consumption of a juice blend of pineapple, black currant, and plum prevented increases
in the proinflammatory cytokines IL-17, IL-6, and TNF-α in healthy overweight individuals consuming
a high-fat meal [56]. In a crossover study of 48 patients with peripheral arterial disease, consuming an
orange and black currant juice blend for 1 month reduced the inflammatory biomarkers CRP and fibrino-
gen relative to a control drink [57].
Black currant is often blended with apple juice. Beyond the additive effect of black currant polyphe-
nols, black currant improves the yield of apple polyphenols during processing. Crushed black currants
were ground with apples at 1:5 (w/w), treated with pectinase, pressed, and further heat treated [58]. The
addition of black currant juice increased apple juice flavonols by 16% and chlorogenic acid by 50%,
likely by inhibiting polyphenol oxidase. However, as reported with other studies, storage in glass bottles
led to significant losses of monomeric anthocyanins by 6 months [58]. Therefore, further attention is
needed to retain black currant anthocyanins during storage in black currant juice blends.
A 7-day intake of 1.5 L of blended black currant and apple juices increased plasma ascorbic acid and
decreased plasma MDA levels in five healthy individuals, whereas lower doses did not [34]. Further, the
same dose reduced erythrocyte superoxide dismutase and catalase and increased plasma 2-aminoadipic
semialdehyde, a marker of lipid oxidation, indicating potential pro-oxidant activities [34].
Black currant juice may also be blended with whey to create functional beverages. Whey provides
protein and antioxidant peptides, as well as additional vitamins and minerals. Black currant and black
currant–whey beverages were evaluated for nutrient and polyphenol content and consumer acceptance
over 12 months of storage [5]. Beverages were prepared from a blend of black currant juice concentrate
and black currant extract. Sensory analysis rated black currant–whey beverages overall as better than
“very good” after production and declining to “very good” after storage up to 12 months [5]. Notably,
black currant odor declined during storage [5]. The whey-blended beverage was also perceived as less
sweet and having less characteristic black currant flavor intensity. In contrast, black currant beverages
prepared from extract and concentrated juices were rated as “excellent.” Whey blends could be further
developed as functional black currant beverages [5].
12.5.2 Use of By-Products from Juice Making

Following black currant juicing, residual press cake containing polyphenols and other currant bioactives
is available for reclamation. Pressed, thawed berries lost 90% of their sugar content in juice but retained
the majority of polyphenol compounds [19]. Nearly all polyphenols remaining in the black currant press
cake could be extracted by ethanol, resulting in an astringent extract with weaker black currant flavors
than its juice [19]. Likewise, press cake obtained from an industrial process was rich in anthocyanins and
flavonols [59]. Black currant press cake can also be used as a source of insoluble dietary fiber. Enzymatic
treatments yielded variable levels of pectic sugars and mannose, with the latter being related to the
presence of seeds in press cake [60]. Therefore, black currant could be used as a source of dietary fiber,
polyphenols, and flavoring agents.
12.5.3 Novel Processing Methods

Processing methods can increase the yield of functional components of black currant during juice mak-
ing. Black currant anthocyanins remain stable during commercial juice processing stages, including
pressing, milling, and pectin hydrolysis [30]. Sodium bisulfite addition also increases recovery of antho-
cyanins [30]. Excessive heating during pasteurization can reduce anthocyanin content of black currant
juice, so short-time protocols will minimize thermal degradation of polyphenols.
Pectinases are commonly employed as a processing aid to increase black currant juice yields. Pectinase
treatment of pressed black currants increased polyphenol content of juices 4–10-fold and increased
organic acid content by 1.3-fold [49]. Enzyme-treated juice was more astringent and bitter than pressed
juice, due in part to the increase in phenolics, but also to the loss of pectin, which masks the sensory
contribution of polyphenols [49]. Pectinase treatment retained the sensory differences between berry cul-
tivars [49]. Laaksonen et al. [49] suggested that juice processed without pectinase would be more palat-
able and reduce the amount of sweeteners or diluents required to make black currant juice more pleasing.
Certain pectinases cleave flavonoid glycosides and rhamnosides, generating aglycones [61]. Similarly,
heat treatment increased flavonoid aglycones from black currant juice produced without pectinolysis [61].
Alternatively, α-l-rhamnosidases can be added directly during black currant juice processing to increase
anthocyanin and flavonol aglycones [62]. Thus, the black currant polyphenol profile can be shifted to
more aglycone-rich compounds and higher content based on enzyme treatment. It remains to be dem-
onstrated if this shift will impart improved functionality in vivo. A limitation to this approach may be
higher astringency associated with black currant polyphenols.
12.6 Conclusion
Black currant juice is rich in antioxidant phytochemicals, vitamin C, and a good source of potassium.
The in vitro antioxidant activity of black currant juice appears to be mainly derived from anthocyanin
content, with smaller contributions from proanthocyanidins and flavonols. The phytochemical content of
black currant juices varies considerably due to the use of different cultivars and juice processing meth-
ods. Black currant juice may be fortified with other juice blends or treated with pectinases to increase
its phytochemical content. The bioavailability of antioxidant phytochemicals has been demonstrated
in several studies. Despite apparent low bioavailability of these compounds from black currant juice, a
number of studies have demonstrated the ability of black currant juices or phytochemical consumption
to reduce biomarkers of inflammation and oxidative stress in both healthy individuals and those with
chronic disease. To date, these studies have mainly employed small numbers of individuals, so further
efforts are needed to study the efficacy in broader populations, including those at higher risk for chronic
disease. Preclinical studies also show promising anticarcinogenic and immunomodulatory activity, both
of which require further testing.
REFERENCES
1. Zamora-Ros, R., Knaze, V., Luján-Barroso, L., Slimani, N., Romieu, I., Touillaud, M., Kaaks, R.
et al., Estimation of the intake of anthocyanidins and their food sources in the European Prospective
Investigation into Cancer and Nutrition (EPIC) study. Br. J. Nutr., 106, 1090–1099, 2011.
2. Wedick, N.M., Pan, A., Cassidy, A., Rimm, E.B., Sampson, L., Rosner, B., Willett, W., Hu, F.B., Sun, Q.,
and van Dam, R.M., Dietary flavonoid intakes and risk of type 2 diabetes in US men and women. Am. J.
Clin. Nutr., 95, 925–933, 2012.
3. del Castillo, M.L.R., Dobson, G., Brennan, R., and Gordon, S., Fatty acid content and juice characteris-
tics in black currant (Ribes nigrum L.) genotypes. J. Agric. Food Chem., 52, 948–952, 2004.
4. Iversen, C.K., Black currant nectar: Effect of processing and storage on anthocyanin and ascorbic acid
content. J. Food Sci., 64, 37–41, 1999.
5. Jaworska, G., Sady, M., Grega, T., Bernas, E., and Pogon, K., Qualitative comparison of blackcurrant
and blackcurrant-whey beverages. Food Sci. Technol. Int., 17, 331–341, 2011.
6. Mattila, P.H., Hellstrom, J., McDougall, G., Dobson, G., Pihlava, J.M., Tiirikka, T., Stewart, D., and
Karjalainen, R., Polyphenol and vitamin C contents in European commercial blackcurrant juice
products. Food Chem., 127, 1216–1223, 2011.
7. Zheng, J., Yang, B., Tuomasjukka, S., Ou, S., and Kallio, H., Effects of latitude and weather conditions
on contents of sugars, fruit acids, and ascorbic acid in black currant (Ribes nigrum L.) juice. J. Agric.
Food Chem., 57, 2977–2987, 2009.
8. del Castillo, M.L.R., Dobson, G., Brennan, R., and Gordon, S., Genotypic variation in fatty acid content
of blackcurrant seeds. J. Agric. Food Chem., 50, 332–335, 2002.
9. Sojka, M. and Krol, B., Composition of industrial seedless black currant pomace. Eur. Food Res.
Technol., 228, 597–605, 2009.
10. Hilz, H., Bakx, E.J., Schols, H.A., and Voragen, A.G.J., Cell wall polysaccharides in black currants and
bilberries—Characterisation in berries, juice, and press cake. Carbohyd. Polym., 59, 477–488, 2005.
11. Bakowska-Barczak, A.M., Schieber, A., and Kolodziejczyk, P., Characterization of Canadian black
currant (Ribes nigrum L.) seed oils and residues. J. Agric. Food Chem., 57, 11528–11536, 2009.
12. Savikin, K.P., Dordevic, B.S., Ristic, M.S., Krivokuca-Dokic, D., Pljevljakusic, D.S., and Vulic, T.,
Variation in the fatty-acid content in seeds of various black, red, and white currant varieties. Chem.
Biodivers., 10, 157–165, 2013.
13. Goffman, F.D. and Galletti, S., Gamma-linolenic acid and tocopherol contents in the seed oil of 47
accessions from several Ribes species. J. Agric. Food Chem., 49, 349–354, 2001.
14. Anttonen, M.J. and Karjalainen, R.O., High-performance liquid chromatography analysis of black cur-
rant (Ribes nigrum L.) fruit phenolics grown either conventionally or organically. J. Agric. Food Chem.,
54, 7530–7538, 2006.
15. Borges, G., Degeneve, A., Mullen, W., and Crozier, A., Identification of flavonoid and phenolic antioxi-
dants in black currants, blueberries, raspberries, red currants, and cranberries. J. Agric. Food Chem.,
58, 3901–3909, 2011.
16. Jin, Y., Alimbetov, D., George, T., Gordon, M.H., and Lovegrove, J.A., A randomised trial to investigate
the effects of acute consumption of a black currant juice drink on markers of vascular reactivity and
bioavailability of anthocyanins in human subjects. Eur. J. Clin. Nutr., 65, 849–856, 2011.
17. Mattila, P., Hellström, J., and Törrönen, R., Phenolic acids in berries, fruits, and beverages. J. Agric.
Food Chem., 54, 7193–7199, 2006.
18. Törrönen, R., McDougall, G.J., Dobson, G., Stewart, D., Hellström, J., Mattila, P., Pihlava, J.-M.,
Koskela, A., and Karjalainen, R., Fortification of black currant juice with crowberry: Impact on polyphe-
nol composition, urinary phenolic metabolites, and postprandial glycemic response in healthy subjects.
J. Function. Food., 4, 746–756, 2012.
19. Sandell, M., Laaksonen, O., Jarvinen, R., Rostiala, N., Pohjanheimo, T., Tiitinen, K., and Kallio, H.,
Orosensory profiles and chemical composition of black currant (Ribes nigrum) juice and fractions of
press residue. J. Agric. Food Chem., 57, 3718–3728, 2009.
20. Djordjevic, B., Savikin, K., Zdunic, G., Jankovic, T., Vulic, T., Pljevljakusic, D., and Oparnica, C.,
Biochemical properties of the fresh and frozen black currants and juices. J. Med. Food, 16, 73–81, 2013.
21. Kruger, E., Dietrich, H., Hey, M., and Patz, C.-D., Effects of cultivar, yield, berry weight, temperature
and ripening stage on bioactive compounds of black currants. J. Appl. Bot. Food Qual., 84, 40–46, 2011.
22. Graversen, H., Becker, E., Skibsted, L., and Andersen, M., Antioxidant synergism between fruit juice
and α-tocopherol. A comparison between high phenolic black chokeberry (Aronia melanocarpa) and
high ascorbic blackcurrant (Ribes nigrum). Eur. Food Res. Technol., 226, 737–743, 2008.
23. Pinelo, M., Landbo, A.K., Vikbjerg, A.F., and Meyer, A.S., Effect of clarification techniques and rat
intestinal extract incubation on phenolic composition and antioxidant activity of black currant juice.
J. Agric. Food Chem., 54, 6564–6571, 2006.
24. Slatnar, A., Jakopic, J., Stampar, F., Veberic, R., and Jamnik, P., The effect of bioactive compounds on
in vitro and in vivo antioxidant activity of different berry juices. PLoS One 7, e47880, 2012.
25. Wu, X., Gu, L., Prior, R.L., and McKay, S., Characterization of anthocyanins and proanthocyanidins in
some cultivars of Ribes, Aronia, and Sambucus and their antioxidant capacity. J. Agric. Food Chem.,
52, 7846–7856, 2004.
26. Bordonaba, J.G. and Terry, L.A., Electrochemical behaviour of polyphenol rich fruit juices using dispos-
able screen-printed carbon electrodes: Towards a rapid sensor for antioxidant capacity and individual
antioxidants. Talanta, 90, 38–45, 2012.
27. Bolling, B.W., Chen, Y.Y., and Chen, C.Y.O., Contributions of phenolics and added vitamin C to the
antioxidant capacity of pomegranate and grape juices: Synergism and antagonism among constituents.
Int. J. Food Sci. Technol., 48, 2650–2658, 2013.
28. Zheng, J., Yang, B.R., Ruusunen, V., Laaksonen, O., Tahvonen, R., Hellsten, J., and Kallio, H.,
Compositional differences of phenolic compounds between black currant (Ribes nigrum L.) cultivars
and their response to latitude and weather conditions. J. Agric. Food Chem., 60, 6581–6593, 2012.
29. Hollands, W., Brett, G.M., Radreau, P., Saha, S., Teucher, B., Bennett, R.N., and Kroon, P.A., Processing
blackcurrants dramatically reduces the content and does not enhance the urinary yield of anthocyanins
in human subjects. Food Chem., 108, 869–878, 2008.
30. Woodward, G.M., McCarthy, D., Pham-Thanh, D., and Kay, C.D., Anthocyanins remain stable during
commercial blackcurrant juice processing. J. Food Sci., 76, S408–S414, 2011.
31. Piljac-Zegarac, J., Valek, L., Martinez, S., and Belscak, A., Fluctuations in the phenolic content and
antioxidant capacity of dark fruit juices in refreigerated storage. Food Chem., 113, 394–400, 2009.
32. Casati, C.B., Sanchez, V., Baeza, R., Magnani, N., Evelson, P., and Zamora, M.C., Relationships between
colour parameters, phenolic content, and sensory changes of processed blueberry, elderberry, and black-
currant commercial juices. Int. J. Food Sci. Technol., 47, 1728–1736, 2012.
33. Nielsen, I.L.F., Dragsted, L.O., Ravn-Haren, G., Freese, R., and Rasmussen, S.E., Absorption and excre-
tion of black currant anthocyanins in humans and Watanabe heritable hyperlipidemic rabbits. J. Agric.
Food Chem., 51, 2813–2820, 2003.
34. Young, J.F., Nielsen, S.E., Haraldsdottir, J., Daneshvar, B., Lauridsen, S.T., Knuthsen, P., Crozier,
A., Sandstrom, B., and Dragsted, L.O., Effect of fruit juice intake on urinary quercetin excretion and
biomarkers of antioxidative status. Am. J. Clin. Nutr., 69, 87–94, 1999.
35. Breinholt, V.M., Nielsen, S.E., Knuthsen, P., Lauridsen, S.T., Daneshvar, B., and Sørensen, A., Effects
of commonly consumed fruit juices and carbohydrates on redox status and anticancer biomarkers in
female rats. Nutr. Cancer, 45, 46–52, 2003.
36. Erlund, I., Freese, R., Marniemi, J., Hakala, P., and Alfthan, G., Bioavailability of quercetin from b erries
and the diet. Nutr. Cancer, 54, 13–17, 2006.
37. Kivimaki, A.S., Ehlers, P.I., Siltari, A., Turpeinen, A.M., Vapaatalo, H., and Korpela, R., Lingonberry,
cranberry, and blackcurrant juices affect mRNA expressions of inflammatory and atherothrombotic
markers of SHR in a long-term treatment. J. Funct. Food, 4, 496–503, 2012.
38. Nielsen, I.L.F., Rasmussen, S.E., Mortensen, A., Ravn-Haren, G., Ma, H.P., Knuthsen, P., Hansen, B.F.
et al., Anthocyanins increase low-density lipoprotein and plasma cholesterol and do not reduce athero-
sclerosis in Watanabe heritable hyperlipidemic rabbits. Mol. Nutr. Food Res., 49, 301–308, 2005.
39. Salobir, J., Zontar, T.P., Levart, A., and Rezar, V., The comparison of black currant juice and vitamin E
for the prevention of oxidative stress. Int. J. Vit. Nutr. Res., 80, 5–11, 2010.
40. Boivin, D., Blanchette, M., Barrette, S., Moghrabi, A., and Beliveau, R., Inhibition of cancer cell pro-
liferation and suppression of TNF-induced activation of NF kappa B by edible berry juice. Anticancer
Res., 27, 937–948, 2007.
41. Bishayee, A., Mbimba, T., Thoppil, R.J., Haznagy-Radnai, E., Sipos, P., Darvesh, A.S., Folkesson,
H.G., and Hohmann, J., Anthocyanin-rich black currant (Ribes nigrum L.) extract affords chemopreven-
tion against diethylnitrosamine-induced hepatocellular carcinogenesis in rats. J. Nutr. Biochem., 22,
1035–1046, 2011.
42. Takata, R., Yamamoto, R., Yanai, T., Konno, T., and Okubo, T., Immunostimulatory effects of a
polysaccharide-rich substance with antitumor activity isolated from black currant (Ribes nigrum L.).
Biosci. Biotech. Bioch., 69, 2042–2050, 2005.
43. Rosenblat, M., Volkova, N., Attias, J., Mahamid, R., and Aviram, M., Consumption of polyphenolic-rich
beverages (mostly pomegranate and black currant juices) by healthy subjects for a short term increased
serum antioxidant status, and the serum’s ability to attenuate macrophage cholesterol accumulation.
Food Funct., 1, 99–109, 2010.
44. Moller, P., Loft, S., Alfthan, G., and Freese, R., Oxidative DNA damage in circulating mononuclear
blood cells after ingestion of blackcurrant juice or anthocyanin-rich drink. Mutat. Res. Fund. Mol. M.,
551, 119–126., 2004.
45. Matsumoto, H., Takenami, E., Iwasaki-Kurashige, K., Osada, T., Katsumura, T., and Hamaoka, T.,
Effects of blackcurrant anthocyanin intake on peripheral muscle circulation during typing work in
humans. Eur. J. Appl. Physiol., 94, 36–45, 2005.
46. Lyall, K.A., Hurst, S.M., Cooney, J., Jensen, D., Lo, K., Hurst, R.D., and Stevenson, L.M., Short-term
blackcurrant extract consumption modulates exercise-induced oxidative stress and lipopolysaccharide-
stimulated inflammatory responses. Am. J. Physiol. Reg. I, 297, R70–R81, 2009.
47. Kessler, T., Jansen, B., and Hesse, A., Effect of blackcurrant-, cranberry- and plum juice consumption
on risk factors associated with kidney stone formation. Eur. J. Clin. Nutr., 56, 1020–1023, 2002.
48. Ohguro, H., Ohguro, I., and Yagi, S., Effects of black currant anthocyanins on intraocular pressure in
healthy volunteers and patients with glaucoma. J. Ocul. Pharmacol. Ther., 29, 61–67, 2013.
49. Laaksonen, O., Mäkilä, L., Tahvonen, R., Kallio, H., and Yang, B., Sensory quality and compositional
characteristics of blackcurrant juices produced by different processes. Food Chem., 138, 2421–2429,
2013.
50. Dette, S.S. and Jansen, H., Freeze concentration of black currant juice. Chem. Eng. Technol., 33,
762–766, 2010.
51. Kozák, Á., Békássy-Molnár, E., and Vatai, G., Production of black-currant juice concentrate by using
membrane distillation. Desalination, 241, 309–314, 2009.
52. Sotoft, L.F., Christensen, K.V., Andrésen, R., and Norddahl, B., Full scale plant with membrane based
concentration of blackcurrant juice on the basis of laboratory and pilot scale tests. Chem. Eng. Proc.
Proc. Int., 54, 12–21, 2012.
53. Nualkaekul, S., Salmeron, I., and Charalampopoulos, D., Investigation of the factors influencing
the survival of Bifidobacterium longum in model acidic solutions and fruit juices. Food Chem., 129,
1037–1044, 2011.
54. Nualkaekul, S., Deepika, G., and Charalampopoulos, D., Survival of freeze dried Lactobacillus planta-
rum in instant fruit powders and reconstituted fruit juices. Food Res. Int., 48, 627–633, 2012.
55. West, N.X., Hughes, J.A., Parker, D.M., Moohan, M., and Addy, M., Development of low erosive car-
bonated fruit drinks 2. Evaluation of an experimental carbonated blackcurrant drink compared to a
conventional carbonated drink. J. Dent., 31, 361–365, 2003.
56. Peluso, I., Raguzzini, A., Villano, D.V., Cesqui, E., Toti, E., Catasta, G., and Serafini, M., High fat meal
increase of IL-17 is prevented by ingestion of fruit juice drink in healthy overweight subjects. Curr.
Pharm. Design, 18, 85–90, 2012.
57. Dalgard, C., Nielsen, F., Morrow, J.D., Enghusen-Poulsen, H., Jonung, T., Horder, M., and de Maat,
M.P.M., Supplementation with orange and black currant juice, but not vitamin E, improves inflamma-
tory markers in patients with peripheral arterial disease. Br. J. Nutr., 101, 263–269, 2009.
58. Oszmiański, J. and Wojdyło, A., Effects of black currant and apple mash blending on the phenolics
contents, antioxidant capacity, and colour of juices. Czech. J. Food Sci., 27, 338–351, 2009.
59. Sojka, M., Guyot, S., Kolodziejczyk, K., Krol, B., and Baron, A., Composition and properties of purified
phenolics preparations obtained from an extract of industrial blackcurrant (Ribes nigrum L.) pomace.
J.Hortic. Sci. Biotech., SI, 100–106, 2009.
60. Kosmala, M., Kołodziejczyk, K., Markowski, J., Mieszczakowska, M., Ginies, C., and Renard,
C.M.G.C., Co-products of black-currant and apple juice production: Hydration properties and polysac-
charide composition. LWT—Food Sci. Technol., 43, 173–180, 2010.
61. Laaksonen, O., Sandell, M., Nordlund, E., Heiniö, R.-L., Malinen, H.-L., Jaakkola, M., and Kallio, H.,
The effect of enzymatic treatment on blackcurrant (Ribes nigrum) juice flavour and its stability. Food
Chem., 130, 31–41, 2012.
62. Gonzalez-Barrio, R., Trindade, L.M., Manzanares, P., de Graaff, L.H., Tomas-Barberan, F.A., and
Espin, J.C., Production of bioavailable flavonoid glucosides in fruit juices and green tea by use of fungal
alpha-l-rhamnosidases. J. Agric. Food Chem., 52, 6136–6142, 2004.
Release 25, National Technical Information Service, USDA, Springfield, VA, 2013.
64. New Zealand Blackcurrant Cooperative, Report 766402. Published online at: http://www.nzblackcur-
rants.com/assets/Uploads/July-07-Agriqual-vitamin-results-190707.pdf (accessed April 1, 2014).
65. New Zealand Blackcurrant Cooperative, Mineral Report. Published online at: http://www.nzblackcur-
rants.com/assets/pdfs/18.7.07-Minerals-report-summary-612429-TMP-1-2.pdf (accessed April 1, 2014).
66. Lucozade Ribena Suntory Limited, Ribena Blackcurrant Squash. Published online at: http://www.
ribena.co.uk/products/original/ (accessed April 1, 2014).
67. Knudsen & Sons Inc., Organic Black Currant Nectar. Published online at: http://www.rwknudsenfamily.
com/products/organic-juices/organic-black-currant-nectar (accessed April 1, 2014).
68. Bhagwat, S., Haytowitx, D.B., and Holden, J.M., USDA Database for the Flavonoid Content of Selected
Foods, Release 3.1, USDA, Springfield, VA, 2013.
69. Rothwell, J.A., Urpi-Sarda, M., Boto-Ordoñez, M., Knox, C., Llorach, R., Eisner, R., Cruz, J. et al.,
Phenol-Explorer 2.0: A major update of the Phenol-Explorer database integrating data on polyphenol
metabolism and pharmacokinetics in humans and experimental animals. Database, 31, 2012.
13
Blueberry Juice
William L. Kerr
CONTENTS
13.1 Introduction....................................................................................................................................163
13.3.1 Phytochemicals in Blueberries..........................................................................................165
13.3.2 Blueberry Juice and Processing........................................................................................167
13.3.3 Anthocyanins in Blueberry Juice......................................................................................167
13.3.4 Bioactivity of Anthocyanins..............................................................................................167
13.4 Health Effects.................................................................................................................................168
13.4.1 Vision Improvement..........................................................................................................168
13.4.2 Neurocognition..................................................................................................................168
13.4.3 Urinary Tract Infections....................................................................................................169
13.4.4 Cancer................................................................................................................................169
13.4.5 Cardiovascular Disease.....................................................................................................169
13.6 Conclusion......................................................................................................................................171
References................................................................................................................................................171
13.1 Introduction
The blueberry is a member of the Vaccinium family of berries, which includes the cranberry, bilberry,
and crowberry. Recent estimates are that over 427,000 metric tons (MT) of blueberries are produced
worldwide. Most (295,000 MT) are grown in North America, especially in the United States and Canada
[1]. South America, including Chile, Argentina, and Uruguay, has been developing blueberry production
with some 70,000 MT. Blueberries are also grown in Europe, Australia, and Asia.
Blueberries have a limited harvest period. In North America, the season lasts from May to late July,
extended somewhat by early- and late-blooming cultivars. In the southern hemisphere, the growing sea-
son can be longer. Thus, while fresh market berries are still desirable, there is considerable demand for
processed blueberries. In the United States, for example, about two-thirds are further processed [1].
Of these, about half are quick frozen, while the remaining is turned into juices, purees, or dried fruit. The
perception that blueberries, and other berries, can provide health benefits has helped drive the demand
for processed products, and several blueberry juices and juice blends are becoming available on the
market. Thus, major brands including Minute Maid, Ocean Spray, and Odwalla all have products featur-
ing blueberry juice. This chapter highlights the nutritional and bioactive components of blueberry juice
and how these are affected by formulation and processing.
163

As with many other fruits, blueberries contain a variety of compounds beneficial to human health
(Table 13.1). Among the macronutrients, blueberries have less than 1% fat or protein and are rela-
tively low in caloric value (~84 kcal/cup) [2]. The bulk of the fruit is water (~84%), and approximately
15% of the berry is carbohydrate. A one-cup serving provides 3.6 g of total dietary fiber (TDF) or
10%–18% of the 20–35 g/day recommended for healthy adults. Some variations arise due to culti-
var. For example, lowbush varieties were found to have 4.1–4.4 g/100 g TDF [3], compared to the
TABLE 13.1
Compositional and Nutritional Characteristics of Whole Blueberry and Beverages with Blueberry Juice
Whole Dynamic RW Trader Ocean Odwalla Lakewood
Unit Blueberrya Healthb Knudsenc Joesd Spraye Blueberryf Organicg
Serving Size 148 g 240 mL 240 mL 240 mL 240 mL 330 mL 240 mL
Energy kcal 57 110 100 150 110 210 100
Protein g 0.74 0 0 1 0 0 1
Lipid (fat) g 0.33 0 0 0 0 0 0
Carbohydrate g 14.49 27 24 36 28 50 22
Total sugars g 9.96 26 18 31 28 34 16
Total dietary fiber g 2.4 na na 0 na 1 4
Minerals
Calcium mg 6 na 12 0 na na 12
Iron mg 0.28 na 0.28 0 na na 0.72
Magnesium mg 6 na na na na na 12
Manganese mg 0.336 na na na na na 0.84
Phosphorus mg 12 na na na na na 24
Potassium mg 77 na 180 na 40 530 na
Sodium mg 1 5 10 na 35 15 na
Zinc mg 0.16 na na na na na 0.32
Vitamins
Folate μg 6 na na na na 24.0 16
Niacin mg 0.418 na na na na 10.4 0.64
Pantothenic acid mg 0.124 na na na na 4.0 na
Riboflavin mg 0.041 na na na na 6.8 0.078
Thiamine mg 0.037 na na na na 6.3 0.072
Vitamin A IU 54 na na 0 na na 36
Vitamin B 6 mg 0.052 na na na na 9.0 0.10
Vitamin C mg 9.7 na na 0 80 31.5 18
Vitamin E mg 0.57 na na na na na 0.9
Vitamin K μg 19.3 na na na na na 60
a Adapted from U.S. Department of Agriculture (USDA), USDA National Nutrient Database for Standard Reference,
Release 26, 2013, Published online at: http://www.ars.usda.gov/ba/bhnrc/ndl (accessed April 22, 2014).
b Pure concentrate from organically grown blueberries not reconstituted.
c,d Filtered water (sufficient to reconstitute), blueberry juice concentrate.
e Filtered water, cane sugar, blueberry juice from concentrate, grape juice from concentrate, apple juice from concentrate,
malic acid, natural flavors, ascorbic acid, and sodium citrate.

f Odwalla blueberry B-orange juice, blueberry puree, mango puree, banana puree, apple juice, concord grape juice,
vitamin C, niacinamide, vitamin B6, vitamin B2, calcium pantothenate, vitamin B1, biotin, and vitamin B12. Calculated
from % daily value.
g Juice and puree (fiber) from whole ripe organic blueberries.

Blueberry Juice 165
2.4 g/100 g reported by the USDA. The types of fiber vary with cultivar and maturity, and the total
fiber decreases when the berry fully ripens.
Blueberry juice, particularly without pulp, has little dietary fiber (Table 13.1). Indeed, part of the juice
production may involve enzymes that help dissolve cell wall materials and improve yield, and the juice
may be filtered or clarified to remove the fiber-rich pulp. In some cases, however, the pulp may be left
in the juice as with high-pulp or smoothie-type beverages. In this case, some brands of blueberry juice
report up to 4 g of dietary fiber per serving (Table 13.1).
Ripe blueberries contain fructose (32.9% dry weight), glucose (32.9%), and sucrose (1.8%), although
these vary with variety and the ratio of glucose/fructose increases during ripening. During juicing, most
of the sugars are expressed with the juice. Most beverages are formulated to have 8%–12% soluble sugars
(°Brix) for the best flavor. The sweet flavor is balanced by various organic acids [4]. Lowbush varieties
contain citric acid (36%), malic acid (31%), and quinic acid (20%), while highbush blueberries contain
citric acid (75%) and succinic acid (17%) [5].
Blueberries contain a variety of micronutrients but are particularly rich in vitamin C (9.7 mg/serving),
vitamin K (19.3 μg/serving), and to a lesser extent vitamin B6 (0.052 mg/serving) and vitamin E (0.57 mg/
serving). They also contain relatively high levels of manganese (0.336 mg/serving) and lesser amounts of
iron, magnesium, phosphorous, potassium, and zinc. Being water soluble, most of the sugars, organic acids,
and water-soluble vitamins are expressed with the juice. However, nutrients may remain with the bagasse.
In orange juice, for example, only about a quarter of the available vitamin C is carried over into the juice [6].
Heat pasteurization is known to diminish vitamin C, carotenoids, and vitamin A, while lipids, miner-
als, and amino acids are not significantly affected. Vitamin C loss is minimal in low-pH products such as
fruit juices. With flash pasteurization, juice may have 95% of the ascorbic acid of fresh-squeezed juice.
Deaeration is particularly important to reduce oxygen prior to thermal processing. Bulk storage under
nitrogen, and bottling juice with limited headspace, is beneficial. Changes in vitamin C are more sig-
nificant during juice storage, if subject to high temperatures or long storage times. If refrigerated (<5°C),
only 0%–7% losses occurred during 8 weeks, but up to 30%–40% was lost at 12°C [7]. Again, it is criti-
cal to limit oxygen in the finished product to limit the aerobic degradation of vitamin C.
The nutritional content of blueberry juice, juice blends, and nectars found on the market varies with
manufacturer and formulation (Table 13.1). Pure juices that include the pulp retain much of the nutri-
ent content of fresh blueberries. Clarified juices may have limited amounts of dietary fiber, vitamin C,
vitamin K, and manganese. Blueberry, grape, and apple (or other fruits) juices may be combined with
water, sugars, organic acids, and ascorbic acid to produce juice blends and nectars with a vitamin C
content similar to fresh juice. In other cases, blueberry and other fruit purees are combined to produce
pulpy smoothie-type drinks. These may have added vitamins and minerals, sometimes at levels much
greater than the recommended dietary intake levels. Such products are usually positioned as better-for-
you, nutrient-dense products.

13.3.1 Phytochemicals in Blueberries
Blueberry juice contains a variety of phytochemicals with antioxidant and other bioactivity. Blueberries
contain several phenolic compounds including flavonols, hydroxycinnamic acids, flavon-3-ols, and
procyanidins (Table 13.2). Of particular importance are the plant pigments known as anthocyanins, poly-
phenolic compounds in the class of flavonoids (Figure 13.1). While often in the glycosylated form, they
may also exist as sugar-free anthocyanidins. These reside within cell vacuoles and give rise to a variety
of colors in Vaccinium spp. and similar fruits such as blackberry, raspberry, and cherry. Colors vary as
red, violet, and blue depending on the type of anthocyanin, pH, and the presence of copigments. In blue-
berry, most of the anthocyanins reside in the skin, concentrated in the epidermal layer. Thus, while most
blueberries appear as deep blue, when opened they have an off-white fleshy fruit within. Blueberry skins
have the highest level of anthocyanins, polyphenolics, and antioxidant activity, while the flesh and seeds
have lesser amounts of all polyphenolic compounds [10].
TABLE 13.2
Phytochemicals Found in Blueberry Juice
Class Compound mg/100 g Fresh Weight References
Anthocyanidins Cyanidin 4.79–28.72 [8]
Delphinidin 20.82–47.37 [8]
Malvidin 32.9–69.44 [8]
Peonidin 1.01–19.37 [8]
Petunidin 7.19–18.25 [8]
Flavon-3-ols (−)-Epicatechin 1.11 [8]
Flavonols Myricetin 0.0–2.60 [8]
Quercetin 1.70–7.30 [8]
Proanthocyanidins Monomers 2.07–5.58 [8]
Dimers 1.66–9.48 [8]
Trimers 0.73–7.37 [8]
4–6mers 15.75–26.04 [8]
7–10mers 10.99–17.40 [8]
Polymers 58.37–200.6 [8]
Phenolic acids p-Hydroxybenzoic acid 103.67 [9]
Caffeic acid 1.38–6.32 [9]
p-Coumaric acid 0.40–15.78 [9]
Stilbenes Resveratrol 0.011–1.61 [9]
Pterostilbene 0.148–0.226 [9]
Piceatannol 0.207–0.293 [9]
OH OCH3
OH OH
+ +
HO O HO O
OCH3
Glycoside Glycoside
HO HO
Cyanidin-3-glycoside Malvinidin-3-glycoside
OCH3 OH
OH OH
+ +
HO O HO O
OH OH
Glycoside Glycoside
HO HO
Petunidin-3-glycoside Delphinidin-3-glycoside
OCH3
OH OH
+ +
HO O HO O
Glycoside Glycoside
HO HO
Peonidin-3-glycoside Peonidin-3-glycoside
FIGURE 13.1 Major anthocyanins found in blueberry juice. The attached sugar may include glucoside, galactoside, or
arabiniside forms.
Blueberry Juice 167
13.3.2 Blueberry Juice and Processing

The antioxidant activity of blueberry juice may be significantly diminished by processing [11]. A substantial
amount of anthocyanins may be lost during crushing, enzyme treatments, and pressing. First, the typical
juice yield is 75%–83%, so that not all of the soluble materials are extracted. For blueberries containing
99.9 mg/100 g anthocyanins, only 33.6 mg/100 g were recovered in the pressed juice, while 184 mg/100 g
remained in the press cake, showing that anthocyanins are not as easily extracted as sugars, acids, and
other water-soluble compounds. Less than 22% of anthocyanins in frozen blueberries were recovered in the
pressed juice. Greater anthocyanin levels were measured after pasteurization, possibly as thermal inactiva-
tion of enzymes prevented catalyzed destruction of polyphenolics.
13.3.3 Anthocyanins in Blueberry Juice

Although in lesser amounts, blueberry juice has the same array of anthocyanins and other phenolic
compounds as in the berries from which it is made. The total amount and distribution of anthocyanins
depends on the bush variety, growing conditions, and degree of ripeness of the fruit. For example, the
anthocyanin content of “Crunchie” highbush varieties, expressed as equivalents of cyanidin-3-glucoside
(C3G), was 120 mg C3G/100 g fresh weight, while “Brightwell” varieties had 153 mg C3G/100 g and
wild varieties as much as 558 mg C3G/100 g [12,13]. In addition to overall varietal differences, smaller
berries tend to have more anthocyanins per unit of mass, as there is more fruit skin as compared to
the flesh.
Most blueberries contain delphinidin, malvidin, petunidin, peonidin, and cyanidin, which are attached
to glucose, galactose, arabinose, rhamnose, and xylose [14]. The sugars may also be modified by acyla-
tion with acetic, p-coumaric, caffeic, and several other acids. Most blueberries have higher levels of
delphinidin and malvidin than petunidin, cyanidin, and peonidin; and some highbush varieties contain
pelargonidin [15]. Highbush varieties often have malvidin, delphinidin, and petunidin present in both
3-galactoside and 3-arabinoside forms. Some researchers have reported as many as 49 distinct anthocya-
nins in wild blueberries.
13.3.4 Bioactivity of Anthocyanins

Anthocyanins from blueberry and many other plant materials have been investigated for their ability
to affect human health. It is expected that any impact on health would be related to bioactivity of these
compounds. Most of the focus has been on the antioxidant properties of polyphenolics and how these
affect human tissues and organs. A common theory is that free radicals contribute to cell damage, and the
decreased ability to combat oxidative stress contributes to cell aging, cancer, cardiovascular disease (CVD), and
cognitive impairment. In addition, protection against oxidative stress may limit inflammation of tissues.
Anthocyanins rank relatively high in terms of aqueous phase antioxidant activity as compared to other
polyphenolics. Many studies have examined the antioxidant and related bioactive properties of isolated
anthocyanins or extracts from plants containing anthocyanins. It was found that C3G and cyanidin could
reduce oxidation in linoleic acid models, liposome models containing a free radical initiator, erythrocyte
membranes, and rat liver microsomes [16]. Subsequent work showed that extracts containing pelargo-
nidin, cyanidin, and delphinidin derivatives could inhibit lipid oxidation and scavenge active oxygen
radicals [17]. Some studies suggest that blueberries are a particularly good source of antioxidants, in the
form of anthocyanins, with oxygen radical absorbance capacity (ORAC) values ranging from 13.9 to
44.6 mmol trolox equivalents (TE)/g fresh berries [18].
Less is known about in vivo antioxidant activity of anthocyanins and other flavonoids. In rats, black
raspberry anthocyanins were rapidly absorbed in both stomach and intestines, reaching levels of 7.5%
in the small intestinal tissue [19]. However, these did not as easily pass into the circulatory system.
In human subjects fed with blueberry powder, all anthocyanins were absorbed in their glycosylated
forms at levels between 4.53 and 16.9 ng/mL, and with a corresponding increase in serum antioxidant
capacity [20]. However, a review of several studies suggested that the bioavailability of anthocyanins
is relatively low. Doses of 150 mg to 2 g total anthocyanins, available in extracts or juices, resulted in
10–50 nmol/L in plasma, with peak levels between 1 and 3 h [21]. This may underestimate true bioavail-
ability, as various metabolites may not be measured in the assays. Glucosylated anthocyanidins are better
absorbed than galactosyl or arabinosyl versions [22].
Anthocyanidins are more bioavailable than anthocyaninins but are less stable to subsequent changes.
The food system also influences availability, absorption, and metabolism of inherent anthocyanins.
In juice, water-soluble anthocyanins are mostly present in the juice, while bound and less water-soluble
components remain with the pulp [23]. Anthocyanins were better absorbed from juice than from aqueous
solutions of purified anthocyanins [24]. Some studies suggest that the presence of a solid matrix helps
slow the absorption of anthocyanins [25].
In addition to general antioxidant activity, anthocyanins can interact with very specific biochemi-
cals and structures in cells and tissues, so as to limit inflammation. One theory is that anthocyanins
reduce inflammation and atherosclerosis by interaction with nitric oxide (NO), a signaling molecule that
inhibits vascular smooth muscle contraction. Anthocyanin-rich berry extracts prevented in vitro loss of
endothelium-dependent NO-mediated relaxation in porcine arteries, likely due to their ability to reduce
reactive oxygen species [26]. Blueberry and cranberry anthocyanins were localized in microvascular
endothelial cells, reducing their vulnerability to oxidative stress [27]. The berry anthocyanins reduced
tumor necrosis factor-α and induced upregulation of inflammatory mediators that direct leukocytes to
the sites of damage along the endothelium. Other studies showed that tannins and anthocyanins, and
particularly delphinidin, induced endothelium-dependent relaxation in rat aorta [28].
Anthocyanins may also interact with cytokines, a family of signaling molecules that are involved
with inflammation and immune function. In one study, several red wine anthocyanins were found in the
plasma of human volunteers between 0.5 and 2.5 h after ingestion [29]. The antioxidant capacity of the
plasma increased, and there was a decrease in circulating chemokines involved with recruiting white
blood cells. Anthocyanins from bilberries inhibited nuclear factor kappa-light-chain-enhancer of acti-
vated B, which reduces proinflammatory mediators in adults [30]. In runners given 250 g of blueberries
per day, there were increases in natural killer cells and interleukins in the plasma and decreased levels of
F2-isoprostanes and nucleic acid oxidation products [31]. They concluded that blueberry supplementation
helped decrease oxidative stress and increase anti-inflammatory cytokines. There is also evidence that
some anthocyanins reduce the production of cell adhesion molecules. These proteins are expressed at
the cell surface and help attract leukocytes to the site of inflammation. Blackberry anthocyanins reduced
inflammation in rat lung model, reducing intracellular adhesion molecules and the formation of prosta-
glandins [32].
13.4 Health Effects

Recent studies have provided evidence that blueberry products have specific health-promoting prop-
erties. As blueberry juice does not typically provide dietary fiber or the full vitamin content of fresh
blueberries, most benefits are derived from the polyphenolic compounds extracted with the juice. Most
research has been on the benefits to vision improvement, neurocognition, preventing urinary tract
infections (UTI), and reducing the risk of cancer and CVD.
13.4.1 Vision Improvement

Several studies have indicated that blueberry and bilberry extracts help promote vision health. Rats sup-
plemented with bilberry were better able to regenerate the rhodopsin pigment in photoreceptor cells [33].
Human subjects fed with anthocyanin had improved adaptation to the dark. However, other studies show
less benefit of anthocyanins on vision health [34], and studies using blueberry juice have not been reported.
13.4.2 Neurocognition
Blueberry polyphenols have been researched for the potential to limit cognitive impairment, particu-
larly as related to aging. Blueberries reduced inflammatory mediators in brain microglia cells, which
Blueberry Juice 169
have been implicated in neurodegenerative diseases [35]. Older rats given a dried blueberry supplement
were able to navigate mazes as well as younger rats [36]. Studies on aged rats supplemented with blue-
berry powder found improved neuronal plasticity and cognitive performance related to memory and
spatial tasks [37]. Related studies showed that blueberry-fed rats not only had improved memory and
spatial learning, but also that several anthocyanins could be found in critical brain tissue [38]. Rats fed
with lowbush blueberries for 6 weeks had less brain damage when subject to restricted blood flow [39].
Biochemical studies reinforce the hypothesis that polyphenols improve neural function. A blueberry-
supplemented diet was also found to reverse the age-related decline in hippocampal heat shock proteins
define 70 (HSP70) proteins, which can help protect cells from stress damage [40]. A 6-year study on
16,000 elderly women found that the dietary intake of blueberries and strawberries, and of anthocyanins
in general, was associated with slower rates of cognitive decay [41]. In one human trial, 9 elderly adults
were given 2–2.5 cups of blueberry juice for 12 weeks and showed improved cognitive skills related to
paired associate learning and word recall [42].
13.4.3 Urinary Tract Infections

There have been several studies on the role that the cranberry, blueberry, and bilberry might play in
promoting urinary health. One theory is that proanthocyanidins in the cranberry and blueberry can limit
the ability of urinary bacteria such as Escherichia coli to attach to mucosal surfaces. Studies have shown
that blueberry and cranberry juices inhibit the production of bacterial adhesins that allow them to stick
to surfaces and colonize [43]. Other theories suggest that NO is formed that inhibits bacteria, or that fla-
vonoids reduce inflammation and thus give UTI sufferers relief from symptoms of UTI. While in vitro
studies with blueberries have been promising, clinical trials using blueberry products for the prevention
or treatment of UTI have been limited.
13.4.4 Cancer
A review of epidemiological studies has shown that regular consumption of fruits and vegetables is asso-
ciated with reduced risk of cancer. One large study indicated that higher levels of dietary flavonoids were
correlated with lower risk of lung cancer and other malignant neoplasms [44]. Anthocyanin fractions
from blueberries were able to induce apoptosis in colon carcinoma cells in a dose–response manner, with
the implication that this infers a greater ability to engage in normal programmed cell death [45]. One
study showed that pterostilbene found in blueberries suppresses aberrant crypt foci formation in colon
cancer cells of rats [46]. Protective effects were related to the ability to suppress inflammatory processes.
Proanthocyanidin fractions from blueberries were shown to inhibit the growth of some prostate cancer
cell lines, particularly those that are dependent on androgen signaling [47].
13.4.5 Cardiovascular Disease

Foods containing polyphenolic compounds have also been investigated for their ability to reduce the
risk of CVD. As clarified juice has reduced levels of vitamins and dietary fiber, most benefits would be
realized from the phenolic compounds. Several studies have indicated that polyphenols can affect CVD
biomarkers, reduce low-density lipoproteins (LDL) involved with atherosclerotic plaques, or reduce
vascular inflammation [48]. Mice fed with 1% dried blueberries had fewer atherosclerotic lesions in the
aorta and had lower biomarkers for lipid peroxidation and greater upregulating of antioxidant enzymes
[49]. Blueberries also helped reduce inflammatory cytokines inhibiting signaling processes between
the cell surface and nuclear DNA. In human trials, a 12-year study indicated that men with the high-
est berry consumption had a lower risk of CVD-related death [50]. A study of 34,489 postmenopausal
women found that there was an inverse association between anthocyanindins and coronary heart dis-
ease or CVD mortality [51]. The best outcomes were predicted when subjects consumed strawberries or
blueberries at least once a week. In an 8-week study on adults, a daily intake of a blueberry beverage
reduced several CVD risk factors including blood pressure, plasma oxidized LDL, and serum malondi-
aldehyde levels [52].

The market for blueberry juice has not been large, particularly when compared to orange, apple, and
cranberry juices. While sales in the conventional juice market have been stable, most growth is expected
in the “health and wellness” category [53]. Among the leading trends are Kosher beverages, beverages
that are “all natural” or have no additives or preservatives, and those with environmentally friendly pack-
aging. Juices with reduced sugar and fewer calories have also become popular, growing at a rate of 15%
from 2005 to 2010.
As can be seen in Table 13.1, there are several products that contain blueberry juice. The most typical are
clarified beverages, often made with filtered water and blueberry juice concentrate. These have the widest
consumer appeal. While these contain some of the original polyphenolic compounds, they are reduced
sometimes by more than 50%. In addition, much of the dietary fiber and vitamins are lost. In some cases,
vitamin C may be added back to the product. Other popular products are blends of several juices to a
single strength. Blends with blueberry and pomegranate juice are popular, although combinations with
cranberry or blackberry can also be found. While some of the products feature or highlight “blueberry”
or “pomegranate,” they may contain higher proportions of apple, pear, or grape juice. Another variation
is carbonated beverages with blueberry. These typically contain 70% fruit juice with carbonated water.
Pasteurized in a sealed can, these present as healthier alternatives to soda beverages.
Only a few manufacturers have developed products from whole berries. As noted, the fiber, nutrient, and
phytochemical content is best preserved in this way. Perhaps as the costs of blueberry juice are relatively
high as compared to apple or grape juices, these products sell at a premium. Being cloudy beverages, they
may also show signs of sedimentation over time. While objectionable to some consumers, the product is
not defective, and the cloud can be redispersed by shaking. Another popular category is smoothie-type
beverages. In addition to blueberry, they often contain banana and other purees to make a thick, smooth
product. Typical products contain orange juice, blueberry juice, banana puree, as well as other juices. It is
often common to see such products fortified with high levels of vitamins C and B complex. Thus, they are
geared toward health-conscious individuals looking for nutrient-dense products. A few energy drinks have
also been produced that incorporate blueberry juice, along with other berries, tea, calcium, and vitamin C.
In addition, these products may include caffeine, ginseng, B vitamins, and taurine to boost energy.
Many consumers are also looking for lower-calorie versions of juice products. This is most easily
realized with clarified beverages made from juice concentrates and water. Thus, the product can use
lesser amounts of sugar-containing juice and replace some of the sweetness with nonnutritive sweet-
eners. These come in “light” and “diet” varieties. Light juice drinks typically have 40–60 cal/226 g
serving. This is attained by reducing the juice content to 40%–50% and then enhancing flavor with
fruit acids and nonnutritive sweeteners. At this time, combinations of sucralose and acesulfame K are
most common for sweetening. The blending of sweeteners helps mask the defects of each individually
and better mimics the sweetness of fruit sugars. Diet juice drinks have ~5 cal/serving. While the exact
technology is proprietary, patents have described how juice can be made that is low in sugar and high in
phytochemicals. In short, juice is passed through an ultrafiltration unit to remove low-molecular-weight
sugars and acids [54]. A higher proportion of this stream can be added to a product without increasing
caloric content. The tartness and sweetness are added back through organic acids (e.g., malic, fumaric,
and citric) and sweeteners (e.g., sucralose and acesulfame K).
There has also been academic research in novel processing of blueberry and related juices. For exam-
ple, static high-pressure processing (HPP) at 200–600 MPa was examined as an alternative to heat-based
pasteurization of the juice [55]. HPP resulted in >92% retention of vitamin C and higher levels of total
phenolics. Others have found that at a given temperature, higher pressures result in greater degradation
of anthocyanins, although this was not the case in strawberry and blackberry purees [56]. Blueberry–
whey functional beverages were pasteurized by a continuous high-pressure system, resulting in a product
that was preferred over thermally processed beverages [57].
As polyphenol oxidases (PPO) are a concern in many juices, nonthermal methods of inactivation of
PPO have also been studied. One approach used dense phase CO2 at different processing pressures.
While processing at 48.3 MPa reduced PPO 40%, including 7.5% CO2 provided 35% additional decrease
Blueberry Juice 171
in PPO activity. Other studies have sought to increase the phytochemical components of berries or
extracts by exposure to ultraviolet (UV) radiation. Blueberries that were subjected to UV dosages up to
6.45 kJ/m2 had greater levels of flavonoids including resveratrol, quercetin and kaempferol glycosides,
and to a lesser extent, anthocyanins [58].
An interesting medical application for blueberry juice has also been tested. In patients who underwent
magnetic resonance imaging of the pancreatic and bile duct systems, prior administration of blueberry
juice improved image contrast [59]. An enhanced contrast was also found for imaging of the stomach and
duodenum following an the intake of 450 mL of blueberry juice.
13.6 Conclusion
There is certainly an increased demand for fruit juices and juice-based drinks, and functional beverages,
particularly those made from berries that have perceived high health benefit. Research on the health benefits
specifically derived from the consumption of blueberry juice is somewhat limited. Most studies have
been conducted on whole fruit or juices from other berries, although those that have been conducted with
blueberry juice have been fairly promising. A compositional analysis would suggest that whole blueber-
ries would provide more fiber, nutrients, and phytochemicals than the juice made from those berries.
However, blueberry and other juices still provide more nutrients than many other beverages. The trend
toward low-calorie juice drinks may also pose a paradox. While these tend to be less nutrient dense due
to lower juice levels, the consumption of less overall calories is definitely a bonus to some individuals.
REFERENCES
1. Brazelton, C., World Blueberry Acreage and Production, United States Highbush Blueberry Council-
Industry Relations Committee, Folsom, CA, 2011.
Release 26, 2013. Published online at: http://www.ars.usda.gov/ba/bhnrc/ndl (accessed April 22, 2014).
3. Chen, H.C. and Camire, M.E., Recovery of anthocyanins, pectin, and dietary fiber from cull lowbush
blueberries. J. Food Qual., 20, 199–209, 1997.
4. Kalt, W. and McDonald, J.E., Chemical composition of lowbush blueberry cultivars. J. Am. Soc. Hort.
Sci., 121, 142–146, 1996.
5. Ehlenfeldt, M.K., Meredith, F.I., and Ballington J.R., Unique organic acid profile of rabbiteye vs high-
bush blueberries. Hort. Sci., 29, 321–323, 1994.
6. Nagy S., Vitamin C contents of citrus fruit and their products: A review. J. Agric. Food Chem., 28, 8–18,
1980.
7. Bratley, C.O., Loss of ascorbic acid from tangerines during storage on the market. Proc. Am. Soc. Hort.
Sci., 37, 526–573, 1940.
8. U.S. Department of Agriculture (USDA), USDA Database for the Flavonoid Content of Selected Foods,
Release 3.0. Published online at: http://www.ars.usda.gov/SP2UserFiles/Place/12354500/Data/Flav/
Flav_R03.pdf (accessed April 26, 2014).
9. Sellappan, S., Akoh, C.C., and Krewer G., Phenolic compounds and antioxidant capacity of Georgia-
grown blueberries and blackberries. J. Agric. Food Chem., 50, 2432–2438, 2002.
10. Lee, J., The blueberry: Composition, anthocyanins, and polyphenolics, PhD thesis, Oregan State
University, Corvallis, OR, 2004.
11. Skrede, G., Wrolstad, R.E., and Durst, R.W., Changes in anthocyanins and polyphenolics during juice
processing of highbush blueberries (Vaccinium corymbosum L.). J. Food Sci., 65, 357–364, 2000.
12. Lohachoompol, V., Mulholland, M., Srzednicki, G., and Craske, J., Determination of anthocyanins in
various cultivars of highbush and rabbiteye blueberries. Food Chem., 111, 249–254, 2008.
13. Hosseinian, F.S. and Beta, T., Saskatoon and wild blueberries have higher anthocyanin contents than
other Manitoba berries. J. Agric. Food Chem., 55, 10832–10838, 2007.
14. Cho, M.J., Howard, L.R., Prior, R.L., and Clark, J.R., Flavonoid glycosides and antioxidant capacity of
various blackberry, blueberry, and red grape genotypes determined by high-performance liquid chro-
matography/mass spectrometry. J. Sci. Food Agric., 84, 1771–1782, 2004.
15. Gao, L. and Mazza, G., Quantitation and distribution of simple and acylated anthocyanins and other
phenolics in blueberries. J. Food Sci., 59, 1057–1059, 1994.
16. Tsuda, T., Watanabe, M., Ohshima, K., Norinobu, S., Choi S.-W., Kawakishi, S., and Osawa, T.,
Antioxidative activity of the anthocyanin pigments cyanidin-3-O-β-D-glucoside and cyanidin. J. Agric.
Food Chem., 42, 2407–2410, 1994.
17. Tsuda, T., Shiga, K., Ohshima, K., Kawakishi, S., and Osawa, T., Inhibition of lipid peroxidation and
the active oxygen radical scavenging effect of anthocyanin pigments isolated from Phaseolus vulgaris
L. Biochem. Pharmacol., 52, 1033–1039, 1996.
18. Chin, K., Rosene, R.T., and Ho, C.-T., Antioxidative and cytotoxic components of highbush blueberry
(Vaccinium corymbosum L.), in Phytochemicals and Phytopharmaceuticals, Shahidi, F. and Ho, C-.T.,
Eds, AOCS Press, Champaign, IL, 2000, pp. 271–277.
19. He, J., Wallace, T.C., Keatley, K.E., Failla, M.L., and Giusti, M.M., Stability of black raspberry antho-
cyanins in the digestive tract lumen and transport efficiency into gastric and small intestinal tissues in
the rat. J. Agric. Food Chem., 57, 3141–3148, 2009.
20. Mazza, G., Kay, C.D., Cottrell, T., and Holub, B.J., Absorption of anthocyanins from blueberries and
serum antioxidant status in human subjects. J. Agric. Food Chem., 50, 7731–7737, 2002.
21. Manach, C., Williamson, G., Morand, C., Scalbert, A., and Remesey, C., Bioavailability and bioefficacy
of polyphenols in humans. I. Review of 97 biovailability studies. Am. J. Clin. Nutr., 81, 230S–242S,
2005.
22. Milbury, P.E., Vita, J.A., and Blumberg, J.B., Anthocyanins are bioavailable in humans following an
acute dose of cranberry juice. J. Nutr., 140, 1099–1104, 2010.
23. Mertens-Talcott, S.U., Rios, J., Jilma-Stohlawetz, P., Pacheco-Palencia, L.A., Meihbohm, B., Talcott, S.T.,
and Derendorf H., Pharmacokinetics of anthocyanins and antioxidant effects after the consumption of
anthocyanin-rich acai juice and pulp (Euterpe oleracea Mart.) in human healthy volunteers. J. Agric.
Food Chem., 7796–7802, 2008.
24. Nielsen, I.L.F., Dragsted, L.O., Ravn-Haren, G., Freese, R., and Rasmussen, S.E., Absorption and excre-
tion of black currant anthocyanins in humans and watanabe heritable hyperlipidemic rabbits. J. Agric.
Food Chem., 51, 2813–2820, 2003.
25. McDougall, G.J., Dobson, P., Smith, P., Blake, A., and Stewart, D., Assessing potential bioavailability of
raspberry anthocyanins using an in vitro digestion system. J. Agric. Food Chem., 53, 5896–5904, 2005.
26. Bell, D.R. and Gochenaur, K., Direct vasoactive and vasoprotective properties of anthocyanin-rich
extracts. J. Appl. Physiol., 100, 1164–1170, 2006.
27. Youdim, K.A., McDonald, J., Kalt, W., and Joseph, J.A., Potential role of dietary flavonoids in reducing
microvascular endothelium vulnerability to oxidative and inflammatory insults. J. Nutr. Biochem., 13,
282–288, 2002.
28. Andriambeloson, E., Magnier, C., Haan-Archipoff, G., Lobstein, A., Anton, R., Beretz, A., Stoclet, J.C.,
and Andriantsitohaina, R., Natural dietary polyphenolic compounds cause endothelium-dependent
vasorelaxation in rat thoracic aorta. J. Nutr., 128, 2324–2233, 1998.
29. Garcia-Alonso, M., Minihane, A.-M., Rimbach, G., Rivas-Gonzalo, J.C., and Pascual-Teresa, S., Red
wine anthocyanins are rapidly absorbed in humans and affect monocyte chemoattractant protein 1 lev-
els and antioxidant capacity of plasma. J. Nutr. Biochem., 20, 521–529, 2009.
30. Karlsen, A., Retterstol, L., Laake, P., Paur, I., Kjolsrud-Bohn, S., Sandvik, L., and Blonhoff, R.,
Anthocyanins inhibit nuclear factor-κB activation in monocytes and reduce plasma concentrations of
pro-inflammatory mediators in healthy adults. J. Nutr. Dis., 137, 1951–1954, 2007.
31. McAnulty, L.S., Nieman, D.C., Dumke, C.L., Shooter, L.A., Henson, D.A., Utter, A.C., Milne, G., and
McAnulty, S.R., Effect of blueberry ingestion on natural killer cell counts, oxidative stress, and inflam-
mation prior to and after 2.5 h of running. Appl. Physiol. Nutr. Metab., 36, 976–984, 2011.
32. Wang, D., Wei, X., Yan, X., Jin, T., and Ling, W., Protocatechuic acid, a metabolite of anthocyanins,
inhibits monocyte adhesion and reduces atherosclerosis in apolipoprotein E-deficient mice. J. Agric.
Food Chem., 58, 12722–12728, 2010.
33. Bastide, P., Rouher, F., and Tronche, P., Rhodopsin et anthocyanosides. Bull. Soc. d’ophtalmol.
de France, 9, 801–807, 1968.
34. Muth, E.R., Laurent, J.M., and Jasper, P., The effect of bilberry nutritional supplementation on night
visual acuity and contrast sensitivity. Alt. Med. Rev., 5, 164–173, 2000.
Blueberry Juice 173
35. Lau, F.C., Bielinski, D.F., and Joseph, J.A., Inhibitory effects of blueberry extract on the production
of inflammatory mediators in lipopolysaccharide-activated BV2 microglia. J. Neurosci. Res., 85,
1010–1017, 2007.
36. Joseph, J.A., Shukitt-Hale, B., Denisova, N.A., Bielinski, D., Martin, A., McEwen, J.J., and Bickford,
P.C., Reversals of age-related declines in neuronal signal transduction, cognitive, and motor behavior
deficits with blueberry, spinach, or strawberry dietary supplementation. J. Neurosci., 19, 8114–8121,
1999.
37. Casadesus, G., Shukitt-Hale, B., Stellwagen, H.M., Zhu, X., Lee, H.G., Smith, M.A., and Joseph, J.A.,
Modulation of hippocampal plasticity and cognitive behavior by short-term blueberry supplementation
in aged rats. Nutr. Neurosci., 7, 309–316, 2004.
38. Andres-Lacueva, C., Shukitt-Hale, B., Galli, R.L., Juaregui, O., Lamuela-Raventos, R.M., and
Joseph J.A., Anthocyanins in aged blueberry-fed rats are found centrally and may enhance memory.
Nutr. Neurosci., 8, 111–120, 2005.
39. Sweeney, M.I., Kalt, W., Mackinnon, S.L., Ashby, J., and Gottschall-Pass, K.T., Feeding rats diets
enriched in lowbush blueberries for six weeks decreases ischemia-induced brain damage. Nutr.
Neurosci., 5, 427–431, 2002.
40. Galli, R.L., Bielinski, D.F., Szprengiel, A., Shukitt-Hale, B., and Joseph, J.A., Blueberry supple-
mented diet reverses age-related decline in hippocampal HSP70 neuroprotection. Neurobiol. Aging, 27,
344–350, 2006.
41. Devore, E.E., Hang, J.H., Breteler, M.M.B., and Grodstein, F., Dietary intake of berries and flavonoids
in relation to cognitive decline. Ann. Neurol., 72, 135–143, 2012.
42. Krikorian, R., Shidler, M.D., Nash, T.A., Kalt, W., Vinqvist-Tymchuk, M.R., Shukitt-Hale, B., and
Joseph, J.A., Blueberry supplementation improves memory in older adults. J. Agric. Food Chem., 58,
3996–4000, 2010.
43. Ofek, I., Goldhar, J., and Sharon, N., Anti-Escherichia coli adhesion activity of cranberry and blueberry
juices. Adv. Exp. Med. Biol., 408, 178–183, 1996.
44. Knekt, P., Järvinen, R., Seppänen, R., Heliövaara, M., Teppo, L., Pukkale, E., and Aromaa, A., Dietary
flavonoids and the risk of lung cancer and other malignant neoplasms. Am. J. Epidemiol., 146, 223–230,
1997.
45. Srivastava, A., Akoh, C.C., Fischer, J., and Krewer, G., Effect of anthocyanin fractions from selected
cultivars of Georgia-grown blueberries on apoptosis and phase II enzymes. J. Agric. Food Chem., 55,
3180–3185, 2007.
46. Suh, N., Paul, S., Hao, X., Rimando, A.M., and Reddy, B.S., Pterostilbene, an active constituent of blue-
berries, suppresses aberrant crypt foci formation in the azoxymethane-induced colon carcinogenesis
model in rats. Clin. Cancer Res., 13, 350–355, 2007.
47. Schmidt, B.M., Erdman Jr, J.W., and Lila, M.A., Differential effects of blueberry proanthocyanidins on
androgen sensitive and insensitive human prostate cancer cell lines. Cancer Lett., 231, 240–246, 2006.
48. Folts, J.D., Potential health benefits from the flavonoids in grape products on vascular disease, in
Flavonoids in Cell Function, Buslig, B. and Manthey, J., Eds., Kluwer Academic/Plenum Publishing,
New York, 2002, pp. 95–111.
49. Wu, X., Kang, J., Xie, C., Burris, R., Ferguson, M.E., Badger, T.M., and Nagarajan, S., Dietary blueber-
ries attenuate atherosclerosis in apolipoprotein E-deficient mice by upregulating antioxidant enzyme
expression. J. Nutr., 140, 1628–1632, 2010.
50. Rissanen, T.H., Voutilainen, S., Virtanen, J.K., Venho, B., Vanharanta, M., Mursu, J., and Salonen, J.T.,
Low intake of fruits, berries and vegetables is associated with excess mortality in men: The Kuopio
ischaemic heart disease risk factor study. J. Nutr., 133, 199–204, 2003.
51. Mink, P.J., Scrafford, C.G., Barraj, L.M., Harnack, L., Hong, C.-P., Nettleton, J.A., and Jacobs, D.R.,
Flavonoid intake and cardiovascular disease mortality: A prospective study in postmenopausal women.
Am. J. Nutr., 85, 895–909, 2007.
52. Basu, A., Du, M., Leyva, M.J., Sanchez, K., Betts, N.M., Wu, M., Aston, C.E., and Lyons T.J., Blueberries
decrease cardiovascular risk factors in obese men and women with metabolic syndrome. J. Nutr., 140,
1582–1587, 2010.
53. Agri-Canada, Fruit juices in the United States: Market Indicator Report, Agriculture and Agri-Food
Canada, Ottawa, Ontario, Canada, 2011.
54. Mantius, H.L. and Rose, L., Process for producing sugars and acids-rich juice and phytochemical-rich
juice, U.S. Patent Application Publication 2006/0177560A1.
55. Barba, F.J., Esteve, M.J., and Frigola, A., Physicochemical and nutritional characteristics of blueberry
juice after high pressure processing. Food Res. Int., 50, 545–549, 2013.
56. Buckow, R., Kastell, A., Terefe, N.S., and Versteeg, C., Pressure and temperature effects on degra-
dation kinetics and storage stability of total anthocyanins in blueberry juice. J. Agric. Food Chem.,
58, 10076–10084, 2010.
57. Peck, D.C., The effects of high-pressure throttling versus thermal pasteurization on a blueberry-whey
beverage, PhD thesis, University of Georgia, Athens, GA, 2004.
58. Wand, C.Y., Chen, C.-T., and Wang, S.Y., Changes of flavonoid content and antioxidant capacity in blue-
berries after illumination with UV-C. Food Chem., 117, 426–431, 2009.
59. Papanikolaou, N., Apostolos, K., Thoma, M., and Guortsoyiannis, N., MR cholangiopancreatography
before and after oral blueberry juice administration. J. Comput. Assist. Tomogr., 24, 229–234, 2000.
14
Cherry Juice
Gamze Toydemir, Dilek Boyacioglu, Jules Beekwilder,

Robert D. Hall, and Esra Capanoglu
CONTENTS
14.1 Introduction....................................................................................................................................175
14.4 Health Effects.................................................................................................................................181
14.6 Conclusion......................................................................................................................................182
References................................................................................................................................................182
14.1 Introduction
In recent years, an increasing interest has been placed on a new diet-health paradigm with an emphasis
on the positive aspects of diet and an examination of foods for their protective and disease-preventive
potential. Accordingly, fruits and vegetables have gained an important status in the human diet as
“functional foods,” which are capable of providing additional physiological benefits, including prevent-
ing or delaying the onset of several chronic diseases, due to their phytochemical content [1,2]. The 5 A
Day program was developed in 1989, with a report by National Academy of Sciences, recommending the
daily consumption of five or more servings of fruits and vegetables in order to reduce the risk of certain
chronic diseases [3].
Berry fruits have gained a renewed global interest due to their recognized phytochemical composi-
tion, resulting in an increased industrial demand for these fruits and their products [4]. Among these,
cherries, with the most important being species of the sweet cherry (Prunus avium L.) and sour cherry
or tart cherry (Prunus cerasus L.), have been well documented to be rich sources of several phenolic
antioxidants, including anthocyanins as the major flavonoid group [5]. While sweet cherries are com-
monly consumed as fresh fruit, sour cherries are consumed after processing into various products, such
as canned products, marmalades, frozen fruits, and fruit juices [6]. Among these, cherry juice, which is
known as “sour cherry juice” or “tart cherry juice” in the common literature, is the most popular cherry
product around the world.
Sour cherry is an important fruit in Turkey, which is the world’s leading producer of it, generating
almost 200,000 metric tons (MT) in 2012. The top sour cherry–producing countries, other than Turkey,
include Russia, Poland, Ukraine, and Iran [7]. For the Turkish juice industry, sour cherry is an important
starting material, with 25%–40% of the fruit being processed into juice called nectar [8]. Sour cherry
juice is the second mostly consumed juice in Turkey with 19.7% consumption rate, following peach juice
which has 32.4% of consumption rate [9].
Sour cherry fruit is rarely processed into pure fruit juice because of its strong characteristic sour
taste. Consequently, the juice obtained from sour cherry is not palatable in its natural form and so,
sugar addition is required to obtain a drinkable “nectar” product [8]. Nectars are defined as fruit drinks
containing 25%–99% fruit juice [9]. Figure 14.1 represents the production scheme for sour cherry juice
(nectar), involving several steps such as heating, pressing, and filtration [10].
175
Stalks
Washing and Seperation Mash heating Mash

Fruit
Selection of stalks pressing
First Initial
juice press cake
Final press cake
and seeds
Press cake
extraction
Press outlet Press cake
Clarification Enzymation Pasteurization collecting tanks extract
Processing filtered
Evaporation to Paper filtration
Ultrafiltration concentrated juice to Pasteurization
concentrated juice
final juice (nectar)
Filtration Pasteurized
residue juice
FIGURE 14.1 A schematic representation of industrial-scale sour cherry juice (nectar) processing. (Adapted from
Toydemir, G. et al., J. Funct. Foods, 5, 1402, 2013. With permission.)
Cherry juice, which has been reported to be closely similar to the raw fruit in its polyphenolic com-
position, contains substantial quantities of phenolic antioxidants, including specifically a number of
anthocyanins [11]. In addition, other phenolic compounds, such as flavan-3-ols (proanthocyanidins) and
flavonols, are present [11,12]. Anthocyanin consumption has been linked to the prevention of a broad
range of human illnesses, for example, cancer [13], obesity, and diabetes [14]. This chapter highlights
the nutritional characteristics of cherry juice by focusing on the antioxidant properties and related health
effects, as well as the future trends regarding this promising juice product.

Cherry fruit and its processed juice are rich in nutrients, including a variety of vitamins and minerals
(Table 14.1) [15,16]. The nutrient composition of sour cherry juice exhibits differences due to various fac-
tors such as the cultivar and the maturity level of the starting fruit material, and the processing method
used, among others. The basic juice parameters are discussed in this section while bioactive compounds
in sour cherry juice are included in a subsequent section.
The Brix values (°Brix) of sour cherry juices prepared from 11 varieties of Turkish sour cherries
were between 16.0 and 26.3°Brix [17], while this range was reported to be between 13.2 and 18.2°Brix
elsewhere [18] for 25 different sour cherry juices. The levels of glucose and fructose in five different
cultivars of sour cherry juice samples were between 48.7 and 63.4 and between 39.6 and 52.2 g/L,
respectively [12].
Sour cherry juice is characterized by its high acidity, in which the total acid concentration is nearly
exclusively made up from L-malic acid. L-malic acid levels in sour cherry juice have been reported to be
in the range of 14.3–29.2 g/L [19]. On the other hand, ascorbic acid (43–177 mg/L) [12] and citric acid
(52–115 mg/L) [19] concentrations are known to be quite low.
Sour cherry juice contained 1336–2378 mg/L colloids, mainly composed of pectic substances. Protein
and pectin levels in sour cherry juice are ~5 [17,19] and ~2 g/L [20], respectively. Moreover, the high
Cherry Juice 177
TABLE 14.1
Compositional and Nutritional Characteristics of Sour Cherry and Its Juice
Nutrient Unit Cherry Fruita [15] Unit Cherry Juiceb [16]
Energy kcal 78 kcal 140
Protein g 1.55 g 1
Lipid (fat) g 0.46 g 0
Carbohydrate g 18.88 g 33
Total sugars g 13.16 g 25
Total dietary fiber 2.5 g 0
Minerals
Calcium mg 25 % daily valuec 2
Iron mg 0.50 % daily value 8
Magnesium mg 14 — nr
Phosphorus mg 23 — nr
Potassium mg 268 — nr
Sodium mg 5 mg 30
Zinc mg 0.16 — nr
Vitamins
Vitamin C mg 15.5 % daily value 0
Thiamin mg 0.046 — nr
Riboflavin mg 0.062 — nr
Niacin mg 0.62 — nr
Vitamin B6 mg 0.068 — nr
Vitamin A (RAE) µg 99 % daily value 4
Vitamin E (ATE) mg 0.11 — nr
a The values represented are for a serving size of 1 cup (without pits, 155 g) fresh sour cherry fruit.
b The values represented are for a serving size of 227 g juice.
c Percent daily values are based on a 2000 Cal diet.
Abbreviations: RAE, retinol activity equivalents; ATE, alpha-tocopherol equivalents; nr, not reported.
concentrations of ash and minerals add value to the positive nutritional properties of sour cherry juice.
In addition, sour cherry juice contains potassium (2140–3686 mg/L), calcium (267–368 mg/L), and mag-
nesium (148–172 mg/L) as well as trace elements, namely, boron (4.2–8.0 mg/L), copper (0.5–1.2 mg/L),
iron (2.2–3.4 mg/L), manganese (1.2–1.5 mg/L), and zinc (0.6–1.0 mg/L) [12].

Sour cherry fruit and its commonly consumed juice product are rich sources of phenolic compounds,
including specifically substantial quantities of anthocyanins as the major flavonoid group. They are also
rich in other flavonoids, such as flavan-3-ols (proanthocyanidins), flavonols, as well as hydroxycinna-
mates [11]. The chemical structures of some major phenolics in sour cherry juice are represented in
Figure 14.2.
The total phenolics (TP) in sour cherry fruit correspond to 228.9 mg gallic acid equivalents (GAE)/100 g
serving [5]. The processing of fruit into fruit juice involves several steps, such as heating, pressing, and
filtration (Figure 14.1), which can potentially affect the recovery of phenolic compounds. On the other
hand, Toydemir et al. [11], who studied the entire industrial-scale juice processing, found out that the
majority of the phenolic compounds (anthocyanins in particular) present in sour cherry fruit were well
recovered in the juice fraction.
There are substantial differences in the literature for the measured values of TP and total anthocya-
nins (TA) in sour cherry juice (Table 14.2). This could be linked to the variations in the starting fruit
OH
OH
O
+
HO O CH2OH
O OH
O HO
O OH
OH O
OH
HO OH H
H3C
HO O
HO OH
CH2
HO O O OH
H
HO OH
Cyanidin-3-2G-glucosylrutinoside (–)-Epicatechin
O
HO OH
O OH
OH
HO
OH
Neochlorogenic acid
FIGURE 14.2 Chemical structures of some major phenolic compounds found in sour cherry juice.
material and also to the existing differences in the juice processing steps and parameters. In addition,
lack of standardization of the analytical methods used may yield several orders of magnitude differences
in the detected contents. For instance, in one study, the TP and TA contents of 11 types of sour cherry
juice samples (obtained from different Turkish sour cherry fruit varieties) were reported to range from
1510 to 2550 mg GAE/L and from 350 and 633 mg cyanidin-3-glucoside equivalents (C3GE)/L, respec-
tively [17]. Whereas, in another study, the TP and TA contents of a Turkish sour cherry juice (nectar)
sample were 635 mg GAE/L and 86 mg C3GE/L, respectively, which were substantially lower than those
obtained in a previous study (Table 14.2) [21]. Thus, it seems that findings in the literature cover a wide
has range antioxidant potential of values and show little reproducibility [17].
Sour cherry has an antioxidant potential comparable to some berry fruits, such as strawberry, and has
higher antioxidant potential than that of apple and kiwi fruits [22]. Moreover, sour cherry juice has a higher
antioxidant activity than some other fruit juices containing high amounts of anthocyanins, including red
raspberry, blackberry, and strawberry [23]. However, similar to the findings observed for TP and TA con-
tents, the reported total antioxidant capacity (TAC) levels of different sour cherry juice samples studied also
showed significant differences. Thus, in one study, the TAC levels of five different sour cherry juice samples
were between 27 and 55 mmol trolox equivalents (TE)/L juice [12], while in another study, TAC of 11 dif-
ferent sour cherry juice samples ranged from 20 to 38 mmol TE/L [17]. On the other hand, the antioxidant
potential of a sour cherry juice (nectar) sample was much lower in comparison with the previous examples.
The TAC values of this sample measured by using three different methods (1,1-diphenyl-2-picrylhydrazyl
[DPPH], 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid [ABTS]), and copper-reducing antioxidant
capacity [CUPRAC]), were in between 3.9 and 9.5 mmol TE/L (Table 14.2) [21].
Cherries have been well documented to be among the richest sources of anthocyanins [24]. The
sour cherry varieties generally have higher anthocyanin contents in comparison to the sweet cherry
varieties [5]. Various individual anthocyanin components have been identified in sour cherry and
its products, including cyanidin-3–2G -glucosylrutinoside as the major sour cherry anthocyanin,
followed by cyanidin-3-rutinoside, cyanidin-3-sophoroside, and cyanidin-3-glucoside [11,12].

Cherry Juice 179
TABLE 14.2
Total Phenolics, Anthocyanins, and Antioxidant Capacity of Sour Cherry Juice
Product Constituent Unit Concentration References
Sour cherry juicea TP mg GAE/L 1510–2550 e [17]
TA mg C3GE/L 350–633e [17]
TAC mmol TE/L 20–38e [17]
Sour cherry juice (nectar)b TP mg GAE/L 635 [21]
TA C3GE/L 86 mg [21]
TAC mmol TE/L 4–9f [21]
Sour cherry juicec TP — nr [32]
TA mg/L 546g [32]
TAC — nr [32]
Sour cherry juiced TP mg CE/L 2704–4998h [12]
TA mg C3GE/L 569–858h [12]
TAC mmol TE/L 27–55h [12]
a Juice samples were extracted from different Turkish sour cherry varieties.
b Juice was extracted from a single Turkish sour cherry variety, Kutahya.
c Juice was extracted from a single Croatian sour cherry variety, Maraska.
d Juice samples were extracted from five different sour cherry varieties.
e The concentrations represented were the ranges determined for 11 different sour cherry juice samples.
f The antioxidant capacity of the juice sample was determined using three different in vitro tests in parallel.
g TA was determined using high-performance liquid chromatography, with the calibration of individual standards as a
reference.
h The concentrations represented were the ranges determined for five different sour cherry juice samples.
Abbreviations: TP, total phenolics; TA, total anthocyanins; TAC, total antioxidant activity; GAE, gallic acid equivalents;
C3GE, cyanidin-3-glucoside equivalents; TE, trolox equivalents; CE, catechin equivalents; nr, not
reported.
The levels of two major sour cherry anthocyanins, namely, cyanidin-3–2G -glucosylrutinoside and
cyanidin-3-rutinoside, in three different sour cherry fruit cultivars, were in the range of 17–52 and
9–25 mg C3GE/100 g fresh fruit, respectively [25]. In another study, Kim et al. [26] determined the
values of these two major anthocyanins in sour cherry variety ranging from 89 to 228 and from 16 to
22 mg C3GE/100 g fresh fruit, respectively. Moreover, in a Turkish sour cherry fruit variety, the
reported values of cyanidin-3–2G -glucosylrutinoside and cyanidin-3-rutinoside were 42 and 11 mg
C3GE/100 g fresh fruit, respectively [11]. All these varying results could be linked to the fact that
the fruit variety is a major factor influencing the anthocyanin concentration in the fresh fruit as well
as their processed products.
The concentrations of cyanidin-3–2G-glucosylrutinoside and cyanidin-3-rutinoside in five different
freshly pasteurized sour cherry juice samples, obtained from five different fruit cultivars, were in the
range of 361–515 and 125–213 mg C3GE/L, respectively (Table 14.3) [12]. On the other hand, the values
determined for these two anthocyanin compounds were 119 and 26 mg C3GE/L, respectively, in a sour
cherry juice (nectar) sample (Table 14.3) [11]. Moreover, the anthocyanin components in sour cherry
contributed ~60% to the total antioxidant activity. Among these, cyanidin-3–2G-glucosylrutinoside pro-
vided the highest antioxidant activity, being responsible for approximately 50% and 40% of the total
antioxidant peak area in fruit and its processed juice, respectively [10].
Sour cherry juice has also been identified as an important source of flavonoids other than antho-
cyanins. Flavan-3-ols (procyanidins) and flavonols are the two other main subclasses that have been
identified and quantified in sour cherry juice (Table 14.3) [11,27]. (–)-Epicatechin is the most abun-
dant flavan-3-ol in sour cherry products followed by (+)-catechin [11,12,27]. One of the distinctive
characteristics of sour cherries from other cherry varieties is their short-chain procyanidins with a
low degree of polymerization (DP ≤ 3) [11,27]. There is an apparent correlation between DP values
of procyanidins, which can range from two subunits to several hundred units, and their absorption
TABLE 14.3
Polyphenolic Composition of Sour Cherry Juice
Product Class Flavonoid Unit Concentration References
Sour cherry juice Anthocyanin Cyanidin-3–2 -G mg C3GE/L 119 [21]
(nectar)a glucosylrutinoside
Cyanidin-3-rutinoside mg C3GE/L 26 [21]
Cyanidin-3-sophoroside mg C3GE/L 4 [21]
Cyanidin-3-glucoside mg C3GE/L 3 [21]
Flavan-3-ol (−)-Epicatechin mg/L 123 [21]
(+)-Catechin mg/L 17 [21]
Flavonol Quercetin-3-rutinoside mg/L 14 [21]
Kaempferol-3-rutinoside mg/L 3 [21]
Phenolic acid Neochlorogenic acid mg/L 184 [21]
p-Coumaroylquinic acid mg/L 53 [21]
Chlorogenic acid mg/L 33 [21]
Sour cherry juiceb Anthocyanin Cyanidin-3–2G- mg C3GE/L 361–515c [12]
glucosylrutinoside
Cyanidin-3-rutinoside mg C3GE/L 125–213c [12]
Cyanidin-3-sophoroside mg C3GE/L 37–185c [12]
Cyanidin-3-glucoside — nr [12]
Flavan-3-ol (−)-Epicatechin mg/L 24–336c [12]
(+)-Catechin mg/L 1–14c [12]
Procyanidin B1 mg/L 23–99c [12]
Procyanidin B2 mg/L <0.2–272c [12]
Flavonol Quercetin-3-rutinoside mg/L 18–59c [12]
Quercetin-3–2G- mg/L 11–31c [12]
glucosylrutinoside
Kaempferol-3-rutinoside mg/L 4–13c [12]
Isorhamnetin-rutinoside mg/L 14–33c [12]
Phenolic acid Neochlorogenic acid mg/L 212–998c [12]
p-Coumaroylquinic acid mg/L 191–999c [12]
Chlorogenic acid mg/L 119–268c [12]
a Juice was extracted from a single Turkish sour cherry variety, Kutahya.
b Juice samples were extracted from five different sour cherry varieties.
c The concentrations represented were the ranges determined for five different sour cherry juice samples.
Abbreviations: C3GE, cyanidin-3-glucoside equivalents; nr, not reported.
in the intestinal tract [28]. Khanal et al. [29] pointed out that the long-chain procyanidin oligomers
and polymers were poorly absorbed, while short-chain procyanidins (monomers, dimers, and trimers)
(DP value up to 3) were absorbed more efficiently. Toydemir et al. [11] reported a slight decrease in
DP value, from DP 2.1 to 1.7, as a result of processing of sour cherry fruit into its juice (nectar). This
indicated a loss in longer-chain procyanidins during processing, by the removal of processing waste,
which may provide higher procyanidin bioavailability in juice than in fruit.
Flavonol glycosides, including the rutinosides and glucosides of quercetin and kaempferol, are the
other important group of sour cherry phenolics. Among these, quercetin-3-rutinoside has been identified
as the major flavonol in several studies [11,12,21,30]. Sour cherry also possesses substantial levels of
hydroxycinnamates, which are at relatively higher levels than those in sweet cultivars. Neochlorogenic
acid, p-coumaroylquinic acid, and chlorogenic acid are the predominant phenolic acids in sour cherry
(Table 14.3) [12,21,31].
Cherry Juice 181
14.4 Health Effects

Sour cherry juice contains several phenolic compounds, anthocyanins in particular, whose biological
activities, such as antioxidant, anti-inflammatory, anticancer, antidiabetic, and antiobesity properties,
have recently been investigated in different experimental models.
The antioxidant activities of sour cherry have been studied and confirmed using different biological
samples. Mice, having diet supplementation with sour cherry juice, were shown to increase superoxide
dismutase antioxidative enzyme activity in the liver and blood, and glutathione peroxidase antioxida-
tive enzyme activity in liver tissues, as well as decreasing lipid peroxidation [32]. In a study on human
subjects, 12 older adult volunteers (6 men and 6 women, age 69 ± 4 years) consumed either sour cherry
juice or a placebo (240 mL twice daily for 14 days) in random order. The capacity to resist oxidative dam-
age was determined by measuring the changes in plasma F2-isoprostane levels, a marker of oxidative
damage. This is generated in response to forearm ischemia reperfusion, which is a factor that increases
the production of reactive oxygen species resulting in acute events such as cardiovascular disease (CVD).
Sour cherry juice intake was associated with a reduced I/R-induced F2-isoprostane response, suggesting
that sour cherry juice consumption enhances antioxidant activity in vivo in older adults [33].
Sour cherry juice consumption has been indicated to render cyclooxygenase-2 (COX-2) inhibitory
activity in mice peritoneal macrophages, suggesting an anti-inflammatory effect of sour cherry, which is
most likely linked to the abundance of anthocyanins [32]. COXs are proinflammatory enzymes that have
an important role in processes such as inflammation, carcinogenesis, apoptosis, cell proliferation, and
angiogenesis [34]. Additionally, sour cherry anthocyanins have been reported to play an effective role in
the inhibition of other inflammation related diseases, such as obesity and diabetes. Ataie-Jafari et al. [35]
determined that concentrated sour cherry juice consumption with intake of 40 g/day for 6 weeks led to a
decreased body weight, reduced blood pressure, and reduced hemoglobin A1c (long-term representative
of blood glucose) in type 2 diabetic women and improved blood lipids in diabetic patients with hyperlip-
idemia, which was linked to the substantial content of anthocyanins.
Moreover, sour cherry anthocyanins have also been shown to possess multiple anticarcinogenic
effects, using several in vitro colon, liver, and breast cell culture systems [36]. Anthocyanins in sour
cherry inhibited intestinal tumor development in ApcMin mice and prevented the growth of human
colon cancer cell lines HT 29 and HCT 116, suggesting that these bioactive components have a substan-
tial potential to reduce the risk of colon cancer [37]. Moreover, Bobe et al. [38] investigated the cancer
chemopreventive effect of a therapy including the simultaneous utilization of an anthocyanin-rich sour
cherry extract (as dietary phytochemical source) with an anti-inflammatory drug to ApcMin mice for
19 weeks. This combination resulted in fewer tumors in the small intestine when compared to mice fed
with the drug alone.
Recently, sour cherry juice has been shown to possess substantial effects on aiding recovery and
reducing muscle damage, inflammation, and oxidative stress. The consumption of cherry juice by 20
recreational marathon runners, for 5 days before the day of and for 48 h following a run, was examined
for its effect on markers of muscle damage (creatine kinase, lactate dehydrogenase, muscle soreness,
and isometric strength), inflammation (interleukin-6 [IL-6], C-reactive protein [CRP], and uric acid),
total antioxidant status (TAS), oxidative stress, thiobarbituric acid–reactive species (TBARS), and pro-
tein carbonyls, before and after the race. The results indicated significantly faster recovery in isometric
strength (P = 0.024), reduced inflammation (IL-6, P < 0.001; CRP, P < 0.01; uric acid, P < 0.05) and
oxidative stress (significantly lower TBARS, P < 0.05), and ~10% greater TAS in the cherry juice group
in comparison to the placebo group [39]. In another study, 54 healthy runners (36 male, 18 female; 35.8 ±
9.6 years) ingested 335 mL of sour cherry juice or a placebo cherry drink twice daily for 7 days prior
to and on the day of the run; the results revealed significantly lower increase in pain in the cherry juice
group compared to the placebo group (P < 0.001) [40]. These effects of sour cherry juice have been
attributed to the presence of numerous bioactive phytochemicals, which have been shown to exert anti-
oxidant and anti-inflammatory properties [39,40].

The characteristic sour taste of sour cherry fruit does not support the production of 100% fruit juice from
this fruit. Instead, it requires the addition of sugar to some extent in order to provide a drinkable “nectar”
product [9]. As a result, the final sour cherry nectar drink contains 35% fruit (v/v) (minimum) and 20%
sugar (sucrose) (m/m) (maximum) in it [41]. On the other hand, recent consumer attitudes have been
changing with increasing interest on “low-calorie” foods and on “natural” foods with no additives. Thus,
improved and new formulations are desirable to meet changing market demands, in particular, demand
for beverages having improved nutritional characteristics, such as alternative calorie content, alternative
flavor profiles, and greater use of natural ingredients [42]. This has led to a new research area that has
focused on the production of nectar drinks with no added sugar, possible use of synthetic sweeteners, or
some other fruits. Aspartame and acesulfame K are the artificial sweeteners that have been studied for
use in sour cherry drinks [43]. Additionally, there are newly formulated sour cherry juice products in
Turkey that include apple as a natural sweetener and provides 100% fruit juice [44].
Sour cherry juice processing involves thermal treatment up to 95°C in order to prevent microbial
spoilage and enzymatic degradation (Figure 14.1) [11]. This heat application may result in color loss, loss
of vitamins, and degradation of anthocyanins to some extent [12]. Alternative technologies to process
sour cherry juice that may protect the nutritional components and physical properties at a higher level
are being researched [45]. Altuntas et al. [45] studied the effect of pulsed electric field (PEF) process-
ing on sour cherry juice parameters as a nonthermal food preservation method that is also commonly
applied to a variety of fruit juices. They reported a significant increase in microbial inactivation, includ-
ing Escherichia coli O157:H7, Staphylococcus aureus, Listeria monocytogenes, Erwinia carotovora,
Pseudomonas syringae subs. syringae, Botrytis cinerea, and Penicillium expansum, with increased elec-
tric field strength and treatment time (P ≤ 0.05), while the applied PEF treatment parameters did not lead
to any significant change on the measured other properties of sour cherry juice, including pH, oBrix,
titratable acidity, conductivity, color (L*, a*, and b*), nonenzymatic browning index, metal ion con-
centration, total ascorbic acid, and total anthocyanin content (P > 0.05). This study indicated that PEF
treatment is a promising approach in sour cherry juice production with enhanced microbial inactivation
and with no adverse effect on the other juice parameters [45]. Furthermore, the use of high hydrostatic
pressure (350 MPa), as another nonthermal treatment, combined with mild heat (40°C), has been shown
to be effective for treating sour cherry juice to improve microbial kill, with respect to the most pressure-
resistant strains of S. aureus, E. coli O157:H7, and Salmonella enteritidis [46].
14.6 Conclusion
Sour cherry juice can be regarded as a functional fruit beverage due to its high content of phenolics, in
particular anthocyanins, which are well recovered from the fruit fraction during processing. In addition,
the substantial quantities of procyanidins with a low DP (DP < 3) make this juice superior to others.
Consumption of sour cherry juice has been correlated to specific health-enhancing outcomes, which has
been linked to the presence of various phytochemicals that have been shown to possess antioxidant, anti-
inflammatory, anticancer, antidiabetic, and antiobesity properties, among others. The current research
has focused on the enhancement of the nutritional characteristics of sour cherry juice and has the poten-
tial of introducing new formulations and novel processing techniques in juice processing, which will
meet the recent consumer demands in the market.
REFERENCES
1. Kaur, C. and Kapoor, H.C., Antioxidants in fruits and vegetables—The millennium’s health. Int. J. Food
Sci. Tech., 36, 703–725, 2001.
2. Nicoli, M.C., Anese, M., and Parpinel, M., Influence of processing on the antioxidant properties of fruits
and vegetables. Trends Food Sci. Technol., 10, 94–100, 1999.
Cherry Juice 183
3. National Academy of Sciences, Committee on Diet and Health, National Research Council, Diet and
Health: Implications for Reducing Chronic Disease Risk. National Academy Press, Washington, DC,
1989.
4. Hakkinen, S., Heinonen, M., Karenlampi, S., Mykkanen, H., Ruuskanen, J., and Torronen, R., Screening
of selected flavonoids and phenolic acids in 19 berries. Food Res. Int., 32, 345–354, 1999.
5. Ferretti, G., Bacchetti, T., Belleggia, A., and Neri, D., Cherry antioxidants: from farm to table. Molecules,
15, 6993–7005, 2010.
6. Chaovanalikit, A. and Wrolstad, R.E., Total anthocyanins and total phenolics of fresh and processed
cherries and their antioxidant properties. J. Food Sci., 69, C67–C72, 2004.
7. Food and Agriculture Organization of the United Nations (FAOSTAT), Food and Agricultural
Commodities Production, Countries by Commodity, 2012. Published online at: http://faostat.fao.org/
site/339/default.aspx (accessed March 15, 2014).
8. Akdag, E., Turkey fruit juice like products industrial report, 2011. Published online at: http://www.meyed.
org.tr/userfiles/file/sektor_istatistikleri/meyve_suyu_sektoru_raporu_2011.pdf (accessed September 15,
2012).
9. Euromonitor International, Fruit/vegetable Juice in Turkey, 2012. Published online at: http://www.
marketresearch.com/Euromonitor-International-v746/Fruit-Vegetable-Juice-Turkey- 7067594/ (accessed
November 22, 2012).
10. Toydemir, G., Capanoglu, E., Kamiloglu, S., Boyacioglu, D., De Vos, R.C.H., Hall, R.D., and Beekwilder,
J., Changes in sour cherry (Prunus cerasus L.) antioxidants during nectar processing and in vitro gastro-
intestinal digestion. J. Funct. Foods, 5, 1402–1413, 2013.
11. Toydemir, G., Capanoglu, E., Gomez Roldan, M.V., De Vos, R.C.H., Boyacioglu, D., Hall, R.D., and
Beekwilder, J., Industrial processing effects on phenolic compounds in sour cherry (Prunus cerasus L.)
fruit. Food Res. Int., 53, 218–225, 2013.
12. Bonerz, D., Wurth, K., Dietrich, H., and Will, F., Analytical characterization and the impact of aging
on anthocyanin composition and degradation in juices from five sour cherry cultivars. Eur. Food Res.
Technol., 224, 355–364, 2007.
13. Butelli, E., Titta, L., Giorgio, M., Mock, H.P., Matros, A., Peterek, S., Schijlen, E.G.W.M., Hall, R.D.,
Bovy, A.G., Luo, J., and Martin, C., Enrichment of tomato fruit with health-promoting anthocyanins by
expression of select transcription factors. Nat. Biotechnol., 26, 1301–1308, 2008.
14. Tsuda, T., Horio, F., Uchida, K., Aoki, H., and Osawa, T., Dietary cyanidin 3-O-β-D-glucoside-rich
purple corn color prevents obesity and ameliorates hyperglycemia in mice. J. Nutr., 133, 2125–2130,
2003.
Release 27. Published online at: http://ndb.nal.usda.gov/ndb/foods/show/2226?fgcd=&manu=&lfacet=
&format=&count=&max=35&offset=&sort=&qlookup=cherry (accessed September 21, 2015).
16. Cherry Marketing Institute (CMI), The Red Report: The Science Behind Tart Cherries, 2012. Published
online at: http://www.choosecherries.com/wp-content/uploads/2014/03/TheRedReport.pdf (accessed
April 12, 2014).
17. Damar, I. and Eksi, A., Antioxidant capacity and anthocyanin profile of sour cherry (Prunus cerasus L.)
juice. Food Chem., 135, 2910–2914, 2012.
18. Velioglu, S. and Yildiz, O., Chemical composition of Turkish sour cherry juices. Gıda, 21, 103–107,
1996.
19. Eksi, A., Reicheneder, E., and Kieninger, H., Über die chemische zusammensetzung der sauerkirshc-
muttersaefte aus verschiedenen sorten. Flüssiges Obst, 47, 494–496, 1980.
20. Meyer, A.S., Koser, C., and Adler-Nissen, J., Efficiency of enzymatic and other alternative clarification
and fining treatments on turbidity and haze in cherry juice. J. Agric. Food. Chem., 49, 3644–3650,
2001.
21. Toydemir, G., The effects of nectar processing on sour cherry antioxidant compounds: Changes
in metabolite profile and bioavailability, PhD thesis, Istanbul Technical University, Istanbul,
Turkey, 2013.
22. Lister, C.E., Wilson, P.E., Sutton, K.H., and Morrison, S.C., Understanding the health benefits of
blackcurrants. Acta Hort., 585, 443–449, 2002.
23. Jakobek, L., Seruga, M., Medvidovic-Kosanovic, M., and Novak, I., Anthocyanin content and antioxidant
activity of various red fruit juices. Deut. Lebensm-Rundsch., 103, 58–64, 2007.
24. Pantelidis, G.E., Vasilakakis, M., Manganaris, G.A., and Diamantidis, G., Antioxidant capacity, phenol,
anthocyanin and ascorbic acid contents in raspberries, blackberries, red currants, gooseberries, and
Cornelian cherries. Food Chem., 102, 777–783, 2007.
25. Blando, F., Gerardi, C., and Nicoletti, I., Sour cherry (Prunus cerasus L) anthocyanins as ingredients for
functional foods. J. Biomed. Biotechnol., 5, 253–258, 2004.
26. Kim, D.O., Heo, H.J., Kim, Y.J., Yang, H.S., and Lee, C.Y., Sweet and sour cherry phenolics and their
protective effects on neuronal cells. J. Agric. Food Chem., 53, 9921–9927, 2005.
27. Capanoglu, E., Boyacioglu, D., De Vos, R.C.H., Hall, R.D., and Beekwilder, J., Procyanidins in fruit
from sour cherry (Prunus cerasus) differ strongly in chainlength from those in laurel cherry (Prunus
lauracerasus) and cornelian cherry (Cornus mas). J. Berry Res., 1, 137–146, 2011.
28. Deprez, S., Mila, I., Huneau, J.F., Tome, D., and Scalbert, A., Transport of proanthocyanidin dimer, tri-
mer, and polymer across monolayers of human intestinal epithelial Caco-2 cells. Antioxid. Redox Sign.,
3, 957–967, 2001.
29. Khanal, R.C., Howard, L.R., and Prior, R.L., Procyanidin content of grape seed and pomace, total
anthocyanin content of grape pomace as affected by extrusion processing. J. Food Sci., 74, H174–H182,
2009.
30. Kirakosyan, A., Seymour, E.M., Urcuyo Llanes, D.E., Kaufman, P.B., and Bolling, S.F., Chemical pro-
file and antioxidant capacities of tart cherry products. Food Chem., 115, 20–25, 2009.
31. Robbins, R.J., Phenolic acids in foods: an overview of analytical methodology. J. Agric. Food Chem.,
51, 2866–2887, 2003.
32. Saric, A., Sobocanec, S., Balog, T., Kusic, B., Sverko, V., Dragovic-Uzelac, V., Levaj, B., Cosic, Z.,
Safranko, Z.M., and Marotti, T., Improved antioxidant and anti-inflammatory potential in mice
consuming sour cherry juice (Prunus cerasus cv. Maraska). Plant Food Hum. Nutr., 64, 231–237,
2009.
33. Traustadottir, T., Davies, S.S., Stock, A.A., Su, Y., Heward, C.B., Roberts, L.J., and Harman, S.M.,
Tart cherry juice decreases oxidative stress in healthy older men and women. J. Nutr., 139, 1896–1900,
2009.
34. Mulabagal, V., Lang, G.A., Dewitt, D.L., Dalavoy, S.S., and Nair, M.G., Anthocyanin content, lipid
peroxidation and cyclooxygenase enzyme inhibitory activities of sweet and sour cherries. J. Agric. Food
Chem., 57, 1239–1246, 2009.
35. Ataie-Jafari, A., Hosseini, S., Karimi, F., and Pajouhi, M., Effects of sour cherry juice on blood glucose
and some cardiovascular risk factors improvements in diabetic women, a pilot study. Nutr. Food Sci.,
38, 355–360, 2008.
36. Wang, L.-S. and Stoner, G.D., Anthocyanins and their role in cancer prevention. Cancer Lett., 269,
281–290, 2008.
37. Kang, S.Y., Seeram, N.P., Nair, M.G., and Bourquin, L.D., Tart cherry anthocyanins inhibit tumor devel-
opment in Apc mice and reduce proliferation of human colon cancer cells. Cancer Lett., 194, 13–19,
2003.
38. Bobe, G., Wang, B., Seeram, N.P., Nair, M.G., and Bourquin, L.D., Dietary anthocyanin-rich tart cherry
extract inhibits intestinal tumorigenesis in APCMin mice fed suboptimal levels of sulindac. J. Agric.
Food Chem., 54, 9322–9328, 2006.
39. Howatson, G., McHugh, M.P., Hill, J.A., Brouner, J., Jewell, A.P., van Someren, K.A., Shave, R.E.,
and Howatson, S.A., Influence of tart cherry juice on indices of recovery following marathon running.
Scand. J. Med. Sci. Sports, 20, 843–852, 2010.
40. Kuehl, K.S., Perrier, E.T., Elliot, D.L., and Chesnutt, J.C., Efficacy of tart cherry juice in reducing
muscle pain during running: a randomized controlled trial. J. Inter. Soc. Sports Nutr., 7, 1–6, 2010.
41. Turkish Food Codex (TGK), Fruit juice and similar products Legislation, No: 2006:56, 2006. Published
online at: http://www.resmigazete.gov.tr/eskiler/2006/12/20061230–42.htm (accessed March 3, 2014).
42. Lee, T., Chang, P.K., and Chen, H., Non-nutritive sweetened beverages with LHG juice concentrate. US
Patent Application Publication, Pub. No. US2008/0226796A1, 2008.
43. Ova, G., Zorba, M., and Gur, E., The use of synthetic sweeteners in cherry and orange fruit drinks.
Gıda, 26, 223–228, 2001.
Cherry Juice 185
44. Aroma Bursa Fruit Juice and Food Industry Inc., Aroma 100% Fruit Juice, Sour Cherry-Apple,
Specification No: SP003T, 2015. Published online at: http://www.aroma.com.tr/images/pdf/100de100/
SP003T-100-aroma-visne-elma-suyu.pdf (accessed September 21, 2015).
45. Altuntas, J., Akdemir Evrendilek, G., Sangun, M.K., and Zhang, H.Q., Effects of pulsed electric field
processing on the quality and microbial inactivation of sour cherry juice. Int. J. Food Sci. Technol., 45,
899–905, 2010.
46. Bayindirli, A., Alpas, H., Bozoglu, F., and Hizal, M., Efficiency of high pressure treatment on inactiva-
tion of pathogenic microorganisms and enzymes in apple, orange, apricot and sour cherry juices. Food
Control, 17, 52–58, 2006.
15
Cherry Laurel Syrup (Pekmez)
CONTENTS
15.1 Introduction....................................................................................................................................187
15.4 Health Effects.................................................................................................................................189
15.6 Conclusion..................................................................................................................................... 192
References............................................................................................................................................... 192
15.1 Introduction
Cherry laurel (Laurocerasus officinalis Roem.) belongs to the Rosaceae family and is a popular fruit
(dark purple or black when mature), mainly distributed and cultivated off the coasts of the Black Sea
region of Turkey and is locally called Taflan or Karayemiş [1,2]. It is native to some Asian countries,
the Caucasia, Iran, some Mediterranean countries, Bulgaria, Serbia, and Western Europe [3,4]. Cherry
laurel is mostly consumed as fresh fruit in local markets but may also be dried, pickled, and processed
into syrup (known as pekmez), jam, and marmalade. In addition to its use for food, both fruit and seeds
of cherry laurel are well known as traditional medicine in Turkey and have been used for many years for
the treatment of stomach ulcer, digestive system complaints, bronchitis, eczemas, hemorrhoids, diuretic
agent, hypoglycemia, and hyperglycemia, among others [3,5–9].
From a technological point of view, the traditional process to prepare pekmez is to start from a boiled
fruit pulp in water and pressed to extract the juice. The extracted juice is concentrated by boiling/heating
over low heat until becoming colored and a syrupy liquid, which has a total sugar concentration of
approximately 60°Brix. Pekmez has traditionally been used for centuries in Turkey. Fresh or dried fruits
such as grapes are mainly used for the production of pekmez, but other sugar-rich fruits such as apples,
pears, plums, mulberries, cherry laurels, watermelons, and apricots can also be used in pekmez produc-
tion [10,11]. Among these fruits, pekmez from cherry laurel is becoming increasingly popular because of
its potential and perceived health benefits [1,2,12]. Pekmez from cherry laurel is also diluted with water
(1:5 or 1:10, pekmez/water) and used as refreshing or energy drink in Turkey. This chapter highlights
and compares the nutritional characteristics, bioactives, antioxidant efficacy, health effects, and novel
formulations of cherry laurel fruit and pekmez.

The proximate compositions of cherry laurel and pekmez are summarized in Table 15.1. Significant
differences (P < 0.05) exist among cherry laurel and pekmez. Moisture is the predominant component,
followed by carbohydrate with small amounts of protein and fat. Six sugars are present in cherry laurel
and pekmez; these include monosaccharides (xylose, arabinose, fructose, and glucose), a sugar alcohol
(sorbitol), and trace amounts of the disaccharide sucrose [1]. Monosaccharides, which represent 80.5%
187
TABLE 15.1
Compositional and Nutritional Characteristics of Cherry Laurel and Pekmez (per 100 g)
Nutrient Unit Cherry Laurel Pekmez References
Water g 77.28 52.46 [1]
Protein g 1.51 1.33 [1]
Fat (lipid) g 0.23 0.14 [1]
Carbohydrate g 20.23 44.22 [1]
Ash g 0.75 1.85 [1]
Sugars
Xylose g 0.19 0.48 [1]
Arabinose g 0.08 0.21 [1]
Fructose g 5.16 13.76 [1]
Glucose g 5.88 16.39 [1]
Sorbitol g 4.80 7.43 [1]
Sucrose g tr 0.05 [1]
Total sugars g 16.11 38.32 [1]
Minerals
Calcium mg 15.3 151 [3]
Copper mg 0.08 tr [3]
Iron mg 0.83 4.73 [3]
Magnesium mg 17.9 59.98 [3]
Manganese mg 2.42 5.01 [3]
Phosphorus mg na 21.26 [3]
Potassium mg 222 645 [3]
Selenium μg na na [3]
Sodium mg 5.5 12.82 [3]
Zinc mg 0.19 0.48 [3]
Vitamins
Ascorbic acid mg 204 na [3]
Niacin mg na 0.95 [3]
Pyridoxine mg na 0.14 [3]
Riboflavin mg na 0.05 [3]
Thiamin mg na 0.01 [3]
Note: Unpublished data (mineral and vitamin contents of pekmez).
Abbreviations: tr, trace; na, not available.
of the total sugar in pekmez, can easily pass into the bloodstream without digestion. Glucose is the pre-
dominant sugar with fructose, followed by sorbitol and small amounts of xylose, arabinose, and sucrose
in both cherry laurel and pekmez. Total sugar content varies between 38.22 g/100 g in pekmez and
16.11 g/100 g in cherry laurel.
With respect to minerals, cherry laurel and pekmez are excellent sources of manganese (2.42 and
5.01 mg/100 g, respectively). The Recommended Dietary Allowances (RDA) of manganese is 2.3 and
1.8 mg/day for males and females, respectively. It is interesting to note that cherry laurel fruit is an
excellent source of ascorbic acid (204 mg/100 g) [3], but this vitamin is lost completely during heat
processing in pekmez preparation. Small amounts of niacin, pyridoxine, riboflavin, and thiamin are also
present in pekmez (unpublished data) (Table 15.1).
Harvest time, maturity state, season, age of trees, geographical origin, environmental factors, storage,
processing conditions, and handling conditions, in addition to the variety of cherry laurel, affect the
composition of products [3,13,14].
Cherry Laurel Syrup (Pekmez) 189
TABLE 15.2
Content of Total Antioxidant Activity: Anthocyanins, Carotenoids, and Phenolics
in Cherry Laurel and Pekmez
Phenolics Unit Cherry Laurel Pekmez
ORAC μmol TE/g 3363 19981
Total anthocyanins mg C3GE/100 g 123.8 9.3
Total carotenoids μg/100 g 254 114
Total phenolics mg FAE/100 g 454 1444
Source: Adapted from Alasalvar, C. et al., J. Food Sci., 70, S47, 2005. With permission.
ORAC, oxygen radical absorbance capacity; TE, trolox equivalents; C3GE, cyan-
Abbreviations:
idin 3-glucoside equivalents; FAE, ferulic acid equivalents.

Table 15.2 presents total antioxidant activity, anthocyanins, cartonenoids, and phenolics in cherry laurel
and pekmez. Pekmez contains significantly higher oxygen radical absorbance capacity value and content
of phenolics than cherry laurel, whereas total anthocyanins and carotenoids are lower. Alasalvar et al.
[1] found that significant amount of anthocyanins (92.5%) and carotenoids (44.9%) were lost during heat
processing in the production of pekmez. Studies have also shown that anthocyanins are readily destroyed
by heat during food processing [15,16]. Other than heat, many other factors have been reported as being
responsible for the degradation of anthocyanins during the processing of fruits [16,17].
Cherry laurel and pekmez were examined for their antioxidant activities using different free radical
scavenging activity tests (hydrogen peroxide, superoxide radical anion, and 2,2-diphenyl-1-picrylhydrazyl
[DPPH] radical), together with reducing power and inhibition of oxidation of human low-density lipo-
protein cholesterol [2]. On a fresh weight, pekmez exhibited a significantly (P < 0.01) higher antioxidant
activity than that of cherry laurel in most cases; however, on a dry weight basis, hydrogen peroxide and
DPPH radical scavenging activities and reducing power were significantly higher (P < 0.01) in cherry
laurel than in its pekmez, with some exceptions, thus indicating possible destruction of antioxidative
compounds during pekmez production (Table 15.3).
The content of free and bound phenolic acids in cherry laurel and pekmez are listed in Table 15.4. The
samples studied contained hydroxylated derivatives of benzoic acid (gallic acid, protocatechuic acid,
p-hydroxybenzoic acid, vanillic acid, and syringic acid) and cinnamic acid (chlorogenic acid, caffeic
acid, p-coumaric acid, ferulic acid, and o-coumaric acid) (Figure 15.1) [1]. Chlorogenic acid was the main
phenolic acid in the free form, whereas caffeic acid predominated in the bound form. Total phenolic acids
ranged from 390 to 836 mg/100 g, being highest in pekmez and lowest in cherry laurel.
Ayaz et al. [18] found five phenolic acids (protocatechuic acid, p-hydroxybenzoic acid, vanillic acid,
caffeic acid, and p-coumaric acid) in three cultivated and one wild cherry laurel varieties, among
which vanillic acid was the major compound. Subsequently, Ayaz [19] studied the changes in phe-
nolic acids of cultivated cherry laurel during maturation. In this work, seven phenolic acids (benzoic
acid, 4-hydroxybenzoic acid, p-coumaric acid, vanillic acid, 3,4-dihydroxybenzoic acid, caffeic acid,
and ferulic acid) were identified and quantified, among which benzoic, caffeic, and vanillic acids were
present in the highest amounts, respectively.
15.4 Health Effects

In Turkish folk medicine, cherry laurel fruit and seeds have been used as a home medicine for the
treatment of digestive system complaints, stomach ulcer, bronchitis, eczemas, hemorrhoids, diuretic
agent, hypoglycemia, and hyperglycemia, among others [3,5–9]. Pekmez from cherry laurel is becoming
TABLE 15.3
Free Radical Scavenging Activities, Reducing Power, and Inhibition of Oxidation of Human Low-Density
Lipoprotein Cholesterol by Cherry Laurel and Pekmez
FW DW
100 ppm 200 ppm 400 ppm 100 ppm 200 ppm 400 ppm
Hydrogen Peroxide Scavenging
Cherry laurel 0.8 0.8 3.7 3.5 3.5 16.3
Pekmez 1.0 1.2 2.8 2.1 2.5 5.9
Catechina 93.5 96.0 100 93.5 96.0 100
Superoxide Radical Scavenging
Cherry laurel 18.7 22.3 25.1 82.3 98.2 100
Pekmez 61.9 79.7 92.1 100 100 100
Catechin 90.7 94.0 100 90.7 94.0 100
DPPH Radical Scavenging
Cherry laurel 14.0 20.7 23.4 61.6 91.1 100
Pekmez 13.9 25.4 50.8 29.2 53.4 100
Catechin 97.0 100 100 97.0 100 100
Reducing Power
Cherry laurel 7.0 11.8 20.7 30.8 51.9 91.1
Pekmez 6.2 17.2 39.3 13.0 36.2 82.7
Catechin 622 622 622 622 622 622
Inhibition of Oxidation of Human LDL Cholesterol
Cherry laurel 9.7 — 13.3 42.7 — 58.5
Pekmez 23.4 — 36.5 49.2 — 76.8
Catechin 100 — 100 100 — 100
Source: Adapted from Liyana-Pathirana, C.M.M. et al., Food Chem., 99, 121, 2006. With permission.
a Reference antioxidant compound.
Abbreviations: FW, fresh weight; DW, dry weight; DPPH, 2,2-diphenyl-1-picrylhydrazyl; LDL, low-density lipoprotein.
increasingly popular because of its potential and perceived health benefits. Most of its carbohydrate is in
the form of fructose and glucose (13.76 and 16.39 g/100 g pekmez, respectively) (Table 15.1), which eas-
ily passes into blood without digestion. This is nutritionally important especially for babies, children,
athletes, and in situations demanding a rapid energy source [12]. Research on the health benefits specifically
derived from the consumption of pekmez is not available. Therefore, health effects should be investi-
gated through well-designed human intervention studies. Although not studied as much as some other
fruit juices, there appears to be a significant scope to develop the evidence and a profile for positioning
pekmez in terms of its health benefits.

As mentioned previously, cherry laurel is mostly consumed as fresh fruit in local markets but may
also be dried, pickled, and processed into pekmez, jam, marmalade, and fruit juice products. To
benefit the global consumer, there is a need to develop juice-based products using novel process-
ing techniques for best preserved properties of cherry laurel along with globally acceptable taste as
some cherry laurel varieties have astringent and sour taste. Novel product formulations are gaining
importance every day. Therefore, new formulations and fortification with other juice blends should
also be developed.
Cherry Laurel Syrup (Pekmez) 191
TABLE 15.4
Content of Free and Bound Phenolic Acids in Cherry Laurel and Pekmez (mg/100 g)
Phenolic Acids Kiraz Pekmez
Free Phenolic Acids
Protocatechuic 1.12 8.34
p-Hydroxybenzoic 2.38 nd
Chlorogenic 136 195
Vanillic 12.41 nd
Caffeic nd 74.39
Syringic 56.15 nd
p-Coumaric 49.70 20.47
Bound Phenolic Acids
Gallic 37.57 276
p-Hydroxybenzoic 2.32 28.00
Vanillic 4.96 5.46
Caffeic 74.83 198
Syringic nd 1.98
p-Coumaric 12.89 28.09
Ferulic nd nd
o-Coumaric nd nd
Total Phenolic Acids 390 836
Source: Adapted from Alasalvar, C. et al., J. Food Sci., 70, S47, 2005. With permission.
HO O
HO
HO OH
O
OH
HO O
O
HO OH
OH
Caffeic acid Chlorogenic acid
HO
O OH
O HO OH
HO OH
p-Coumaric acid Gallic acid
HO HO
HO
O O
HO HO
p-Hydroxybenzoic acid Protocatechuic acid
FIGURE 15.1 Chemical structures of major phenolic acids present in pekmez.

15.6 Conclusion
Nutritionally, pekmez contains high amount of sugars combined with a low content of protein and fat. It
is a good source of manganese and iron. Pekmez is also a good source of bioactive compounds (particu-
larly, phenolic acids) and possesses high antioxidant activities. Further research is warranted to identify
other bioactive compounds present and to develop and introduce cherry laurel juice to the market.
REFERENCES
1. Alasalvar, C., Al-Farsi, M., and Shahidi, F., Compositional characteristics and antioxidant components
of cherry laurel varieties and pekmez. J. Food Sci., 70, S47–S52, 2005.
2. Liyana-Pathirana, C.M., Shahidi, F., and Alasalvar, C., Antioxidant activity of cherry laurel
(Laurocerasus officinalis Roem.) and its concentrated juice. Food Chem., 99, 121–128, 2006.
3. Kolayli, S., Küçük, M., Duran, C., Candan, F., and Dinçer, B., Chemical and antioxidant properties
of Laurocerasus officinalis Roem. (Cherry laurel) fruit grown in the Black Sea region. J. Agric. Food
Chem., 51, 7489–7494, 2003.
4. Sahan, Y., Cansev, A., Celik, G., and Cinar, A., Determination of various chemical properties, total phe-
nolic contents, antioxidant capacity and organic acids in Laurocerasus officinalis fruits. Acta Hortic.,
939, 359–366, 2012.
5. Baytop, T., Therapy with Medicinal Plants in Turkey (Past and Present), Istanbul University Publication
No. 3255, Istanbul, Turkey, 1984.
6. Yeşilada, E., Sezik, E., Honda, G., Takaishi, Y., Takeda, Y., and Tanaka, T., Traditional medicine in
Turkey IX: Folk medicine in north-west Anatolia. J. Ethnopharmacol., 64, 195–210, 1999.
7. Colak, A., Özen, A., Dincer, B., Güner, S., and Ayaz, F.A., Diphenolases from two cultivars of cherry
laurel (Laurocerasus officinalis Roem.) fruits at an early stage of maturation. Food Chem., 90, 801–807,
2005.
8. Halilova, H. and Ercisli, S., Several physico-chemical characteristics of cherry laurel (Laurocerasus
officinalis Roem.) fruits. Biotechnol. Biotechnol. Eq., 24, 1970–1973, 2010.
9. Turan, M.I., Turkoglu, M., Dundar, C., Celik, N., and Suleyman, H., Investigating the effect of Prunus
laurocerasus fruit extract in type II diabetes induced rats. Int. J. Pharmacol., 9, 373–378, 2013.
10. Aksu, M.İ. and Nas, S., Dut pekmezi üretim tekniği ve çesitli fiziksel—kimyasal özellikleri (Mulberry
pekmez manufacturing technique and physical and chemical properties). Gıda, 21, 83–88 (in Turkish),
1996.
11. Tosun, I. and Ustun, N.S., Nonenzymic browning during storage of white hard grape pekmez (Zile
pekmezi). Food Chem., 80, 441–443, 2003.
12. Batu, A., Kuru üzüm ve pekmezin insan sağlığı ve beslenmesi açısından önemi (The importance of
raisin and “pekmez” on human health and nutrition). Gıda, 18, 303–307 (in Turkish), 1993.
13. Ayaz, F.A., Kadioğlu, A., Reunanen, M., and Var, M., Sugar composition in fruits of Laurocerasus
officinalis Roem. and its three cultivars. J. Food Comp. Anal., 10, 82–86, 1997.
14. Ustun, N.S. and Tosun, I., A research on composition of wild cherry laurel (Laurocerasus officinalis
Roem.). J. Food Technol., 1, 80–82, 2003.
15. Markakis P., Anthocyanins and their stability in foods. CRC Crit. Rev. Food Technol., 4, 437–456, 1974.
16. Mazza, G. and Miniati, E., Anthocyanins in Fruits, Vegetables, and Grains, CRC Press, Boca Raton,
FL, 1993.
17. Shahidi, F. and Naczk, M., Phenolics in Food and Nutraceuticals, CRC Press, Boca Raton, FL, 2004.
18. Ayaz, F.A., Kadioğlu, A., Reunanen, M., and Var, M., Phenolic acid and fatty acid composition in the
fruits of Laurocerasus officinalis Roem. and its cultivars. J. Food Comp. Anal., 10, 350–357, 1997.
19. Ayaz, F.A., Changes in phenolic acids of cherry laurel (Laurocerasus officinalis “Oxygemmis”) fruit
during maturation. Acta Biol. Cracov. Bot., 43, 23–26, 2001.
16
Coconut Juice
Melinda Phang and Manohar Garg
CONTENTS
16.1 Introduction................................................................................................................................... 193
16.3.1 Phytohormones................................................................................................................. 195
16.3.2 Micronutrients for Antioxidant Efficacy.......................................................................... 195
16.4 Health Effects................................................................................................................................ 196
16.4.1 Hypolipidemic and Antiatherosclerotic Effects............................................................... 196
16.4.2 Hypoglycemic Effects...................................................................................................... 197
16.4.3 Antibacterial Activity....................................................................................................... 197
16.4.4 Antidote Effects................................................................................................................ 197
16.4.5 Hormone-like Effects....................................................................................................... 197
16.4.6 Effects of Vitamins........................................................................................................... 198
16.4.7 Effects of Minerals and Electrolytes................................................................................ 198
16.4.8 Effects of Phytohormones................................................................................................ 198
16.6 Conclusion..................................................................................................................................... 200
References............................................................................................................................................... 200
16.1 Introduction
The coconut (Cocos nucifera) provides food and drink for millions of people, especially those in tropical
and subtropical regions. Due to its many uses, such as providing food, drink, oil, medicine, and mate-
rial for domestic utensils, it is often called the “tree of life” [1]. The coconut palm is noninvasive, and
its cultivation and spread from its natural habitat is largely man-made. In the nineteenth century, the
arrival of Europeans in the Pacific signaled the commercialization of the plant, and coconut oil was the
first vegetable oil to appear in the world trade. Such demand triggered the establishment of large coconut
plantations in the European colonies around the world. However, after the World War II, its importance
declined with the emergence of alternative vegetable oils such as soybean, groundnut, sunflower, and
canola [1]. Today, Indonesia, the Philippines, and India are the largest coconut-producing countries [2].
The coconut palm continues to be a multivariate unique source for the development of industrial prod-
ucts as well as medicine.
The coconut fruit is a fibrous drupe consisting of a thin hard skin (exocarp), thicker layer of fibrous
mesocarp (husk), hard endocarp (shell), white endosperm (kernel), and a large cavity filled with liquid
(juice or water). The edible part of the coconut fruit (coconut meat and coconut juice) is the endosperm
tissue. Initially, the cellular endosperm is translucent and jelly-like, but it later hardens at maturity to
become white flesh (coconut meat). Unlike other plants, (e.g., wheat and corn), the cellularization process
of the endosperm does not fill up the entire embryo sac cavity [3]. In a coconut fruit, the cavity is filled
with what is commonly known as “coconut juice” or sometimes referred to as “coconut water.” With
193
its many applications, it is one of the world’s most versatile natural products. In this chapter, the term
“coconut juice” is used and exclusively focuses on the health benefits of coconut juice where information
pertaining to whole or dried fruits or coconut oils are not discussed.

Coconut juice is a refreshing and nutritious beverage that is consumed worldwide. In a healthy, undam-
aged coconut, the juice is sterile and a ready source of clean drinking water, especially after a natural
disaster (cyclones and flooding). The composition of coconut juice changes during the course of the rip-
ening of the fruits. The juice of a very young coconut (~3–5 months, before formation of endosperm) is
like tasteless astringent water, while mature coconut juice has a slightly salty taste. During the early days
of a coconut, only inverted sugars are present, and their concentration in the coconut juice increases to a
maximum by the fifth and sixth month. Therefore, the ideal time to harvest a coconut for drinking its juice
is at age 6–7 months when the endosperm begins to form. At this time, the coconut juice has maximum
sweetness and low acidity, serving as a pleasant and refreshing drink.
Coconut juice is traditionally used as a growth supplement in plant tissue cultures; the wide applications of
coconut juice can be justified by its unique chemical composition of sugars, vitamins, and minerals (Table 16.1).
TABLE 16.1
Compositional and Nutritional Characteristics of Coconut Juice (per 100 g)
Unit Coconut Juice
Water g 94.99
Energy kcal 19
Protein g 0.72
Lipid (fat) g 0.20
Carbohydrate g 3.71
Total sugars g 2.61
Minerals
Calcium mg 24
Copper mg 0.04
Iron mg 0.29
Magnesium mg 25
Manganese mg 0.14
Phosphorus mg 20
Potassium mg 250
Selenium µg 1.0
Sodium mg 105
Zinc mg 0.10
Vitamins
Folate (DFE) µg 3
Niacin mg 0.080
Pantothenic acid mg 0.043
Riboflavin mg 0.057
Thiamin mg 0.030
Vitamin B6 mg 0.032
Vitamin C mg 2.4
Source: Adapted from the U.S. Department of Agriculture (USDA), USDA National Nutrient
Database for Standard Reference, Release 26, 2013, Published online at: http://www.
nal.usda.gov/fnic/foodcomp/cgi-bin/list_nut_edit.pl/ (accessed June 20, 2014).
Abbreviation: DFE, dietary folate equivalents.
Coconut Juice 195
It is rich in potassium (250 mg/100 g) and contains calcium, magnesium, phosphorus, and sodium.
Coconut juice also contains B vitamins including niacin (0.08 mg), folic acid (3 µg), pantothenic acid
(0.043 mg), riboflavin (0.057 mg), pyridoxine (0.032 mg), thiamin (0.03 mg), and vitamin C (2.4 mg) per
100 g of juice [4,5] (Table 16.1).

16.3.1 Phytohormones
The bioactive compounds present in coconut juice are not fully characterized. A number of growth-
promoting factors, known as phytohormones, widely used in the plant tissue culture industry have been
identified (Table 16.2). Since the discovery of auxin in 1926, several types of phytohormones mainly
including auxins, cytokinins, abscisic acid, gibberellins, and ethylene have been reported (Figure 16.1). A
high-performance liquid chromatography (HPLC) method has been developed for the simultaneous deter-
mination of various classes phytohormones, including indole-3-acetic acid, indole-3-butyric acid, abscisic
acid, gibberellic acid, zeatin, N6-benzyladenine, naphthaleneacetic acid, and 2,4-dichlorophenoxyacetic
acid in young coconut juice [6,9]. There is limited information in the literature as to whether the benefi-
cial health properties of coconut juice can be attributed to the presence phytohormones, therefore, merits
further investigations.
16.3.2 Micronutrients for Antioxidant Efficacy

Micronutrients such as minerals and vitamins in coconut juice play a vital role for providing antioxi-
dants in the human body. They can act as electron donors to directly quench free radicals or indirectly
as a part of metalloenzymes (enzymes that require a catalytic metal ion for their biological activity) such
as glutathione peroxidase (selenium) or superoxide dismutase (zinc and copper) to catalyze the removal
of oxidizing species [10]. The free amino acid, l-arginine, is also present in coconut juice (30 mg/dL),
which is known to significantly reduce free radical generation [11]. Other components found in coconut
water include nitrogenous compounds, organic acids, phytohormones, and enzymes [5]. Each of these
components play different functional roles in plant and human systems due to their distinct chemical
properties.
TABLE 16.2
Phytohormones in Young Coconut Juice
Class Phytohormone Concentration (nM) References
Auxin Indole-3-acetic acid 150.6 [6]
Abscisic acid 65.5 [6]
Cytokinins N6-Isopentenyladenine 0.26 [7]
Dihydrozeatin 0.14 [7]
trans-Zeatin 0.09 [7]
Kinetin 0.31 [7]
ortho-Topolin 3.29 [7]
Dihydrozeatin-O-glucoside 46.6 [7]
trans-Zeatin-O-glucoside 48.7 [7]
trans-Zeatin riboside 76.2 [7]
Kinetin riboside 0.33 [7]
trans-Zeatin riboside-5′-monophosphate 10.2 [7]
Gibberellins Gibberellin-1 16.7 [8]
Gibberellin-2 37.8 [8]
O H O
CI
O OH
OH
HO
CH3H CH2 N
HO O H
Gibberellin-3 Indole-3-acetic acid
O OH CH3
OH
O O
O
HO
Abscisic acid Jasmonic acid
CH3
HO
NH
N N
N N
H
Zeatin (cytokinin)
FIGURE 16.1 Common phytohormones found in coconut juice.
16.4 Health Effects

The parts of a coconut, like coconut kernels and coconut juice, have numerous medicinal properties
including antibacterial, antifungal, antiviral, antiparasitic, antidermatophytic, antioxidant, cardioprotec-
tive, hypoglycemic, and hypolipidemic effects.
16.4.1 Hypolipidemic and Antiatherosclerotic Effects

Elevated plasma cholesterol, particularly low-density lipoprotein (LDL) and very-low-density lipopro-
tein (VLDL) cholesterols are major risk factors for atherosclerotic disease, while increasing the levels
of high-density lipoprotein (HDL) cholesterol is associated with a reduced risk [12]. The effects of
coconut juice on the formation of hyperlipidemia and atherosclerosis have been reported in animal
studies. In 8-week-old quails, a high-fat diet supplemented with coconut juice (20 mL) daily for 12 weeks
resulted in an increase in serum HDL cholesterol levels by 46.2% and liver total cholesterol levels by
26.3% compared to those without coconut juice supplementation [13]. More recent studies by Sandhya
and Rajamohan [14] showed that coconut juice feeding significantly reduced hyperlipidemia in choles-
terol fed rats and that the hypolipidemic effect of coconut juice (4 mL/100 g body weight) was similar to
that of the lipid lowering drug lovastatin (0.1/100 g diet) [15]. In this study, coconut juice supplementa-
tion reduced the levels of serum total cholesterol, VLDL + LDL cholesterol, triacylglycerols (TAG), and
increased HDL cholesterol in experimental rats (P < 0.05). Coconut juice supplementation decreased
activities of hepatic lipogenic enzymes and increased lipoprotein lipase activity, hepatic bile acid, fecal
bile acids, neutral sterols, and 3-hydroxy-3-methylglutaryl-CoA reductase, the rate-limiting enzyme in
cholesterol synthesis [15].
Coconut juice also contains several biologically active components, l-arginine, ascorbic acid, cal-
cium, magnesium, and potassium (Table 16.1), which have beneficial effects on lipid levels. Several
animal studies have shown that dietary calcium, magnesium, and potassium decreases serum choles-
terol levels and results in less extensive atherosclerotic lesions [16–18]. Calcium supplementation has
Coconut Juice 197
also been shown to decrease intestinal absorption of fat and lowering cholesterol concentration, while a
potassium-enriched diet decreased cholesterol ester deposition during hypercholesterolemia [16].
16.4.2 Hypoglycemic Effects

Hyperglycemia is a condition characterized by an abnormal postprandial increase in blood glucose
level and has been associated with the onset of type 2 diabetes [19]. Coconut juice has been reported
to reduce blood glucose levels as well as to exert antioxidant activity in diabetic rats [20]. Coconut
juice supplementation (4 mL/100 g body weight) in diabetic rats for 45 days resulted in a significant
reduction in blood glucose, glycated hemoglobin levels, improved insulin levels, and increased levels
of carbohydrate-metabolizing enzymes. This study suggested that coconut juice exerts antidiabetogenic
activity via modulating glucose and insulin levels as well as the enzymes involved in carbohydrate
metabolism and its protective effects on pancreatic tissue [20].
Dietary amino acids are important modulators of insulin and glucose homeostasis [21]. l-arginine
possesses antiglycation and antiperoxidative potential in diabetes [22]. A recent study also reported
that l-arginine has the ability to regenerate pancreatic β-cells and reduce pancreatic damage in diabetic
rats [23]. Magnesium and potassium, both present in coconut juice, have been reported to possess anti-
hyperglycemic potential [24,25].
16.4.3 Antibacterial Activity

The increased tendency of microorganisms to develop resistance to antibiotics presents a negative impact
on human health. Antimicrobial peptides have been isolated and identified in coconut juice. A study
reported that green coconut is able to synthesize different antimicrobial peptides in their water, with
activity against human pathogenic bacteria such as Staphylococcus aureus [26]. Crude water extract
from the coconut husk fiber also showed inhibitory action against acyclovir-resistant herpes simplex
virus type 1 (HSV-1-ACVr) [27].
16.4.4 Antidote Effects

The use of coconut juice to counteract poisons has been a common practice in India and Africa and is
used as an immediate remedy for drug overdose [28]. Coconut juice enhances the dilution effect, which
increases the rate and degree of gastrointestinal absorption of various chemicals [29]. Due to its rich min-
eral content, coconut juice aids in quick absorption of the drug and makes their peak concentration in the
blood easier by its electrolytic effect [30]. Coconut juice has been reported to eliminate mineral poisoning
and ameliorate drug induced over dosage toxicity in a dose-dependent manner [30]. In addition, coconut
juice is enriched with B vitamins, which play a role in the maintenance of liver cell integrity [31].
16.4.5 Hormone-like Effects

Coconut juice has been used in Thailand for several decades to alleviate symptoms in postmenopausal
women associated with the decrease in estrogen levels. These estrogenic characteristics were also dem-
onstrated to accelerate the healing process of cutaneous wounds in ovariectomized rats, a model often
used for menopause. Morphological changes in those receiving 100 mL/kg body weight of coconut juice
daily for 1 week included better wound healing and less scaring [32]. With a 2-week supplementation, the
accelerated wound healing was characterized by reduced wound depth and width, increased epidermal
thickness, and more abundant collagen fibers [33].
In addition, it has been postulated that low estrogen levels may be associated with an increased risk
of Alzheimer’s disease [34,35]. Studying the brains from the ovariectomized rats displaying similar
features to postmenopausal women with Alzheimer’s disease, it was found that the estrogenic proper-
ties in coconut juice significantly reversed some pathologies in the rat brain associated with hormonal
imbalance [36,37].
16.4.6 Effects of Vitamins

Coconut juice is rich in B complex vitamins that are essential for the normal functioning of the human
body (Table 16.1). The B complex vitamins are water soluble and are required as coenzymes for enzy-
matic reactions essential for cellular function [38]. Vitamin B6 (pyridoxal, pyridoxine, and pyridoxamine)
serves as a coenzyme in various enzymatic reactions including the transamination and decarboxylation
reactions. Vitamin B6 is required as a coenzyme of γ-cystathionase [39], which catalyses the cleavage
of cystathionine, and releasing α-ketobutyrate and cystein [40]. The α-ketobutyrate molecule is subse-
quently converted into succinyl-CoA and fed to the tricarboxylic acid cycle, while cystein is involved in
the biosynthesis of protein and glutathione [41]. Indeed, a deficiency in vitamin B6 can affect the inflam-
matory process and renal function [38].
Coconut juice also contains vitamin B9 (folate), which was identified in the 1930s as the nutrient
required in treating anemia during pregnancy [42]. Later studies also showed that folic acid prophy-
laxis decreased the risk of premature deaths in malnourished populations [43]. Folate can also pre-
vent mitochondrial toxicity induced by methanol metabolites. Furthermore, the active form of folate,
5-methyltetrahydrofolate, is believed to be one of the central methyl donors required for mitochondrial
protein and nucleic acid synthesis [10]. Studies have reported that low circulating blood levels of vitamin
B6 and folate are associated with an increased risk of atherosclerosis and other vascular diseases [44].
Conversely, another study reported that higher plasma levels of vitamin B6 and folate may reduce the risk
for breast cancer [45]. In addition to B complex vitamins, coconut juice also contains vitamin C (ascorbic
acid), which possesses antioxidant activities [10].
16.4.7 Effects of Minerals and Electrolytes

Minerals are crucial for enzyme activation, bone formation, hemoglobin function, gene expression, and
the metabolism of carbohydrates, lipids, and amino acids. Coconut juice contains a variety of miner-
als (Table 16.1), which contribute to the nutritional and therapeutic value of coconut juice. It serves as
an effective rehydration drink to replenish the electrolytes of the body excreted through sweat such as
sodium, potassium, and magnesium. The concentration of these electrolytes in coconut juice generates
an osmotic pressure similar to that observed in blood [46]. Furthermore, coconut juice does not affect
hemostasis (plasma coagulation) and, therefore, can be used as short-term intravenous hydration and
resuscitation fluid [47]. In a clinical setting, coconut juice has administered as an oral rehydration aid
to replenish fluids and electrolytes in patients suffering severe dehydration due to diarrhea and is a use-
ful alternative to glucose saline in cases of gastroenteritis and urinary diseases [48,49]. In the sporting
arena, coconut juice has been reported to provide hydrating effects similar to those of carbohydrate–
electrolyte sport drinks [50,51]. Coconut juice also contains vitamin C, sugars, lipids, free amino acids,
phytohormones (auxin, 1, 3-diphenylurea, and cytokinin), enzymes (acid phosphatase, catalase, dehy-
drogenase, diastase, peroxidase, and RNA polymerases), and growth-promoting factors [3]. There is
increasing scientific evidence that supports the role of coconut juice in health and medicinal applications.
The high content of potassium has been reported to lower blood pressure [11] and the ethanolic extract
of the endocarp was found to possess vasorelaxant and antihypertensive effects [52]. Another study also
reported that coconut juice had cardioprotective effects in myocardial infarction-induced rats, and this
was probably attributed to the rich content of mineral ions in coconut juice [53].
16.4.8 Effects of Phytohormones

Phytohormones are a group of naturally occurring organic compounds that play a pivotal role in regu-
lating plant growth. Coconut juice contains the phytohormones: auxin, gibberellins, abscisic acid, and
various cytokinins [9]. The cytokinins found in coconut juice support cell division and thus promote
rapid growth. Of the cytokinins, only kinetin, trans-zeatin and their derivatives are known to possess
medicinal values (Table 16.2) [3].
Kinetin was previously assumed to be an unnatural and synthetic compound, until 1996 when it was
detected in freshly extracted cellular DNA from human cells and in plant cell extracts [54] and recently
Coconut Juice 199
in coconut juice [55]. Kinetin is known for its ability to retard senescence in plants [56], and more
recently, its strong antiaging effects on human skin cells have been reported [57]. Kinetin was shown to
delay the onset of several cellular and biochemical characteristics associated to cellular aging in human
skin fibroblast cultures [57]. Consequently, skin care products containing kinetin were developed to treat
photodamaged skin [58].
Emerging evidence revealed that oxidative DNA damage plays an important role in cancer develop-
ment and that dietary antioxidants are effective against oxidative damage [59]. Kinetin has demonstrated
to protect DNA against oxidative damage by inhibiting the formation of 8-oxo-2′deoxyguanosine, a com-
mon marker of oxidative damage in DNA. Kinetin inhibited 8-oxo-dG formation in a dose-dependent
manner with a maximum of 50% protection observed at 100 μM kinetin [60]. In another study, kinetin
was found to inhibit oxidative and glycooxidative protein damage generated as well as preventing struc-
tural changes induced by glucose, ribose, arabinose, and glyoxal [61]. The antioxidative properties of
kinetin suggest that it may also offer preventative effects against the oxidative damage of unsaturated
fatty acids in cell membranes [62].
trans-Zeatin was the first naturally occurring cytokinin isolated from a plant source (Zea mays) in
1963 [63] and was later identified to be present in coconut juice [64]. In 1975, the presence of both
trans-zeatin and trans-zeatin riboside was verified in coconut juice [65]. Recent studies have shown that
trans-zeatin can be a potential drug to treat neural diseases. The only currently approved compound
for the treatment of Alzheimer’s disease is acetylcholinesterase, which enhances cholinergic transmis-
sion by reducing the enzymatic degradation of acetylcholine. Acetylcholinesterase degrades the neural
compounds that mediate neural transmission, and thus by blocking its action, synaptic transmission can
be improved. Some studies have found that trans-zeatin possesses an inhibitory effect on acetylcholines-
terase, and it can be used to treat neural dysfunctions, such as Alzheimer’s disease or dementia [66,67].
Another study reported the antioxidative and protective effects of zeatin against amyloid β-protein-
induced neurotoxicity. Zeatin can prevent amyloid β-protein formation, which plays a causal role in the
development and progress of Alzheimer’s disease [68]. Coconut juice also significantly reduced histo-
pathological changes in the brain where a significant reduction in neuronal cell death was observed [37].
Like kinetin, trans-zeatin also exhibits antiaging and youth-preserving effects on human fibroblast
cells. Long-term treatment of human skin fibroblasts cells in the presence of zeatin resulted in the pre-
vention of cell enlargement, reduction of intracellular debris, prevention of actin polymerization, and
enhancement of cellular ability to decompose hydrogen peroxide and oxidative stresses [69].

Coconut juice has been extensively studied since its introduction to the scientific community in the
1940s. It is a naturally, refreshing, and nutritious beverage, which is widely consumed due to its health-
promoting properties. The chemical components, which contribute to its bioactivity, are essential to the
plant industry, biotechnology, and biomedical fields.
The recent discovery of other medicinal values of coconut juice signifies great potential in improving
human health. Cytokinins are currently the most important components present in coconut juice and the
potential anticancer properties of specific cytokinins encourage novel perspectives in the treatment of
certain cancers.
The chemical composition of coconut juice is affected by several factors as the juice of different coco-
nut varieties contains varying concentration of compounds. The chemical contents of the juice also vary
during the different stages of maturity [70]. Furthermore, soil and environmental conditions also affect
the chemical profile of the coconut juice where its physical properties have been reported to be affected
by varying nitrogen and potassium applications [71]. Hence, it would be ideal to conduct studies to deter-
mine the factors that produce the desirable chemical composition for a specific purpose. Breeding studies
can also be carried out to produce coconut juice enriched with specific chemical compounds.
In the view that coconut juice supports cell growth, it may be used in the application to promote
growth of human tissues including hair follicles, therefore may be used in the cosmetic industry in hair
formulation and rejuvenating topical treatments.
In addition, coconut juice is widely used in the plant tissue culture industry as a growth-promoting
component in tissue culture medium. As the coconut is able to synthesize different antimicrobial pep-
tides in its juice, application in pathogen inhibition may lead to genetic engineering–derived antibiotic
production or development of novel disinfectants for hospital environments.
16.6 Conclusion
Coconut juice or water is very low in fat and calories, and is a refreshing liquid found inside young green
coconuts that is rich in electrolytes such as potassium and unique nutrients including vitamins, minerals,
enzymes, free amino acids, cytokinins, and phytohormones with potential antiaging, anticarcinogenic,
and antithrombotic effects. A nutritionally well-balanced fluid has the potential to be a more ideal drink
than any other brand of soft drink beverages in dehydration conditions and as a “natural sports drink.”
Coconut juice also contains several naturally occurring bioactive enzymes such as acid phosphatase, cat-
alase, dehydrogenase, diastase, peroxidase, and RNA polymerases that can aid in digestion and metabo-
lism. Health claims on coconut juice including lowering blood pressure, antiaging, and antithrombotic
and anticarcinogenic potentials for use as a sports drink and rehydration fluid in diarrheal patients merit
further substantiation using randomized controlled intervention trials.
REFERENCES
1. Chan, E. and Elevitch, C., Species profiles for Pacific Island Agroforestr, 2006. Published online at:
http://www.agroforestry.org/images/pdfs/Cocos-coconut.pdf (accessed September 12, 2013).
2. DebMandal, M. and Mandal, S., Coconut (Cocos nucifera L.: Arecaceae): In health promotion and
disease prevention. Asian Pac. J. Trop. Med., 4, 241–247, 2011.
3. Yong, J.W., Ge, L., Ng, Y.F., and Tan, S.N., The chemical composition and biological properties of
coconut (Cocos nucifera L.) water. Molecules, 14, 5144–5164, 2009.
4. Santoso, U., Kubo, K., Ota, T., Tadokora, T., and Maekawa, A., Nutrient composition of kopyor c oconuts.
Food Chem., 57, 299–304, 1996.
Release 26, 2013. Published online at: http://www.nal.usda.gov/fnic/foodcomp/cgi-bin/list_nut_edit.pl/
(accessed June 20, 2014).
6. Ma, Z., Ge, L., Lee, A.S., Yong, J.W., Tan, S.N., and Ong, E.S., Simultaneous analysis of different classes
of phytohormones in coconut (Cocos nucifera L.) water using high-performance liquid chromatography
and liquid chromatography-tandem mass spectrometry after solid-phase extraction. Anal. Chim. Acta,
610, 274–281, 2008.
7. Ge, L., Yong, J.W.H., Tan, S.N., and Ong, E.S., Analysis of cytokinin nucleotides in coconut (Cocos
nucifera L.) water using capillary zone electrophoresis-tandem mass spectrometry after slid-pahse
extraction. Electrophoresis, 1133, 322–331, 2006.
8. Ge, L., Peh, C.Y.C., Yong, J.W.H., Tan, S.N., Hua, L., and Ong, E.S., Analyses of gibberellins by capillary
electrophoresis-mass spectrometry combined with solid-phase extraction. J. Chromatogr., 242–249, 2007.
9. Wu, Y. and Hu, B., Simultaneous determination of several phytohormones in natural coconut juice
by hollow fiber-based liquid-liquid-liquid microextraction-high performance liquid chromatography.
J. Chromatogr. A., 1216, 7657–7663, 2009.
10. Shenkin, A., The key role of micronutrients. Clin. Nutr., 25, 1–13, 2006.
11. Loki, A.L. and Rajamohan, T., Hepatoprotective and antioxidant effect of tender coconut water on
carbon tetrachloride induced liver injury in rats. Indian J. Biochem. Biophys., 40, 354–357, 2003.
12. Gotto, A.M., Jr. and Brinton, E.A., Assessing low levels of high-density lipoprotein cholesterol as a risk
factor in coronary heart disease: A working group report and update. J. Am. Coll. Cardiol., 43, 717–724,
2004.
13. Zhao, G., Dai, Y., and Wang, Y., Effects of coconut juice on the formation of hyperlipidemia and athero-
sclerosis. Zhonghua Yu Fang Yi Xue Za Zhi, 29, 216–218, 1995.
14. Sandhya, V.G. and Rajamohan, T., Beneficial effects of coconut water feeding on lipid metabolism in
cholesterol-fed rats. J. Med. Food., 9, 400–407, 2006.
Coconut Juice 201
15. Sandhya, V.G. and Rajamohan, T., Comparative evaluation of the hypolipidemic effects of coconut
water and lovastatin in rats fed fat-cholesterol enriched diet. Food Chem. Toxicol., 46, 3586–3592, 2008.
16. Tobian, L., Jahner, T.M., and Johnson, M.A., High K diets markedly reduce atherosclerotic cholesterol
ester deposition in aortas of rats with hypercholesterolemia and hypertension. Am. J. Hypertens., 3,
133–135, 1990.
17. Vaskonen, T., Mervaala, E., Seppanen-Laakso, T., and Karppanen, H., Diet enrichment with calcium
and magnesium enhances the cholesterol-lowering effect of plant sterols in obese Zucker rats. Nutr.
Metab. Cardiovasc. Dis., 11, 158–167, 2001.
18. Altura, B.T., Brust, M., Bloom, S., Barbour, R.L., Stempak, J.G., and Altura, B.M., Magnesium dietary
intake modulates blood lipid levels and atherogenesis. Proc. Natl. Acad. Sci. USA, 87, 1840–1844, 1990.
19. Shaw, J.E., Sicree, R.A., and Zimmet, P.Z., Global estimates of the prevalence of diabetes for 2010 and
2030. Diabetes Res. Clin. Pract., 87, 4–14, 2010.
20. Preetha, P.P., Devi, V.G., and Rajamohan, T., Hypoglycemic and antioxidant potential of coconut water
in experimental diabetes. Food Funct., 3, 753–757, 2012.
21. Tremblay, F., Lavigne, C., Jacques, H., and Marette, A., Role of dietary proteins and amino acids in the
pathogenesis of insulin resistance. Annal. Rev. Nutr., 27, 293–310, 2007.
22. Al-Shabrawey, M. and Smith, S., Prediction of diabetic retinopathy: Role of oxidative stress and rel-
evance of apoptotic biomarkers. EPMA J., 1, 56–72, 2010.
23. Salil, G., Nevin, K.G., and Rajamohan, T., Arginine rich coconut kernel protein modulates diabetes in
alloxan treated rats. Chem. Biol. Interact., 189, 107–111, 2011.
24. Abayomi, A.I., Adewoye, E.O., Olaleye, S.B., and Salami, A.T., Effect of magnesium pre-treatment on
alloxan induced hyperglycemia in rats. Afr. Health Sci., 11, 79–84, 2011.
25. Chatterjee, R., Yeh, H.C., Shafi, T., Selvin, E., Anderson, C., Pankow, J.S., Miller, E., and Brancati, F.,
Serum and dietary potassium and risk of incident type 2 diabetes mellitus: The Atherosclerosis Risk in
Communities (ARIC) study. Arch. Intern. Med., 170, 1745–1751, 2010.
26. Mandal, S.M., Dey, S., Mandal, M., Sarkar, S., Marai-Neto, S., and Franco, O.L., Identification and
structural insights of three novel antimicrobial peptides isolated from green coconut water. Peptides, 30,
633–637, 2001.
27. Esquenazi, D., Wigg, W.D., Miranda, M.M., Rodrigues, H.M., Tostes, J.B., Rozental, S., da Silva, A.J.,
and Alviano, C.S., Antimicrobial and antiviral activities of polyphenolics from Cocos nucifera Linn.
(Palmae) husk fiber extract. Res. Microbiol., 153, 647–652, 2002.
28. Biplab, D., Sabita, P.D., Bhattacharjee, P.R., and Goswami, B.B., Coconut: The tree of life. Indian J.
Pharm. Educ., 16, 23–30, 2002.
29. Henderson, M.L., Picchioni, A.L., and Chin, L., Evaluation of oral dilution as a first aid measure in
poisoning. J. Pharm. Sci., 55, 1311–1313, 1966.
30. Effiong, G.S., Ebong, P.E., Eyong, E.U., Uwah, A.J., and Ekong, U.E., Amelioration of chloramphenicol
induced toxicity in rats by coconut water. J. Appl. Sci. Res., 6, 331–335, 2010.
31. Ravichandran, V. and Selvam, R., Increased lipid peroxidation in kidney of vitamin B-6 deficient rats.
Biochem. Int., 21, 599–605, 1190.
32. Radenahmad, N., Vongvatcharanon, U., Withyachumnarnkul, B., and Connor, J.R., Serum lev-
els of 17β-estradiol in ovariectomized rats fed young-coconut-juice and its effect on wound healing.
Songklanagarind J. Sci. Technol., 897–910, 2006.
33. Radenahmad, N., Saleh, F., Sayoh, I., Sawangjaroen, K., Subhadhirasakul, P., Boonyoung, P., Rundorn,
W., and Mitranun, W., Young coconut juice can accelerate the healing process of cutaneous wounds.
BMC Complement. Altern. Med., 12, 252, 2012.
34. Campbell, L., Emmerson, E., Davies, F., Gilliver, S.C., Krust, A., Chambon, P., Ashcroft, G.C., and
Hardman, M.J., Estrogen promotes cutaneous wound healing via estrogen receptor beta independent of
its antiinflammatory activities. J. Exp. Med., 207, 1825–1833, 2010.
35. Hardman, M.J., Emmerson, E., Campbell, L., and Ashcroft, G.S., Selective estrogen receptor modula-
tors accelerate cutaneous wound healing in ovariectomized female mice. Endocrinology, 149, 551–557,
2008.
36. Radenahmad, N., Saleh, F., Sawangjaroen, K., Vongvatcharanon, U., Subhadhirasakul, P., Rundorn, W.,
Withyachumnarnkul, B., and Connor, J.R., Young coconut juice, a potential therapeutic agent that could
significantly reduce some pathologies associated with Alzheimer’s disease: Novel findings. Br. J. Nutr.,
2011, 105, 738–746, 2011.
37. Radenahmad, N., Saleh, F., Sawangjaraoen, K., Rundorn, W., Withyachumnarnkul, B., and Connor,
J.R., Young coconut juice significantly reduces histopathological changes in the brain that are induced
by hormonal imbalance: A possible implication to postmenopausal women. Histol. Histopathol., 24,
667–674, 2009.
38. Depeint, F., Bruce, W.R., Shangair, N., Mehta, R., and O’Brien, P.J., Mitochondrial function and toxic-
ity: Role of B vitamins on the one-carbon transfer pathways. Chem. Biol. Interact., 163, 113–132, 2006.
39. Matsuo, Y. and Greenberg, D.M., A crystalline enzyme that cleaves homoserine and cystathionine.
I. Isolation procedure and some physicochemical properties. J. Biol. Chem., 230, 545–560, 1958.
40. Carroll, W.R., Stacy, G.W., and Du Vigneaud, V., Alpha-ketobutyric acid as a product in the enzymatic
cleavage of cystathionine. J. Biol. Chem., 180, 375–382, 1949.
41. Stipanuk, M.H., Dominy, J.E., Lee, J.I., and Coloso, R.M., Mammalian cysteine metabolism: New
insights into regulation of cysteine metabolism. J. Nutr., 136(Suppl.), 1652S–1659S, 2006.
42. Goh, Y.I. and Koren, G., Folic acid in pregnancy and fetal outcomes. J. Obstet. Gynaecol., 28, 3–13,
2008.
43. Metz, J., Stevens, K., Krawitz, S., and Brandt, V., The plasma clearance of injected doses of folic acid as
an index of folic acid deficiency. J. Clin. Pathol., 14, 622–625, 1961.
44. Robinson, K., Arheart, K., Refsum, H., Brattstrom, L., Boers, G., Ueland, P., Rubba, P. et al., Low cir-
culating folate and vitamin B6 concentrations: Risk factors for stroke, peripheral vascular disease, and
coronary artery disease. European COMAC Group. Circulation, 97, 437–443, 1998.
45. Zhang, S.M., Willet, W.C., Selhub, J., Hunter, D.J. Giovannucci, E.L., Holmes, M.D., Colditz, G.A.,
and Hankinson, S.E., Plasma folate, vitamin B6, vitamin B12, homocysteine, and risk of breast cancer.
J. Natl. Cancer Inst., 95, 373–380, 2003.
46. Fernandes, J.C.B., de Oliveira Neto, G., Rohwedder, J.J.R., and Kubota, L.T., Simultaneous determina-
tion of chloride and potassium in carbohydrate electrolyte beverages using an array of ion-selective
electrodes controlled by a microcomputer. J. Braz. Chem. Soc., 11, 349–354, 2000.
47. Pummer, S., Heil, P., Maleck, W., and Petroianu, G., Influence of coconut water on hemostasis. Am. J.
Emerg. Med., 19, 287–289, 2001.
48. Chavalittamrong, B., Pidatcha, P., and Thavisri, U., Electrolytes, sugar, calories, osmolarity and pH of
beverages and coconut water. Southeast Asian J. Trop. Med. Public Health, 13, 427–431, 1982.
49. Adams, W. and Bratt, D.E., Young coconut water for home rehydration in children with mild gastroen-
teritis. Trop. Geogr. Med., 44, 149–153, 1992.
50. Saat, M., Singh, R., Sirisinghe, R.G., and Nawawi, M., Rehydration after exercise with fresh young
coconut water, carbohydrate-electrolyte beverage and plain water. J. Physiol. Anthropol. Appl. Human
Sci., 21, 93–104, 2002.
51. Ismail, I., Singh, R., and Sirisinghe, R.G., Rehydration with sodium-enriched coconut water after
exercise-induced dehydration. Southeast Asian J. Trop. Med. Public Health, 38, 769–785, 2007.
52. Bankar, G.R., Nayak, P.G., Bansal, P., Paul, P., Pai, K.S., Singla, R.K., and Bhat, V.G., Vasorelaxant and
antihypertensive effect of Cocos nucifera Linn. endocarp on isolated rat thoracic aorta and DOCA salt-
induced hypertensive rats. J. Ethnopharmacol, 134, 50–54, 2011.
53. Anurag, P. and Rajamohan, T., Cardioprotective effect of tender coconut water in experimental myocar-
dial infarction. Plant Foods. Hum. Nutr., 58, 1–12, 2003.
54. Barciszewski, J., Siboska, GH.E., Pedersen, B.O., Clark, B.F., and Rattan, S.I., Evidence for the presence
of kinetin in DNA and cell extracts. FEBS Lett., 393, 197–200, 1996.
55. Ge, L., Yong, J.W., Goh, N.K., Chia, L.S., Tan, S.N., and Ong, E.S., Identification of kinetin and kinetin
riboside in coconut (Cocos nucifera L.) water using a combined approach of liquid chromatography-
tandem mass spectrometry, high performance liquid chromatography and capillary electrophoresis.
J. Chromatogr. B, 829, 26–34, 2005.
56. Sobieszczuk-Nowicka, E., Wieczorek, P., and Legocka, P., Kinetin affects the level of chloroplast
polyamines and transglutaminase activity during senescence of barley leaves. Acta Biochim. Pol., 56,
255–259, 2009.
57. Rattan, S.I. and Clark, B.F., Kinetin delays the onset of ageing characteristics in human fibroblasts.
Biochem. Biophys. Res. Commun., 201, 665–672, 1994.
58. McCullough, J.L. and Weinstein, G.D., Clinical study of safety and efficacy of using topical kinetin
0.1% (Kinerase R) to treat photodamaged skin. Cosmetic Dermatol., 15, 29–32, 2002.
Coconut Juice 203
59. Uttara, B., Singh, A.V., Zamboni, P., and Mahajan, R.T., Oxidative stress and neurodegenerative dis-
eases: A review of upstream and downstream antioxidant therapeutic options. Curr. Neuropharmacol.,
7, 65–74, 2009.
60. Olsen, A., Siboska, G.E., Clark, B.F., and Rattan, S.I., N(6)-Furfuryladenine, kinetin, protects against
Fenton reaction-mediated oxidative damage to DNA. Biochem. Biophys. Res. Commun., 265, 499–502,
1999.
61. Verbeke, P., Siboska, G.E., Clark, B.F., and Rattan, S.I., Kinetin inhibits protein oxidation and glycoxi-
dation in vitro. Biochem. Biophys. Res. Commun., 276, 1265–1270, 2000.
62. Leshem, Y.Y., Plant senescence processes and free radicals. Free Radic. Biol. Med., 5, 39–49, 1988.
63. Letham, D.S., Zeatin, a factor inducing cell division isolated from zea mays. Life Sci., 8, 569–573, 1963.
64. Letham, D.S., Regulators of cell division in plant tissues—II. Phytochemistry, 5, 269–286, 1966.
65. Van Staden, J.J., Identification of zeatin and zeatinriboside in coconut milk. Physiol. Plant., 34, 106–109,
1975.
66. Heo, H.J., Hong, S.C., Cho, H.Y., Hong, B., Kim. H.K., Kim, E.K., and Shin, D.H., Inhibitory effect of
zeatin, isolated from Fiatoua villosa, on acetylcholinesterase activity from PC12 cells. Mol. Cells, 13,
113–117, 2002.
67. Kim, M.J., Choi, S.J., Lim, S.T., Kim, H.K., Kim, Y.J., Yoon, H.G., and Shin, D.H., Zeatin supple-
ment improves scopolamine-induced memory impairment in mice. Biosci. Biotechnol. Biochem., 72,
577–581, 2008.
68. Choi, S.J., Jeoung, C.H., Choi, S.G., Chun, J.Y., Kim, Y.J., Lee, J., Shin, D.H., and Heo, H.J., Zeatin pre-
vents amyloid beta-induced neurotoxicity and scopolamine-induced cognitive deficits. J. Med. Food.,
12, 271–277, 2009.
69. Rattan, S.I. and Sodagam, L., Gerontomodulatory and youth-preserving effects of zeatin on human skin
fibroblasts undergoing aging in vitro. Rejuvenation Res., 8, 46–57, 2005.
70. Jackson, J.C., Gordon, A., Wizzard, G., McCook, K., and Rolle, R., Changes in chemical composition of
coconut (Cocos nucifera) water during maturation of the fruit. J. Sci. Food Agric., 84, 1049–1052, 2004.
71. Neto, M.F., de Holanda, J.S., Folegatti, M.V., Gheyi, H.R., Pereira, W.E., and Cavalcante, L.F., Quality
of green fruits of “anão verde” coconut in relation to doses of nitrogen and potassium via fertigation.
Rev. Bras. Eng. Agríc. Ambient., 11, 453–458, 2007.
17
Cranberry Juice
Monique Lacroix and Khanh Dang Vu
CONTENTS
17.1 Introduction................................................................................................................................... 205
17.4 Health Effects................................................................................................................................ 209
17.4.1 Antimicrobial Effects....................................................................................................... 209
17.4.2 Antimutagenic and Anticarcinogenic Properties..............................................................210
17.4.3 Cardiovascular Health.......................................................................................................211
17.6 Conclusion......................................................................................................................................212
References................................................................................................................................................212
17.1 Introduction
Cranberry juice is processed from cranberry fruit and is of growing public interest as a functional
food because of the potential health benefits linked to its phytochemicals [1]. There are two main cran-
berry species, the North American cranberry (Vaccinium macrocarpon) and the European cranberry
(Vaccinium oxycoccos) [2]. According to Statistics Canada [3], cranberry sales were US$94 million in
2013, accounting for more than 11% to the Canadian total sales of all fruits, which is US$825 million.
Cranberry fruits used for processing are normally stored in containers under frozen conditions after har-
vesting. About 95% of the cranberry fruits are processed into products such as juice (pure or cocktail),
sauce, sweetened, and dried. The remaining 5% are sold for fresh consumption. Approximately 1.5 kg of
fresh fruits produces 1 L of juice. Cranberry juice cocktail is approximately 26%–33% pure cranberry
juice, sweetened with fructose or an artificial sweetener [4,5].
Cranberry juice contains several subclasses of phytochemicals such as phenolic acids, flavonoids,
anthocyanins, and tannins [1]. These phytochemicals have different biological properties such as anti-
oxidant, antiradical, antimicrobial, and anticancer effects, and therefore cranberry juice is considered
as functional food product for health improvement [1]. This chapter highlights cranberry juice in terms
of its nutritional characteristics, bioactive and antioxidant/antiradical properties, health-promoting
effects, and finally, trends in developing novel products based on cranberry juice as well as the future
prospects.

There are several factors that can affect the nutritional characteristics and quality of cranberry juice.
For example, different cultivars, production regions, fertilization, as well as different pretreatment and
processing methods of fruits can cause changes in the nutritional composition of cranberry juice.
Table 17.1 presents the compositional and nutritional characteristics of unsweetened cranberry juice [6].
It can be observed that cranberry juice contains mostly water (87.13 g/100 g) and provides low calorie
205
TABLE 17.1
Compositional and Nutritional Characteristics of Cranberry Juice (per 100 g)
Nutrition Unit Cranberry Juice (Unsweetened)
Water g 87.13
Energy kcal 46
Protein g 0.39
Lipid (fat) g 0.13
Carbohydrate g 12.20
Total sugars g 12.10
Minerals
Calcium mg 8
Iron mg 0.25
Magnesium mg 6
Phosphorus mg 13
Potassium mg 77
Sodium mg 2
Zinc mg 0.10
Vitamins
Folate (DFE) μg 1
Niacin mg 0.091
Riboflavin mg 0.018
Thiamin mg 0.009
Vitamin A (RAE) μg 2
Vitamin B12 μg 0.00
Vitamin B6 mg 0.052
Vitamin C mg 9.3
Vitamin K μg 5.1
Database for Reference, Release 26, 2013, Published online at http://ndb.nal.usda.
gov/ndb/foods/show/8395 (accessed July 7, 2014).
DFE, dietary folate equivalents; RAE, retinol activity equivalents; ATE,
Abbreviations:
(46 kcal/100 g) and high carbohydrate (12.20 g/100 g). It also contains high potassium (77 mg/100 g)
and vitamin C (9.3 mg/100 g) [6]. Six ounces (177.4 mL) of cranberry juice cocktail (Ocean Spray®), on
average, contain 96 calories, 24 g carbohydrate, 0.17 g crude fiber, 0.02 g benzoic acid, 0.28 g citric acid,
0.30 g malic acid, and 0.45 g quinic acid [7,8].
Cranberry juice has a short shelf life due to the rapid color deterioration [9]. The anthocyanins in
the juice are not stable during storage. The process conditions have also an important influence on the
juice color. The heat treatment conditions, the concentration of ascorbic acid, the pH, the activity of the
polyphenol oxidases and/or glycosidases enzymes, the presence of color derivative cofactors such as
cinnamic acids or flavonoids, and some metals such as copper and iron have an important influence on
the concentration of anthocyanins and color retention during the process [10]. Pasteurization conditions
(time and temperature) are considered the most important factors affecting the stability of anthocyanins.
A lower pH and the presence of ascorbic acids can protect better anthocyanins during pasteurization
treatment and during storage. It is, however, known that ascorbic acid is not stable and it is estimated
that more than 25% of ascorbic acid is degraded during pasteurization treatment [11]. Since these factors
Cranberry Juice 207
affect the nutritional quality of cranberry juice during storage, an advanced method or technology should
be considered to improve the quality of cranberry juice.

Table 17.2 presents polyphenols present in cranberry fruit, cranberry juice, and cranberry juice cocktail
based on the USDA [12,13] database. It can be observed that cranberry juice contains high quantity of
(+)-catechin (0.92 mg/100 g), myricetin (4.41 mg/100 g), and quercetin (16.41 mg/100 g), while it con-
tains nondetectable concentration of other flavones (apigenin and luteolin), anthocyanindins (cyanidin,
delphinidin, and pelargonidin), and proanthocyanidins (monomers, dimers, trimers, 4–6mers, 7–10mers,
and polymers). In the case of cranberry juice cocktail, due to the mixture of cranberry juice with seltzer
or soda water, this cocktail contains less of (+)-catechin (0.19 mg/100 g), myricetin (0.51 mg/100 g), and
quercetin (1.27 mg/100 g), but the cocktail contains some other flavones, anthocyanindins, and proan-
thocyanidins that are not available in cranberry juice (Table 17.2).
More than 150 phytochemicals have been identified in cranberries [14]. The most important com-
pounds are flavonoids and phenolic acids (Figure 17.1). Among them, anthocyanins are responsible for
the bright red color of the juice. Flavonols are responsible for their secondary yellowish pigments, and
proanthocyanidins in cranberry juice are associated with the protection of urinary tract infection. The
presence of catechins, organic acids, and resveratrol contributes to the sourness and the astringency
that is unique to cranberry juice. Simple phenolics and benzoates are also present in cranberry juice
and possess strong odor and aroma. These compounds provide potent antioxidant and antimicrobial
properties [15]. Pectins in cranberry juice are responsible for gel formation after cooking during sauce
preparation [14]. Flavonoids and proanthocyanins are more stable during the juice production process.
It is estimated that the retention of flavonoids is between 40% and70% as compared to 2%–30% for
anthocyanins, and the level of proanthocyanins could be relatively constant if pasteurized under high-
temperature short-time commercial conditions [14].
As mentioned earlier, cranberry juice is a rich source of phytochemicals. The compounds responsible
for the beneficial effect on human health are phenolic compounds including anthocyanins, flavonoids
(ellagitannins and proanthocyanidins), stilbenes, and phenolic acids [16,17]. Over the past 15 years, these
compounds and other secondary plant metabolites found in cranberry juice (flavanols, proanthocyanidins,
TABLE 17.2
Polyphenols in Cranberry Juice (mg/100 g)
Cranberry
Class Components Cranberry Fruit Cranberry Juice Juice Cocktail References
Flaval-3-ols (+)-Catechin 0.39 0.92 0.19 [12]
Flavonols Myricetin 6.78 4.41 0.51 [12]
Quercetin 15.09 16.41 1.27 [12]
Kaempferol 0.09 na 0.01 [12]
Flavones Apigenin na na 0.01 [12]
Luteolin na na 0.03 [12]
Anthocyanidins Cyanidin 41.81 na 0.38 [12]
Delphinidin 42.10 na 0.03 [12]
Pelargonidin 7.66 na 0.03 [13]
Proanthocyanidins Monomers 7.26 na 0.56 [13]
Dimers 25.93 na 2.71 [13]
Trimers 18.93 na 1.59 [13]
4–6mers 70.27 na 4.58 [13]
7–10mers 62.90 na 3.84 [13]
Polymers 234 na 8.33 [13]
OH
O
O
OH
HO OH
OH HO
Gallic acid p-Coumaric acid
OH
OH OH
HO O HO O
OH OH
OH O OH O
Quercetin Kaempferol
OH
OH OH
HO O+ HO O+
OH OH
OH OH
OH OH
Cyanidin Delphinidin
FIGURE 17.1 Common phenolic compounds found in cranberry juice.
and cinnamic acid derivatives) have attracted a great deal of attention mainly because of their antioxi-
dant, antibacterial, and antimutagenic role in preventing several diseases [18].
Cranberry phenolics have been shown to have free radical scavenging (FRS) properties against
superoxide radical (O•−2), hydrogen peroxide (H2O2), hydroxyl radicals (•OH), and singlet oxygen
(1O2). They can also inhibit lipid peroxidation, as well as protein and lipid oxidation in liposomes [19].
The anthocyanin pigments are responsible for the brilliant red color of cranberry juice and are among
the principal antioxidant constituents [15]. Other phytochemicals found in cranberry juice (tannins, fla-
vonols, flavanols, and benzoic and cinnamic acid derivatives) have also attracted a great deal of attention
because of their antioxidant activity [15]. The condensed tannins (proanthocyanidins), trimmers, and
dimers in cranberry juice were shown to be effective antioxidants in various food systems including
emulsion, liposomes, and low-density lipoprotein (LDL).
Côté et al. [20] and Caillet et al. [21] have evaluated the effect of juice processing on cranberry antioxi-
dant and antiradical properties. The activities were evaluated on whole fruit, macerated and depectinized
mash, initial press juice, and juice concentrate. The FRS and the antioxidant properties via the inhibi-
tion of lipid peroxidation (LPI) activities of each sample and phenolic extracts (water-soluble phenols,
flavonoids, and anthocyanins) were evaluated. Results showed that the water-soluble phenols showed
the greatest FRS (68.2 mmol trolox equivalents [TE]/mg phenol) and LPI (13.4 mmol TE/mg phenol)
activities. Results also showed that the polarity of the phenols, the pH, and the juice process had an influ-
ence on the antioxidant properties of flavonoids, water-soluble phenols, and anthocyanins. The milling,
heating, and maceration steps increased the FRS capacity of flavonoids and anthocyanins but decreased
their LPI capacity. The evaporation of the juice did not have any significant effect on the FRS capacity
but lowered the LPI capacity of all phenolic extracts evaluated. Before and after each step of the juice
process, the FRS capacity of the cranberry extracts was greater than their LPI capacity. These results
were consistent in classifying the FRS capacity of the extracts in the following order of polarity: water-
soluble phenols > anthocyanins > flavonoids. The content of the total phenolic compounds during the
Cranberry Juice 209
process was also in the order of anthocyanins > flavonoids > water-soluble phenols. The cranberry phe-
nolic compounds in the cranberry pomace also showed promising antioxidant activities. According to
Lee et al. [22], concentrated cranberry juice powder (0.32%) was also effective in retarding thiobarbituric
acid–reactive substances (TBARS) formation. Its antioxidant activity was similar to rosemary extract
(0.04%). The use of this ingredient in food system could be effective to protect against lipid oxidation.
According to Piljac-Žegarac et al. [23], the storage has also substantial effects on total phenolic and anti-
oxidant capacity of fruit juices. Important fluctuations have been observed, but cranberry juice exhibited
the greatest storage stability in terms of antioxidant property.
17.4 Health Effects

Cranberry juice has been known for its potential in preventing urinary infections mostly caused by
Escherichia coli. It has been established that tannic components in cranberry juice can inhibit bacterial
adherence to the epithelial cells of the urinary tract [1]. According to Heinonen [15], a dose of 36 mg of
cranberry proanthocyanidins can prevent the adhesion of E. coli bacteria to the urinary tract.
As previously mentioned, phenolic compounds in cranberry juice have potential health and nutritional
benefits. One of the most important aspects about the nutrients’ benefit is the bioavailability. Nutrients’
bioavailability is influenced not only by the molecular species but also by interactions between food
components [24]. Anthocyanins are poorly absorbed (<1%) [25]. According to Manach et al. [25], the
level of intact anthocyanin’s released in urine is, however, fivefold higher after ingestion of cranberry
juice than wine or strawberry juice. It is also interesting to note that anthocyanins can reach the brain,
while catechins can reach liver and kidney and flavonols reaching lungs, liver, kidney, and heart [26,27].
Proanthocyanidins are also poorly absorbed. Their oligomers and their polymers are absorbed in small
quantities in intestinal epithelium, and it is suggested that gut microflora degrades polymeric proantho-
cyanidins in aromatic acids, which may be absorbed under this form [28,29]. Flavonols are the most
available flavonoids in cranberry (10%). It is suggested that their absorption depends on the structure, the
presence of glycosidic linkage, and the food composition [27]. The simple phenolic compounds seem to
be the most well-absorbed phenols in cranberry. According to Pederson et al. [30], more than 600 mg gal-
lic acid equivalents (GAE)/L of total phenolics were found in human plasma after ingestion of 500 mL of
cranberry juice [30]. Studies on resveratrol showed that this compound was not detected in human plasma
[31]. It is, however, suggested that resveratrol could be converted to sulfonated and glucuronated deriva-
tives within 30 min after ingestion [32].
17.4.1 Antimicrobial Effects

As described previously, cranberry juice is rich in phenolic acids, flavonoids, including anthocyanins and
tannins. It has long been consumed for the prevention of urinary tract infections. E. coli is the pathogen
responsible for most of the urinary tract infections that affect more than 150 million people worldwide
per year [33]. Urinary tract infection involves a specific interaction between adhesion, positioned at the
distal end of bacterial pili, and its receptor on the surface of the host tissue and having a maximum
binding at pH 5 [34].
Sobota [35] found that cranberry juice played an important role in inhibiting the adherence of the bac-
terial strain to uroepithelial cells ex vivo. It was also demonstrated that this reaction was dose dependent.
A daily consumption of 250 mL of cranberry juice was able to inhibit 45% the adhesion to uroepithelial
cells, and a consumption of 750 mL was able to inhibit the adhesion by 63% [36]. Liu et al. [33] found
that cranberry juice decreased nanoscale adhesion forces between P-fimbriated E. coli and uroepithe-
lial cells. Proanthocyanins could be active constituents responsible for these antimicrobial properties;
however, these compounds are poorly absorbed [37]. Until today, the compounds responsible for the
antiadhesion of E. coli remain unknown.
Other studies have demonstrated that cranberry juice can prevent adhesion of the stomach pathogen
Helicobacter pylori [38]. A few studies have been carried out to understand the mechanism of action of
cranberry against this pathogen. However, an interesting study demonstrated that cranberry induces H.
pylori to develop a coccoid, thereby inhibiting its growth bacteriostatically [39]. Anthocyanins are also
able to inhibit the secretion of toxins by H. pylori [40]. According to the same authors, the most inten-
sively studied virulence factors of H. pylori are the production of cytotoxin and the production of these
toxins correlate with gastric ulceration and cancer development.
In an in vitro study, Huttumen et al. [41] found that cranberry juice had a strong activity against the
adhesion of Streptococcus pneumonia on human bronchial cells surface. The adhesion inhibition was
estimated as more than 90% at a concentration of 8.7 mg/g of soluble solids, and pneumococcal growth
was inhibited totally at a concentration of 86 mg/g. This research indicated that cranberry juice has a
potential against pneumococcal infections.
Côté et al. [42] evaluated the antibacterial potential of frozen cranberries and cranberry juice process-
ing intermediaries (mash, macerated, depectinized mash, and pomace) and final products (initial pressed
juice, clarified juice, and juice concentrate) against seven pathogenic bacterial strains. The results showed
that the juice process reduced the ability of flavonoids and anthocyanins to inhibit microbial growth
probably due to the degradation during the process. However, the pomace still kept good antimicro-
bial activity. Flavonoids were most efficient against Pseudomonas aeruginosa, Staphylococcus aureus,
Listeria monocytogenes, and Salmonella typhimurium showing a low minimal inhibitory concentration
of 45.50 μg phenol/well tested. Phenolic compounds in cranberry also have a good antimicrobial
property against L. monocytogenes [43]. According to these authors, apolar fraction containing flavo-
noids were the most antimicrobially effective. Lian et al. [44] have evaluated the efficiency of different
commercial juices including two commercial cranberry juices and found that Lakewood pure organic
cranberry juice and Spray cranberry cocktail juice possessed potent antimicrobial effects on S. aureus.
Both juices had the same amount of phenolic compounds, and it was estimated that the antimicrobial
property was associated with the presence of C18-bound phenolic materials. O’May and Tufenkji [45]
also suggested that proanthocyanins can block pathogens swarming motility of P. aeruginosa. Further
studies have to be carried out on other pathogens to verify if this mechanism of action could be extended
to other pathogens.
17.4.2 Antimutagenic and Anticarcinogenic Properties

Caillet et al. [46] evaluated cancer chemopreventive properties of different cranberry polyphenols
fractions of different polarity from cranberry juices, frozen cranberries, and pomace containing antho-
cyanins and water-soluble and apolar phenolic compounds. Cranberry fractions were screened for their
ability to induce the phase II xenobiotic detoxification enzyme quinone reductase (QR). The results
showed that there was no cytotoxicity against the cells used in the test. All samples stimulated the QR
activity except the less polar fraction of anthocyanin-rich extract from the pomace, which inhibited the
QR activity. The QR induction for all samples varied with the concentration, and there was an opti-
mal concentration for which the QR induction was maximal. The technological process to manufacture
cranberry juice had little influence on the overall QR inducer potencies of cranberry fractions, whereas
the ability of phenols in fractions to stimulate the QR activity has been reduced significantly (P ≤ 0.05)
during the technological process. The phenolic compounds of the most polar fraction (rich in phenolic
acids) and those of the less polar fraction (rich in proanthocyanidins) showed stronger induction than
those observed with phenols from intermediate fractions.
Two colon cancer cell lines HT-29 and LS-513 were treated with cranberry extracts from juice, fruits,
puree, depectinized puree, and pomace. This study demonstrated that all extracts were able to inhibit the
growth of the two cancer cells lines with an IC50 (concentration needed to inhibit the growth of 50% of
cancer cells) varying from 3.8 to 179.2 μg GAE/mL [47]. Proanthocyanidins can also induce a rapid apop-
tosis of lung cancer cells. It was proposed that purified proanthocyanidins be used as a single drug or in a
combinational treatment for ovarian cancer since these compounds can target both ovarian cancer viability
and angiogenesis in vitro [48]. Hochman et al. [49] demonstrated in vivo in mice that cranberry juice
injected intraperitoneally can inhibit the growth of lymphoma tumor in mice and enhanced the generation
of antilymphoma antibodies. He and Liu [50] and Sun and Liu [51] further demonstrated that flavonoids
and total phenols can also have antiproliferative effects against breast cancer cells in vitro. Neto [17]
suggested that cranberry extracts can act through induction of apoptosis in tumor cells, the reduction of
Cranberry Juice 211
ornithine decarboxylase activity, the decrease of expression of matrix metalloproteinases associated with
tumor metastasis, and anti-inflammatory activities including inhibition of cyclooxygenases.
17.4.3 Cardiovascular Health

Cranberry bioactives have been demonstrated to inhibit the oxidation of lipoproteins in vitro [52]. The
consumption of 500 mL of cranberry juice by volunteer females demonstrated a significant increase in
the antioxidant property of the blood samples after 1–2 h of consumption [30]. This antioxidant property
can decrease the level of oxidized LDL by around 10% [53].

Figure 17.2 illustrates the industrial process for the production of cranberry juice. The initial step to
make the juice includes the washing step, then reducing the size of cranberry fruits by milling/grinding.
The mash is heated and macerated under minimal agitation. At the same time, enzymes may be added
to increase yields of extraction of raw juice by pressing. Then, raw juice is filtered by a cross-flow mem-
brane filtration to obtain a clear juice. Finally, the clarified juice is concentrated by evaporation to obtain
a juice concentrate (can be at different °Brix values depending on objectives) [54].
Cranberry fruits
Sanitizing
Grinding/milling
Pectinases Maceration (55°C)
Pressing Pomace
Raw juice
Clarification
Clarified juice
Evaporating under vacuum
Concentrated juice
FIGURE 17.2 Flowchart of cranberry juice production from fruits on an industrial scale. (Adapted from Caillet, S. et al.,
Food Res. Int., 44, 902, 2011. With permission.)
Heat treatment conditions and the presence of oxygen are the most important factors involved in
d egradation of anthocyanins and the color of cranberry juice. The development of anaerobic pasteuriza-
tion and packaging or cold pasteurization and the use of cofactors could be promising approaches for
controlling the color and the stability of anthocyanins in cranberry juice [14].
The polyphenol content of cranberry juice could serve as effective ingredients for the protection
against lipid oxidation in food systems. However, further studies are needed to elucidate sensory percep-
tion and microbial evolution during storage of foods supplemented with cranberry concentrated juice.
There are some new products based on cranberry juice on the market. For example, Ocean Spray
has developed different types of products based on cranberry juice such as cranberry, cranberry grape,
cranberry/strawberry, and cranberry/blueberry/blackberry [55]. Cystex cranberry liquid supplement is
a complex that contains natural cranberry concentrate, D-mannose, bromelain (from pineapple), vita-
min C, and inulin [56]. This product can help prevent urinary tract infections.
It should be mentioned that even though there are different commercial products based on cranberry
juice, the bioavailability/bioabsorption of cranberry polyphenols during consumption should be further
evaluated, and new formulations developed. For example, many polyphenols are available in cranberry
fruits but are not available in cranberry juice (Table 17.2). Thus, the method of juice extraction from
cranberry fruits should be further improved or new formulations of cranberry products with highly
controlled-release properties developed to ensure the high bioavailability/bioabsorption of polyphenols
to render health benefits.
The antimicrobial properties of polyphenols of cranberry juice can open a market for the development
of new products to prevent dental or stomach infection and even ingredient formulations to prevent food
contamination with pathogens such as L. monocytogenes. In addition, with the increase of the resistant
pathogens to antibiotics, the cranberry juice could be a good alternative to antibiotics in the future.
Finally, a key for the future will be to identify marker molecules and their bioactivity in vivo to permit
the development of efficient nutraceutical products for protection against cardiovascular disease (CVD),
cancer development, and protection against infection with pathogens as well as resistant pathogen to
antibiotics.
17.6 Conclusion
Cranberry juice is rich in functional components especially phenolic compounds. The content of phenolic
compounds of cranberry juice varies depending on fruit cultivar and during storage of juice as well as
their biological activities. The water-soluble phenols in cranberry juice showed the highest antioxidant
property. Concentrated cranberry juice shows promising antioxidant properties and could be exploited
more for the development of new nutraceutical products or food ingredients. However, studies should be
done to develop formulations that do not change the sensory properties of foods supplemented with cran-
berry juice concentrate. Polyphenols from cranberry juice also have antimicrobial effects. This property
could be beneficial to protect food products against contamination but could also protect humans against
contamination of the urinary tract, stomach, and even against the development of caries and dental
plaque biofilms. Polyphenols of cranberry juice can protect against the development of CVD and also
have anticancer properties. Their efficacy was demonstrated in vitro but also in vivo in mice. Further
clinical studies should be done in order to demonstrate their efficiency and bioavailability.
REFERENCES
1. Côté, J., Caillet, S., Doyon, G., Sylvain, J.F., and Lacroix, M., Bioactive compounds in cranberries and
their biological properties. Crit. Rev. Food Sci. Technol., 50, 666–679, 2010.
2. Girard, K.K. and Sinha, N., Cranberry, blueberry, currant, and gooseberry, in Handbook of Fruits and
Fruit Processing, Hui, Y.H., Ed., Blackwell Publishing Ltd., Oxford, UK, 2006, pp. 369–390.
3. Statistics Canada, Fruit and vegetable production, 2013. Published online at: http://www.statcan.gc.ca/
daily-quotidien/140129/dq140129d-eng.htm (accessed May 28, 2014).
Cranberry Juice 213
4. MedlinePlus, Cranberry. Published online at: http://www.nlm.nih.gov/medlineplus/druginfo/natural/

958.html (accessed May 28, 2014).
5. Food Reference, Cranberries: Cranberry Trivia. Published online at: http://www.foodreference.com/
html/fcranberries.html (accessed May 28, 2014).
6. U.S. Department of Agriculture (USDA), USDA National Nutrient Database for Reference, Release 26,
2013. Published online at http://ndb.nal.usda.gov/ndb/foods/show/8395 (accessed July 7, 2014).
7. Hughes, B.G. and Lawson, L.D., Nutritional content of cranberry products. Am. J. Hosp. Pharm.,
46, 1129–1132, 1989.
8. Sigma-Aldrich, Cranberry (Vaccinium macrocarpon). Published online at: http://www.sigmaaldrich.
com/life-science/nutrition-research/learning-center/plant-profiler/vaccinium-macrocarpon.html
(accessed May 28, 2014).
9. Francis, F.J., Quality as influenced by color. Food Qual. Pref., 6, 149–155, 1995.
10. Francis, F.J. and Servadio, G.J., Relationship between color of cranberries and color and stability of
juice. Am. Soc. Hort. Sci., 83, 406–415, 1963.
11. Starr, M.S. and Francis, F.J., Oxygen and ascorbic acid effect on the relative stability of four anthocy-
anin pigments in cranberry juice. Food Technol., 22, 1293–1295, 1968.
12. U.S. Department of Agriculture (USDA), USDA Database for the Flavonoid Content of Selected Foods,
Release 2.1, USDA, Beltsville, MD, 2007.
13. U.S. Department of Agriculture (USDA), USDA Database for the Proanthocyanidin Content of Selected
Foods, USDA, Beltsville, MD, 2004.
14. Pappas, E. and Schaich, K.M., Phytochemicals of cranberries and cranberry products: Characterization,
potential health effects and processing stability. Crit. Rev. Food Sci. Nutr., 49, 741–781, 2009.
15. Heinonen, M., Antioxidant activity and antimicrobial effect of berry phenolics—A finish perspectives.
Mol. Nutr. Food Res., 51, 684–691, 2007.
16. Paredes-López, O., Cervantes-Ceja, M.L., Vigna-Pérez, M., and Hernández-Pérez, T., Berries:
Improving human health and healthy aging, and promoting quality life—A review. Plant Foods Hum.
Nutr., 65, 299–308, 2010.
17. Neto, C.C., Cranberry and its phytochemicals: A review of in vitro anticancer studies. J. Nutr.,
137(Suppl.), 186S–193S, 2007.
18. Côté, J., Caillet, S., Doyon, G., Sylvain, J.F., and Lacroix, M., Bioactive compounds in cranberries and
their biological properties. Crit. Rev. Food Sci. Technol., 50, 666–679, 2010.
19. Wang, S.Y. and Jiao, H., Scavenging capacity of berry crops on superoxide radicals, hydrogen peroxide,
hydroxyl radicals, and singlet oxygen. J. Agric. Food Chem., 48, 5677–5684, 2000.
20. Côté, J., Caillet, S., Doyon, G., Dussault, D., Salmieri, S., Lorenzo, G., Sylvain, J.F., and Lacroix, M.,
Effects of juice processing on cranberry antioxidants properties. Food Res. Int., 44, 2907–2914, 2011.
21. Caillet, S., Côté, J., Doyon, G., Sylvain, J.F., and Lacroix, M., Antioxidant and antiradical properties of
cranberry juice and extracts. Food Res. Int., 44, 1408–1413, 2011.
22. Lee, C.H., Reed, J.D., and Richards, M.P., Ability of various polyphenolic classes from cranberry to
inhibit lipid oxidation in mechanically separated turkey and cooked ground pork. J. Muscle Foods, 17,
248–266, 2006.
23. Piljac-Žgarac, J., Valek, L., Martinez, S., and Belščak, A., Fluctuations in the phenolic content and
antioxidant capacity of dark fruit juices in refrigerated storage. Food Chem., 113, 394–400, 2009.
24. Parada, J. and Aguilera, J.M., Food microstructure affects the bioavailability of several nutrients.
J. Food Sci., 72, R21–R32, 2007.
25. Manach, C., Williamson, G., Morand, C., Scalbert, A., and Remesy, C., Bioavailability and bioeffi-
cacy of polyphenols in humans. Review of 97 bioavailability studies. Am. J. Clin. Nutr., 81(Suppl.),
230S–242S, 2005.
26. Ohnishi, R., Ito, H., Kasajima, N., Kaneda, M., Kariyama, R., Kumon, H., Hatano, T., and Yoshida,
T., Urinary excretion of anthocyanins in humans after cranberry juice ingestion. Biosci. Biotechnol.
Biochem., 70, 1681–1687, 2006.
27. Erlund, I., Review of the flavonoids quercetin, hesperetin and naringenin. Dietary sources, bioactivities,
bioavailability and epidemiology. Nutr. Res., 24, 851–874, 2004.
28. Deprez, S., Brezillon, C., Rabot, S., Philippe, C., Mila, I., Lapierre, C., and Scalbert, A., Polymeric pro-
anthocyanidins are catabolized by human colonic microflora into low molecular weight phenolic acids.
J. Nutr., 130, 2733–2738, 2000.
29. Deprez, S., Mila, I., Huneau, J.F., Tome, D., and Scalbert, A., Transport of proanthocyanin dimer, trimer
and polymer across monolayers of human intestinal epithelial Caco-2 cells. Antioxid. Redox. Signal., 3,
957–967, 2001.
30. Pederson, C.B., Kyle, J., Jenkinson, A.M., Gardner, P.T., McPhail, D.B., and Duthie, G.G., Effects of
blueberry and cranberry juice consumption on the plasma antioxidant capacity of healthy female volun-
teers. Eur. J. Clin. Nutr., 54, 405–408, 2000.
31. Zhang, K. and Zuo, Y., GC-MS determination of flavonoids and phenolic and benzoic acids in human
plasma after consumption of cranberry juice. J. Agric. Food Chem., 52, 222–227, 2004.
32. Walle, T., Hsieh, F., DeLegge, M.H., Oatis, J.E., and Walle, UK, High absorption but very low bioavail-
ability of oral resveratrol in humans. Drug Metab. Dispos., 32, 1377–1382, 2004.
33. Liu, Y., Pinzón-Arango, P.A., Gallardo-Moreno, A., and Camesano, T.A., Direct adhesion force
measurements between E. coli and human uroepithelial cells in cranberry juice cocktail. Mol. Nutr.
Food Res., 54, 1744–1752, 2010.
34. Klinth, J.E., Castelain, M., Uhlin, B.E., and Axner, O., The influence of pH on the specific adhesion of
P Piliated Escherichia coli. PLoS One, 7, 6, e38548, 1–8, 2012.
35. Sobota, A.E., Inhibition of bacterial adherence by cranberry juice: Potential use for the treatment of
urinary tract infections. J. Urol., 131, 1013–1016, 1984.
36. Di Martino, P., Agniel, R., David, K., Templer, C., Gaillard, J.L., Denys, P., and Botto, H., Reduction
of Escherichia coli adherence to uroepithelial bladder cells after consumption of cranberry juice:
A double-blind randomized placebo-controlled cross-over trial. World J. Urol., 24, 21–27, 2006.
37. LaPlante, K., Sarkisian, S.A., Woodmansee, S., Rowley, D.C., and Seeram, N.P., Effects of cranberry
extracts on growth and biofilm production of Escherichia coli and Staphylococcus species. Phytother.
Res., 26, 1371–1374, 2012.
38. Vattem, D.A., Lin, Y.T., Ghaedian, R., and Shetty, K., Cranberry synergies for dietary management of
Helicobacter pylori infections. Proc. Biochem., 40, 1583–1592, 2005.
39. Matsushima, M., Suzuki, T., Masui, A., Kasai, K., Kouchi, T., Takagi, A., Shirai, T., and Mine, T.,
Growth inhibitory action of cranberry on Helicobacter pylori. J. Gastroenterol. Hepatol., 23(Suppl. 2),
S175–S180, 2008.
40. Kim, S.H., Park, M., Woo, H., Tharmalingam, N., Lee, G., Rhee, K.J., Eom, Y.B., Han, S.I., Seo, W.D.,
and Kim, J.B., Inhibitory effects on anthocyanins on secretion of Helicobacter pylori CagA and VacA
toxins. Int. J. Med. Sci., 9, 838–842, 2012.
41. Huttumen, S., Toivanen, M., Arkko, S., Ruponen, M., and Tikkanen-Kaukanen, C., Inhibition activ-
ity of wild berry juice fractions against Streptococcus pneumonia binding to human bronchial cells.
Phytother. Res., 25, 122–127, 2011.
42. Côté, J., Caillet, S., Doyon, G., Dussault, D., Sylvain, J.F., and Lacroix, M., Antimicrobial effect of
cranberry juice and extracts. Food Control, 22, 1413–1418, 2011.
43. Côté, J., Caillet, S., Dussault, D., Sylvain, J.F., and Lacroix, M., Effect of juice processing on cranberry
antibacterial properties. Food Res. Int., 44, 2922–2929, 2011.
44. Lian, P.Y., Maseko, T., Rhee, M., and Ng, K., The antimicrobial effects of cranberry against
Staphylococcus aureus. Food Sci. Technol. Int., 18, 179–186, 2012.
45. O’May, C. and Tufenkji, N., The swarming motility of Pseudomonas aeruginosa is blocked by cranberry
proanthocyanidins and other tannin-containing materials. Appl. Environ. Microbiol., 77, 3061–3067,
2011.
46. Caillet, S., Lorenzo, G., Côté, J., Doyon, G., Sylvain, J.-F., and Lacroix, M., Cancer chemopreventive
effect of fractions from cranberry products. Food Res. Int., 320–330, 2012.
47. Vu, K.D., Carlettini, H., Bouvet, J., Côté, J., Doyon, G., Sylvain, J.F., and Lacroix, M., Effect of different
cranberry extracts and juices during cranberry juice processing on the antiproliferative activity against
two colon cancer cell lines. Food Chem., 132, 959–967, 2012.
48. Kim, K.K., Singh, A., Singh, R., DeMartino, A., Brard, L., Vorsa, N., Lange, T., and Moore, R., Anti-
angiogenic activity of cranberry proanthocyanidins and cytotoxic poperties in ovarian cancer cells.
Int. J. Oncol., 40, 227–235, 2012.
49. Hochman, N., Houri-Haddad, H., Koblinski, J., Wahl, L., Roniger, M., Bar-Sinai, A., Weiss, E.I., and
Hochman, J., Cranberry juice constituents impair lymphoma growth and augment the generation of
antilymphoma antibodies in syngeneic mice. Nutr. Cancer, 60, 511–517, 2008.
Cranberry Juice 215
50. He, X. and Liu, R.H., Cranberry phytochemicals: Isolation, structure elucidation and their antiprolifera-
tive and antioxidant activities. J. Agric. Food Chem., 54, 7069–7074, 2006.
51. Sun, J. and Liu, R.H., Cranberry phytochemical extracts induce cell cycle arrest and apoptosis in human
MCF-7 breast cancer cells. Cancer Lett., 241, 124–134, 2006.
52. Reed, J.D., Cranberry flavonoids, artherosclerosis and cardiovascular health. Crit. Rev. Food Sci. Nutr.,
42, 301–316, 2002.
53. Ruel, G., Pomerleau, S., Couture, P., Lamarche, B., and Couillard, C., Changes in plasma antioxidant
capacity and oxidized low-density lipoprotein levels in men after short-term cranberry juice consump-
tion. Metabol. Clin. Exp., 54, 856–861, 2005.
54. Caillet, S., Côté, J., Doyon, G., Sylvain, J.-F., and Lacroix, M., Effect of juice processing on the cancer
chemopreventive effect of cranberry. Food Res. Int., 44, 902–910, 2011.
55. Ocean Spray, Cranberry and cranberry blends. Published available at: http://www.oceanspray.com/
Products/Juices/By-Flavor/Cranberry-and-Cranberry-Blends.aspx (accessed May 30, 2014).
56. Cystex, How do I prevent UTI? Published online at: http://www.cystex.com/how-do-i-prevent-utis/
cystex-liquid-cranberry-is-clinically-proven-to-promote-urinary-health/ (accessed May 22, 2014).
18
Date Syrup
Sami Fattouch, Karima Dhaouadi, and Manel Belkhir
CONTENTS
18.1 Introduction....................................................................................................................................217
18.4 Health Effects................................................................................................................................ 223
18.6 Conclusion..................................................................................................................................... 227
References............................................................................................................................................... 227
18.1 Introduction
Dates (Phoenix dactylifera L.) are largely produced in the hot desert regions (arid and semiarid
regions) of the world. During the last decades, world production of dates increased largely from
1.9 million tons in 1963 to 6.7 million tons in 2003. In 2012, the production was estimated at
7 million tons worth US$3.43 billion, with the major producing countries such as Egypt (1,352,950
metric tons [MT]), Saudi Arabia (1,078,300 MT), Iran (1,023,130 MT), United Arab Emirates
(775,000 MT), and Algeria (710,000 MT) [1]. Among the producing countries of this “Saharian”
typical fruit, Tunisia is the world’s leading producer of the Deglet Nour variety even though this
country accounts for only 2.7% of the overall world date production [1]. Date fruit is marketed all
over the world as a high-value confectionery. In addition to direct consumption of the whole date as
a fresh, delicious fruit, dates are traditionally used to prepare a wide range of products such as date
powder, date juice concentrates (syrup and liquid sugar), fermented date products (wine, alcohol,
vinegar, and organic acids), and date pastes for various uses (e.g., bakery and confectionary). These
date products can be incorporated into an array of foods, including different types of bread, marma-
lade, sweet candy, chocolate, and animal feed [2].
The term “syrup” comes from the ancient languages of Latin, sirupus, and Persian, sherbet, and is
described as a viscous liquid consisting of a dissolved sugar–water solution with little tendency toward
crystallization [3]. Commonly, syrup can refer to a viscous solution prepared from the saps of some
trees, especially date palm and maple tree. In this case, the sap is heated until reaching a high °Brix
to obtain what is locally called legmi syrup from date palm sap and maple syrup from maple tree sap.
Typically, in North Africa and in Arabian countries, fruit syrups (commonly known as rub) are more
frequently used than sap syrups. The most locally produced fruit syrups using old traditional processes
are from date (Phoenix dactylifera), barbary fig (Opuntia ficus-indica), and carob (Ceratonia siliqua)
known as rub al-tamr or dibs, rub el-hendy, and rub el-kharoub, respectively. Since date palm is native
to the Mediterranean region and Persian Gulf area, local populations and tribes cultivated this tree for
centuries (for the last 7000 years) and consumed its fruit-derived products, including syrup, as typi-
cal beverage, which is used in traditional medicine although ignoring their chemical compositions and
217
nutritional values [4]. Hence, this chapter addresses all issues related to date syrup chemistry and nutri-
tional and functional properties. In addition, an overview of the production of date syrup and its applications
in different areas including food and drink processing is provided.

Date fruit constitutes an important nutritional product, which contains high fiber, sugar, protein, vita-
mins, and minerals content [5]. Dates contain up to 70% total sugars, from which more than 50% is
inverted sugar, and have a water activity (aw) of 0.6 with a pH of 5.5 [6]. Processing date into concentrated
beverage, date syrup, is convenient for industrial use, especially in the Gulf area where it is the most
commonly derived date product. This thick dark-brown syrup resembles dark-colored honey, presenting
the same viscosity but having a very peculiar organoleptic flavor, and contains different nutrients. Date
syrup is a common traditionally derived date product generally used for softening and preserving dates.
Early records revealed that date syrup, turned into a nutritious drink, might have been an overland export
article between the Gulf area and China in the past. Very limited records are available regarding the
compositional and nutritional characteristics of date syrups and other by-products (such as press cakes
and seeds) [5,7]. Analytical data suggest that this syrup has roughly the same qualities as the date juice,
but it is more concentrated [8]. As for their starting fruit pulps, date syrups are claimed to be excellent
sources of natural micronutrients, which may improve human health and nutrition [9,10]. Data on the
nutritional composition of date syrup are summarized in Table 18.1, emphasizing the characteristics and
quality richness of this date-derived beverage.
Date syrup contains mostly sugars, including sucrose, glucose, and fructose as well as other soluble
matters. It is a high-energy dense food rich in carbohydrate and a good source of minerals. The syrup is
TABLE 18.1
Compositional and Nutritional Characteristics of Date Syrups (per 100 g Fresh Weight)
Unit [8] [11]a [12] [13] [14]
Water % nr 20.56–34.33 nr 23.72 16.00
Energy kcal nr 260–327b nr nr 339b
Protein g 1.02 0.95–1.09 0.50 nr 0.83
Lipid (fat) g nr 0.62–2.84 nr nr 1.98
Ash g 2.03 1.23–1.76 nr 2.63 6.80
Total dietary fiber g nr 0.01–0.18 nr nr nr
Pectin content g nr nr nr nr 1.46
Total sugars g 70.81 62.73–74.24 nr nr 79.45
Reducing sugar g 68.42 nr nr nr 74.87
Fructose g 30.93 nr 37.00 45.70 nr
Glucose g 33.32 nr 35.00 43.30 nr
Sucrose g 3.97 nr 5.00 0.50 nr
Minerals
Calcium mg 37.70 nr 750 157.30 338
Iron mg 9.30 nr nr nr 7.80
Magnesium mg nr nr nr 85.20 143
Potassium mg 217 nr nr 1310.9 202.8
Sodium mg 70.40 nr nr 157.30 13.00
Vitamins
Vitamin C mg nr nr nr nr 0.185
a The values are given as a range from minimum to maximum (syrups are from three date varieties).
b Calculated on the basis of total carbohydrate/protein/fat content.
Date Syrup 219
Sorting/checking of dates
Washing with water
Drying at 40°C (30 min)
Destoning
Slicing/grinding the pulp
Mixing with water

(Ratio 1:3, w/v)
Date syrup
Macerate (30 min)
Boiling2
Boiling1 (30 min)
until
~70°Brix
Filtration
Press cake Pooled juices
FIGURE 18.1 Diagram of the traditional process of date syrup. (Note: numbers 1 and 2 indicate the “boiling step 1” and
“boiling step 2’’ in the process.)
particularly rich in potassium, which is believed to be helpful in improving kidney function and
controlling blood pressure (Table 18.1). It also contains a very complex mixture of other saccharides,
amino and organic acids, polyphenols, and carotenoids [2]. The traditional processing method is remark-
ably based on heating at nonexcessive temperatures, even for a long time (step Boiling2 in Figure 18.1). To
produce date syrup, all soluble solids are extracted from the fruit. Then, the juice is filtered and concen-
trated to 70°Brix with a pH of 4.8–5.5. The process of date syrup preparation has been reported in many
works with slight specific variations. Al-Farsi et al. [15] prepared syrups from different date varieties fol-
lowing two extractions with water (1:1, w/v) at 60°C and then filtration through a Whatman No. 41 filter
paper. The syrup was then obtained by concentrating the date juice to 72°Brix using a rotary evaporator
at 70°C. Iranian date syrup is concentrated to about 75–80°Brix with pH adjusted to 2.8–3.0 using citric
acid. The “soft” processing, starting either from fresh or frozen fruit pulp, is thought to preserve many
nutrients present in the starting fruit material. Generally, in thermal processing, heat is used for two
purposes: (1) to destroy spoilage organisms and to inactivate troublesome enzymes and (2) to concentrate
the by-product and, therefore, reduce water activity, a critical parameter involved in food quality deterio-
ration during storage [16]. Inappropriately, the heating (boiling) remains the key step to be optimized in
order to reduce the loss of nutritional value. The quantity of added water is controlled on the basis of the
subsequent process steps and products. When the final product is concentrated, such as date spread, date
syrup, or liquid date sugar, water acts only as a carrier to facilitate separation and is finally removed by
evaporation. After refining, juice with soluble solid content of 20%–25% can be obtained, which needs
to be concentrated. The most common soluble solid content of date syrup is 75% [4].
In recent years, frequent consumption of high-energy drinks has been discouraged due to their adverse
effects on diabetic and obese patients as well as for overweight children. To the best of our knowledge,
neither epidemiological nor clinical works have been done on this subject; thus, we may depend on
the available data for other high-calorie beverages. Several long-term intervention studies have found
that chronic consumption of sugar-sweetened beverages leads to increased energy intake and weight
gain [17]. This nutritional concern should stimulate scientists to investigate the impact of the frequent
consumption of date syrup in both traditional and modern cultures [18].

In addition to its nutritional content, date syrup is rich in bioactive compounds. In our recent works
[19–22], we studied fruit-derived syrups, particularly their polyphenolic content and related biological
potential. The phenolic profile of the aqueous-acetone extracts of Tunisian syrups from date [19], barbary
fig [20], azarole [21], and carob [22] was qualitatively and quantitatively studied. Several phenolic com-
pounds were identified by comparison with standard reference compounds or previously reported data in
the literature. The functional potentials of these extracts were also evaluated in terms of antioxidant and
antimicrobial activities in comparison with well-established reference substances. Polyphenolic com-
pounds extracted from date syrup were fractionated and analyzed by reversed phase–high-performance
liquid chromatography–diode array detection (RP-HPLC-DAD) and electrospray ionization–mass spec-
trometric (ESI-MS) techniques. Table 18.2 shows the content of phenolic compounds in date syrup.
The major compounds identified by Dhaouadi et al. [19] were attributed to coumaric acid, an ester of
gallic acid, an ester of vanillic acid, cinnamic acid, 3,4-dicaffeoylquinic acid, caffeoylshikimic acid,
caffeoylsinapylquinic acid, and caffeic acid (Figure 18.2). The major polyphenolic compounds in date
syrup were caffeoylsinapylquinic acid (72%), cinnamic acid (9%), and coumaric acid (6%). These results
are consistent with those reported by Mansouri et al. [24], who worked on the extracts of different date
varieties. The peculiarity is that the presence of a high content of free cinnamic acid is not commonly
found in dates. Abbès et al. [23] also analyzed Tunisian traditionally prepared date syrup using HPLC-
DAD method and confirmed the presence of cinnamic acid in the syrup phenolic extracts (Table 18.2).
These investigators also provided supplementary data about additional phenolics detected in date syrup,
namely, catechin, sinapic acid, syringic acid, vanillic acid, and ferulic acid (Figure 18.2), but did not
detect dicaffeoylquinic acid, gallic acid and vanillic acid esters, caffeoylshikimic acid, and caffeoylsina-
pylquinic acid that have been reported by Dhaouadi et al. [19]. The differences between these reports
may be related to the extraction methods employed. Some protocols comprise a prechromatographic
step to remove sugars and other polar compounds from the crude polyphenolic extract [23]. Whereas,
in other protocols, phenolic compounds are extracted with cold (–20°C) aqueous-acetone (3:7, v/v) that
eliminates interfering compounds (precipitates proteins and peptides), thus circumventing preparative
chromatographic extractions (solid-phase extraction and solid-phase microextraction), a way to reduce
the number of extraction steps and, consequently, might preserve the bioavailable native forms of the
phenolics [19]. According to Macheix et al. [25], the presence of coumaric acid is probably associated
TABLE 18.2
Polyphenolics in Date Syrup (per 100 g Fresh Weight)
Unit [19] [23]
Catechin mg nd 2.51
3,4-Dicaffeoylquinic acid mg 13.86 nd
5-O-Caffeoylshikimic acid mg 17.70 nd
Caffeic acid mg 9.81 2.26
Caffeoylsinapylquinic acid mg 285 nd
Cinnamic acid mg 35.79 19.04
Coumaric acid mg 23.03 79.43
Gallic acid mg 6.79 nd
Vanillic acid mg 2.55 nd
Ferulic acid mg nd 12.42
Sinapic acid mg nd 10.89
Syringic acid mg nd 3.19
Vanillic acid mg nd 30.02
Total Phenolics mg 395 160
Note: Values are given with respect to standard compounds analyzed in same condition as the extract.
Date Syrup 221
O OH O OH O OH O OH O OH
H3C H3C CH3

HO O O O
OH OH OH OH
Cinnamic acid Coumaric acid Caffeic acid Ferulic acid Sinapic acid
O OH O OH O OH
H3C H3C CH3

HO OH O O O
OH OH OH
Gallic acid Vanillic acid Syringic acid
OH
OH
OH OH OH
O
OH O
OH
O O
HO O HO O HO
OH O
OH HO O
OH O
H
OH OH OH
Catechin Dicaffeoylquinic acid Caffeoylshikimic acid
FIGURE 18.2 Chemical structures of polyphenols present in date syrup.
with fruit ripening and intense extraction conditions. Additionally, the presence of 5-O-caffeoylshikimic
(dactyliferic acid) and a shikimate ester is suggested to be a contributor to browning reactions that
occur during date fruit ripening. Using colorimetric methods, the flavonoid and carotenoid content were
also estimated in Tunisian date syrups that were 194.5 mg catechin equivalents (CE) per 100 g fresh
weight (FW) and 0.018 mg/100 g FW, respectively [23].
The antioxidant activity of functional compounds, particularly flavonoids, phenolic acids, and carot-
enoids, has been attributed to various mechanisms such as prevention of continued hydrogen abstraction,
decomposition of peroxides, prevention of chain initiation, binding of transition metal ion catalysts,
reductive capacity, and radical scavenging [21]. Antioxidants are considered as beneficial to human health
since they decrease the risk of degenerative diseases and certain types of cancers by reduction of oxidative
stress and inhibition of oxidation of macromolecules. The antioxidant activity of date syrup was deter-
mined by different methods. Al-Farsi et al. [15] analyzed different date syrups using the oxygen radical
absorbance capacity (ORAC) method and found that antioxidant activities ranged from 84 to 106 μmol
trolox equivalents (TE)/100 g FW of syrup. In our recent report [19], 2,2-diphenyl-1-picrylhydrazyl
(DPPH) and 2,2′-azino-bis-(3-ethylbenz-thialzoline-6-sulfonic acid) (ABTS) radical-scavenging assays
were used. DPPH and ABTS radicals are stable free radicals, which have been widely used as a tool to
determine the antiradical activity of plant extracts [19,26]. Date syrup polyphenolic extract exhibited a
strong DPPH-scavenging effect of about 101.3 μmol TE/100 g FW [19]. The DPPH radical is known to
involve electron transfer reaction. This radical is believed to accept an electron and become a stable dia-
magnetic molecule. It has been shown that many antioxidant molecules such as ascorbic acid (vitamin C),
tocopherol (vitamin E), and numerous flavonoids reduce DPPH due to their hydrogen-donating ability,
a property that the date syrup extract may also share. The free radical scavenging potential of date syrup
extracts was also assessed by means of their ability to quench the cationic ABTS radical generated
experimentally in the assay system. The ABTS-scavenging ability of the date syrup extract was 521 μmol
TE/100 g FW. The difference between ABTS-/DPPH-based antioxidant activities has been observed for
other plant extracts and thought to be correlated with the concentration and chemical structures of indi-
vidual polyphenols, which could selectively scavenge anionic or cationic free radicals.
Moreover, the capacity of the Tunisian Deglet Nour date syrup extract to reduce Fe(III) was evaluated
using the ferric-reducing antioxidant power (FRAP) method, which was found to be equivalent to 38 μg
TE/100 g FW [19]. For all antioxidant evaluation tests, the high repeatability of the assays was verified,
and no significant difference (P > 0.05) was found between different syrup extracts.
The effect of concentration temperature on the antioxidant activity, carotenoid, phenolics (includ-
ing tannin, nontannin, and flavonoid), and 5-hydroxymethyl-2-furfuraldehyde (5-HMF) content of date
syrups prepared by date juice concentration was investigated [27]. Following date juice concentration
at relatively high temperatures (60°C–100°C), all date syrups showed slight decreases in the analyzed
contents, except for total phenolics and 5-HMF amounts that increased significantly. Statistical analysis
of data showed that concentration at 100°C significantly enhanced the antioxidant activities that were
correlated with the 5-HMF contents.
Several studies were focused on the effect of the innovative methods for the extraction of date syrup,
particularly, involving a pretreatment step using pectinase/cellulase mixtures on the nutritional and
organoleptic properties [9,23,28]. The enzymatic treatment of dates’ flesh during juice extraction is sup-
posed to increase the extraction yield as the water/flesh ratio increased, thus, facilitating the highest
recovery of total soluble solids. The collected reports permitted to conclude that enzymatic pretreat-
ment led to a syrup highly desirable than cane syrup. El-Sharnouby et al. [28] recommended the use of
pectinase/cellulase mixture to produce concentrated date syrup from “tamr” (date) fruits for use in food
product development. Furthermore, the investigators noticed that the produced syrup was suitable for
manufacturing different food products. When compared to the traditionally made date syrup, qualita-
tive and quantitative analyses demonstrated that the produced pectinase/cellulase preextracted syrup
showed the lowest phenolic and flavonoid content and, inversely, the highest carotenoids. Additionally,
the antioxidant activity of the date syrups was significantly (P < 0.05) diminished by the enzymatic
extraction method. “Deglet Nour” variety obtained by the traditional method presented the highest
total antioxidant activity among all the analyzed date syrups. For this variety, the antioxidant potential
was decreased from 136 to 118 mg ascorbic acid/g of syrup when enzymes mixture was used for date
juice extraction [27]. Using the hydrogen peroxide–scavenging assay, the IC50 (sample concentration
needed to decrease the initial hydrogen peroxide concentration by 50%) values (mg/mL) of the enzyme-
treated date syrup were significantly lower (P < 0.05) than the control without enzyme pretreatment.
Similar trends were obtained when the reducing power of the syrups was assessed using the FRAP
assay. The Fe2+ chelating method confirmed the significant reducing effect on this antioxidant property
when date syrup extraction was conducted together with enzymatic pretreatment. For the “Deglet Nour”
variety, the percentage of metal chelation was decreased from 72.59% to 45.37%.
Moreover, according to Dhaouadi et al. [19], the polyphenolic extract of date syrup exhibited a very
strong antibacterial potential. The highest antibacterial activity was recorded against Staphylococcus
epidermidis, a Gram-positive bacterium frequently found on the skin and mucous membranes of humans
and animals. This bacterium is responsible for infections of skin, nasal (such as sinusitis), and urinary
tract. It has the ability to produce biofilms that allow them to adhere to surfaces of medical implants. The
date syrup extracts showed both bacteriostatic and bactericide activities with minimum inhibitory and
bactericidal concentrations (MIC and MBC) that ranged from 50 to 500 µg/mL. In contrast, no inhibi-
tory effect was observed against Escherichia coli, a Gram-negative bacterium that makes up about 80%
of our intestinal flora and includes some pathogenic strains. In addition, the polyphenolic extract did not
show antifungal activity using the yeast Candida albicans. The inhibitory effect of plant extracts against
microbial pathogens was recurrently attributed to their phenolic composition [26,29]. Using the standard
disk diffusion technique, varying degrees of bacterial sensitivity were observed, suggesting a differen-
tial intrinsic tolerance of microorganisms and/or the particular nature and combination of the phenolic
compounds present in the date syrup extract. Several reports explained the antimicrobial effect phenolic
Date Syrup 223
compounds by bacterial growth inhibition as they adsorb to cell membranes, interaction with enzymes
and effectors, or deprivation of substrates and metal ions [30]. Therefore, Dhaouadi et al. [19] assumed
that structural diversity of the bioavailable phenolics in the date syrup extracts plausibly influenced their
exhibited antimicrobial potentials.
18.4 Health Effects

Although few scientific studies have been conducted specifically on the date fruit, epidemiological studies
have shown that their high consumption decreases the risk of cardiovascular disease (CVD), some cancers,
and chronic diseases [31]. A number of medicinal uses are directly or indirectly ascribed to the consump-
tion of dates. The fruit is rich in tannins, making it a good astringent remedy for intestinal troubles. The
roots are used for treating toothache [32]. Dates contain about 43% of soluble and 57% of insoluble fibers.
The latter plays an important role in bowel regularity and preventing constipation. By retaining water in
the colon, they increase the volume and stool weight, reducing the transit time and facilitating the removal.
Furthermore, studies have shown that soluble fiber plays a role in reducing cholesterol and in the normal-
ization of blood glucose and insulin levels. Therefore, dates can help reduce the risk of CVD. Dates are
indicated in folk remedies for the treatment of various infectious diseases. In studies of fruit extracts, the
immunomodulatory activity of dates has been shown by Praveen [33]. The traditionally made date syrup
preserved some of the properties of date fruit since fiber content was found to be relatively important in
this product. El Hadrami and Al-Khayri [34] presented a review of the available information regarding
the importance of the date palm and its by-products. These authors emphasized that, in addition to the
use of fresh fruits for human consumption, a number of date-derived by-products, including jam, jelly,
juice, syrup, and fermented beverages (vinegar and alcohol), also have various uses. Date by-products
are also taken to relieve fever, liver and abdominal aches, cystitis, gonorrhea, and edema. The syrup
contains a high concentration of phytochemicals, especially carotenoids and phenolic compounds that
are well known to protect body cells from damage caused by free radicals. According to El Hadrami and
Al-Khayri [34], the formulations produced from date palm fruits, including syrup, are administered for
use against colds, sore throat, and bronchial cough, as well as to help relieve fever and abdominal aches.
Date syrup contains shikimate derivatives, which can be used to synthesize (6S)-6-fluoroshikimic acid, an
antibiotic, which inhibits the aromatic biosynthetic pathway. Shikimic acid is used as a base material for
the production of Oseltamivir (Tamiflu), which is an antiviral drug approved to prevent or slow the spread
of influenza virus between cells in the body by stopping the virus from chemically cutting ties with its host
cell. In addition, date syrup contains coumaric and cinnamic acids, which have antioxidant properties and
are believed to reduce the risk of stomach cancer by reducing the formation of carcinogenic nitrosamines
[35]. Additionally, cinnamic acid is used in flavors, synthetic indigo, and a range of pharmaceuticals; it is
also a precursor to the sweetener aspartame via enzyme-catalyzed amination to phenylalanine [36].
In our recent works [19–22], we studied fruit-derived syrups, particularly their polyphenolic content
and related biological potentials. The phenolic profile of the aqueous-acetone extracts of the Tunisian
traditionally made syrups from date, barbary fig, azarole, and carob was qualitatively and quantita-
tively studied. The functional properties of these extracts were also evaluated in terms of antioxidant
and antimicrobial activities in comparison with well-established reference substances. In addition, the
effect of the extracts on the in vitro viability of human tumorigenic and nontumorigenic cells was also
investigated. When neuroblastoma cells were treated for 1, 3, 6, and 24 h with different concentrations
of the date syrup polyphenolic extract, viability decreased significantly in a dose-dependent manner of
the time, compared to the control cells [19]. In the same way, and to see the effect of these date syrup
polyphenols on noncancerous cells, 3T3 fibroblasts were used as a model. These cells are present in
connective tissues and are sometimes called supporting cells. They include resident dermal cells that
ensure consistency and flexibility. For both cell lines, the obtained results showed that final concentra-
tions of syrup extract in the medium culture were higher than 45 µg/mL and decreased cell viability
by 70%–80% of the treated cells for 1 and 3 h. Following incubation for 1, 3, or 6 h in the presence of
the extracts at 65 µg/mL final concentration, fibroblasts maintained viability ≥60%. However, after 6 h
incubation, the viability of SH-5YSY neuroblastoma cells reached 23%, and practically no viability was
detected after 24 h. Dhaouadi et al. [19] noticed a dissimilar behavior of cancerous and noncancerous
cells when treated with date syrup polyphenolic extract. Following 6 h of incubation, the viability of
neuroblastoma cells was significantly decreased more than that of fibroblasts. The higher susceptibil-
ity of SH-5YSY neuroblastoma cells compared to 3T3 fibroblast cells suggests a particularly pro-
nounced cytotoxicity of the date syrup phenolic extract against cancer cells. Obtained data emphasize
the potential use of the date syrup polyphenols for curative purposes. This distinction is central in many
scientific studies to selectively target cancer cells in a therapeutic treatment. Similarly, Lantto et al. [37]
used SH-SY5Y cells and fibroblast CV1-P cell lines and found that curcumin reduced the viability of
tumorigenic SH-SY5Y cells and increased their p53 content more efficiently in comparison with the
CV1-P nontumorigenic cells.

The natural concentrated date syrup with no added preservatives has up to 1-year shelf life [6] and blends
easily into batters for sweet breads, cakes, muffins, cookies, and other bakery products, providing sweet-
ness and moisture retention, which helps retard spoilage. Because it is readily available and relatively
low priced, date syrup has become a popular product worldwide and constitutes an economical source
of carbohydrates both for human use and in microbial fermentations [38]. This syrupy liquid brings
its sweetness and flavor to baked goods and to all types of liquid foods and beverage a pplications [11].
It can be used for several purposes, for example as a sweetening and flavoring agent, including a sweetener
in tea and hot chocolate, topping on ice creams, bread making, spreading in breads, and mixing with
cold and hot milk [39].
From a technological point of view, the traditional process to prepare a fruit syrup is to start from a
boiled fruit pulp in water and pressed to extract the juice. The extracted juice is concentrated by cooking
over low heat until colored and obtains syrupy liquid, which has a total sugar concentration of approxi-
mately 70°Brix [7]. This old and popular process provides a mean of keeping fruits far beyond their
normal storage life. This explains why these food by-products are widely consumed in North Africa and
Arabian countries throughout the year. This date-derived syrup is preferentially consumed during the
cold periods for their high energetic sugar content. In addition, several usages as a sweetener or for char-
acteristic odor have been noted. The syrup is poured on cooked dough (asseeda) on specific occasions,
such as the celebration of Islamic special occasions and festivities [19]. Early records reveal that date
syrup in the past may have been an overland export article as a product even more concentrated than the
date itself and easily turned into nutritious drinks.
The traditional method starting from pressured date juice produces only 15% of extracted syrup, while
this amount reaches 60% in industrial operations by employing boiling and evaporation of date juice or
enzymatic treatment with pectinase and cellulase [9,28]. The general method for date syrup production
includes pretreatment of date, if necessary, by adding water and mixing juice extraction, filtration, reex-
traction of the press cake, and concentration [40]. In all processes, the preservation of the taste-producing
substances should be greatly considered. Juice extraction can be carried out by batch and continuous
methods [4]. Several reports indicated the effective use of ultrasonic waves to extract date syrup along
with microbial count reduction. The sonication was found to significantly decrease the microbial count
in the product in comparison with the classical method. The ultrasonic waves permitted to increase the
extraction yield in a shorter time with a better physical quality of the product [41,42]. Extraction, clari-
fying with active charcoal and concentration of date juice with microwave at 40% or 70% power level,
improved the color of date syrup compared to local traditionally produced syrup. The obtained syrup
was characterized with higher viscosity and total soluble solids and contained higher percentages of
glucose and fructose.
During recent decades, especially in countries where the date palm is cultivated on a large scale, date
syrup is becoming central for the development of broadenening industrial food products as a base for
fermented (vinegar and alcohol) or nonfermented (soft drinks) beverages, in confectionery and bakery
products. Date syrup is mixed with tahini or other pastries [2]. Its uses on bread have become increasingly
accepted. According to Sidhu et al. [43], the concentrated date syrup used in pan bread formulations did
Date Syrup 225
not adversely affect the baking loss but rather gave a significantly higher specific loaf volume. At 100%
replacement of sucrose with date syrup, compression force was decreased, a parameter that indicates
a softer texture of test bread samples than the control. Sensory analysis showed that the panelists gave
slightly higher scores for the formulated bread samples regarding texture, flavor, and overall acceptabil-
ity than for the control bread.
Date syrup could also substitute honey by up to 75%. Furthermore, date syrup could replace invert
sugar used in the wheat and sorghum biscuits up to 50% without affecting quality. For millet biscuits,
not more than 10%–15% of replacement could be tolerated. More than 15% substitution may lead to mil-
let sticky dough, which is difficult to handle. In cakes, up to 17% sugar replacement could be achieved
without significant quality changes in the cake.
Date syrup was used to replace sugar in an original formulation of Gaz, a popular Iranian traditional
delicious sweet [44]. This product is made from egg white, rosewater, sugar, pistachio, or almond. This
substitution caused formation of softer texture of samples and, consequently, reduction of the chewiness.
Study of sensory characteristics of the innovative formulated Gaz samples also indicated that samples
with 50%, 25%, and 75% date syrup had more acceptability than the control sample in terms of general
acceptance.
Date syrups are able to disguise the bitter or strong taste of some foods or food ingredients. Generally,
syrups excel at soothing sore throats, coughs, and many digestive upsets [45]. However, the high sugar
content of date syrups makes them inappropriate to treat nutritive imbalances or deep-seated chronic
disorders such as diabetes [3]. Although there have been limited attempts to produce a date soft drink,
which has to face the strong competition for this type of product, a typical process was successfully
developed including extra clarification steps of the diluted juice and reinforcing of the flavor and acidity
if a soft drink is desired. In the United States, several attempts have been made to introduce date syrup
along similar lines as other fruit juice concentrates as from raisin, fig, and prune for use in the prepara-
tion of cakes, cookies, and sweet breads, among others. Furthermore, Ghafari et al. [46] examined the
effect of using bleached date syrup as a substitute for glucose syrup in the formulation of original beer
(nonalcohol). The obtained results showed that 25% date syrup gave a minimum viscosity nonalcoholic
beer, whereas the maximum viscosity was obtained when using 50% date syrup. In addition, vinegar
made from dates is a traditional product in the Middle East. Moreover, diluted date syrup was success-
fully tested as raw material for the production of caramel, which is usually used as a color additive in
food processing.
Several attempts have been made to use date syrup as a sweetening and flavoring agent for dairy prod-
ucts such as date-flavored buffalo skim milk and date-flavored yogurt [47]. Similarly, a date juice milk
beverage was produced from date syrup and powdered or fresh milk. The pH adjustment (>6, to prevent
curdling) and ultrahigh treatment gave rise to drinks with a shelf life of 3 months at room temperature.
The ratio of the two 20°Brix diluted date syrup and milk was 4:6. When using date syrup in ripple ice
cream, early crystallization at the low temperature remained to be a problem that was not found experi-
mentally with date spread. Nevertheless, date syrup was successfully used as a sugar replacement, up to
15%, in ice cream making without affecting overrun or viscosity. Generally, syrups as liquids are more
easily used in mixed drinks and beverages than granulated sugar. Multiple hydrogen bonds between
sugar and water are responsible for the viscous consistency of the solution, which should not be close to
a supersaturation point (65%–67% by weight) [45].
Date syrup offers opportunities for the food industry to create novel products, such as yogurt drinks.
Yogurt manufacturers more widely prefer using liquid sugar because of the efficiency of its handling
[47]. Its incorporation into yogurt after fermentation has the potential to be used in the manufacture of
flavored yogurt [47]. However, the commercial applications of these innovative products are limited.
For safety reasons, the syrup should be incorporated into the yogurt mix prior to heat treatment in order
to eliminate osmophilic yeasts and molds, preventing postpasteurization contamination and better tex-
tural quality in the final product. In some cases, incorporation of sugar into fermented milk after incuba-
tion is required. In this situation, sugar should be added as pasteurized liquid sugar or flavored sweetened
syrups, and extra care must be taken to avoid any contamination.
Fructose, which is 60% sweeter than sucrose and 150% more than glucose, is mainly used in food and
beverage industries at relatively high concentrations, known as high-fructose syrup (HFS). These syrups
are produced from different raw materials including cornstarch, sugarcane, and sugar beet. Since date
syrup is rich in reduced sugars, particularly fructose and glucose, several trials aimed to valorize dis-
carded dates and produce low-cost HFS by means of enzymatic isomerization, which converts glucose
into fructose [48].
Among common viscous carbohydrate-based liquids, syrups are important industrial ingredients in
many foods and beverages where they can be used as food for yeast (rice of selected baked goods) as a
sweetening agent or for browning [49]. During the last decades, several works have been carried out in
order to set up innovative techniques and processes and develop value-added products and substances
using date syrup. Alanazi [50] tried to produce tablet binder (glue that hold powders together to form
granules and tablet) starting from date syrup to substitute two well-known binders, namely, sucrose
syrup and starch paste. Interestingly, the authors noticed a better flavoring and masking taste effect from
an evaluation by human volunteers, thus demonstrating the usefulness of date syrup as sweetener and
flavoring the tablets in addition to its use as a binder. Other products from date syrup are yeasts (rich in
protein), organic acids, vitamins, and also baker’s yeast [49]. Roukas and Kotzekidou [51] investigated
the pretreatment of date syrup with sulfuric acid, tricalcium phosphate, hydrochloric acid, potassium
ferrocyanide, and ethylenediaminetetraacetic acid to determine the ability of enhancing citric acid pro-
duction. It was found that 2% tricalcium phosphate was most effective. Optimum pH for citric acid
production was 6.5. Adding 4% methanol to date syrup treated with 2% tricalcium phosphate increased
citric acid concentration. Fat can also be produced from date juice and syrup by means of microorgan-
isms like Penicillium lilacinum, P. soppi zaluski, and Aspergillus nidulans [4].
The quality richness of date syrup inspired the scientists to incorporate it in medium cultures.
Alkhateeb [14] investigated the replacement of the commonly used sucrose, as carbon source, by date
palm syrup for micropropagation of date palm “cv. Suckary.” The results indicated that date syrup, used
at 6% was taken up from the media, permitted the production of high number of somatic embryos, long
shoot, and improved the germination of the somatic embryos in comparison with a control experiment.
Nevertheless, the supplementation of 10% of date syrup to the medium caused severe reduction in a
number of somatic embryos, which was attributed to osmotic stress. Date syrups and date pits were sug-
gested to be a suitable substrate for the cultivation of microorganisms and were reported to have positive
influence as nutrients for the cultivation of Lactococcus lactis [52,53]. The effects of different carbon
sources on biomass and yield indicated that date syrup could be evaluated as a favorable carbon source in
baker’s yeast Saccharomyces cerevisiae production. Hence, fermentation of date extracts to ethanol and
vinegar in batch and continuous membrane reactors and date syrup and waste was tried for the produc-
tion of ethanol [53]. Date syrup was also experimented with for the microbial production of biomass and
citric acid [53–55], particularly in immobilized cells techniques. During the fermentation process of date
waste syrup, the citric acid production by Aspergillus niger reached 98.42 g/L [54].
Starting from different date fruit by-products, including syrup, as substrates, Elsanhoty et al.
[56] investigated the optimization of the medium components using Plackett–Burman design for
the production of carotenoids by the Lactobacillus plantarum QS3 during the course of fermentation.
The data so obtained indicated that, when date syrup at 5% sugar concentration was used alone, it
resulted in 16.21 mg carotenoids per kilogram of dry cell. Whereas, there was an increase in carotenoids
production (54.89 mg/kg dry cell) when date syrup was used as a carbon source and supplementation
of the Man, Rogosa, and Sharpe (MRS) medium with salts and organic nitrogen after the optimization
of pH and temperature [56]. Date syrup was also used as an additional carbon source in the medium
culture of Streptomyces mobaraensis to produce Bleomycin, a glycopeptide-derived antibiotics, which
exhibits a strong antitumor activity and has been widely employed for the treatment of several malig-
nancies, including non-Hodgkin’s lymphoma, squamous cell carcinoma, and testicular tumors [57].
The study concluded that 40 g/mL of date syrup in the complex medium enhanced the production of
Bleomycin by 73%. Omar et al. [58] reported the isolation of a Bacillus megaterium strain that has
been optimized for growth and poly(3-hydroxybutyrate) (PHB) accumulation in medium enriched with
5% (w/v) date syrup. Polyhydroxyalkanoates (PHAs) are nontoxic, biocompatible, and biodegradable
materials. PHAs have the potential to replace petroleum-based plastics as biomedical materials for
use in surgical pins, sutures, staples, blood vessel replacements, bone replacements and plates, medi-
cal implants, and drug delivery devices owing to their superior biodegradability and biocompatibility.
Date Syrup 227
According to Omar et al. [58], using date syrup as a substrate in the culture medium enhanced the
growth of the B. megaterium strain, which reached a cell density of about 3 g/L with a PHB content
of the cells of 50% (w/w).
18.6 Conclusion
Date-derived syrup is commonly consumed as a typical beverage in North Africa and many other Arab
and Mediterranean countries. This concentrated extra-sweet solution has a reduced water activity that
fits conservative purposes avoiding microbial and chemical alterations that are induced in the presence
of free water. The syrup as well as its value-added derived products has many applications in food
and nutraceutical industries. In addition, employing of innovative bioprocessing technologies may lead
to the emergence of new bioindustries for valorization of discarded date fruit by-products and waste.
The potential downstream emerging industries from low-priced date syrup and derivatives should be
promoted for local as well as export markets. As being new branded products, marketing campaigns
are needed to promote these novel industries. Additionally, in-depth investigations should follow about
the health effects of this functional date-derived product to elucidate the molecular mechanisms and to
identify the bioactive compounds. While industrial developments for new drugs or biomolecules derived
from date syrup may need time, the immediate major therapeutic interest in this syrup could be for its
use in disease risk reduction and preventive practice.
REFERENCES
1. FAOSTAT, Agro-Statistics Database 2012. Published online at: http://faostat.fao.org/site/339/default.
aspx (accessed July 11, 2014).
2. Ashraf, Z. and Hamidi-Esfahan, Z., Date and date processing: A Review. Food Rev. Int., 27, 101–133,
2011.
3. Varzakas, T. and Chryssanthopoulos, C., Nutritional and health aspects of sweeteners, in Sweeteners:
Nutritional Aspects, Applications, and Production Technology, Varzakas, T., Labropoulos, A., and
Anestis, S., Eds., CRC Press/Taylor & Francis Group, Boca Raton, FL, 2012, pp. 329–365.
4. Barreveld, W.H., Date Palm Products, FAO Agricultural Services Bulletin, No. 101, Rome, Italy, 1993.
5. Alasalvar, C. and Shahidi, F., Composition, phytochemicals, and beneficial health effects of dried fruits:
An overview, in Dried Fruits Phytochemicals and Health Effects, Alasalvar, C. and Shahidi, F., Eds.,
Wiley-Blackwell, Oxford, UK, 2013, pp. 1–18.
6. Varzakas, T. and Labropoulos, A., Other sweeteners, in Sweeteners: Nutritional Aspects, Applications,
and Production Technology, Varzakas, T., Labropoulos, A., and Anestis, S., Eds., CRC Press/Taylor &
Francis Group, Boca Raton, FL, 2012, pp. 175–208.
7. Gabsi, K., Trigui, M., Barrington, S., Helal, A.N., and Taherian, A.L., Evalution of rheological proper-
ties of date syrup. J. Food Eng., 117, 162–172, 2013.
8. Mostafa, A.M. and Ahmed, A.A., Libyan date syrup (rub-at-tamr). J. Food Sci., 46, 1162–1166, 1981.
9. Al-Hooti, S., Sidhu, J.S., Al-Saqer, J.M., and Al-Othman, A., Chemical composition and quality of date
syrup as affected by pectinase/cellulase enzyme treatment. Food Chem., 79, 215–220, 2002.
10. Abbès, F., Bouaziz, M.A., Blecker, C., Masmoudi, M., Attia, H., and Besbes, S., Date syrup: Effect of
hydrolytic enzymes (pectinase/cellulase) on physico-chemical characteristics, sensory and functional
properties. LWT—Food Sci. Technol., 44, 1827–1834, 2011.
11. Al-Farsi, M., Clarification of date juice. Int. J. Food Sci. Technol., 38, 241–245, 2003.
12. Jamshidi Mokhber, M., Alemzadeh, I., and Vossoughi, M., Optimization of HFDS production from date
syrup. Arch. SID IJE Trans. B: Appl., 21, 127–134, 2008.
13. Nasehi, S.M., Ansari, S., and Sarshar, M., Removal of dark colored compounds from date syrup using
activated carbon: A kinetic study. J. Food Eng., 111, 490–495, 2012.
14. Alkhateeb, A., Comparison effects of sucrose and date palm syrup on somatic embryogenesis of date
palm (Phoenix dactylifera L). Am. J. Biotechnol. Biochem., 4, 19–23, 2008.
15. Al-Farsi, M., Alasalvar, C., Al-Abid, M., Al-Shoaily, K., Al-Amry, M., and Al-Rawahy, F., Compositional
and functional characteristics of dates, syrups, and their by-products. Food Chem., 104, 943–947, 2007.
16. Yousif, A.K., Morton, I.D., and Mustafa, A.I., Processing, evaluation and water relations of date paste.
J. Trop. Sci., 31, 147–158, 1991.
17. Eshak, E.S., Iso, H., Mizoue, T., Inoue, M., Noda, M., and Tsugane, S., Soft drink, 100% fruit juice, and
vegetable juice intakes and risk of diabetes mellitus. Clin. Nutr., 32, 300–308, 2013.
18. Slining, M.M., Mathias, K.C., and Popkin, B.M., Trends in food and beverage sources among US
children and adolescents: 1989–2010. J. Acad. Nutr. Diet., 113, 1683–1694, 2013.
19. Dhaouadi, K., Raboudi, F., Estevan, C., Barrajon, E., Vilanova, E., Hamdaoui, M.H., and Fattouch, S.,
Cell viability effects and antioxidant and antimicrobial activities of Tunisian date syrup (Rub El Tamer)
polyphenolic extracts. J. Agric. Food Chem., 59, 402–406, 2011.
20. Dhaouadi, K., Raboudi, F., Funez, L., Pamies, D., Estevan, C., Barrajon, E., Hamdaoui, M.H., and
Fattouch, S., Polyphenolic extract of barbary-fig (Opuntia ficus-indica) syrup: RP–HPLC–ESI–MS anal-
ysis and determination of antioxidant, antimicrobial and cancer-cells cytotoxic potentials. Food Anal.
Method., 6, 45–53, 2013.
21. Belkhir, M., Rebai, O., Dhaouadi, K., Congiu, F., Tuberoso, C.I.G., Amri, M., and Fattouch, S.,
Comparative analysis of Tunisian wild Crataegus azarolus (yellow azarole) and Crataegus monogyna
(red azarole) leaf, fruit, and traditionally derived syrup: Phenolic profiles, antioxidant and antimicrobial
activities of the aqueous-acetone extracts. J. Agric. Food Chem., 61, 9594–9601, 2013.
22. Dhaouadi, K., Belkhir, M., Akinocho, I., Raboudi, F., Pamies, D., Barrajón, E., Estevan, C., and
Fattouch, S., Sucrose supplementation during traditional carob syrup processing affected its chemical
characteristics and biological activities. LWT—Food Sci. Technol., 57, 1–8, 2014.
23. Abbès, F., Kchaou, W., Blecker, C., Ongenac, M., Lognay, G., Attia, H., and Besbes, S., Effect of pro-
cessing conditions on phenolic compounds and antioxidant properties of date syrup. Ind. Crop. Prod.,
44, 634–642, 2013.
24. Mansouri, A., Embarekb, G., Kokkalouc, E., and Kefalasa, P., Phenolic profile and antioxidant activity
of the Algerian ripe date palm fruit (Phoenix dactylifera). Food Chem., 89, 411–420, 2005.
25. Macheix, J.J., Fleuriet, A., and Billot, J., Fruit Phenolics, CRC Press, Boca Raton, FL, 1990.
26. Fattouch, S., Caboni, P., Coroneo, V., Tuberoso, C.I.G., Angioni, A., Dessi, S., Marzouki, N., and
Cabras, P., Comparative analysis of polyphenolic profiles and antioxidant and antimicrobial activities
of Tunisian pome fruit pulp and peel aqueous acetone extracts. J. Agric. Food Chem., 56, 1084–1090,
2008.
27. Abbès, F., Besbes, S., Brahim, B., Kchaou, W., Attia, H., and Blecke, C., Effect of concentration tem-
perature on some bioactive compounds and antioxidant proprieties of date syrup. Food Sci. Technol.
Int., 19, 323–33, 2013.
28. El-Sharnouby, G.A., Al-Eid, S.M., and Al Otaibi, M.M., Utilization of enzymes in the production of
liquid sugar from dates. Afr. J. Biochem. Res., 3, 41–47, 2009.
29. Belkhirr, M., Rebai, O., Dhaouadi, K., Sioud, B., Amri, M., and Fattouch, S., Antioxidant and antimi-
crobial activities of Tunisian azarole (Crataegus azarolus L.) leaves and fruit pulp/peel polyphenolic
extracts. Int. J. Food Prop., 16, 1380–1393, 2012.
30. Denyer, S.P., Mechanisms of action of antibacterial biocides. Int. Biodeterior. Biodegrad., 36, 227–245,
1995.
31. Ishurd, O. and Kennedy, J.F., The anticancer activity of polyscharide prepared from Libyan dates
(Phoenix dactylifera L.). Carbohydr. Polydr. Polm., 59, 531–535, 2005.
32. Al-Shahib, W. and Marshall, R.J., The fruit of the date palm: It’s possible use as the best food for the
future. Int. J. Food Sci. Nutr., 54, 247–259, 2003.
33. Praveen, K.V., Antioxidant and antimutagenic properties of aqueous extract of date fruit (Phoenix
dactylifera L. Arecaceae). J. Agric. Food Chem., 50, 610–617, 2002.
34. El Hadrami, A. and Al-Khayri J.M., Socioeconomic and traditional importance of date palm. Emir. J.
Food Agric., 24, 371–385, 2012.
35. Ferguson, L.R., Shuo-Tun, Z., and Harris, P.J., Antioxidant and antigenotoxic effects of plant cell wall
hydroxycinnamic acids in cultured HT-29. Mol. Nutr. Food Res., 49, 585–693, 2005.
36. Dorothea, G., Cinnamic acid, in Ullmann’s Encyclopedia of Industrial Chemistry, Verlag Chemie
GmbH, Eds., Wiley-VCH Verlag GmbH & Co, Weinheim, Germany, 2000, pp. 231–239.
37. Lantto, T.A., Colucci, M., Zavadova, V., Hiltunen, R., and Raasmaja, A., Cytotoxicity of curcumin,
resveratrol and plant extracts from basil, juniper, laurel and parsley in SH- SY5Y and CV1-P cells. Food
Chem., 117, 405–411, 2009.
Date Syrup 229
38. Mehaia, M.A. and Cheryan, M., Fermentation date extracts to ethanol and vinegar in batch and continu-
ous membrane reactors. Enzyme Microb. Technol., 13, 257–261, 1991.
39. Gohari-Ardebili, A.A., Survey on effects of replacement of sugar with date syrup on physico-chemical
characteristics of soft ice cream, MSc thesis, Ferdosi University, Mashhad, Iran, 2004.
40. Al-Hooti, S.N. and Sidhu J.S., Functional foods from date fruits, in Asian Functional Foods, Shi, J., Ho,
C.-T. and Shahidi, F., Eds., CRC Press/Taylor & Francis Group, Boca Raton, FL, 2005, pp. 508–516.
41. Entezari, M.H., Hagh Nazary, S., and Haddad Khodaparast, M.H., The direct effect of ultrasound on the
extraction of date syrup and its microorganisms. J. Ultrason. Sonochem., 11, 379–384, 2004.
42. Al-Mutairi, S.K. and Al-Jasser, M.S., Effect of microwave concentration on the quality of dibs. Am. J.
Food Technol., 7, 609–621, 2012.
43. Sidhu, J.S., AL-Saqer, J.M., AL-Hooti, S.N., and AL-Othman, A., Quality of pan bread made by replac-
ing sucrose with date syrup produced by using pectinase/cellulase enzymes. Plant Food. Hum. Nutr.,
58, 1–8, 2003.
44. Raiesi Ardali, F., Nilchian, Z., Ahmadi, S., Shariati, M.A., and Akbarian, M., Replacing sugar by date
syrup in gaz and investigation of texture properties. Int. J. Sci. Eng., 6, 11–14, 2014.
45. Labropoulos, A. and Anestis, S., Syrups, in Sweeteners: Nutritional Aspects, Applications, and
Production Technology, Varzakas, T., Labropoulos, A., and Anestis, S., Eds., CRC Press/Taylor &
46. Ghafari, Z., Hojjatoleslamy, M., Shokrani, R., and Shariaty, M.A., Use of date syrup as a sweetener
in non alcoholic beer: Sensory and rheological assessment. J. Microbiol. Biotechnol. Food Sci., 3,
182–184, 2013.
47. Gad, A.S., Kholif, A.M., and Sayed, A.F., Evalution of the nutritional value of functional yogurt result-
ing from combination of date palm syrup and skim milk. Am. J. Food Technol., 5, 250–259, 2010.
48. Khosravanipour Mostafazadeh, A., Sarshar, M., Javadian, Sh., Zarefard, M.R., and Amirifard
Haghighi, Z., Separation of fructose and glucose from date syrup using resin chromatographic method:
Experimental data and mathematical modeling. Sep. Purif. Technol., 79, 72–78, 2011.
49. Alemzadeh, I. and VosoughiInd, M., Date syrup and baker’s yeast production. Eng. Chem. Res., 41,
128–130, 2002.
50. Alanazi, F.K., Utilization of date syrup as a tablet binder, comparative study. Saudi. Pharm. J., 18,
81–89, 2010.
51. Roukas, T. and Kotzekidou, P., Pretreatment of date syrup to increase citric acid production. Enzyme
Microb. Technol., 21, 273–276, 1997.
52. Khiyami, M., Aboseide, B., and Pometto, A., Influence of complex nutrient sources: Date syrup and date
pits on Lactococcus lactis growth and nisin production. J. Biotechnol., 136, 717–742, 2008.
53. Gupta, N. and Kushwaha, H., Date palm as a source of bioethanol producing microorganisms, in Date
Palm Biotechnology, Mohan, J.S., Al-Khayri, M.J., and Dennis, V.J., Eds., Springer, Berlin, Germany,
2011, pp. 711–727.
54. Acourene, S. and Ammouche, A., Optimization of ethanol, citric acid, and α-amylase production from
date wastes by strains of Saccharomyces cerevisiae, Aspergillus niger, and Candida guilliermondii.
J. Ind. Microbiol. Biotechnol., 39, 759–766, 2012.
55. Mostafa, Y.S. and Alamri, S.A., Optimization of date syrup for enhancement of the production of citric
acid using immobilized cells of Aspergillus niger. Saudi. J. Biol. Sci., 19, 241–246, 2012.
56. Elsanhoty, R.M., Al-Turki, I.A., and Ramadan, M.F., Screening of medium components by Plackett–
Burman design for carotenoid production using date (Phoenix dactylifera) wastes. Ind. Crops Prod.,
36, 313–320, 2012.
57. Radwan, H.H., Alanazi, F.K., Taha, E.I., Dardir, H.A., Moussa, I.M., and Alsarra, I.A., Development of
a new medium containing date syrup for production of bleomycin by Streptomyces mobaraensis ATCC
15003 using response surface methodology. Afr. J. Biotechnol., 9, 5450–5459, 2010.
58. Omar, S., Rayes, A., Eqaab, A., Voß, I., and Steinbuchel, A., Optimization of cell growth and poly
(3-hydroxybutyrate) accumulation on date syrup by a Bacillus megaterium strain. Biotechnol. Lett.,
23, 1119–1123, 2001.
19
Dragon Fruit Juice
Lee-Fong Siow
CONTENTS
19.1 Introduction....................................................................................................................................231
19.4 Health Effects................................................................................................................................ 233
19.6 Conclusion..................................................................................................................................... 235
References............................................................................................................................................... 235
19.1 Introduction
Dragon fruit (Hylocereus spp.), commonly known as the pitaya/pitahaya/strawberry pear, is a tropical
fruit native to Mexico and Central and South America [1] and widely grown in Malaysia, Vietnam,
Taiwan, and Thailand. It has a unique vine-like outer scale, therefore it is named the “dragon” fruit. It is
a member of the Cactaceae (cactus) family and order of Caryophyllales [2]. Two varieties of dragon fruit
that are commonly consumed in Malaysia are Hylocereus polyrhizus (pink-skin and red-flesh dragon
fruit) and H. undatus (pink-skin and white-flesh dragon fruit) (Figure 19.1). The pulp is juicy, has a mild
taste, and is low in calories and contains numerous tiny black seeds. The fruit weights 150–600 g and is
commonly consumed as a cut fruit or juice or added into salads. More specifically, red-flesh dragon fruit
puree or juice is used in making jelly because of its attractive red color and the tiny seeds that improve
the overall exotic esthetic value of dragon fruit jelly. Red-flesh dragon fruit juice is increasingly popular
in restaurants and specific health food stores in Malaysia. A recent study showed the prebiotic properties
of the juice for both red-flesh and white-flesh dragon fruits [3]. This chapter highlights the bioactives and
antioxidant efficacy of dragon fruit juices and the novel products derived from them.

Proximate composition and nutritional characteristics of red-flesh and white-flesh dragon fruit juices are
given in Table 19.1. Although no ascorbic acid (vitamin C) has been detected in both red-flesh and white-
flesh dragon fruit juices [5], a more recent study by Liaotrakoon et al. [6] reported 4.90 and 9.85 mg
ascorbic acid/100 g for red-flesh and white-flesh dragon fruit purees, respectively. The amount of
vitamin C in the red-flesh and white-flesh dragon fruit purees is lower than those of the other fruit juices
such as apple juice (25.75–77.00 mg/100 g) [7], orange juice (8.87–67.36 mg/100 mL) [8], or other cactus
juices (21.8–25.6 mg/100 mL) [5]. Low acidity of the dragon fruit juice and its high sugar-to-acid ratio
provide a poor sensory quality when the juice is consumed alone. Therefore, dragon fruit juice is normally
mixed with other juices and consumed as a juice blend.
The content of oligosaccharides in red- and white-flesh dragon fruit juice is 89.6 and 86.2 g/kg fruit,
respectively [3]. The potassium content (3.2 g/L) in red-flesh dragon fruit juice is relatively high com-
pared to other minerals such as magnesium (0.312 g/L) and calcium (0.023 g/L) [5,9]. Citric acid and
231
(a) (b)
FIGURE 19.1 Dragon fruits: (a) White-flesh dragon fruit (Hylocereus undatus)—outer skin with a scaly texture. (b) Red-
flesh dragon fruit (Hylocereus polyrhizus)—cut dragon fruits, interspersed with tiny black seeds.
TABLE 19.1
Compositional and Nutritional Characteristics of Red-flesh and White-Flesh Dragon Fruit Juices
Nutrient Unit Red-Flesh Juice White-Flesh Juice References
Water % 88.5 — [4]
Protein % 0.93 — [4]
Lipid (fat) % 0.40 — [4]
Ash % 0.59 — [4]
Carbohydrate % 9.44 — [4]
Glucose g/L 55.4 46.6 [5]
Fructose g/L 19.2 18.4 [5]
Total dietary fiber % 0.12 — [4]
Total soluble solids °Brix 10.7 9.4 [5]
Minerals
Calcium mg/L 23.2 30.6 [5]
Magnesium mg/L 312 265 [5]
Potassium mg/L 3284 3995 [5]
Sodium mg/L 733 33.0 [5]
Organic Acids [5]
Ascorbic mg/L nd nd [5]
Citric mg/L 579 132 [5]
Isocitric mg/L 10.0 19.2 [5]
Lactic mg/L 18.5 153 [5]
Malic mg/L 4.8 7.2 [5]
lactic acid (2.4 to 2.5 g/L) are organic acids found in red-flesh dragon fruit juice [9,10]. Previous studies
have shown the antioxidant properties of both red-flesh [11,12] and white-flesh dragon fruits [13] juices,
indicating the potential of dragon fruit juice as a functional beverage.

The antioxidant properties of red-flesh dragon fruit juice have previously been reported, as shown
in Table 19.2. The purplish-red color of the red-flesh dragon fruit and its juice is contributed by
nitrogen-containing pigments, betalains [9,12]. Betalains and anthocyanins (active compounds that con-
tribute to red, purple, or blue depending on pH) have never been found to coexist in the same plant [14].
Dragon Fruit Juice 233
TABLE 19.2
Antioxidant Properties of Red-Flesh Dragon Fruit Juice
Concentration of Juice Extracted Concentration of Juice Extracted
Antioxidant Properties Unit in Acetone [11] in Ethanol [33]
Total phenolics mg GAE/100 g 42.4 ± 0.04 481 ± 0.01
Total flavonoids mg CE/100 g 7.21 ± 0.02 288 ± 0.04
Betacyanin mg BE/100 g 10.3 ± 0.22 nd
EC50 (DPPH) µmol AAE/g 22.4 ± 0.29 3.27 ± 0.05a
EC50 (ABTS+) µmol TE/g 28.3 ± 0.83 332 ± 0.21
a Unit in µg fresh weight/µg DPPH.
Abbreviations: AAE, ascorbic acid equivalents; ABTS, 2,2′-azino-bis-(3-ethylbenz-thialzoline-6-sulfonic acid); BE, betanin
equivalents; CE, catechin equivalents; DPPH, 2,2-diphenyl-1-picryhydrzyl; EC50, efficient concentration;
GAE, gallic acid equivalents; nd, not determined; TE, trolox equivalents.
Betalains are more water-soluble than anthocyanins and have a wider pH stability ranging from pH 3
to 7, making it suitable for use in low acid and neutral food as anthocyanins are usualy unstable at
low pH [15]. Betalains are water soluble and comprise of betacyanin (red purple) and betaxanthins
(yellow) [9], with maximum absorptivity at 535 and 480 nm, respectively [16]. Betacyanin is respon-
sible for red-purple color, while betaxanthin is responsible for yellow [17] color. Minimum betaxanthins
are present in red-flesh dragon fruit, and there are at least six identified betacyanins in the Hylocereus
genus, namely, betanin, isobetanin, phyllocactin, isophyllocactin, hylocerenin, and isohylocerenin [15].
Structures of betanin and phyllocactin and their respective C15 epimers are shown in Figure 19.2. All the
seven identified betacyanins contribute to the deep purple pulp or juice [18]. Betanin has been reported to
decompose into cyclo-dopa 5-O-β-glucoside and betalamic acid, resulting in the loss of the purplish-red
color [19]. Betanidin and isobetanidin (the C-15 diastereoisomer) are the simplest betacyanins [20,21].
Betacyanins consist of monoacylglycosides [2,22] and show a stable purple color over a wide pH range
and shift of hue angle to purplish blue at pH 1.0–1.5 and pH 7.0–8.0 [5]. The highest chroma of red-flesh
dragon fruit juice was reported at pH 6.5, and the chroma increased with lower acidity. At higher and
lower acidities, a slight darkening was observed in the juice [5].
Cai et al. [23] reported that various betacyanins and betaxanthins show free radical scavenging capa-
bilities. Betacyanin is of commercial interest not only as an aesthetic exotic natural food colorant but also
because it has antioxidant properties, which protect against oxidative stress-related disorders [9]. The
betacyanin content in red-flesh dragon fruit juice is 525.3 mg/L, much higher than that of other cactus
juices, namely, Opuntia ficus-indica cv. “Rossa” and O. ficus-indica cv. “Gialla” [5]. Betaxanthins of
5.3 mg/L has been reported in the juice of red-flesh dragon fruit [5]. Table 19.2 shows the antioxidant
properties of red-flesh dragon fruit juice. Phenolic compounds, namely, hydroxycinnamates, have been
reported in the pulp of red-flesh and white-flesh dragon fruits [13]. In another study [24], in addition to
betalains, antioxidant capacity of the genus of Hylocereus is contributed by their biosynthetic precursors
followed by other nonbetalain phenolics, namely, gallic acid and acetylcoumarin.
19.4 Health Effects

The juice from red-flesh dragon fruit plays an important role in contributing health benefits via the
presence of various bioactive compounds. The antioxidant properties of betalains are structure depen-
dent. The number of hydroxy and amino residues in betaxanthins improves their free radical scavenging
activity. In betacyanins, glycosylation reduces the antioxidant property, while acylation increases their
antioxidant property [25]. Betalains exhibit antiradical, antioxidant, anti-inflammatory, and anticarcino-
genic properties [26], while betacyanins are recognized as antiradical agents [27]. One of the betacya-
nins, betanin, inhibits lipid peroxidation and heme decomposition [28]. These phenolic compounds show
antiproliferative activity or cytotoxicity in human oral cancer cells [29], melanoma cells [30], and lung
metastasis induced by B16F10 melanoma cells [31].
R1 R2
C
N H
11 13 O
O
C N C
H
OH
HO
R1— H, R2-glutamic acid; trivial name: Vulgaxanthin I
R1— R2-proline; trivial name: Indicaxanthin
(a)
R3O
5
3
R4O 6 2 O
+
N C
O–
14
O 15 17 O
C N C
H
HO OH
R3-β-glucose, R4-H; trivial name: Betanin

R3-6΄-O-(malonyl)-β-glucose, R4-H; trivial name: Phyllocactin
R3-6΄-O-(3˝-hydroxy-3˝-mathylglutaryl)-β-glucose, R4-H; trivial name: Hylocerenin
(b)
O 6 O
C N C
H
HO OH
(c)
FIGURE 19.2 Chemical structures of (a) betaxanthins, (b) betacyanins, and (c) betalamic acid found in dragon fruit
juices.
The soluble fiber of the juice from red-flesh dragon fruit regulates blood sugar in humans, while
ucilage improves cholesterol metabolism [32]. Oligosaccharides of red-flesh dragon fruit juice are good
m
sources of prebiotics that are able to stimulate the growth of probiotics [3]. This has been confirmed by
Kunnika and Pranee [33] that demonstrated juice of red-flesh dragon fruit is a potential source of prebi-
otics, specifically for Lactobacillus acidophilus La5, Bacillus lactis Bb 12, and Escherichia coli ATCC
29922. The presence of lactic acid bacteria (LAB) in fermented red-flesh dragon fruit juice has previ-
ously been reported, and the LAB was reported to belong to the genus Enterococcus [34].

Both red-flesh [35] and white-flesh [36] dragon fruits are processed into juices. Dragon fruit juices are
cloudy and viscous. They exhibit non-Newtonian and pseudoplastic behavior [4]. Arrhenius model has
been reported as the best model to describe the effect of temperature on dragon fruit juice [4]. Enzymatic
clarification of the red-flesh or white-flesh dragon fruit juices using pectinases improved the stability
and appearance of the juices. A previous study reported that application of enzyme increases juice
recovery, juice physicochemical characteristics, and nutritional components such as protein and total
polyphenols [35]. Addition of 0.25% ascorbic acid, pH 4.0, and pasteurization at 65°C for 30 min was
selected as the best processing conditions to retain betacyanin content in red-flesh dragon fruit juice
[37]. The optimum enzymatic clarification of white-flesh dragon fruit juice was at 0.06% pectinase,
49°C for 40 min [36]. Consumer acceptance test revealed that drink reconstituted from red-flesh dragon
fruit concentrate was better accepted in terms of sweetness, flavor, and overall acceptability compared
to its juice counterpart [37]. In the aforementioned studies, the tiny seeds of dragon fruit were removed
from the juice to improve its stability as tri-unsaturated triacylglycerols were mainly found in the oil
from seeds of dragon fruit.
Since both red-flesh and white-flesh dragon fruits are potential functional foods, several studies have
been conducted to dry the fruit juices into a powder to facilitate the use of dragon fruits in various
food applications. Lee et al. [38] reported inlet air temperatures of 110°C and 120°C as the minimum
conditions to obtain spray dried white and red dragon fruit powders, respectively. The spray dried pow-
der stored at 43, 54, or 75% relative humidity (RH) at 25°C for 25 days resulted in structural changes
correlating to the depression of glass transition temperature to below storage temperature. At 33% RH,
no visible structural changes were observed. These spray dried red and white dragon fruit powders are
excellent natural food colorant and functional food ingredients [38].
Oligosaccharides of red-flesh and white-flesh dragon fruit juice stimulate the growth of Lactobacilli
and Bifidobacteria [3], therefore oligosaccharides of red-flesh and white-flesh dragon fruits may serve
as potential prebiotic. A recent study reported LAB of the genus of Enterococcus could be isolated from
fermented red-flesh dragon fruit juice, indicating the probiotic potential of red-flesh dragon fruit juice.
19.6 Conclusion
Both red-flesh and white-flesh dragon fruit juices are invaluable in their nutritional, antioxidative, and
antiproliferative activities. Future studies should focus on the extraction of the bioactive compounds
in both red-flesh and white-flesh dragon fruit juice and examination of the stability of the compounds in
various food matrices. Other potential studies include developing fermented dragon fruit juice or juice
blend consisting of dragon fruit juice, as dragon fruit juice alone may be bland in taste without much
“kick.’’ The presence of seeds increases the esthetic value of the overall juice.
REFERENCES
1. Benzing, D.H., Vascular Epiphytes, General Biology, and Related Biota, Cambridge University Press,
Cambridge, UK, 1990.
2. Stintzing, F.C., Schieber, A., and Carle, R., Betacyanins in fruits from red-purple pitaya, Hylocereus
polyrhizus (Weber) Britton Rose. Food Chem., 77, 101–106, 2002.
3. Wichienchot, S., Jatupornpipat, M., and Rastall, R.A., Oligosaccharides of pitaya (dragron fruit) flesh
and their prebiotic properties. Food Chem., 120, 850–857, 2010.
4. Chuah, T.G., Ling, H.L., Chin, N.L., Choong, T.S.Y., and Fakhru’l-Razi, A., Effects of temperatures on
rheological behavior of dragon fruit (Hylocereus sp.) juice. Int. J. Food Eng., 4, 1–28, 2008.
5. Stintzing, F.C., Schieber, A., and Carle, R., Evaluation of color properties and chemical quality param-
eters of cactus juices. Eur. Food Res. Technol., 216, 303–311, 2003.
6. Liaotrakoon, W., De Clercq, N., Van Hoed, V., Van de Walle, D., Lewille, B., and Dewettinck, K.,
Impact of thermal treatment on physicochemical, antioxidative and rheological properties of white-flesh
and red-flesh dragon fruit (Hylocereus spp.) purees. Food Bioprocess Technol., 6, 1365–1365, 2013.
7. Campeanu, G., Neata, G., and Darjanschi, G., Chemical composition of the fruits of several apple culti-
vars growth as biological crop. Not. Bot. Hortic. Agrobo., 37, 161–164, 2009.
8. Inga, K., Maria, M., Mirosawa, S., and Anna, G., Effect of storage on the content of polyphenols,
vitamin C and the antioxidant activity of orange juices. J. Food Comp. Anal., 20, 313–322, 2007.
9. Le Bellec, F., Vaillant, F., and Imbert, E., Pitahaya (Hylocereus spp.): A new fruit crop, a market with a
future. Fruits, 61, 237–250, 2006.
10. Vaillant, F., Perez, A., Davila, I., Dornier, M., and Reynes, M., Colorant and antioxidant properties of
red pitahaya (Hylocereus sp.). Fruits, 60, 1–7, 2005.
11. Wu, L.C., Hsu, H.W., Chen, Y.C., Chiu, C.C., Lin, Y.I., and Ho, J.A.A., Antioxidant and antiproliferative
activities of red pitaya. Food Chem., 95, 319–327, 2006.
12. Rebecca, O.P.S., Boyce, A.N., and Chandran, S., Pigment identification and antioxidant properties of red
dragon fruit (Hylocereus polyrhizus). Afr. J. Biotechnol., 9, 1450–1454, 2010.
13. Mahattanatawee, K., Manthey, J.A., Luzio, G., Talcott, S.T., Goodner, K., and Baldwin, E.A., Total
antioxidant activity and fiber content of select Florida-grown tropical fruits. J. Agric. Food Chem., 54,
7355–7363, 2006.
14. Stafford, H.A., Anthocyanins and betalains: Evolution of the mutually exclusive pathways. Plant Sci.,
101, 91–98, 1994.
15. Stintzing, F.C. and Carle, R., Betalain-emerging prospect for food scientist. Trends Food Sci. Technol.,
18, 514–525, 2007.
16. Herbach, K.M., Stintzing, F.C., and Carle, R., Betalain stability and degradation-structural and
chromatic aspects. J. Food Sci., 71, R41–R50, 2006.
17. Woo, K., Ngou, F., Ngo, L., Soong, W., and Tang, P., Stability of betalain pigment from red dragon fruit
(Hylocereus polyrhizus). Am. J. Food Technol., 6, 140–148, 2011.
18. Wybraniec, S. and Mizrahi, Y., Fruit flesh betacyanin pigments in Hylocereus cacti. J. Agric. Food
Chem., 50, 6086–6089, 2002.
19. Stintzing, F.C., Schieber, A., and Carle, R., Identification of betalains from yellow beet (Beta vulgaris L.)
and cactus pear (Opuntia ficus-indica [L.] Mill.) by high-performance liquid chromatography-
electrospray ionization mass spectrometry. J. Agric. Food Chem., 50, 2302–2307, 2002.
20. Minale, L., Piattelli, M., De Stefano, S., and Nicolaus, R.A., Pigments of centrospermae-VI Acylated
betacyanins. Phytochemistry, 5, 1037–1052, 1966.
21. Schwartz, S.J. and von Elbe, J.H., Quantitative determination of individual betacyanin pigments by
high-performance liquid chromatography. J. Agric. Food Chem., 28, 540–543, 1980.
22. Wybraniec, S., Platzner, I., Geresh, S., Gottlieb, H.E., Haimberg, M., Mogilnitzki, M., and Mizrahi, Y.,
Betacyanins from vine cactus Hylocereus polyrhizus. Phytochemistry, 58, 1209–1212, 2001.
23. Cai, Y.Z., Sun, M., and Corke, H., Antioxidant activity of betalains from plants of the Amaranthaceae.
J. Agric. Food Chem., 51, 2288–2294, 2003.
24. Esquivel, P., Stintzing, F.C., and Carle, R., Phenolic compound profiles and their corresponding antioxi-
dant capacity of purple pitaya (Hylocereus sp.) genotypes. Z. Naturforsch C., 62, 636–644, 2007.
25. Stintzing, F.C. and Carle, R., Functional properties of anthocyanins and betalains in plants, food, and in
human nutrition. Trends Food Sci. Technol. 15, 19–38, 2004.
26. Harlev, E., Nevo, E., Solowey, E., and Bishayee, A., Cancer preventive and curative attributes of plants
of the Cactaceae family: A review. Planta Med., 79, 713–722, 2013.
27. Pedreno, M.A. and Escribano, J., Correlation between antiradical activity and stability of betanine from
Beta vulgaris L. roots under different pH, temperature and light conditions. J. Sci. Food Agric., 81,
627–631, 2001.
28. Kanner, K., Harel, S., and Granit, R., Betalains—A new class of dietary cationized antioxidants.
J. Agric. Food Chem., 49, 5178–5185, 2001.
29. Seeram, N.P., Adams, L.S., Hardy, M.L., and Heber, D., Total cranberry extract versus its phytochemi-
cal constituents: Antiproliferative and synergistic effects against human tumor cell lines. J. Agric. Food
Chem., 52, 2512–2517, 2004.
30. Rodriguez, J., Yanez, J., Vicente, V., Alcaraz, M., Benavente-Garcia, O., Castillo, J., Lorente, J., and
Lozano, J.A., Effects of several flavonoids on the growth of B16F10 and SK-MEL-1 melanoma cell lines:
Relationship between structure and activity. Melanoma Res., 12, 99–107, 2002.
31. Menon, L.G., Kuttan, R., and Kuttan, G., Inhibition of lung metastasis in mice induced by B16F10
melanoma cells by polyphenolic compounds. Cancer Lett., 95, 221–225, 1995.
32. Luz-Fernandez, M., Lin, E.C.K., Trejo, A., and McNamara, D.J., Prickly pear (Opuntia sp.) pectin alters
hepatic cholesterol metabolism without affecting cholesterol absorption in guinea pigs fed a hypercho-
lesterolemic diet. J. Nutr., 124, 817–824, 1994.
33. Kunnika, S. and Pranee, A., Influence of enzyme treatment on bioactive compounds and color stability
of betacyanin in flesh and peel of red dragon fruit Hylocereus polyrhizus (Weber) Britton and Rose.
Int. Food Res. J., 18, 1437–1448, 2011.
34. Ong, Y.Y., Tan, W.S., Rosfarizan, M., Chan, E.S., and Tey, B.T., Isolation and identification of lactic acid
bacteria from fermented red dragon fruit juices, J. Food Sci., 77, M560–M564, 2012.
35. Aliaa, A.R.N., Mazlina, M.K.S., and Taip, F.S., Effects of commercial pectinases application on selected
properties of red pitaya juice. J. Food Process Eng., 34, 1523–1534, 2011.
36. Aliaa, A.R.N., Mazlina, M.K.S., Taip, F.S., and Abdullah, A.G.L., Response surface optimization for
clarification of white pitaya juice using a commercial enzyme. J. Food Process Eng., 33, 333–347, 2010.
37. Wong, Y.M. and Siow, L.F., Effects of heat, pH, antioxidant, agitation and light on betacyanin stability
using red-fleshed dragon fruit (Hylocereus polyrhizus) juice and concentrate as models. J. Food Sci.
Technol., 52, 3086–3092, 2015.
38. Lee, K.H., Wu, T.Y., and Siow, L.F., Spray drying of red (Hylocereus polyrhizus) and white (Hylocereus
undatus) dragon fruit juices: Physicochemical and antioxidant properties of the powder. Int. J. Food Sci.
Technol., 48, 2391–2399, 2013.
20
Goji Berry Juice
Patricia Navarro, Luis Noguera-Artiaga, Santiago López-Miranda,

Ángel A. Carbonell-Barrachina, and Antonio J. Pérez-López
CONTENTS
20.1 Introduction................................................................................................................................... 239
20.4 Health Effects................................................................................................................................ 243
20.4.1 Antioxidant and Antiaging and Enhancement of Sleep Quality and Well-Being........... 243
20.4.2 Stimulation of Metabolism............................................................................................... 243
20.4.3 Hypoglycemic and Hypolipidemic Effects...................................................................... 243
20.4.4 Prevention of Hepatic Diseases........................................................................................ 243
20.4.5 Immunomodulatory Activity............................................................................................ 244
20.4.6 Antitumor Activity........................................................................................................... 244
20.4.7 Neuroprotection Effects................................................................................................... 244
20.4.8 Radioprotective Activity.................................................................................................. 244
20.4.9 Cardiovascular Protection and Antiosteoporosis Effects................................................. 244
20.6 Conclusion..................................................................................................................................... 246
References............................................................................................................................................... 246
20.1 Introduction
For some years, the goji berry (also known as the wolfberry), has been a traditional food and medicine in
East Asia, has become increasingly popular in Europe and North America. A lot of products are commer-
cialized under the name goji (derived from the Chinese name gouqi) on the health food market. Goji is a
relatively new name given to Lycium barbarum and L. chinense, two close species with a long tradition
of use as medicinal and food plants in East Asia, particularly in China [1]. L. barbarum is a solanaceous
defoliated shrubbery that grows in China, Tibet, and other parts of Asia, and its fruits are 1–2 cm long,
bright-orange-red ellipsoid berries. The fruits are collected in the summer and autumn, dried in the shade
till the skin shrinks, and are then exposed to the sun until the outer skin becomes dry and hard, but the
pulp is still soft. The fruits are either dried, or the fresh fruits may be squeezed for their juice that is then
concentrated to preserve it for future use in making various beverages [2]. This chapter highlights the
nutritional characteristics, antioxidant activity, health effects, and novel formulations of goji berry juice.

The goji berry is normally mixed with other fruits, in order to yield a more acceptable flavor, and con-
sumed as a juice. It is recognized for its high content of polysaccharides and protein, higher than those of
the raspberry, blueberry, or black currant, but it is difficult to know its exact nutritional composition from
239
TABLE 20.1
Compositional and Nutritional Characteristics of Traditional and Organic Goji Berry Juices (per 100 g
Fruit or Juice)
Nutrient Unit Traditional Organic References
Water g 85–95 na [3,4]
Energy kcal 45–58 67 [4–7]
Protein g 0.22–0.4 0.66 [4,5]
Lipid (fat) g 0–0.2 nd [4]
Carbohydrate g 10.2–11.5 16.6 [4–8]
Total dietary fiber g 0.1–2.2 na [4,8]
Vitamins
Niacin mg 0.73 0.15 [4,5]
Riboflavin mg 6.6 58.5 [4,5]
Thiamin mg 0.025 0.022 [4,5]
Vitamin A (RAE) µg 16 3 [4,5]
Vitamin C mg 2.6 0.5 [4,5]
Vitamin E (ATE) mg 2.6 0.5 [4,5]
Carotenoids
Zeaxanthin mg/g 0.045 na [9]
β-Cryptoxanthin mg/g na na [9]
β-Carotene mg/g na na [9]
Lutein mg/g na na [9]
Abbreviations: nd, not detected; na, not available; RAE, retinol activity equivalents; ATE, alpha-tocopherol equivalents.
reputable government websites such as the U.S. Department of Agriculture (USDA). Table 20.1 shows
compositional and nutritional characteristics of traditional and organic goji berry juices [3–9].
Polysaccharides represent quantitatively the most important group of substances in goji berry juice. The
polysaccharide fraction consists of a complex mixture of highly branched and only partly characterized
polysaccharides and proteoglycans. The glycosidic part consists of arabinose, glucose, galactose, man-
nose, rhamnose, xylose, and galacturonic acid. A second major group is carotenoids with zeaxanthin as the
predominant constituent [9]. Goji berry juice is an excellent source of riboflavin (6.6–58.5 mg/100 g) and
reasonable source of other vitamins such as niacin, thiamin, and vitamins (A, C, and E) (Table 20.1) [4,5].
The Recommended Dietary Allowances (RDA) of riboflavin is 1.3 and 1.1 mg/day for males and females,
respectively. Free amino acids are presented with proline as major constituent in goji berry juice [10].
As can be seen from Table 20.2, goji berry and its juice are rich in potassium (1460 mg/100 g and
187 mg/100 mL, respectively) and iron (5.5 mg/100 g and 0.3 mg/100 mL, respectively). The mineral
content can be translated into high values of the RDA for some of these minerals as established by the
Commission of the European Community [11]. For example, the RDA values for potassium and copper
are 14.6 and 13.4%, respectively.

Berries comprise the major proportion of the fruits included in the diet of Europeans and are consumed
normally as fresh or processed fruits mainly as juice. Their presence in the diet makes an important
contribution to the intake of health-promoting compounds, such as vitamins, minerals, dietary fibers,
and antioxidants (e.g., phenolics, carotenoids, and polysaccharides) [12].
Total antioxidant activity is determined by synergistic interactions between different compounds and
the special action of each one, so it is necessary to compare more than one method to correctly assess the
Goji Berry Juice 241
TABLE 20.2
Mineral Content and Percentage of RDA Values in Goji Products
Berry (mg/100 g)/ Juice (mg/100 mL)/ Capsule (µg/Capsule)/
Mineral RDA (%RDA) (%RDA) (%RDA) Mixture (mg/100 g)
Calcium 800 50/1.2 15/0.8 405/0.1 415
Copper 1 0.7/13.4 0.02/0.7 0.5/0.1 1.1
Iron 14 5.5/7.9 0.3/1.2 52/0.9 109
Magnesium 375 90/5 8/0.9 751/0.5 280
Manganese 2 0.9/9 0.07/1.3 3/0.4 7
Phosphorus 700 184/5.3 24/1.4 282/0.1 425
Potassium 2000 1460/14.6 187/3.7 7080/0.9 1310
Sodium — 550 22 2254 122
Zinc 10 1.3/2.6 0.07/0.3 10/0.3 4
Sources: Adapted from Llorent-Martínez, E.J. et al., Microchem. J., 110, 444, 2013. With permission.
Abbreviation: RDA, Recommended Dietary Allowances.
antioxidant activity of a sample or a food. The most widely used methods in fruit juices are (1) oxygen rad-
ical absorbance capacity (ORAC) assay, based on the inhibition of the peroxyl-radical-induced oxidation
initiated by thermal decomposition of azo compounds [13]; (2) 2,2-diphenyl-1-picrylhydrazyl (DPPH)
method, which uses DPPH•, a free radical that measures the ability of a compound to donate an electron
[14]; (3) ferric-reducing antioxidant potential (FRAP) method developed to measure the ability to reduce
ferric complex with the molecule tripyridyl-S-triazine (TPTZ) to its ferrous form at low pH [15]; and
(4) finally by 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) method; antioxidants are
added prior to the generation of the ABTS radical, and the inhibition in the radical formation is evaluated
[16]. The antioxidant capacity of commercial goji berry juice has been reported and compared with that
of other berry juices, using the methods previously described [17]. Antioxidant activity in goji berry juice
(ranges from 5.1 to 14.7 mmol trolox equivalents [TE]/L) was similar to that of cranberry juice (ranges
from 8.1 to 18.6 mmol TE/L) and was double that of pomegranate juice (ranges from 3.2 to 7.2 mmol
TE/L) for each of the studied methods (Table 20.3). These experimental values certainly suggest that goji
berry juice has higher antioxidant activity than other juices commonly accepted as being healthy drinks.
Unfortunately, the number of references describing the content of bioactive compounds in goji berry
juice is very low [17,10]. It is widely claimed that this juice is rich in polysaccharides, but this statement
is mainly based on the composition of fresh berry [18] or dried berry [19] or products, such as extracts.
The functional components, such as polysaccharides, carotenoids, and phenolic compounds, have been
identified, quantified, and linked with health effects after consumption of this berry and its products [19].
There are around 50 references on goji berry in the literature, but in reality, only few of them describe
the bioactive compositions of the goji berry juice [10,17].
It is important to mention that juice production significantly changes the composition, especially the
contents of bioactive compounds because most of them are heat sensitive. For instance, Nuncio-Jáuregui
TABLE 20.3
Antioxidant Activity of Different Berry Juices (mmol TE/L)
Methods Goji Blueberry Cranberry Açai Pomegranate
ABTS 8.4 16.6 10.9 9.3 4.7
DPPH 10.6 17.4 13.7 17.4 5.4
FRAP 5.1 14.4 8.1 10.9 3.2
ORAC 14.7 34.3 18.6 15.1 7.2
Sources: Adapted from Horszwald, A. and Andlauer, W., J. Berry Res., 1, 189, 2011. With permission.
Abbreviations: TE, trolox equivalents; ABTS, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid; DPPH, 2,2 diphenyl-
1-picrylhydrazyl; FRAP, ferric reducing antioxidant potential; ORAC, oxygen radical absorbance capacity.
et al. [20] reported that the manufacturing of pomegranate juice causes changes in the bioactive com-
pounds; the punicalagins increase, while ellagic acid and anthocyanins decrease. This type of study has
not yet been conducted in goji berry juice, and it is urgently needed to clearly prove the potential health
effects of goji berry juice.
In any case, it is necessary to briefly comment that the polysaccharide fraction of goji berry juice
consists of a complex mixture of highly branched and not fully characterized polysaccharides and pro-
teoglycans [10]. L. barbarum polysaccharide (LBP) content is considered as the main indicator of the
medicinal efficacy of Lycium products, including juice [21]. For instance, Wang et al. [22] found a con-
tent of crude polysaccharides of 57.2 mg/kg. The glycosidic part consists of mainly six monosaccharides,
namely, glucose, galactose, mannose, rhamnose, xylose, and galactose [10].
Carotenoids are responsible for the red color of goji berry juice and are present in high amounts
(0.03%–0.5%) [23]. The amounts of these compounds increase along the ripening of berry; consequently,
juices from ripe fruits can contain significantly higher amounts of carotenoids than those prepared using
unripe berries. Goji berry juice is an excellent source of carotenoids, not only because of their high con-
tent but also because of their unique profile. Zeaxanthin and its esters are the predominant carotenoids
in goji berry juice; zeaxanthin palmitate represents as much as 56% of total carotenoids in L. barbarum
products, including juice, and 49% in L. chinense [23].
Phenolics are often related with the antioxidant capacity of most juices. The main phenolics found
in goji berry juice are phenolic acids (e.g., chlorogenic, dicaffeoylquinic, caffeic, and p-coumaric) and
flavonoids (e.g., quercetin diglucoside, rutin, and kaempferol-3-O-rutinoside) [22]. Figure 20.1 shows the
most important bioactive compounds (zeaxanthin, dicaffeoylquinic acid, and quercetin-3-O-rutinoside)
found in goji berry juice.
H3C OH
H3C CH3 CH3 CH3
CH3 CH3 H3C CH3

HO CH3
Zeaxanthin
O
HO OH
O O
HO OH
O O
OH
HO OH
Dicaffeoylquinic acid
OH
HO
O OH
O
HO OH
O O OH
O OH
HO
OH O
OH
CH 3
Quercetin-3-O-rutinoside
FIGURE 20.1 Chemical structures of major bioactive compounds found in goji berry juice.
20.4 Health Effects

20.4.1 Antioxidant and Antiaging and Enhancement of Sleep Quality and Well-Being
Several studies have shown that goji berry juice helps prevent oxidant stress-related conditions in humans.
Among all of the bioactive substances, the most well-studied components are a group of water-soluble
glycoconjugates (e.g., LBPs) [24,25]. Various animal and in vitro cell culture studies have demonstrated
the efficacy of LBP identified as the major active antioxidant that protects against various peroxidation-
related conditions, including lipid peroxidation [26].
LBPs may exert their effects in other ways, such as acting as bioactive fibers or prebiotics, by con-
tributing to the synthesis and release of antioxidants by probiotic bacteria and inhibiting inflammation.
Thus, the gastrointestinal effects of LBP may lead to antioxidant action within the gastrointestinal tract
before absorption of the LBP or its degradation compounds. Absorption of LPB and the active constitu-
ents of goji in the gut are important to goji’s effects in vivo. The bioavailability and kinetic behavior of
LBP is important for understanding its in vivo bioactivity. Clinical studies have shown that goji berry
juice significantly increases feelings of well-being and improved neurologic/psychologic performance,
including improvement of the quality of sleep and gastrointestinal function [25]. The antioxidant effects
of goji berry juice may be associated with mechanisms that underline the physiologic effects of this fruit.
With regard to sleep quality, alterations in the metabolism of reactive oxygen species (ROS) result in
prolonged sleep deprivation. Other observed improvements after consumption of goji berry juice may
result from its antioxidant effects. We suggest that these antioxidant actions of goji berry juice are at least
partially responsible for its clinical benefits since free radical oxidation plays a role in the development of
various diseases. Goji berry juice may be useful in preventing or reducing the development, severity, or
symptoms of disease conditions. As elderly people demonstrate age-associated decreases in glutathione
peroxidase (GSH-Px) and superoxide dismutase (SOD) levels, goji berry juice may have antiaging effects
that are consistent with the traditionally recognized effects of goji berry. Easing of menstrual complaints
were found in female subjects in a randomized trial after consumption of a standardized goji berry juice,
while the placebo group showed no significant change [27–29]. At the same time, the LBP enhanced
semen quantity by 68% and sperm motility by 40% in semicastrated rats [30].
20.4.2 Stimulation of Metabolism

Goji berry intake was effective in controlling the waist circumference in humans and may reduce the
risks of metabolic syndrome [24]. The goji berry may stimulate the metabolic rate through adrenocorti-
cal hormone control, and these effects may be related to changes in waist circumference produced by
daily consumption of the goji berry in the form of juice [31].
20.4.3 Hypoglycemic and Hypolipidemic Effects

There are several clinical and experimental reports showing an antidiabetic effect for goji berry juice.
Diabetes is associated with significant oxidative stress, and increasing evidence in both experimental
and clinical studies suggests that oxidative stress caused by hyperglycemia plays a major role in the
pathogenesis of diabetes mellitus [32]. In vitro and in vivo experiments on the hypoglycemic effects have
proven that LBP alleviates insulin resistance by preserving the β cell mass and increasing insulin secre-
tion and glucose utilization. The underlying mechanisms of these results included β cell proliferation and
increasing activity of key enzymes of glucose metabolism [28,33]. LBP has reduced serum total choles-
terol and triacylglycerols (TAG) contents and increased high-density lipoprotein (HDL) levels in hyper-
lipidemic rabbits. These results clearly suggest a hypolipidemic effect of the goji berry and its juice [34].
20.4.4 Prevention of Hepatic Diseases

Carbon tetrachloride (CCl4) at a high dose often rapidly causes cellular necrosis, oxidative stress, and
inflammation, which leads to acute liver injury and failure [35]. Alanine transaminase (ALT) is a cellular
enzyme that indicates the presence of injury in the liver. Under healthy circumstances, ALT is contained
within the cell. When cellular injury occurs, the enzyme is released from the cytoplasm into the blood-
stream, suggesting liver damage [36]. LBP has significantly reduced the serum ALT level, which was
elevated by CCl4 injection, indicating the protective effect of LBP on liver injury [25]. The protection of
LBP is of significant clinical application because several liver diseases, including fatty liver diseases and
cirrhosis, share similar pathological mechanisms with CCl4 intoxication [37].
20.4.5 Immunomodulatory Activity

Lymphocyte proliferation is a crucial event in the activation cascade of both the cellular and humoral
immune responses. As is well known, dendritic cells are the most powerful antigen presenting cells of
the immune system, which are closely related to the occurrence and development of tumor [32]. Zhu
et al. [38] investigated the effects of LBP on the phenotypic and functional maturation of murine bone
marrow–derived dendritic cells (BMDC) in vitro. It was found that LBP present in the juice were capable
of promoting both the phenotypic and functional maturation of dendritic cells.
20.4.6 Antitumor Activity

He et al. [39] reported that the antitumor activity by LBP may come from the induction of cell-cycle
arrest and apoptosis and inhibition of some signaling pathways, which play a protective effect against
carcinogenesis by eliminating abnormal excess tumor cells. These results demonstrated that LBP could
inhibit tumor cell growth by suppressing insulin-like growth factor-1 (IGF-1)-induced angiogenesis via
PI3 K/HIF-1α/VEGF signaling pathways.
20.4.7 Neuroprotection Effects

In order to develop the ideal neuroprotective agents, the protective effects of LBP pretreatment on an
experimental stroke model were studied [40]. It was suggested that LBP might be used as a prophylactic
neuroprotectant in patients at a high risk for ischemic stroke. Future efforts may focus on the isolation
and the elucidation of the neuroprotective compounds from the goji berry. LBP represents a potential
neuroprotective agent, which deserves to be further explored to prevent neurodegeneration in Alzheimer’s
disease cases.
20.4.8 Radioprotective Activity

The possible antioxidant effects of LBP against membrane damage induced by free radicals generated
during γ-irradiation were examined [40]. The results showed that LBP significantly protected the liver cells
against irradiation-induced loss of protein thiols and inactivation of SOD, GSH-Px, and catalase (CAT) in
a dose-dependent manner. Moreover, LBP was more effective than α-tocopherol in inhibiting irradiation-
induced oxidative injury. The results of the aforementioned studies indicated that the radioprotective effect
of LBP present in the juice may come from both antioxidative and cytoprotective mechanisms [27].
20.4.9 Cardiovascular Protection and Antiosteoporosis Effects

Using rats, Lu and Zhao [41] investigated the effects of LBP on the development of cardiovascular
disease (CVD). The results showed that LBP possessed a typical protective effect on DOX-induced
acute cardiotoxicity via suppressing oxidative stress. Zhu et al. [29] evaluated the effects of LBP against
osteoporosis in estrogen-deficient ovariectomized rats, and it was found that LBP could enhance bone
mineral density and bone mineral content in ovariectomized rats. To summarize, LBP present in juice
had antiosteoporotic effects, which seemed to be partially mediated by enhancing the synthesis of Col5a2
and Alpl gene products.
LBPs present in the juice have various important bioactivities, such as antioxidant, immunomodula-
tion, antitumor, neuroprotection, radioprotection, antidiabetes, hepatoprotection, antiosteoporosis, and
antifatigue. Most of these bioactivities were investigated in vitro or in mouse models, and the high-order
structure of this active component as well as the relationship between bioactivities and chemical struc-
ture is still not well established. Further research on the exact order structure, the bioactive effects on
human subjects, and the structure–bioactivity relationship of LBP is required, which would allow a bet-
ter understanding of the functional effects about this macromolecule present in goji berry juice.

In recent years, interest of consumers by consumption of exotic fruits has increased rapidly due to the
health benefits that they present [11]. New functional beverages such as fortified water, teas, energy and
sports drinks, or dairy products have increased their convenience, providing an image of healthy drink
and supported by an innovative and fun marketing strategy [42].
One of the most representative products of this emerging fever by Traditional Chinese Medicine in
Western countries is the goji berry. The fruits of goji have been used for thousands of years by Traditional
Chinese Medicine as a medicinal against different diseases and health problems, as well as functional
food [2,43]. Concentrated extracts and teas of goji berries have a long history of use as an ingredient in
various types of traditional drinks and recipes [2]. The bioactive compounds of the goji berry with special
interest as functional food ingredients are polysaccharides [28], carotenoids such as zeaxanthin [2],
flavonoids [43], and minerals [11].
Traditional recipes indicate that goji berry can be incorporated into formulas or recipes at equivalent
to 6–18 g/100 g dried material. In case of decoction, references indicate that 5–15 g/100 mL of goji juice
is equivalent to 25–120 g of fresh berry. There are few scientific studies about the use of goji berry used
as a single component or a major component in a recipe [18].
Actually, it is possible to find products made from goji berry, used as 100% of its composition or as an
ingredient of a more complex recipe. The products are highly varied, being beverages the most common
products, such as juice, energy drink, tea, and dairy. It is also common to find goji berry as dehydrated
fruit and dry extract in capsules. Many goji berry products are sold in health food stores, in particular,
through the Internet and websites related to health, wellness, and longevity [18].
One of the most important commercial examples of goji drink marketed is GoChi®, a liquid dietary
supplement based on reconstituted juice, which is clinically tested. The format of this product has been
standardized to contain, in a daily 120 mL serving, the equivalent of 150 g of fresh goji berry, the amount
customarily consumed in Traditional Chinese Medicine [2]. Wu et al. [44] analyzed the antioxidant
capacity of various commercial juices with concentrations between 90% and 100% goji obtaining values
ranging between 2025 and 570 µmol of TE/30 mL of product.
Combinations of juices using goji fruit as a main or secondary ingredient of a complex formula-
tion beverage are varied. There are goji berry juices that are enriched with other natural b ioactive
compounds that supplement its healthy properties. A commercial example of this product is goji juice
enriched with Oxi3®, a combination of red grape, lycopene, and resveratrol extract. Goji berry juice can
also be used to enrich other types of juices. Navarro et al. [13] investigated a mandarin orange juice
enriched with goji berry juice and pomegranate juice. A disadvantage that can be present in goji berry
juice is lower concentration of some of its bioactive compounds in relation to the concentration in fresh
berry or other preparations such as capsules. Llorent-Martinez et al. [11] observed that the concentra-
tion of minerals in commercial goji juice was significantly lower than those in the berry and extract
capsules.
Dairy products enriched with goji berry juice also used as functional drinks. Scientists at the Nestlé®
Research Center have shown that supplementation with milk enriched with goji berry juice (Wolfberry-
Lacto) had a positive effect on immune function in elderly [45]. Other commercial examples of product
based on vegetable milk enriched with goji berry juice are the almond milk with goji berry juice, from
Cold off the Press®, and organic Kamut® drink enriched with goji berry juice, from Amandín®.
It has been demonstrated that polysaccharides of goji berry can reduce muscular and oxidative stress
caused by intense physical exercise [46]. This has attracted interest from companies of sport products
giving rise to the proliferation of energy drinks as based on goji berry as 180 Red® or Ironclad®.
The other beverage based on goji berry with a significant presence in markets is tea. There are many
commercial examples in which traditional teas are enriched with other fruits and herbs that complement
the beneficial effects announced by trademarks. It is the case of green tea superfruit Lipton® with goji
and raspberry. This brand offers a green tea enriched with concentrated goji juice.
Goji solid extracts in capsules are also present on the market. This is the reason why in recent years,
there is a significant effort to improve methods of extraction and purification of bioactive compounds
present in goji berry fruit in order to use them in food and pharmaceutical industries. Works are mainly
focusing on improving the extraction and purification of goji berry polysaccharides.
Hot water extraction, combined with new methods of extraction such as microwave, ultrasound, and
supercritical fluids, is having recent application for extraction of polysaccharides of goji fruit. After
their deproteinization, goji berry polysaccharides can be purified by precipitation with ethanol, frac-
tional precipitation, exchange chromatography, affinity chromatography, and gel filtration [28]. Studies
performed by Liu et al. [47] showed that the combined use of microwave and enzyme-assisted extrac-
tion can be a method that improves the effectiveness of extraction of goji polysaccharides. Chao et al.
[48] demonstrated that goji polysaccharides can be successfully extracted by using ultrasonic assisted
by supercritical water. Superfine grinding technology is an emerging technology that can show great
potential in nutraceuticals and functional foods. Recently, superfine grinding drew much attention in
the extraction of bioactive compounds because it saves time, solvents, and energy. Therefore, it has
been widely used in the extraction of polysaccharides from different raw materials with high extraction
efficiency [49]. Superfine grinding can reduce the molecular size of the goji berry polysaccharides and
improve its antioxidant capacity, so this technology has great potential for application in the functional
food and medical industries [50].
One of the major areas with huge commercial interest of products are concerned with goji berry, espe-
cially due to the emergence of fraudulent products produced with related species such as L. chinense or
L. ruthencium, whose growing and production is cheaper than L. barbarum. Ten different g enotypes that
can substitute or be used as adulterants of commercial products have been found, and the differentiation
of genotypes by morphologic characteristics is very difficult [18]. For this reason, different identification
systems are being proposed through molecular markers [50].
20.6 Conclusion
The consumption of exotic fruits, especially as juices and beverages, has increased in recent years. The
key products are due to increased consumer interest in products that prevent diseases and/or maintain
good health. In this sense, several studies have shown that goji beery juice helps to prevent oxidative
stress-related conditions in humans. The goji berry and its juice are rich in antioxidants, such as polyphe-
nols and other important bioactive compounds. Thus, juices and beverages from goji berry fruits have a
great potential for future development, due to their proximity to consumers, their ease of use, availability,
and the varied combinations, which can be made with other functional foods. However, most of the state-
ments on the bioactive compounds and the healthy effects of the goji berry need to be researched further
and especially clinically studied to fully prove its health benefits.
REFERENCES
1. Potterat, O., Goji (Lycium barbarum and L. chinense): Phytochemistry, pharmacology and safety in the
perspective of traditional uses and recent popularity. Planta Med., 76, 7–19, 2010.
2. Amagase, H. and Farnsworth, N.R., A review of botanical characteristics, phytochemistry, clinical
relevance in efficacy and safety of Lycium barbarum fruit (Goji). Food Res. Int., 44, 1702–1717, 2011.
3. Ionica, M.E., Nour, V., and Trandafır, I., Polyphenols content and antioxidant capacity of goji fruits
(Lycium chinense) as affected by the extraction solvents. SouthWest. J. Hortic. Biol. Environ., 3,
121–129, 2012.
4. Rabenhorst®, Rabenhorst Goji Juice. Published online at: http://www.rabenhorst.de/english/products/
super-pure-juices/goji-juice (accessed February 10, 2015).
5. Caoh, Absolute goji. Published online at: http://www.caoh.org/pure-goji.html (accessed February 10,
2015).
6. Myfitnesspal, Goji berry juice. Published online at: http://www.myfitnesspal.com.mx/food/calories/
1208616 (accessed February 10, 2015).
7. Waitrose, The Berry Co goji berry 100% juice drink. Published online at: http://www.waitrose.com/
shop/ProductView-10317–10001–55275 (accessed February 10, 2015).
8. Fatsecret, Black cherry goji juice. Published online at: http://www.fatsecret.com/calories-nutrition/
northland/black-cherry-goji-juice?frc=True (accessed February 12, 2015).
9. Ezeyes, Zeaxanthin. Published online at: http://www.ezeyes.info/ezeyes_Zeaxanthin_intake.aspx
(accessed February 12, 2015).
10. Potterat, O. and Hamburger, M., Goji juice: A novel miraculous cure for longevity and well-being?
A review of composition, pharmacology, health-related claims and benefits. Schweiz Z. Ganzheitsmed,
20, 399–405, 2008.
11. Llorent-Martínez, E.J., Fernández-de Córdova, M.L., Ortega-Barrales, P., and Ruiz-Medina, A.,
Characterization and comparison of the chemical composition of exotic superfoods. Microchem. J., 110,
444–451, 2013.
12. Seeram, N.P., Berry fruits: Compositional elements, biochemical activities, and the impact of their
intake on human health, performance, and disease. J. Agric. Food Chem., 56, 627–629, 2008.
13. Navarro, P., Nicolas, T.S., Gabaldon, J.A., Mercader-Ros, M.T., Calín-Sánchez, A., Carbonell-
Barrachina, A.A., and Pérez-López, A.J., Effects of cyclodextrin type on vitamin C, antioxidant activity,
and sensory attributes of a mandarin juice enriched with pomegranate and goji berries. J. Food Sci., 76,
S319–S324, 2011.
14. Calín-Sánchez, A., Figiel, A., Hernández, F., Melgarejo, P., Lech, K., and Carbonell-Barrachina, A.A.,
Chemical composition, antioxidant capacity, and sensory quality of pomegranate (Punica granatum L.)
arils and rind as affected by drying method. Food Bioprocess Technol., 6, 1644–1654, 2013.
15. Zaouay, F., Mena, P., Garcia-Viguera, C., and Mars, M., Antioxidant activity and physico-chemical
properties of Tunisian grown pomegranate (Punica granatum L.) cultivars. Ind. Crop Prod., 40, 81–89,
2012.
16. Carbonell-Barrachina, A.A., Calín-Sánchez, A., Bagatar, B., Hernandez, F., Legua, P., Martínez-Font, R.,
and Melgarejo, P., Potential of Spanish sour-sweet pomegranates (cultivar C25) for juice industry. Food
Sci. Technol. Int., 18, 129–138, 2012.
17. Horszwald, A. and Andlauer, W., Characterisation of bioactive compounds in berry juices by traditional
photometric and modern microplate methods. J. Berry Res., 1, 189–199, 2011.
18. Donno, D., Beccaro, G.L., Mellano, M.G., Cerutti, A.K., and Bounous, G., Goji berry fruit (Lycium spp.):
Antioxidant compound fingerprint and bioactivity evaluation. J. Funct. Foods 18, 1070–1085, 2015.
19. Zhong, Y., Shahidi, F., and Naczk, M., Phytochemicals and health benefits of goji berries, in Dried
Fruits: Phytochemicals and Health Effects, Alasalvar, C. and Shahidi, F., Eds., Wiley-Blackwell,
Oxford, UK, 2013, pp. 133–144.
20. Nuncio-Jáuregui, N., Calín-Sánchez, A., Vázquez-Araújo, L., Perez-López, A.J., Frutos-Fernández,
M.J., and Carbonell-Barrachina, A.A., Processing pomegranates for juice and impact on bioactive com-
ponents, in Processing and Impact on Active Components in Food, Preedi, V., Ed., Academic Press,
Amsterdam, the Netherlands, 2014, pp. 629–636.
21. Wong, C.K., Leung, K.N., Fung, K.P., and Choy, Y.M., Immunomodulatory and anti-tumour polysac-
charides from medicinal plants. J. Int. Med. Res., 22, 299–312, 1994.
22. Wang, C.C., Chang, S.C., and Chen, B.H., Chromatographic determination of polysaccharides in Lycium
barbarum Linnaeus. Food Chem., 116, 595–603, 2009.
23. Peng, Y., Ma, C., Li, Y., Leung, K.S.Y., Jiang, Z.H., and Zhao, Z., Quantification of zeaxanthin dipal-
mitate and total carotenoids in Lycium fruits (Fructus lycii). Plant Foods Hum. Nutr., 60, 161–164,
2005.
24. Amagase, H. and Nance, D.M., Effect of standardized Lycium barbarum (goji) juice, GoChi® intake on
resting metabolic rate and waist circumference: Randomized, placebo-controlled, double-blind clinical
studies. FASEB. J., 23, LB419, 2009.
25. Xiao, J., Liong, E.C., Ching, Y.P., Chang, R.C.C., So, K.F., Fung, M.L., and Tipoe, G.L., Lycium bar-
barum polysaccharides protect mice liver from carbon tetrachloride-induced oxidative stress and necro-
inflammation. J. Ethnopharmacol., 139, 462–470, 2012.
26. Li, X.M., Protective effect of Lycium barbarum polysaccharides on streptozotocin-induced oxidative
stress in rats. Int. J. Biol. Macromol., 40, 461–465, 2007.
27. Amagase, H., Antioxidants in goji berry juice (Lycium barbarum) and effects of processing steps, in
Processing and Impact on Antioxidants in Beverages, Preedy, V., Ed., Academic Press, Amsterdam, the
Netherlands, 2014, pp. 155–163.
28. Jin, M., Huang, Q., Zhao, K., and Shang, P., Biological activities and potential health benefit effects of
polysaccharides isolated from Lycium barbarum L. Int. J. Biol. Macromol., 54, 16–23, 2013.
29. Zhu, M., Jinggang, M., ChangSheng, H., Haiping, X., Ning, M., and Caijiao, W., Extraction, charac-
terization of polysaccharides from Lycium barbarum and its effect on bone gene expression in rats.
Carbohydr. Polym., 80, 672–676, 2010.
30. Luo, Q.L.Z., Huang, X., Yan, J., Zhang, S., and Cai, Y.-Z., Lycium barbarum polysaccharides: Protective
effects against heat-induced damage of rat testes and H2O2-induced DNA damage in mouse testicular
cells and beneficial effect on sexual behavior and reproductive function of hemicastrated rats. Life Sci.,
79, 613–621, 2006.
31. Amagase, H., Comparison of Lycium barbarum containing liquid dietary supplements to caffeinated
beverages on energy/caloric metabolism activity and salivary adrenocortical hormone levels in healthy
human adults. FASEB. J., 24, 540.13, 2010.
32. Jin, M., Zhao, K., Huang, Q., Xu, C., and Shang, P., Biological activities and potential health benefit
effects of polysaccharides isolated from Lycium barbarum L. Carbohydr. Polym., 89, 713–722, 2012.
33. Zhu, J., Liu, W., Yu, J., Zou, S., Wang, J., Yao, W., and Gao, X., Characterization and hypoglycemic
effect of a polysaccharide extracted from the fruit of Lycium barbarum L. Carbohydr. Polym., 98, 8–16,
2013.
34. Luo, Q., Cai, Y., Yan, J., Sun, M., and Corke, H., Hypoglycemic and hypolipidemic effects and antioxi-
dant activity of fruits extracts from Lycium barbarum. Life Sci., 76, 137–149, 2004.
35. Weber, L.W., Boll, M., and Stampfl, A., Hepatotoxicity and mechanism of action of haloalkanes: Carbon
tetrachloride as a toxicological model. Crit. Rev. Toxicol., 33, 105–136, 2003.
36. Ozer, J., Ratner, M., Shaw, M., Bailey, W., and Schomaker, S., The current state of serum biomarkers of
hepatotoxicity. Toxicology, 245, 194–205, 2008.
37. Larrey, D., Drug-induced liver diseases. J. Hepatol., 32, 77–88, 2000.
38. Zhu, J., Zhao, L.H., Zhao, X.P., and Chen, Z., Lycium barbarum polysaccharides regulate phenotypic
and functional maturation of murine dendritic cells. Cell Biol. Int., 31, 615–619, 2007.
39. He, N., Yang, X., Jiao, Y., Tian, L., and Zhao, Y., Characterisation of antioxidant and antiproliferative
acidic polysaccharides from Chinese wolfberry fruits. Food Chem., 133, 978–989, 2012.
40. Yang, D., Li, S.Y., Yeung, C.M., Chang, R.C., and So, K.F., Lycium barbarum extracts protect the brain
from blood-brain barrier disruption and cerebral edema in experimental stroke. PLoS One, 7, 3, 2012.
41. Lu, S.P. and Zhao, P.T., Chemical characterization of Lycium barbarum polysaccharides and their
reducing myocardial injury in ischemia/reperfusion of rat heart. Int. J. Biol. Macromol., 47, 681–684,
2010.
42. Gruenwald, J., Novel botanical ingredients for beverages. Clin. Dermatol., 27, 210–216, 2009.
43. Wu, S., Wang, Y., Gong, G., Li, F., Ren, H., and Liu, Y., Absortion and desorption properties of mac-
rosporous resins for flavonoids from the extract of Chinese wolfberry (Lycium barbarum L.). Food
Bioprod. Process., 93, 148–155, 2015.
44. Wu, X., Beecher, G., Holden, J., Haytowitz, D., Gedhardt, S., and Prior, R., Lipophilic and hydrophilic
antioxidant capacities of common foods in the United States. J. Agric. Food Chem., 52, 4026–4037,
2004.
45. Vidal, K., Bucheli, P., Gao, Q., Moulin, J., Shen, L., Wang, J., Blum, S., and Benyacoub, J.,
Immunomodulatory effects of dietary supplementation with a milk-based wolfberry formulation in
healthy elderly: A randomized, double-blind, placebo-controlled trial. Rejuv. Res., 15, 89–97, 2012.
46. Shan, X., Zhou, J., Ma, T., and Chai, Q., Lycium barbarum polysaccharides reduce exercise-induced
oxidative stress. Int. J. Mol. Sci., 12, 1081–1088, 2011.
47. Liu, Y., Gong, G., Zhang, J., Jia, S., Li, F., Wang, Y., and Wu, S., Response surface optimization of
ultrasound-assisted enzymatic extraction polysaccharides from Lycium barbarum. Carbohydr. Polym.,
110, 278–284, 2014.
48. Chao, Z., Ri-fu, Y., and Tai-Qiu, Q., Ultrasound-enhanced subcritical water extraction of polysaccha-
rides from Lycium barbarum L. Sep. Purif. Technol., 120, 141–147, 2013.
49. Hu, J., Chen, Y., and Ni, D., Effect of superfine grinding on quality and antioxidant property of fine
green tea powders. LWT Food Sci. Technol., 45, 8–12, 2012.
50. Zhang, M., Wang, F., Liu, R., Tang, X., Zhang, Q., and Zhang, Z., Effects of superfine grinding on physi-
cochemical and antioxidant properties of Lycium barbarum polysaccharides. LWT Food Sci. Technol.,
58, 594–601, 2014.
21
Golden Berry and Selected Tropical
(Açai, Acerola, and Maqui) Juices
Coralia Osorio, Maria Elisa Schreckinger, Prerna Bhargava, Woo Young Bang,
Daniel A. Jacobo-Velázquez, and Luis Cisneros-Zevallos
CONTENTS
21.1 Introduction....................................................................................................................................251
21.4 Health Effects................................................................................................................................ 256
21.4.1 Golden Berry Juice........................................................................................................... 256
21.4.2 Açai Juice......................................................................................................................... 258
21.4.3 Acerola Juice.................................................................................................................... 262
21.4.4 Maqui Juice...................................................................................................................... 262
21.4.5 Selected Tropical Juices: Relationship between Chronic Disease, Food, and Medicine.......264
21.6 Conclusion..................................................................................................................................... 266
References............................................................................................................................................... 266
21.1 Introduction
South America is a continent with one of the highest biodiversities in the world, with a wide variety of
tropical and nontropical fruits, frequently consumed among native people but often remain unknown in
the rest of the world [1]. Fruits from South America, such as golden berry (Physalis peruviana L.), açai
(Euterpe oleraceae Mart.), acerola (Malpighia emarginata DC.), and maqui (Aristotelia chilensis [Mol.]
Stuntz), have been recognized not only by their attractive sensory properties but also by their func-
tional properties [2–4]. Phytochemicals such as vitamin C, phytosterols, carotenoids, and polyphenolic
substances (flavonoids and nonflavonoids) are able to prevent lifestyle-related diseases (such as cancer,
obesity, diabetes, and cardiovascular, among others) upon regular consumption [5]. These fruits are usu-
ally freshly consumed but their juices are also an important source of phytochemicals that preserve the
functional properties if they are stored appropriately [6]. This chapter highlights the presence of health-
promoting components in some worldwide-recognized tropical fruit juices, their influence in the control
of harmful factors related to cardiovascular disease (CVD), and their potential as functional beverages
due to the synergy of those components.

Golden berry (Figure 21.1) is a tropical species native to South America, mainly Colombia, Peru, and
Ecuador, but it is also grown in Egypt, South Africa, India, New Zealand, and Australia. The fruit is
also known as “aguaymanto,” “uchuva,” “uvilla,” or “cape gooseberry” and is eaten fresh or used to
251
FIGURE 21.1 Golden berry fruit (Physalis peruviana) and its juice.
make jams, juices, sauces, and preserves. It is an excellent source of provitamin A (3000 IU of carotene
per 100 g), vitamin C, vitamin B complex, and minerals. The golden berry fruit has a unique and sweet
tangy flavor when used alone or when mixed with other dried fruit and nuts, which is a special feature
to its use in “haute cuisine.” The sweetened dried fruit has been included in the U.S. market and des-
ignated as “ultrahigh-fiber superfruit” [7]. The yield of juice from golden berry is approximately 70%
of the berry weight. Application of enzymes leads to juice with higher pulp content, acidity, and total
soluble solids [8]. Sugar content in the juice is 4.90 g/100 g, mainly composed of sucrose and fructose.
The ascorbic acid level in golden berry juice (46 mg/100 g) is comparable with that of orange juice
(50 mg/100 g) (Table 21.1). The juice contains 0.20 g/100 g lipid. Other fat-soluble compounds such as
sterols, vitamin E (α-, β-, and γ-tocopherols), and β-carotene were detected in the juice (Table 21.2), and
these serve as a source of health-promoting components for functional drinks [9].
The other fruits, such as açai, acerola, and maqui, could be classified as anthocyanin-rich sources.
Among them, açai has been cataloged as a “superfruit” and is attractive to consumers due to its high
content of nutritional and health-promoting phytochemical constituents. The açai palm fruit is exten-
sively consumed in the Amazon region and is economically valuable because of the multifaceted uses
that exist for multiple parts of the plant [2]. The açai fruit juice is considered a good supplement for foods
and dietetics because of its protein [10], calcium (118 mg/100 g), vitamins A and C, and omega-3 and
omega-6 fatty acid contents (Tables 21.1 and 21.2) [11].
Acerola, also known as West Indian, Haiti, or Barbados cherry, belongs to an evergreen small tree.
Brazil is the largest grower of acerola, but it is also cultivated in Southern Texas, Mexico, Caribbean,
and India [12,13]. It is a drupaceous round fruit with a thin epicarp that turns red upon ripening, being
extremely fragile and perishable with a shelf life of 2–3 days when it is mature. Acerola is a natural source
of vitamin C and it is commercially utilized as juice, marmalade, gelatin, ice cream, frozen concentrate,
jelly, gum, nutraceutical supplement, liquor, and yogurt [12,14]. The nutritional characteristics of the
juice vary with the maturity stages of acerola, which are used to prepare the juice. It has been reported
that the juice from immature acerola has a higher antioxidant activity compared to the mature fruits [15].
The juice from an immature acerola was found to have significant levels of fructose and glucose, whereas
no significant amounts were detected in juices from mature acerola. According to this study, 87% of the
Golden Berry and Selected Tropical (Açai, Acerola, and Maqui) Juices 253
TABLE 21.1
Compositional and Nutritional Characteristics of Selected Tropical Fruit Juices (per 100 g)
Nutrient Unit Golden Berry [8,9,42] Açai [11,26,57] Acerola [15,30] Maqui [21,36]
Water g 92.70 96.94 94.30 86.60
Energy kcal 26.16 12.31 69.60 70.35
Protein g 0.44 0.21 5.7 14
Lipid (fat) g 0.20 1.26 4.80 1.51
Carbohydrate g 5.65 0.033 0.40 0.19
Ash g 1.01 0.39 0.20 0.44
Total sugars g 4.90 na 0.30 na
Minerals
Calcium mg 8.0 118 10 482 DW
Iron mg 1.2 4.4 DW 0.50 10.6 DW
Phosphorus mg 55.3 na 9 na
Potassium mg na na 97 1863 DW
Vitamins
Niacin mg 1.7 na 0.400 na
Riboflavin mg 0.03 na 0.060 na
Thiamin mg 0.1 na 0.020 na
Vitamin A IU 1002 DW 509 na
Vitamin C mg 46 <0.1 DW 1600 na
Vitamin E (ATE) mg 90.5 na na na
Abbreviations: na, not available; ATE, alpha-tocopherol equivalents; DW, dry weight.
total acid content was ascorbic acid, and the rest included tartaric and malic acids. Moreover, the content
of malic acid was 32% in juice made from ripe fruits and 12% in immature ones. Differences of about
41% in vitamin C content of juice from immature acerola (1.90 g/100 g) to mature acerola (0.97 g/100 g)
were also noticed [16]. The compositional and nutritional characteristics of acerola juice are shown in
Tables 21.1 and 21.2. Detailed information about acerola juice is also provided in Chapter 7.
Maqui is a fruit-bearing shrub that thrives in the temperate forests of central to southern Chile and
western Argentina, and it is also known as “Chilean wineberry.” Maqui yields a small edible purple/
black berry averaging 5 mm in diameter with typically 3–4 seeds and is typically consumed fresh or
used to make jam, tea, wine, and juice [17,18]. The fruit juice is a rich source of polyphenols (~993.21 mg
gallic acid equivalents [GAE]/100 mL) exhibiting many health benefits [19,20]. Due to these properties,
maqui juice is often incorporated into functional foods and beverages. The nutritional characteristics
of concentrated maqui juice with no sugar added are given in Table 21.1. The pH of maqui juice (3.35)
is similar to that reported for the fresh fruit (3.7), indicating that the concentrated juice preserves the
original acidic properties [20]. In addition, maqui fruit contains a variety of minerals of importance, for
instance, calcium, iron, and potassium, in amounts of 4823, 106, and 18633 mg/kg of fruit dry weight
(DW), respectively [21].

Golden berry fruit is recognized not only for its sensorial properties but also because it has been linked
to a variety of health benefits, such as antioxidant, anti-inflammatory [22], and anticancer [23] activi-
ties, among others. Quercetin is the main phenolic compound, followed by myricetin and kaempferol
[24]. The antioxidant activity of golden berry juice was assessed and evaluated by the 1,1-diphenyl-2-
picrylhydrazyl (DPPH) test. Fresh juice shows a 78% decrease in comparison to a control solution, while
the enzyme-treated juice reflects an 82% decrease [8].
TABLE 21.2
Phytochemicals Present in Selected Tropical Fruit Juices
Golden Berry Açai Acerola Maqui
Phytochemicals Unit [9,42] [11,60,61] [15,28,30] [31,36]
Total Anthocyanins mg C3GE/L nd 729 53.50 4.56
Cyanidin-3-glucoside mg/L nd 75.1 na na
Cyanidin-3-rutinoside mg/L nd 202 na na
Cyanidin-3-rhamnoside mg/100 g DW nd na 359 na
Delphinidin-3-glucoside mg/L nd na na 344
Delphinidin-3-sambubioside- mg/L nd na na 388
5-glucoside
Quercetin-3-rutinoside mg/L na na 1.71 na
Total Phenolics mg GAE/100 mL 6.30 210 256 741
Ellagic Acid mg/L na 55.4 na na
Proanthocyanidins mg/g DW nd 12.89 na na
Total Sterols g/kg 55.6 na na na
Δ5-Avenasterol g/kg 12.5 na na na
Campesterol g/kg 12.2 na na na
Lanosterol g/kg 6.55 na na na
Stigmasterol g/kg 6.23 na na na
Fatty Acids
SFA %a 22.7 26.1 mg/g DW na na
MUFA %a 31.9 60.6 mg/g DW na na
PUFA %a 44.4 13.3 mg/g DW na na
Carotenoids
β-Carotene g/kg 4.32 <5.0 IU, 100 g DW na na
Tocopherols
α-Tocopherol g/kg 28.3 na na na
β-Tocopherol g/kg 15.2 na na na
γ-Tocopherol g/kg 45.5 na na na
a Percentage from total fats present in the juice.
C3GE, cyanidin-3-glucoside equivalents; DW, dry weight; GAE, gallic acid equivalents; MUFA,
Abbreviations:
monounsaturated fatty acids; na, not available; nd, not detected; PUFA, polyunsaturated fatty acids; SFA,
saturated fatty acids.
The juice of the açai fruit is dark purple and exhibits DPPH radical scavenging properties, attribut-
able to its high anthocyanin and phenolic content, among which 3-glucosides of cyanidin, malvidin,
delphinidin, and peonidin, p-hydroxybenzoic, and hydroxycinnamic acids have been found [25]. Other
antioxidant phytochemicals include dihydrokaempferol, chrysoeriol, and velutin, a flavonoid with potent
anti-inflammatory effects. The juice is obtained by cold pressing the thin pulp of the ovoid fruit (berry).
The stone is about 80% of the fruit volume [26]. Antioxidant properties of açai, acerola, and maqui juices
are reported in Table 21.3.
It has been reported that frozen single-strength acerola juice has DPPH, oxygen radical absorbance
capacity (ORAC), and total phenolic values of 307 µmol trolox equivalents (TE)/100 mL, 8529 µmol
TE/L, and 256 mg GAE/100 mL, respectively [14,27]. A positive correlation also existed attributing
the antioxidant activity to the presence of ascorbic acid, anthocyanins, and some other phytochemicals.
However, anthocyanins made a significant contribution to the antioxidant activity of acerola juice with
528 mg cyanidin-3-glucoside equivalents (C3GE)/g fresh weight (FW), with cyanindin-3-rhamnoside
and pelargonidin-3-rhamnoside being the major constituents [27]. Mezadri et al. [28] evaluated the
antioxidant activity of homemade juices prepared by crushing or squeezing in comparison to the com-
mercially available juices. This study reported a high correlation coefficient for total phenolic index
and antioxidant activity assays (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) [ABTS], DPPH,
TABLE 21.3
Antioxidant Properties of Selected Tropical Fruit Juices
Antioxidant Assay Açai [61] Acerola [28] Maqui [36]
ABTS 12.8 μmol TE/mL 91.17 μmol TE/mL 10.61 μmol TE/mL
DPPH 18.3 IP% 125.6 μmol TE/mL na
ORAC 19.5 μmol TE/mL 85.3 μmol TE/mL na
FRAP 3.8 μmol FE/mL na na
Inhibition of LDL oxidation (peroxides) 21.7% na na
TBARS, inhibition of LDL oxidation (malondialdehyde) 13.0% na na
ABTS, 2,2′-azino-bis(3-ethylbenzothiazline-6-sulfonic acid); DPPH, 2,2-diphenyl-1-picrylhydrazyl; FE,
Abbreviations:
ferric ions converted to the ferrous form; FRAP, ferric-reducing antioxidant capacity; IP%, inhibition per-
cent; LDL, low-density lipoprotein; na, not available; ORAC, oxygen radical absorbance capacity; TBARS,
thiobarbituric acid reactive substances; TE, trolox equivalents.
and ORAC), indicating that antioxidant activity is due to the synergistic effects of the ascorbic acid and
polyphenols. The study also highlights that the processing technique can influence the amount of ascor-
bic acid extracted and would be higher in unripe acerola juice, in agreement with the data reported by
Righetto et al. [15]. Thus, the antioxidant activity was measured by different methods and it was reported
that crushed fruit pulp juices had higher DPPH, ABTS, and ORAC values (126, 91.76, and 85.39 mmol
TE/L juice, respectively) than the commercially available ones. Ascorbic acid content ranged from 6.32
to 9.20 g/kg of pulp and 9.44 to 17.97 g/L of juice. Total polyphenol content ranged from 6430 to 7510 mg
GAE/kg of pulp and 8050 to 10590 mg GAE/L of juice. Major polyphenols were reported as procyanidin
B1, chlorogenic acid, (−)-epigallocatechin gallate, (−)-epicatechin, and rutin [28]. In addition to that, the
total phenolic content in immature acerola juice was 9.2 mg of GAE/g, whereas in mature acerola juice
it was 1.35 mg GAE/g [15,29,30].
Wang et al. [31] reported that maqui juice, prepared by blending 200 g of berries with 200 mL water,
had a total phenolic content of 7413 mg GAE/L juice, which was very high in comparison to other berry
juices [32]. Among them, anthocyanin value averaged at 4558 mg C3GE/L, which exceeds greatly the
values reported for blueberry juice (45.2 mg C3GE/L), cranberry juice (77 mg C3GE/L), and strawberry
juice (110 to 270 mg of pelargonidin-3-glucoside equivalents (P3GE)/L) [31,33–35]. In beverages contain-
ing lemon juice and maqui, total phenolic values ranged between 87.80 and 280 mg GAE/100 mL [36],
and in isotonic beverages containing maqui, values ranged from 73.91 to 80.97 mg GAE/100 mL [37].
The anthocyanin composition of juice has been reported as being composed of 41.0% delphinidin-3-
sambubioside-5-glucoside, 24.5% delphinidin-3,5-diglucoside, 17.0% cyanidin-3-sambubioside-5-glucoside,
5.2% delphinidin-3-sambubioside, 8.2% delphinidin-3-glucoside, 3.9% cyanidin-3-glucoside, and 0.23%
cyanidin-3-sambubioside [31], with delphinidin being the major anthocyanidin in this juice. Meanwhile, the
anthocyanin distribution in the maqui juice was similar to the methanolic extract, which contained 33.7%
delphinidin-3-sambubioside-5-glucoside and 17.2% delphinidin-3,5-diglucoside as its major constitu-
ents [38]. However, ethanol extract from the maqui fruit showed different ratios with the juice prepared
with water, having delphinidin-3-glucoside as its main anthocyanin, which accounted for 30%, followed
by delphinidin-3-sambubioside-5-glucoside with 14.3%, and delphinidin-3,5-diglucoside with 12.8% [39].
Cyanidin-3-sambubioside in the ethanol extract accounted for 0.5%, which was twice the level (0.23%) in
the maqui juice prepared with water [31]. In general, methanol, ethanol, and mixtures with water serve as
efficient solvents for the extraction of anthocyanins and other flavonoids [40]. It has also been reported that
fermentation can play a beneficial role in effective extraction of anthocyanins in the juice [31]. Maqui juice
also contains a huge amount of flavonoids at 2409 μg (+)-catechin equivalents (CE) of condensed tannins or
proanthocyanidins/mL and 372 μg (+)-CE of hydrolysable tannins/mL [31].
The aforementioned results strongly indicate that maqui juice is a rich source of phenolic com-
pounds that include a large amount of anthocyanins. Therefore, maqui juice exhibits strong antioxidant
properties, mainly attributed to the presence of the aforementioned polyphenols [19,41]. The antioxidant
activity of maqui juice was evaluated together with aqueous ethanolic extract of the fruit and its fractions.
This study showed IC50 (effective concentration [scavenging 50% in the reaction present radicals]) values
of 4.7 and 5.9 ppm for maqui juice against DPPH and thiobarbituric acid reactive substance (TBARS)
formation and IC50 values of 45.3 and 62.4 ppm against superoxide and hydroxyl radical formation,
respectively. In addition, maqui juice had ORAC and ferric-reducing antioxidant capacity (FRAP) values
of 22699 µmol TE/g extract and 9930 µmol CE/g extract, respectively. This study also indicated a cor-
relation between antioxidant activity and total phenolic content, thus suggesting the significant role of
phenolic compounds in the antioxidant properties of maqui juice [41]. The antioxidant activity of maqui
juice was evaluated in comparison with different berry juices (strawberry and blackberry juice) using the
ABTS assay and expressing the results as total reactive antioxidant potential (TRAP) and total antioxi-
dant reactivity (TAR) indices. It was found that maqui had the highest antioxidant activity for both tests
and also a good correlation among total phenol content and antioxidant activity measured by these latter
assays. This study also demonstrated that maqui juice could inhibit in vitro copper-induced oxidation of
low-density lipoprotein (LDL) and protect human endothelial cells against hydrogen peroxide–induced
intracellular oxidative stress. When compared to other berry juices, blackberry juice showed a lower
LDL oxidation inhibition compared to strawberry and maqui juice and there were no differences in
protection against intracellular oxidative stress among the three berries [19]. Recently, there have been
a couple of reports evaluating the antioxidant properties of novel beverages containing lemon juice and
maqui. For instance, a beverage containing lemon juice and 2.5% or 5% (w/v) ground maqui (LM1)
possessed high in vitro antioxidant activity mainly due to the presence of polyphenols in maqui. The
antioxidant activity measured by the ABTS method showed values of 10.94 and 17.25 μmol TE/mL for
2.5 and 5% of LM1, respectively. These values were higher than the lemon juice control (2.03 μmol TE/
mL) and similar to the maqui controls (10.61 and 16.6 μmol TE/mL for 2.5 and 5% of LM1, respectively),
indicating the strong in vitro antioxidant activity attributable to the presence of maqui. In addition, the
antioxidant activity of the LM1 beverage did not exceed 20% loss at 4°C and 30% at 25°C during the
70 days of storage, thus indicating a good stability. This protective effect was attributed to the effect of
vitamin C in protecting anthocyanins and other bioactive compounds in maqui [36].
Comparative antioxidant activities using similar units are presented in Table 21.3 for açai, acerola, and
maqui juices. In addition, the chemical structures of the main bioactive compounds present in selected
tropical fruit juices discussed in this chapter are shown in Figure 21.2.
21.4 Health Effects

21.4.1 Golden Berry Juice
The effect of feeding golden berry juice on rats exhibiting hypercholesterolemia was comparatively
analyzed by monitoring the changes in the blood profile, by evaluating the liver and kidney functions
using enzymatic tests, and by histopathological examination of these tissues of male albino rats fed with
different diets. The rats were divided into four groups according to their diet, one group was the nega-
tive control with the basal diet, another group received the basal diet enriched with cholesterol, and the
other two groups received the high-cholesterol diet (HCD) simultaneously with two different amounts of
golden berry juice (Table 21.4). Thus, juice administration in rats showed lower levels of total cholesterol
(TC), triacylglycerols (TAG), and LDL cholesterol, as well as higher levels of high-density lipoprotein
(HDL) cholesterol in comparison with animals fed with HCD and cholesterol-free diet. Juice admin-
istration also induced a decrease in the activity of glutamic pyruvic transaminase compared with the
positive control group after 2 months of administration. There was a remarkable decrease in total serum
protein, albumin, and globulin for groups treated with golden berry juice. Histological examination of
the liver and kidney were conducted, thus demonstrating that juice consumption may provide beneficial
effects in reversing HCD-associated detrimental changes, such as to protect the liver in response to
oxidative stress as well as to alleviate the magnitude of fatty liver development in response to HCD [42].
Golden berry fruit juice exhibited a mild anti-inflammatory activity compared with methylprednisolone
(a known anti-inflammatory drug), when the juice was applied on the rabbit’s eye, and the irritation
produced was evaluated by comparing with the effect of a saline solution using a Draize test. Pterygium
is one of the most frequently occurring diseases among afflictions of the human visual system, induced
CH3
H3C CH3 H 3C
H3C
CH3
CH3 CH3 CH3
CH3
β-Carotene
CH3
HO
CH3 CH3 CH3
H3C O CH3
CH3
CH3
α-Tocopherol
CH3
HO
H3C CH3 CH3
O CH3
CH3
CH3
β-Tocopherol
HO
CH3 CH3 CH3
H3C O CH3
CH3
CH3
γ-Tocopherol
OH
HO O+
OH
O
OH OH
O
HO
OH
OH
Cyanidin-3-glucoside
H3C CH3 CH3 CH3
OH
CH3
Vitamin A
OH
HO O O
H
HO OH
Vitamin C (ascorbic acid)
FIGURE 21.2 Chemical structures of the most abundant bioactives present in golden berry juice (β-carotene,
α-tocopherol, β-tocopherol, and γ-tocopherol), açai juice (cyanidin-3-glucoside and vitamin A), acerola juice (vitamins A
and C and cyanidin-3-O-rhamnoside), and maqui juice (delphinidin-3-sambubioside-5-glucoside and delphinidin-3-
glucoside). (Continued)
OH
HO
O+ OH
O
HO O OH
HO CH3
OH
Cyanidin-3-O-rhamnoside
HO HO OH
HO HO OH
O O
HO O O OH
O
HO O OH
HO
O+ OH
HO
HO
Delphinidin-3-sambubioside-5-glucoside
OH
OH
+
HO O
OH
O
OH OH
O
HO
OH
OH
Delphinodin-3-glucoside
FIGURE 21.2 (Continued) Chemical structures of the most abundant bioactives present in golden berry juice
(β-carotene, α-tocopherol, β-tocopherol, and γ-tocopherol), açai juice (cyanidin-3-glucoside and vitamin A), acerola
juice (vitamins A and C and cyanidin-3-O-rhamnoside), and maqui juice (delphinidin-3-sambubioside-5-glucoside and
delphinidin-3-glucoside).
by sunlight exposure. This causes a proliferation of fibrovascular tissue that invades the cornea from the
exposed conjunctiva. Pardo et al. [43] histopathologically analyzed the eyes of two rabbits with signs of
pterygium (chronic inflammation, granulose tissue, vascular congestion, and fibrosis), one treated with a
vehicle and the other with golden berry juice. Those eyes treated with the juice presented a lesser content
in the tissues responsible for the irritation in the scleral–corneal limbus, suggesting an antipterygium
effect of golden berry fruit juice that may be related to its inhibition of fibroblast growth.
21.4.2 Açai Juice

The atheroprotective effects of açai freeze-dried fruit and frozen fruit juice were studied in an apoli-
poprotein E (Apo E)-deficient mouse model. Apo E animals were fed with a diet enriched with fruit
juice and after 24 weeks were sacrificed, and plasma and tissue samples (aorta, macrophage, heart,
and liver) were collected and analyzed. Thus, the fruit pulp juice showed an atheroprotective effect in
the hyperlipidemic Apo E-deficient mouse model due to reduced lipid peroxidation, by increasing the
levels and activities of two antioxidant enzymes (glutathione reductase and glutathione peroxidase) and
also by inhibiting proinflammatory cytokines (interleukin-6 [IL-6] and tumor necrosis factors-α [TNF-
α]) through regulating inflammatory mediators [44]. The antioxidant potential and hypocholesterolemic
TABLE 21.4
Potential Positive Health Effect of Tropical Fruit Juices Evaluated in Cell Lines and Animal Studies
Experimental
Fruit Juices Model Study Details Experimental Findings References
Golden Berry
Golden berry Male albino rats 60 days (4 groups and 5 rats each). Group 4 (−) control, Golden berry juice consumption had significant [42]
juice (weighing group 3 (HCD), and groups 1 and 2 (HCD plus golden hypocholesterolemic activities in rats fed with HCD. TC, LDL,
between 120 berry juice [5% and 15%]). and TAG concentrations in the golden berry diet groups were
and 140 g) fed significantly lower than those in the positive control group.
with HCD
Golden berry Male albino A rabbit eye inflammation model was developed, which Golden berry fruit juice exhibited a mild anti-inflammatory [43]
juice rabbits involves two groups. One with transplanting an auricular activity compared with methylprednisolone, a known
(weighing 1500 cartilage fragment obtained from the same rabbit’s ear. The anti-inflammatory drug. An interesting dose-dependent
to 2000 g) other with a thermal injury in the limbus with a soldering cytostatic effect on cultured fibroblasts was also established.
iron. Cytostatic activity was evaluated by measuring and
comparing growth rates of cultured fibroblasts exposed and
not exposed to various fruit juice concentrations.
Açai
Açai juice (blend ApoE−/− mice The mice were fed with a CD or CD formulated to contain The mean lesion areas in the aorta for ApoE−/− mice fed AJP [44]
containing (female and 4 5% freeze-dried AJP for 20 weeks. were 58% less (P < 0.001) compared to that for CD-fed mice.
freeze-dried weeks old) of HDL cholesterol was higher in AJP-fed mice. Biomarkers of
and frozen pulp the C57BL/6 lipid peroxidation, including F2-isoprostanes and isomers of
as the genetic hydroxyoctadecadienoic acids, and hydroxyeicosatetraenoic
predominant background acids, were significantly lower in serum and in the liver of
ingredients) AJP-fed mice.
Golden Berry and Selected Tropical (Açai, Acerola, and Maqui) Juices
Pasteurized açai Nine-week-old Rats were fed a standard AIN-93 M diet (control) or a Açai supplementation caused a hypocholesterolemic effect by [45,46]
pulp female Fischer hypercholesterolemic diet that contained 25% soy oil and reducing TC and HDL cholesterols. Serum levels of carbonyl
rats (weighing 1% cholesterol. The test diet was supplemented with 2% proteins and total, free, and protein sulfhydryl groups were
~145 g) açai pulp for control (group CA) and hypercholesterolemic reduced by açai ingestion in animals receiving the standard or
rats (group HA) for 6 weeks. The levels of protein carbonyl hypercholesterolemic diet. The hypocholesterolemic effect
and sulfhydryl groups, superoxide dismutase and observed can be attributed to the differential gene expression
paraoxonase activities, and lipid profiles of the serum were in the rats (enhanced expression of the ABCG5 and ABCG8
measured. transporters). These alterations directly increased the rate of
biliary sterol excretion and increased uptake of LDL
cholesterol by the liver via the upregulation of LDL-R.
(Continued)
259
260
Experimental
Acerola
Acerola juice n = 42 (healthy Control (standard diet) Upon examination, acerola juice supplements used in the [50,59]
Swiss male n=6 cafeteria diet showed antigenotoxic activities that significantly
mice) Cafeteria dieta; animals were divided into six groups (n = 6) suppressed DNA damage in different tissue profiles of the
and fed 0.1 mL/10 g/day of brain, liver (except unripe acerola), and kidney.
G1: Water Acerola juice or rutin-supplemented diet significantly decreased
G2: Unripe acerola juice micronucleated polychromatic erythrocytes, displaying
G3: Ripe acerola juice antimutagenic potential as compared to other diets.
G4: Industrial acerola juice
G5: 1 mg/kg/day of vitamin C
G6: 200 mg/kg/day of rutin for 30 days daily.
Acerola juice n = 36 (Swiss Control (standard diet) Acerola juice affects the absorption of ascorbic acid inside [50]
albino mice) n=6 plasma and reduced its excretion in urine.
Cafeteria dieta; animals were divided into five groups (n = 6): Adipose index, TNF-α, and pJNK decreased (P < 0.5) in mice
G1: Distilled water with all subgroups in comparison to subgroup 1.
G2:1 mg/mL vitamin C TAG levels reduced in subgroups 3, 4, and 5.
G3: 0.1 mL/10 g of industrial acerola juice The IL-10/TNF-α ratio increased in subgroups 2, 3, 4, and 5
G4: 0.1 mL/10 g of unripe acerola juice (P < 0.05).
G5: 0.1 mL/10 g of ripe acerola juice for 13 weeks. Subgroups 2, 3, 4, and 5 (P < 0.05) increased lipolytic activities.
Acerola juice n = 60 (male Rats were assigned into 4 subgroups of 15 each: Levels of glycemia, TC, TAG, and LDL significantly (P < 0.05) [51]
offspring from G1: Nondiabetic control (water) decreased and an increase was observed in HDL levels in
nondiabetic and G2: Diabetic control (water) subgroup 4 in comparison to subgroup 2.
diabetic Wistar G3: Nondiabetic treated with acerola juice Treatment with acerola prevents hyperglycemia and
rats) G4: Diabetic treated with acerola juice for 30 consecutive dyslipidemias.
days.
Maqui
ANC of maqui HFD obese Mice were divided into two groups: Oral administration of ANC improved hyperglycemia and [53]
diabetic G1: A control group gavaged with a vehicle solution, either insulin resistance in HFD obese diabetic mice.
C57BL/BJ male water or a mixture of labrasol/water. D3S5G, an abundant anthocyanin in maqui, is partially
mice G2: Treated groups gavaged with one of the following responsible for the in vivo antidiabetic effects of ANC.
treatments: ANC, labrasol/water (66/34 + ANC), D3S5G,
and metformin.
(Continued)
Experimental
MeOH extract of Male Wistar rats The rats were divided into three groups: control, I/R + The MeOH extract of maqui showed cardioprotective effects on [54]
maqui vehicle, and I/R + extract. Vehicle (0.9% NaCl) or extract acute ischemia/reperfusion performed in rat heart in vivo.
(100, 10, and 1 ppm/kg body weight of rat) was
administered by intravenous injection 10 min before
ischemia.
MaquiBrightTM Blink-suppressed Sustained suppression of blinking in the rat was achieved by Maqui extract and delphinidin 3,5-O-diglucoside suppressed [56]
(maqui extract) dry eye mode in keeping the rat stationary on a swing. Treatments were ROS formation from lacrimal gland tissue and preserved tear
and its female repeatedly administered orally once a day to rats during the secretion.
anthocyanins 8-week-old swing procedure. Lacrimal function was evaluated on
Sprague- day 11.
Dawley rats
Maqui juice HUVEC exposed Cells were preincubated with increasing concentrations of Maqui juice protects human endothelial cells against H2O2- [19]
to H2O2 as a maqui juice for 30 min before addition of H2O2. induced intracellular oxidative stress.
model of
vascular
oxidative stress
Maqui phenolic LPS-stimulated Cells were pretreated with or without 100 µmol/L (C3GE or Anthocyanin- and proanthocyanidin-enriched fractions from [39]
extract and RAW 264.7 ECE) of phenolic extract or enriched fractions for 1 h maqui decreased inflammation in vitro by inhibiting
enriched macrophages before the addition of LPS. LPS-induced iNOS/NO and COX-2/PGE2 pathways in
fractions macrophages.
MEB and major Murine Cells were preincubated with MBE, D3G5G, and D3S5G for MBE and its anthocyanidins suppress the light-induced [55]
anthocyanins photoreceptor 1 h before being exposed from underneath to 2500 lux of photoreceptor cell death by inhibiting ROS production.
(D3G5G and cells (661W) white fluorescent light for 24 h at 37°C.
D3S5G) of
maqui
a Cafeteria diet consists of mortadella, marshmallow, cheese chips, chocolate wafers, Nuvilab®, Guaraná soft drink, Doritos®, hotdog sausage, peanut candy, calf’s foot jelly, cola soft drink,
and bacon chips.
Abbreviations: AJP, açai juice powder; ANC, anthocyanin-rich formulation; Apo E, apolipoprotein E; C3GE, cyanidin-3-glucoside equivalents; CD, AIN-93G control diet; COX-2, cyclo-
oxygenases-2; D3G5G, delphinidin 3,5-O-diglucoside; D3S5G, delphinidin 3-O-sambubioside-5-O-glucoside; ECE, epicatechin equivalents; HCD, high-cholesterol diet;
HDL, high-density lipoprotein; HFD, high-fat diet; HUVEC, human umbilical vein cells; IL-10, interleukin-10; iNOS, inducible nitric oxide; LDL, low-density lipoprotein;
LDL-R, low-density lipoprotein receptor; LPS, lipopolysaccharide; MEB, aqueous ethanol extract; NO, nitric oxide; PGE2, Prostaglandins E2; pJNK, phosphorylated c-Jun
N-terminal kinase; ROS, reactive oxygen species; TAG, triacylglycerols; TC, total cholesterol; TNF-α, tumor necrosis factors-α.
261
effects of açai pulp ingestion in rats fed a standard or hypercholesterolemic diet has been studied show-
ing that the consumption of açai improves the antioxidant status and has a hypocholesterolemic effect in
that model [45]. Later, it was confirmed that açai fruit pulp exhibits a hypocholesterolemic effect in a rat
model of dietary-induced hypercholesterolemia through an increase in the expression of an ATP-binding
cassette, subfamily G transporters and LDL receptor, ATP-binding cassette subfamily G member 5 and
8 (LDL-R, ABCG5, and ABCG8)] genes [46] (Table 21.4). The bioavailability of açai anthocyanins
in healthy nonsmoking human volunteers upon the consumption of fruit juice and pulp in moderate
amounts caused a significant increase in the antioxidant activity of plasma, due to the presence of antho-
cyanins, indicating the clinical antioxidant potential of açai fruit (Table 21.5) [47].
21.4.3 Acerola Juice

Clein [48] in 1956 recommended that acerola juice was a good substitute for orange juice in infants with
an orange juice allergy, favoring their normal growth and development. The study comprised of 100
infants between 1 and 6 months of age with or without an allergy to orange juice (Table 21.5). They were
tested for their bone growth and vitamin C availability. It was observed that there was higher bioavail-
ability of ascorbic acid due to the presence of acerola [48]. In another human intervention, synthetic
ascorbic acid juice was compared to acerola juice. This work suggested that the presence of cyanidin-3-
α-O-rhamnoside and pelargonidin-3-α-O-rhamnoside in acerola juice enhanced the absorption of ascor-
bic acid in plasma and reduced its excretion through urine, thus increasing the bioavailability of ascorbic
acid [49]. In an animal study investigating the effects of acerola juice on obesity, a mouse model was used
in which the animals were fed with cafeteria diet (including mortadella, marshmallow, cheese chips,
chocolate wafer, water, and Guaraná soft drink, among others) for 13 weeks. Then the diet was supple-
mented with water (control), acerola juice (produced industrially or homemade with ripe or immature
fruits), and synthetic vitamin C. Since obesity is generally considered to be low-grade inflammation, a
few inflammatory markers were measured in addition to the adipose index. The diet supplemented with
acerola juice showed decreased TAG levels and adipose index but increased levels of IL-10/ TNF-α
ratio. Thus, diets that included acerola juice increased lipolysis in mice [50]. In another study, using an
animal model, male adult offsprings of Wistar mice from control and diabetic rat dams were fed with
acerola juice and checked for their plasma lipid profiles, glycemic index (GI), and hyperinsulinemia lev-
els. It was concluded that acerola juice had preventive effects against hyperglycemia and dyslipidemias
in mice from diabetic rat dams [51]. Motohashi et al. [52] reported the antitumor effect of concentrates
of acerola extracts in oral squamous cell carcinoma cells (HSC-2) and submandibular gland carcinoma
cells (HSG). They noted that acerola extracts attenuated the induction of 4-(methylnitrosamino)-1-(3-
pyridyl)-1-butanone (NKK), an important initiator of tumorigenesis in lung cancer.
21.4.4 Maqui Juice

As mentioned earlier, maqui juice is a rich source of phenolic compounds, especially anthocyanins,
which have been reported to have a positive impact on metabolic syndrome–related conditions, which
include inflammation, diabetes, and CVD. For instance, the phenolic extract and enriched fractions from
maqui were evaluated for their anti-inflammatory properties using RAW 264.7 macrophage cell line
(Table 21.4). In this study, the cells were pretreated with or without 100 µmol/L (cyanidin-3-glucoside or
epicatechin equivalents) of phenolic extract or enriched fractions for 1 h before the addition of a lipopoly-
saccharide (LPS). It was found that the anthocyanin- and proanthocyanidin-rich fractions from maqui
decreased inflammation in vitro by inhibiting LPS-induced inducible nitric oxide (iNOS)/nitric oxide
(NO) and cyclooxygenase/prostaglandin E2 (COX-2)/(PGE2) pathways in macrophages [39]. In another
study, the antidiabetic effect of an anthocyanin-rich formulation (ANC) of maqui was evaluated using
a murine model of type II diabetes, finding that oral administration of ANC improved hyperglycemia
and insulin resistance in high-fat diet (HFD) obese diabetic mice [53]. Delphinidin-3-sambubioside-
5-glucoside, a major anthocyanin in maqui juice [31], was identified as one of the main bioactive
anthocyanins responsible for the antidiabetic effect of ANC in vivo. Furthermore, this study showed
TABLE 21.5
Health Effects of Tropical Fruit Juices in Clinical Studies
Fruit Juices Subjects Study Details Experimental Findings References
Açai
Açai pulp and clarified n = 12 (healthy, Subjects were dosed at 7 mL/kg of body weight after a Plasma antioxidant capacity was significantly increased [47]
juice nonsmoking washout phase and overnight fast. Plasma was repeatedly by the açai pulp. Anthocyanins from açai are
volunteers, sampled over 12 h and urine over 24 h after consumption. bioavailable in healthy human volunteers upon the
21–31 years old, An acute four-way crossover clinical trial was carried out consumption of fruit juice and pulp in moderate
with a BMI range compared to applesauce and a non-antioxidant beverage as amounts.
of 17.8–25.9 controls.
Acerola
Acerola juice Japanese men, A single oral dose of ascorbic acid was given at a Bioavailability of ascorbic acid was maximum for [49]
nonsmokers from concentration of 50, 100, 200, and 500 mg after overnight 200 mg.
age groups 22 to 26 fasting. Water was ingested for reference. A 100 mL acerola When acerola juice was given, it facilitated the
juice comprising of 50 mg ascorbic acid was given. The absorption of ascorbic acid in plasma and decreased
samples were collected from 0 to 6 h after the oral dose. excretion through urine.
Acerola juice n = 30 (infants, Randomly divided into two groups: Acerola juice was mixed with apple juice and was given [48]
1–6 months) G1: Infants were given acerola juice. to the infants.
G2: Infants were clinically allergic to orange juice or had X-ray scans affirmed normal bone growth.
eczema, pylorospasm, or severe colic disorders and were No allergic reactions observed with acerola juice.
given acerola juice. Normal growth and development was observed in
infants with acerola juice consumption.
Abbreviation: BMI, body mass index.
263
that ANC significantly increased insulin-stimulated glucose uptake in muscle cells and enhanced the
insulin-stimulated downregulation of gluconeogenic enzyme glucose-6-phosphatase in liver cells. These
results suggest that the antidiabetic effect of ANC observed in the animal study may result from an
improved insulin-mediated glucose metabolism in skeletal muscle and liver [53]. In addition, the metha-
nolic extract from maqui fruit, shown to have a similar distribution of anthocyanins with the one in the
juice [31], exhibited cardioprotective effects on acute ischemia/reperfusion performed in rat heart in vivo
[54]. Thus, male Wistar rats were administered with a vehicle (0.9% NaCl solution) or extract (100, 10,
and 1 ppm/kg body weight of rat) by intravenous injection 10 min before induction of damage to the
myocardium by ischemia and reperfusion. The mechanism by which the maqui extract could protect
animals from heart damage is by diminishing lipid oxidation and reducing the concentration of TBARS
[54]. Anthocyanins from maqui berry showed beneficial effects on the other diseases in addition to the
metabolic syndrome–related conditions. For example, fruit extract and the two main anthocyanins in
the fruit juice, delphinidin-3-sambubioside-5-glucoside and delphinidin-3,5-diglucoside, suppressed the
light-induced cell death in the murine photoreceptor cells (661 W) by inhibiting reactive oxygen species
(ROS) production [55]. Recently, it was proven that maqui extract and delphinidin-3,5-diglucoside are
also able to restore tear secretion in a rat dry eye model by suppressing ROS formation in lacrimal gland
tissue, thus suggesting that maqui juice may have a potential use in eye diseases including dry eye and
light-induced photoreceptor cell death [56].
21.4.5 Selected Tropical Juices: Relationship between

Chronic Disease, Food, and Medicine
Figures 21.3 and 21.4 summarize the relationship between chronic diseases, food, and medicine. It is
well known that a healthy diet like the “Mediterranean diet” can prevent a predisease state, allowing
returning to a normal healthy state. However, it is unknown if a disease state can be reversed by eating
* DNA damage in brain,

liver, and kidney [50,59]
* Diabetes [51]
** Atherosclerosis [44]
** Hypercholesterolemia [45,46]
Reversal by eating well Reversal by eating well? *** Diabetes [53]
**** High cholesterol [42]
Food leading cause * Genotoxicity [50,59]
Cafeteria diet [50,59] * Inflammation [50]
High cholesterol [45,46] ** High cholesterol [44–46]
Regular diet [44] ** High lipid peroxidation [44]
High cholesterol diet [42] ** Protein carbonyl, sulphydryl groups
Normal Predisease [45,46] Disease
state State state
* Acerola juice [50,59] * Acerola juice [51]
** Açai juice [45,46] ** Açai juice [44]
*** Maqui [19,39]
*** Maqui [53]
**** Golden berry juice [42]
* Antigenotoxic [50,59] * Decrease in diabetic markers [51]

* Anti-inflammation [50] - Reduced hyperglycemia
- Reduced dyslipidemia
** Hypocholesterolemic effect [45,46]
** Less lesions of aorta [44] Conventional
*** Protection against oxidative stress [19] - High HDL, lower lipid peroxidation
*** Anti-inflammation [39] medicinal drug
*** Decrease in diabetic markers [53]
**** Hypocholesterolemic activity [42] - Improved hyperglycemia
- Improved insulin resistance
Prevention Intervention/treatment/therapeutics
FIGURE 21.3 Relationship between chronic diseases, food, and medicine in human health (based on in vitro and in vivo
studies). Golden berry, acerola, açai, and maqui juices exert potential preventive effects in human health, while acerola,
açai, and maqui juices exert potential therapeutic effects in different chronic diseases. Preventive or therapeutic effects
associated with *acerola juice, **acai juice, ***maqui juice, and ****golden berry juice.
Reversal by eating well Reversal by eating well?
Food leading cause

Normal Predisease Disease
state state state
* Acerola juice [48,49]

** Açai juice [47]
* Increased absorption of vitamin C in plasma [49]

* Decreased vitamin C excretion from urine [49]
* Vitamin C absorption in plasma, no allergic response [48]
Conventional
** Antioxidant status increased in plasma [47]
medicinal drug
** Anthocyanins are bioavailable in plasma [47]
FIGURE 21.4 Relationship between chronic diseases, food, and medicine in human health (based on clinical studies).
Limited studies on acerola and açai juices showed increasing antioxidant status in plasma indicating potential preventive
effects in human health. Preventive or therapeutic effects associated with *acerola juice and **acai juice.
well as an intervention or therapeutic treatment; thus traditionally, a disease state is treated with conven-
tional medicinal drugs.
The in vitro and in vivo studies with golden berry juice and juices of açai, acerola, and maqui in this
chapter showed their potential to exert preventive effects on human health as well as studies with açai,
acerola, and maqui juices demonstrating the potential to exert a therapeutic effect for different diseases.
Similarly, we presented limited clinical studies done with acerola and açai juices confirming the need to
further explore the preventive and therapeutic potential of these tropical fruit juices.

The use of anthocyanin-rich juices as a source of natural pigments for the food industry has increased
during the last decade, not only because of their color properties but also because of the presence of phy-
tochemicals with functional activities. Thus, the use of clarified (centrifuged) or natural anthocyanin-
rich açai juice for the coloring and flavoring of white yogurt to obtain a novel antioxidant-rich dairy food
has been studied [26]. One of the key points in this regard is the instability of anthocyanins to processing
and storage, since they are sensitive to factors such as temperature, light, pH, and oxygen, among oth-
ers. This obstacle has been overcome by using spray-drying, a technique that has been widely used in
the microencapsulation of food ingredients susceptible to deterioration by external agents. Tonon et al.
[57] studied anthocyanin degradation in the spray-dried açai juice produced with different carrier agents
(maltodextrin 10DE and gum Arabic). The reaction followed a first-order kinetic model, being negatively
accelerated by the increase of temperature and water activity.
New formulations take advantage of the synergistic effect of functional fruit mixtures in beverages,
usually marketed as the claim of “boost fruit-flavored beverages.” Some of the main advantages of these
beverages made from natural ingredients are to revitalize the mind and body by the presence of minerals,
help build skin tissue and muscles, and promote good health. On this frame, Gironés-Vilaplana et al. [37]
designed new isotonic drinks with lemon juice and berries: maqui, açai, and buckthorn. Quality param-
eters such as color parameters, minerals, phytochemicals, antioxidant activity, and biological activities
(in vitro alpha-glucosidase and lipase inhibitory effects) were tested in the samples and compared to the
commercially available isotonic drinks. As a result, new isotonic beverages were more effective than
commercial isotonic drinks in terms of their antioxidant activity, total polyphenol content, minerals,
and biological activities while also possessing good organoleptic properties in terms of color and other
relevant parameters.
A nectar based on a mixture of tropical fruits, papaya pulp, and passion fruit juice, enriched with
the vitamin C present in açai pulp, was developed [58], optimizing the formulation by using a sensory
consumer test (22 nontrained panelists) and the response surface methodology (RSM). Finally, from
11 formulations, some of showed good sensory acceptance suggesting a commercial potential. In gen-
eral, consumer interest leans toward healthier and more natural juices and fruit-based drinks, which
are refreshing and exhibit attractive sensory properties. Thus, beverages formulated with the juice or
pulp of a mixture of fruits are “ready-to-drink” products, which present several advantages, such as the
combination of different flavors (taste and aroma), the sum of their today properties, and their favorable
health-promoting components.
21.6 Conclusion
Golden berry, açai, acerola, and maqui fruit juices could serve as a raw material source for many novel
beverages. In addition to their high vitamin content and well-known antioxidant activity, the main
functional activities of these fruits are related to hypercholesterolemic and atheroprotective effects in
hyperlipidemic environments. These findings show the great potential of beverages developed from the
aforementioned tropical fruits on CVD prevention, which today is one of the leading causes of death
worldwide.
REFERENCES
1. Patil, B.S., Jayaprakasha, G.K., Osorio Roa, C., and Mahattanatawee, K., Preface, in Tropical and
Subtropical Fruits: Flavors, Color, and Health Benefits, Patil, B.S., Jayaprakasha, G.K., Osorio Roa, C.,
and Mahattanatawee, K., Eds., ACS Symposium Series 1129, American Chemical Society, Washington,
DC, 2013, pp. ix–xi.
2. Schreckinger, M.E., Lotton, J., Lila, M.A., and González de Mejía, E., Berries from South America:
A comprehensive review on chemistry, health potential, and commercialization. J. Med. Food., 13,
1–14, 2010.
3. Contreras-Calderón, J., Calderón-Jaimes, L., Guerra-Hernández, E., and García-Villanova, B.,
Antioxidant capacity, phenolic content and vitamin C in pulp, peel and seed from 24 exotic fruits from
Colombia. Food Res. Int., 44, 2047–2053, 2011.
4. Beserra Almeida, M.M., Machado de Sousa, P.H., Campos Arriaga, A.M., Matias do Prado, G., de
Carvalho Magalhães, C.E., Arraes Maia, G., and Gomes de Lemos, T.L., Bioactive compounds and
antioxidant activity of fresh exotic fruits from northeastern Brazil. Food Res. Int., 44, 2155–2159, 2011.
5. Wang, L. and Bohn, T., Health-promoting food ingredients and functional food processing, in Nutrition,
Well-Being and Health, Bouayed, J., Ed., InTech, Rijeka, Croatia, 2012, pp. 201–224.
6. Tzika, E.D., Papadimitriou, V., Sotiroudis, T.G., and Xenakis, A., Antioxidant properties of fruits and
vegetables shots and juices: An electron paramagnetic resonance study. Food Biophys., 3, 48–53, 2008.
7. Pszczola, D.E., The inside of inclusions. Food Technol., 62, 50–62, 2008.
8. Ramadan, M.F. and Mörsel, J.-T., Impact of enzymatic treatment on chemical composition, physico-
chemical properties and radical scavenging activity of golden berry (Physalis peruviana L.) juice. J. Sci.
Food Agric., 87, 452–460, 2007.
9. Ramadan, M.F., Bioactive phytochemicals, nutritional value, and functional properties of cape goose-
berry (Physalis peruviana): An overview. Food Res. Int., 44, 1830–1836, 2011.
10. Araujo, C.L., Bezzera, I.W.L., Dantas, I.C., Lima, T.V.S., Oliveira, A.S., and Miranda, M.R.A., Biological
activity of proteins from pulps of tropical fruits. Food Chem., 85, 107–110, 2004.
11. Schauss, A.G., Wu, X., Prior, R.L., Ou, B., Paterl, D., Huang, D., and Kababick, J.P., Phytochemical
and nutrient composition of the freeze-dried Amazonian palm berry, Euterpe oleraceae Mart. (Acai).
J. Agric. Food Chem., 54, 8598–8603, 2006.
12. De Assis, S.A., Pedro Fernandes, F., Martins, A.B.G., and de Faria Oliveira, O.M.M., Acerola, impor-
tance, culture conditions, production, and biochemical aspects. Fruits, 63, 93–101, 2008.
13. Alves, R.E., Filgueiras, H.A.C., Mosca, J.L., and Menezes, J.B., Brazilian experience on the handling
of acerola fruits for international trade: Harvest and postharvest recommendations. Acta Hortic.,
485, 31–36, 1997.
14. Delva, L. and Schneider, R.G., Acerola (Malpighia emarginata DC.): Production, postharvest handling,
nutrition, and biological activity. Food Rev. Int., 29, 107–126, 2013.
15. Righetto, A., Netto, F., and Carraro, F., Chemical composition and antioxidant activity of juices from
mature and immature acerola (Malpighia emarginata DC.). Food Sci. Technol. Int., 11, 315–321, 2005.
16. Lima, V.L., Mélo, E.A., Maciel, M.I.S., Prazeres, F.G., Musser, R.S., and Lima, D.E., Total phenolic and
carotenoid contents in acerola genotypes harvested at three ripening stages. Food Chem., 90, 565–568,
2005.
17. Hoffmann, A.E., Flora Silvestre de Chile en Zona Araucana, Fundación Claudio Gay, Santiago, Chile,
1991 (in Spanish).
18. Muñoz, M., Barrera, E., and Meza I., El Uso Medicinal y Alimenticio de Plantas Nativas y Naturalizadas
en Chile, Museo Nacional de Historia Natural, Santiago de Chile, Chile, 1981 (in Spanish).
19. Miranda-Rottmann, S., Aspillaga, A.A., Pérez, D.D., Vasquez, L., Martínez, A.L., and Leighton, F.,
Juice and phenolic fractions of the berry Aristotelia chilensis inhibit LDL oxidation in vitro and protect
human endothelial cells against oxidative stress. J. Agric. Food Chem., 50, 7542–7547, 2002.
20. Araneda, X., Quilamán, E., Martínez, M., and Morales, D., Elaboración y evaluación de jugo de
maqui (Aristotelia chilensis [Mol.] Stuntz) por arrastre de vapor. Sci. Agropecuaria, 5, 149–156, 2014
(in Spanish).
21. Damascos, M.A., Arribere, M., Svriz, M., and Bran, D., Fruit mineral contents of six wild species of the
North Andean Patagonia, Argentina. Biol. Trace Elem. Res., 125, 72–80, 2008.
22. Wu, S.-J., Tsai, J.Y., Chang, S.P., Lin, D.L., Wang, S.S., Huang, S.N., and Ng, L.T., Supercritical carbon
dioxide extract exhibits enhanced antioxidant and anti-inflammatory activities of Physalis peruviana.
J. Ethnophamarcol., 108, 407–413, 2006.
23. Wu, S.-J., Ng, L.T., Lin, D.-Y., Huang, S.N., Wang, S.S., and Lin, C., Physalis peruviana extract induces
apoptosis in human Hep G2 cells through CD95/CD95L system and the mitochondrial signaling trans-
duction pathway. Cancer Lett., 215, 199–208, 2004.
24. Häkkinen, S.H., Kärenlampi, S.O., Heinonen, I.M., Mykkänen, H.M., and Riitta, A.T., Content of the fla-
vonols quercetin, myricetin, and kaempferol in 25 edible berries. J. Agric. Food Chem., 47, 2274–2279,
1999.
25. Del Pozo-Insfran, D., Brenes, C.H., and Talcott, S.T., Phytochemical composition and pigment stability
of açai (Euterpe oleracea Mart.). J. Agric. Food Chem., 52, 1539–1545, 2004.
26. Coïsson, J.D., Travaglia, F., Piana, G., Capasso, M., and Arlorio, M., Euterpe oleracea juice as a func-
tional pigment for yogurt. Food Res. Int., 38, 893–897, 2005.
27. De Brito, E.S., De Araujo, M.C.P., Alves, R.E., Carkeet, C., Clevidence, B.A., and Novotny, J.A.,
Anthocyanins present in selected tropical fruits: Acerola, jambolão, jussara, and guajiru. J. Agric. Food
Chem., 55, 9389–9394, 2007.
28. Mezadri, T., Villaño, D., Fernández-Pachón, M., García-Parrilla, M., and Troncoso, A., Antioxidant
compounds and antioxidant activity in acerola (Malpighia emarginata DC.) fruits and derivatives.
J. Food Compos. Anal., 21, 282–290, 2008.
29. Gil, M.I., Tomás-Barberán, F.A., Hess-Pierce, B., Holcroft, D.M., and Kader, A.A., Antioxidant activ-
ity of pomegranate juice and its relationship with phenolic composition and processing. J. Agric. Food
Chem., 48, 4581–4589, 2000.
30. Freitas, C.D., Maia, G., Costa, J.D., Figueiredo, R.D., and Souza, P., Acerola: produção, composição,
aspectos nutricionais e produtos. R. Bras. Agrociência, 12, 395–400, 2006 (in Portuguese).
31. Wang, J., Yousef, G., Rogers, R., González de Mejia, E., Raskin, I., and Lila, M.A., Maqui berry
(Aristotelia chilensis) juices fermented with yeasts: Effects on phenolic composition, antioxidant capac-
ity, and iNOS and COX-2 protein expression, in Emerging Trends in Dietary Components for Preventing
and Combating Disease, Patil, B.S., Jayaprakasha, G.K., Murthy, K.N.C., and Seeram, N.P., Eds., ACS
Symposium Series 1093, American Chemical Society, Washington, DC, 2012, pp. 95–116.
dant activity of various red fruit juices. Dtsch. Lebensmitt. Rundsch., 103, 58–64, 2007.
33. Su, M.-S. and Chien, P.-J., Antioxidant activity, anthocyanins, and phenolics of rabbiteye blueberry
(Vaccinium ashei) fluid products as affected by fermentation. Food Chem., 104, 182–187, 2007.
34. Garzón, G.A. and Wrolstad, R., Comparison of the stability of pelargonidin-based anthocyanins in
strawberry juice and concentrate. J. Food Sci., 67, 1288–1299, 2002.
35. Huopalahti, R., Järvenpää, E., and Katina, K., A novel solid-phase extraction-HPLC method for the
analysis of anthocyanin and organic acid composition of Finnish cranberry. J. Liq. Chromatogr. Relat.
Technol., 23, 2695–2701, 2000.
36. Gironés-Vilaplana, A., Mena, P., García-Viguera, C., and Moreno, D.A., A novel beverage rich in anti-
oxidant phenolics: Maqui berry (Aristotelia chilensis) and lemon juice. LWT-Food Sci. Technol., 47,
279–286, 2012.
37. Gironés-Vilaplana, A., Villaño, D., Moreno, D.A., and García-Viguera, C., New isotonic drinks with
antioxidant and biological capacities from berries (maqui, açai, and blackthorn) and lemon juice. Int. J.
Food Sci. Nutr., 64, 897–906, 2013.
38. Escribano-Bailón, M.T., Alcalde-Eon, C., Muñoz, O., Rivas-Gonzalo, J.C., and Santos-Buelga, C.,
Anthocyanins in berries of maqui (Aristotelia chilensis [Mol.] Stuntz). Phytochem. Anal., 17, 18–14,
2006.
39. Schreckinger, M.E., Wang, J., Yousef, G., Lila, M.A., and González de Mejía, E., Antioxidant capacity
and in vitro inhibition of adipogenesis and inflammation by phenolic extracts of Vaccinium floribundum
and Aristotelia chilensis. J. Agric. Food Chem., 58, 8966–8976, 2010.
40. Macheix, J.-J., Fleuriet, A., and Billot, J., The main phenolic fruits, in Fruit Phenolics, Macheix, J.-J.,
Fleuriet, A., and Billot, J., Eds., CRC Press, Boca Raton, FL, 1990, pp. 1–15.
41. Céspedes, C.L., Valdez-Morales, M., Avila, J.G., El-Hafidi, M., Alarcón, J., and Paredes-López, O.,
Phytochemical profile and the antioxidant activity of Chilean wild black-berry fruits, Aristotelia
chilensis (Mol.) Stuntz (Elaeocarpaceae). Food Chem., 119, 886–895, 2010.
42. Ramadan, M.F., Hassan, N.A., Elsanhoty, R.M., and Sitohy, M.Z., Golden berry (Physalis peruviana)
juice rich in health-beneficial compounds suppresses high-cholesterol diet-induced hypercholesterol-
emia in rats. J. Food Biochem., 37, 708–722, 2013.
43. Pardo, J.M., Fontanilla, M.R., Ospina, L.F., and Espinosa, L., Determining the pharmacological activity
of Physalis peruviana fruit juice on rabbit eyes and fibroblast primary cultures. Invest. Ophthalmol. Vis.
Sci., 49, 3074–3079, 2008.
44. Xiea, C., Kanga, J., Burrisa, R., Ferguson, M.E., Schauss, A.G., Nagarajana, S., and Wua, X., Açaí juice
attenuates atherosclerosis in Apo E deficient mice through antioxidant and anti-inflammatory activities.
Atherosclerosis, 216, 327–333, 2011.
45. Oliveira de Souza, M., Silva, M., Silva, M.E., Oliveira, R.P., and Pedrosa, M.L., Diet supplementation
with açai (Euterpe oleracea Mart.) pulp improves biomarkers of oxidative stress and the serum lipid
profile in rats. Nutrition, 26, 804–810, 2010.
46. Oliveira de Souza, M., Souza e Silva, L., Lopes de Brito Magalhães, C., Barros de Figueiredo, B.,
Caldeira Costa, D., Silva, M.E., and Pedrosa, M.L., The hypocholesterolemic activity of açaí (Euterpe
oleracea Mart.) is mediated by the enhanced expression of the ATP-binding cassette, subfamily G trans-
porters 5 and 8 and low-density lipoprotein receptor genes in the rat. Nutr. Res., 32, 976–984, 2012.
47. Mertens-Talcott, S.U., Rios, J., Jilma-Stohlawetz, P., Pacheco-Palencia, L., Meibohm, B., Talcott, S.T.,
and Derendorf, H., Pharmacokinetics of anthocyanins and antioxidant effects after the consumption of
anthocyanin-rich açai juice and pulp (Euterpe oleracea Mart.) in human healthy volunteers. J. Agric.
Food Chem., 56, 7796–7802, 2008.
48. Clein, N.W., Acerola juice - the richest known source of vitamin C: A clinical study in infants. J. Pediatr.,
48, 140–145, 1956.
49. Uchida, E., Kondo, Y., Amano, A., Aizawa, S., Hanamura, T., Aoki, H., Nagamine, K., Koizumi,
T., Maruyama, N., and Ishigami, A., Absorption and excretion of ascorbic acid alone and in acerola
(Malpighia emarginata) juice: Comparison in healthy Japanese subjects. Biol. Pharm. Bull., 34,
1744–1747, 2011.
50. Dias, F.M., Leffa, D.D., Daumann, F., de Oliveira Marques, S., Luciano, T.F., Possato, J.C., de Santana,
A.A., Neves, R.X., Rosa, J.C., and Oyama, L.M., Acerola (Malpighia emarginata DC.) juice intake
protects against alterations to proteins involved in inflammatory and lipolysis pathways in the adipose
tissue of obese mice fed a cafeteria diet. Lipids Health Dis., 4, 13–24, 2014.
51. Barbalho, S.M., Damasceno, D.C., Spada, A.P.M., Palhares, M., Martuchi, K.A., Oshiiwa, M., Sazaki,
V., and Silva, V.S.D., Evaluation of glycemic and lipid profile of offspring of diabetic Wistar rats treated
with Malpighia emarginata juice. Exp. Diabetes Res., 2011, 173647, 2011.
52. Motohashi, N., Wakabayashi, H., Kurihara, T., Fukushima, H., Yamada, T., Kawase, M., Sohara, Y.,
Tani, S., Shirataki, Y., and Sakagami, H., Biological activity of Barbados cherry (acerola fruits, fruit of
Malpighia emarginata DC.) extracts and fractions. Phytother. Res., 18, 212–223, 2004.
53. Rojo, L.E., Ribnicky, D., Logendra, S., Poulev, A., Rojas-Silva, P., Kuhn, P., Dorn, R., Grace, M.H.,
Lila, M.A., and Raskin, I., In vitro and in vivo anti-diabetic effects of anthocyanins from maqui berry
(Aristotelia chilensis). Food Chem., 131, 387–396, 2012.
54. Céspedes, C.L., El-Hafidi, M., Pavon, N., and Alarcon, J., Antioxidant and cardioprotective activities of
phenolic extracts from fruits of Chilean blackberry Aristotelia chilensis (Elaeocarpaceae), maqui. Food
Chem., 107, 820–829, 2008.
55. Tanaka, J., Kadekaru, T., Ogawa, K., Hitoe, S., Shimoda, H., and Hara, H., Maqui berry (Aristotelia
chilensis) and the constituent delphinidin glycoside inhibit photoreceptor cell death induced by visible
light. Food Chem., 139, 129–137, 2013.
56. Nakamura, S., Tanaka, J., Imada, T., Shimoda, H., and Tsubota, K., Delphinidin 3,5-O-diglucoside, a
constituent of the maqui berry (Aristotelia chilensis) anthocyanin, restores tear secretion in a rat dry eye
model. J. Funct. Foods, 10, 346–354, 2014.
57. Tonon, R.V., Brabet, C., and Hubinger, M.D., Anthocyanin stability and antioxidant activity of spray-
dried açai (Euterpe oleracea Mart.) juice produced with different carrier agents. Food Res. Int., 43,
907–914, 2010.
58. Matsuura, F.C.A.U., Folegatti, M.I.dS., Cardoso, R.L., and Costa Ferreira, R., Sensory acceptance of
mixed nectar of papaya, passion fruit and acerola. Sci. Agric., 61, 604–608, 2004.
59. Leffa, D.D., da Silva, J., Daumann, F., Dajori, A.L.F., Longaretti, L.M., Damiani, A.P., de Lira, F.,
Campos, F., Ferraz, A.dB.F., and Côrrea, D.S., Corrective effects of acerola (Malpighia emarginata
DC.) juice intake on biochemical and genotoxical parameters in mice fed on a high-fat diet. Mutat. Res-
Fund. Mol. Mech. Mutagen, 770, 144–152, 2014.
60. Pacheco-Palencia, L.A., Hawken, P., and Talcott, S.T., Phytochemical, antioxidant and pigment stability
of açai (Euterpe oleraceae Mart.) as affected by clarification, ascorbic acid fortification and storage.
Food Res. Int., 40, 620–628, 2007.
61. Seeram, N.P., Aviram, M., Zhang, Y., Henning, S.M., Feng, L., Dreher, M., and Heber, D. Comparison
of antioxidant potency of commonly consumed polyphenol-rich beverages in the United States. J. Agric.
Food Chem., 56, 1415–1422, 2008.
22
Grape Juice
Gian Carlo Tenore
CONTENTS
22.1 Introduction................................................................................................................................... 271
22.4 Health Effects................................................................................................................................ 276
22.6 Conclusion..................................................................................................................................... 278
References............................................................................................................................................... 278
22.1 Introduction
The grape (Vitis vinifera) is characterized by a high concentration and a great variety of phenolic
compounds, mainly flavonoids (catechin, epicatechin, quercetin, anthocyanin, and procyanidin), and res-
veratrol (3,5,4′-trihydroxystilbene), which are especially abundant in red grape products [1]. White and
red grape juices are regarded as being among the richest food sources of antioxidants and such profile is
ascribed to the technological process employed for their production [2]. In fact, some have affirmed that
such processing may increase polyphenolic levels in juices by promoting extraction processes from all por-
tions of the grape [3]. It should be emphasized that thermal treatments employed before (hot break process,
mainly used in the United States) and/or after (pasteurization and concentration) grape extraction may lead
to considerable changes in the polyphenolic profile of commercial grape juices compared to those of fresh
grapes [3]. In addition, hydrolysis mechanisms would enhance the accumulation of individual phenolic
compounds, mainly glucosides, galactosides, and aglycones and to a lesser extent rutinosides and other gly-
cosidic combinations linked to the flavonol nucleus in the juice, which could be better absorbed in the small
intestine and reach the bloodstream [4]. Particularly, higher amounts of polyphenolic compounds naturally
occurring in red grapes would make the consumption of red grape juice (RGJ) unique health benefits,
such as improvement of the endothelial function, increase in the serum antioxidant capacity, protection
of low-density lipoprotein (LDL) against oxidation, decrease in the native plasma protein oxidation, and
reduction of platelet aggregation [1,2]. Several reports on the pharmaceutical usefulness of grape and wine
phytochemicals have appeared [5–7]. Nevertheless, further studies are needed to legitimize grape extracts
as useful dietary supplements. Therefore, investigation of their radical scavenging properties is of interest,
primarily to discover promising new sources of natural antioxidants, functional foods, food supplements,
and nutraceuticals. This chapter intends to focus on the nutritional and bioactive properties of grape juices,
with special regard to RGJ and its potential nutraceutical applications.

Compositional and nutritional characteristics of red and white grape juices are reported in
Table 22.1 [8]. Both grape juices have low caloric value and contain low content of minerals and vita-
mins, except vitamin C. Consuming 100 g of both juices supplies around 50% of the recommended
271
TABLE 22.1
Compositional and Nutritional Characteristics of Red and White Grape Juices (per 100 g)
Nutrient Unit Red Grape Juice White Grape Juice
Energy kcal 57 55
Water g 85.30 84.12
Protein g 0.00 0.00
Lipid (fat) g 0.00 0.00
Minerals
Calcium mg 7 6
Iron mg 0.13 0.18
Magnesium mg 6 7
Phosphorus mg 6 5
Potassium mg 33 35
Sodium mg 9 8
Zinc mg 0.03 0.04
Vitamins
Niacin mg 0.142 0.135
Vitamin A (RAE) µg tr tr
Vitamin B12 µg 0.00 0.00
Vitamin D µg 0.0 0.0
Vitamin K µg 0.2 0.3
Source: Adapted from U.S. Department of Agriculture (USDA), National Nutrient Database for Standard
Reference, Release 27, 2014, Published online at: http://ndb.nal.usda.gov (accessed January 19, 2015).
Abbreviations: DFE, dietary folate equivalents; RAE, retinol activity equivalents; ATE, alpha-tocopherol equiva-
lents; tr, trace.
dietary allowance for adult males and females. They are low in sodium (8–9 mg/100 g) and high in
total sugars (14.11–14.54 g/100 g).

Despite their low nutritional profile, both grape juices are very rich sources of phenolic compounds,
mainly polyphenols, with much scientific literature which has thus far ascribed several beneficial effects
on health. Particularly, most biological data are referred to RGJ as being much healthier than its white
counterpart and used as the most widespread grape juice product. Both pedoclimatic and technological
parameters concur in making grape juice a much richer source of antioxidant than other grape products,
such as wine. It has long been known that the increased biosynthesis of polyphenols, especially flavo-
nols, is greatly influenced by exposure to sunlight and temperature, so it is expected that the grapes that
are grown in warmer and sunnier areas have a higher level of flavonols. For example, Southern Italy,
among the main world grape juice producers, is characterized by high levels of exposure to sunlight,
which is known to exert a marked influence on the polyphenolic content of grapes. Thus, it has been
Grape Juice 273
TABLE 22.2
Polyphenols in Commercial Red Grape Juice
Polyphenols mg/100 mL References
Anthocyanins
Delphinidin-3-O-glucoside 34.78 [10]
Cyanidin-3-O-glucoside 21.63 [10]
Petunidin-3-O-glucoside 25.63 [10]
Peonidin-3-O-glucoside 31.24 [10]
Malvidin-3-O-glucoside 114.63 [10]
Delphinidin-3-O-acetylglucoside 10.38 [10]
Cyanidin-3-O-acetylglucoside 5.08 [10]
Malvidin-3-(6-O-coumaroyl)glucoside (cis isomer) 12.38 [10]
Malvidin-(6-O-caffeoyl)glucoside 52.99 [10]
Peonidin-3-(6-O-coumaroyl)glucoside (trans isomer) 4.32 [10]
Malvidin-3-(6-O-coumaroyl)glucoside (trans isomer) 14.66 [10]
Flavonols
Myricetin-3-O-glucoside 93.1 [10]
Quercetin-3-O-glucuronide 75.6 [10]
Quercetin-3-O-glucoside 79.8 [10]
Laricitrin-3-O-galactoside 13.3 [10]
Kaempferol-3-O-glucoside 5.6 [10]
Laricitrin-3-O-rhamnose-7-O-trihydroxycinnamic acid 18.2 [10]
Kaempferol-3-O-caffeoylate 16.8 [10]
Isorhamnetin-3-O-glucoside 24.5 [10]
Syringetin-3-O-galactoside 19.6 [10]
Stilbenes
cis-Resveratrol 0.04 [1]
trans-Resveratrol 0.5 [1]
Total Proanthocyanidins 15.88 [31]
shown that sun-exposed grapes can contain up to 10-fold more total phenolics than grapes cultivated in
the shade [9]. Some have also affirmed that special processing may increase flavonol levels in juice by
promoting extraction processes from all portions of the grape [3]. In addition, hydrolysis would enhance
the accumulation in juice of individual phenolic compounds, mainly glucosides, galactosides, and agly-
cones, as well as, to a lesser extent, rutinosides and other glycosidic combinations linked to the flavonol
nucleus, which could be better absorbed in the small intestine and reach the bloodstream [4].
Tenore et al. [10] evaluated the anthocyanin content in RGJ (Table 22.2), which resulted in a generally
higher amount than those reported elsewhere for other grape products [1–3,11,12]. In V. vinifera fruit
products [13,14], the compound malvidin-3-O-glucoside was found as the main monomeric anthocyanin
in the sample tested. One in vitro study demonstrated that malvidin (Figure 22.1) is cytotoxic to human
leukemia cells [15]. Malvidin stopped the cell cycle in the G(2)/M phase and induced apoptosis. At a con-
centration of 40 ppm malvidin, the growth of leukemia cells was halved. For this experiment, research-
ers used malvidin extracted from black rice. It is not known if malvidin has the same protective action
OCH3
OH
O+
HO
OCH3
OH
OH
FIGURE 22.1 Malvidin in red grape juice.

in humans. Nevertheless, a German study found a low absorption of malvidin-3-glucoside in humans

after consumption of red wine or RGJ [13]. The study suggested that not malvidin-3-glucoside but rather
an unidentified anthocyanin metabolites and/or other polyphenols in red wine might be responsible for
the observed antioxidant activity and health effects in humans consuming red wine. Among fruits and
vegetables commonly consumed, grapes and their associated products are regarded as the most important
source of our dietary anthocyanins. These compounds have been shown to contribute to the strong pro-
tection of RGJ and wine against LDL oxidation [4]. Recent studies have demonstrated that the long-term
intake of anthocyanins, which were administered as food matrix or enriched fractions, changed the mark-
ers for the oxidative status in some tissues and affected antioxidant enzyme expression levels and activi-
ties when compared with animals that did not receive dietary polyphenols [16]. Thus, considering the
dietary intake of anthocyanins (approximately 100 mg/diet) [17] and their potential health benefits, juice
samples could be regarded as a valuable source of anthocyanins suitable for use as dietary supplement.
The higher concentration of flavonols in RGJ [10] than other grape products [3,18] has been highlighted
(Table 22.2). The main flavonol glycosides in RGJ and wines were quercetin derivatives [19,20], namely,
quercetin-3-O-glucoside and quercetin-3-O-glucuronide. Results suggested that the rates of hydrolysis
of the flavonol glycosides in grape juice and wine were different and depended on the type of flavonol
aglycone and also the nature of the glycoside moiety [21]. Quercetin (Figure 22.2), the most abundant
flavonoid in nature, is reported to exert many beneficial health effects. It shows anti-inflammatory action
by its direct antioxidant action and inhibition of inflammatory mediators and enzymes, such as lipoxy-
genase [22]. Studies have demonstrated that flavonoid-rich diets protect against myocardial infarction
and stroke. As many other flavonoids, quercetin inhibits oxidation of LDL cholesterol, lowers blood pres-
sure, and reduces the risk of heart disease [23]. In addition, studies have shown that quercetin reduces
cancer risk of prostate, ovary, breast, gastric, and colon cells. Numerous in vitro studies have shown that
quercetin induces apoptosis of cancer cells through different mechanisms [24,25].
In agreement with the aforesaid statement with regard to the peculiar technological production process,
resveratrol (Figure 22.3) in RGJ is generally present at a higher concentration (Table 22.2.) than that reported
in red wines [26]. Resveratrol (mainly trans isomer) is a powerful antioxidant but its antioxidant strength
is weaker than those of quercetin and epicatechin. In vitro studies have shown that resveratrol inhibits
oxidative damage caused by the heavy metal cadmium [27]. The antioxidant activity of resveratrol reduces
damage to endothelial cells exposed to nitrite and protects skin cells against damage caused by ultraviolet
(UV) radiation [28]. The antioxidant action of resveratrol helps to prevent damage to DNA, but it also influ-
ences the transcription of genes responsible for redox metabolism and inhibits proliferation of cancer cells
[29]. Resveratrol appears to decrease tumor promotion activity by inhibiting the enzyme cyclooxygenase-1,
which converts arachidonic acid to substances that promote tumor growth [30]. In vitro experiments provide
support for resveratrol to serve as a candidate preventive agent against prostate cancer, but in vivo effects of
resveratrol and its mechanisms of action on prostate cancer prevention remain largely unknown.
OH
OH
HO O
OH
OH O
FIGURE 22.2 Quercetin in red grape juice.
OH
HO HO
HO OH OH
(a) (b)
FIGURE 22.3 cis-Resveratrol (a) and trans-resveratrol (b) in red grape juice.
Grape Juice 275
Similarly, proanthocyanidins are reported to occur in RGJ at higher levels (Table 22.2.) than in red
wine, probably due to the technological production process that allows a massive extraction of these com-
pounds from grape seeds in which they are mainly located [31]. It was recently demonstrated that grape
proanthocyanidins (Figure 22.4) are able to inhibit proliferation in colorectal cancer cell lines [31]. The
authors concluded from their in vitro studies that the chemopreventive action of grape proanthocyanidins
could be mediated via an upregulation of proteins involved in cell cycle progression. Therefore, these
proteins may be useful targets against colon and intestinal cancers. In addition, a significant inhibition
(up to 66%) against cholesterol uptake in HepG2 cell lines was also noted [31].
Experimental results have revealed good antioxidant activity for RGJ when compared with those of
authentic standards chosen as widely employed food preservatives and strong hydrophilic or lipophilic
antioxidants (Figure 22.5) [10]. It is accepted that flavonoids and their metabolites may interact with
OH
OH
HO O
OH
OH
n
OH O
HO
OH
FIGURE 22.4 Proanthocyanidins in red grape juice.
1.8
1.6
1.4
1.2 FRAP
DPPH
mmoI TE/100 mL
0.8
0.6
0.4
0.2
0
RGJ Vit. E Vit. C BHT Na2S2O5
FIGURE 22.5 Antioxidant activities (FRAP and DPPH radical scavenging capacity) of red grape juice before and after
lyophilization versus antioxidant standards. Values are expressed ± SD (P < 0.001). Standard solutions were of 1 mg/mL
concentration. Abbreviations: FRAP, ferric reducing antioxidant power; DPPH, 1,1-diphenyl-2-picrilhydrazyl; RGJ, red
grape juice; BHT, butylhydroxytoluene; Na2S2O5, sodium metabisulfite. (Adapted from Tenore, G.C. et al., J. Agric. Food
Chem., 60, 9680, 2012. With permission.)
plasma proteins as well as the polar surface region of phospholipid bilayers in lipoproteins and cell
membranes [32]. Because of the nature of these interactions, flavonoids may have the ability to protect
against free radical attack in both aqueous and lipid environments, thus providing an effective anti-
oxidant defense in biological systems. Grape juice is a rich source of antioxidant flavonoids, mainly
catechin, epicatechin, quercetin, and anthocyanins. In vitro studies showed that grape juice has sig-
nificant antioxidant activity and can inhibit the oxidation of LDL [33,34]. Human studies have shown
promising results but were limited by either short duration of supplementation, the confounding effects
of medications or other antioxidants, or the measurement of only a few indices of antioxidant status [33].
22.4 Health Effects

Scientific literature accounts for a large number reports on the beneficial effects of grape products
on human health. Special attention has been focused on protection against cancer and cardiovascular
disease (CVD) that may derive from a moderate intake of wine [35]. Generally, it is established that
an oxidation process is involved in the initial development steps of these diseases. Indeed, reactive
oxygen species (ROS), naturally formed during normal metabolism, can damage biological molecules
such as protein, lipid, or DNA. Human metabolism counts on an antioxidant defense system involving
enzymes and proteins to prevent these effects. However, the defenses can be overwhelmed under cer-
tain circumstances, thus leading to the occurrence of harmful effects. It is accepted that the intake of
antioxidant substances reinforces the defenses against ROS. Therefore, the role of antioxidant nutrients
such as vitamins (A, E, and C) or selenium and other diet-derived antioxidants such as polyphenols is
attracting considerable attention from health, food, and nutrition groups. The health-protective proper-
ties of wines are attributed to their antioxidant activities, for example, their capability to eliminate free
radicals. Consequently, numerous papers have been focused on the determination of the antioxidant
activity of wines [36] as well as on their polyphenolic content, probably responsible for the antioxidant
action. In light of these considerations, RGJ can be regarded as much more effective against oxidative
stress–related pathologies. Actually, recent works have contributed to clarify the influence of grape
polyphenols on cardiovascular functions, highlighting not only protective effects but also harmful
risks from excessive consumption of polyphenolic fractions in grape products and, interestingly, those
from RGJ [10].
Myocardial mitochondria are an important source of oxidative stress. The mitochondrial electron
transport chain, under conditions of reductive stress, is capable of generating ROS and reactive nitrogen
species (RNS) that play a crucial role in the pathophysiology of a variety of CVD including conges-
tive heart failure, valvular heart disease, cardiomyopathy, hypertrophy, atherosclerosis, and ischemic
heart disease [37–39]. Recently, grape seed proanthocyanidins, a group of polyphenolic bioflavonoids
ubiquitously found in the lignified portions of grape clusters, were found to possess cardioprotective
abilities by functioning as in vivo antioxidants and by virtue of their ability to directly scavenge ROS
including hydroxyl and peroxyl radicals [40–42]. However, their pro-oxidant toxicity at higher doses
(100–500 µg/mL), particularly their ability to cause apoptosis in cardiomyocytes induced by ROS gen-
eration, has also been reported [43–45]. The effects on cardiomyocytes by directly testing the whole
grape juice are unknown. Tenore et al. [10] examined the effect on free radical and manganese super-
oxide dismutase levels in cardiac-derived H9C2 myocytes exposed to increasing doses (0.01–1 µg) of
lyophilized RGJ (LioRGJ) (Table 22.3). Recent studies have reported that grape seed proanthocyanidin
extract (GSPE) possess potent antioxidant activity against exogenous H2O2, hydroxyl radical, and super-
oxide and may chelate iron, when tested on cardiomyocyte culture [40–42]. Similarly, data reported in
Table 22.3 demonstrate that antioxidants in the juice sample at a maximum sample dose of 0.01 µg were
able to directly scavenge free radicals (with the exception of RNS) without interfering with cell anti-
oxidant defensive system involving enzymes and proteins for cardioprotection. Nevertheless, exposure
to increasing concentrations of LioRGJ resulted in pro-oxidant effects as demonstrated by the increase
in ROS, RNS, and antioxidant enzyme levels at a sample dose of 0.05 µg. These results suggest what is
already stated in the literature for GSPE that higher doses of antioxidants occurring in the juice sample
may cause apoptotic cell injury via effector caspase-3 activation and subsequent induction of ROS and
Grape Juice 277
TABLE 22.3
Effect of Lyophilized RGJ on Physiological and Induced Oxidative Stress in Lysate of H9C2
Cardiomyocytes
0.01 μg 0.05 μg Dox 1 Dox 1 μM + Dox 1 μM +
Unit Control LioRGJ LioRGJ μM 0.01 μg LioRGJ 0.05 μg LioRGJ
TBARS μM/μg protein 0.0043a 0.0025b 0.0047a 0.0068c 0.0021a 0.0065c
NO2− nmol/μg protein 0.0010a 0.0039b 0.0080c 0.0065d 0.0040b 0.0240e
MnSOD U/μg protein 0.0100a 0.0100a 0.0180b 0.0200d 0.0100a 0.0350e
Source: Adapted from Tenore, G.C. et al., J. Agric. Food Chem., 60, 9680, 2012. With permission.
Note: a–dMeans followed by the same letter, within a row, are not significantly different (P > 0.05).
Abbreviations: LioRGJ, lyophilized red grape juice; TBARS, thiobarbituric acid reactive substances; MnSOD, manga-
nese superoxide dismutase.
RNS generation [43–45]. Among many known regulators and effectors of apoptosis, caspases are a
family of cytoplasmic proteases that play an important role in the execution phase of apoptosis. Two
groups of caspases can be identified: upstream initiator caspases that cleave and activate other caspases
and downstream effector caspases, including caspase-3, caspase-6, and caspase-7, that cleave a variety
of cellular substrates or inactivating enzymes. Caspase-3 is a central executioner in apoptosis [46]. It has
also been suggested that flavonoid compounds can affect protein kinase C (PKC) activities [47]. It is,
thus, plausible that LioRGJ, at higher doses, may cause cell death via the caspase-3-mediated apoptotic
pathway by activating PKC isoforms that induce apoptosis (e.g., PKC-δ) or inhibit isoforms that protect
against apoptosis (e.g., PKC-ε) [47]. Interestingly, Tenore et al. [10] also assessed the potential protec-
tive effects of RGJ on cardiac-induced oxidative stress. It is well known that the clinical use of anthra-
cyclines, especially doxorubicin, in the treatment of many neoplastic diseases is limited by acute and
chronic dose-related, cumulative, and essentially irreversible cardiotoxicities. The available laboratory
evidence shows that doxorubicin and its metabolites induce generation of ROS that could interfere with
iron metabolism and trigger the intrinsic mitochondria-dependent apoptotic pathway in cardiomyocytes
[48,49]. Nevertheless, to date, only few single chemicals or raw extracts have proven to be able to reduce
the deleterious action of doxorubicin [50,51], but no experiments with whole grape juice have been con-
ducted. In this regard, H9C2 cardiomyocytes were exposed to 1 μM doxorubicin and a combination of
doxorubicin and different doses of LioRGJ for 72 h (Table 22.3). Sample aliquot of 0.01 µg provided an
appreciable radical scavenging activity as indicated by the decrease in the free radical levels (especially
ROS species, about 31%) and the unchanged antioxidant defense system activity. Interestingly, the asso-
ciation of doxorubicin with higher LioRGJ doses (from 0.01 to 0.05 µg) led to the enhancement of cardiac
cell oxidative stress, probably due to sample pro-oxidant effects, as indicated mainly by the increase in
RNS and antioxidant enzyme levels.

Grape juice, especially RGJ, appears as a concentrated source of polyphenolic compounds that may
reasonably be regarded as a useful tool for an effective antioxidant protection through diet. European
Legislation may take into account the possibility of including these beverages into the category of novel
foods with healthy properties [52].
Experimental results have shown a good antioxidant stability of RGJ to lyophilization that may be
regarded as a suitable process for the formulation of food supplements [10]. In fact, LioRGJ may be
proposed for the formulation of antioxidant nutraceutical products that could effectively allow for
the prevention and care of CVD induced by ROS. Particularly, these LioRGJ-based nutraceutical prod-
ucts may be used in support of a well-balanced dietary regimen and proper physical activity in order
to optimize health and individuals under doxorubicin therapy, in order to contrast the pharmacological
side effects, consisting, mainly in cardiovascular oxidative stress. However, it is difficult to draw direct
comparison of the aforementioned in vitro studies with animal models. The blood levels of antioxidants
from LioRGJ have not been measured in these experiments, and it would be difficult to extrapolate what
sort of oral dosage would be required to achieve the equivalent levels of such antioxidants that cells in
our experiments were exposed to. Thus, further studies are needed to optimize its dosage in order to
avoid harmful pro-oxidant effects.
22.6 Conclusion
Scientific literature indicates grape juice, mainly RGJ, as a functional beverage for protection of human
organism in terms of prevention and care from oxidative stress–related pathologies. In fact, despite its
low nutritional profile, RGJ would ensure the consumption of up to 10-fold higher polyphenolics present
in an equivalent volume of the same grape variety of wine. Nutraceutical applications may allow formu-
lation of pharmaceuticals based on LioRGJ in order to optimize conditions of healthy people through
diet and/or via pharmacological therapies by contrasting specific drug side effects due to oxidative stress
induced to cardiovascular system. Further studies are needed in order to establish the appropriate dosage
to avoid pro-oxidant effects.
REFERENCES
1. Dani, C., Oliboni, L.S., Vanderlinde, R., Bonatto, D., Salvador, M., and Henriques, J.A.P., Phenolic con-
tent and antioxidant activities of white and purple juices manufactured with organically or c onventionally
produced grapes. Food Chem. Toxicol., 45, 2574–2580, 2007.
2. Dávalos, A., Bartolomé, B., and Gómez-Cordovés, C., Antioxidant properties of commercial grape
juices and vinegars. Food Chem., 93, 325–333, 2005.
3. Bermúdez-Soto, M.J. and Tomás-Barberán, F.A., Evaluation of commercial red fruit juice concentrates
as ingredients for antioxidant functional juices. Eur. Food Res. Technol., 219, 133–141, 2004.
4. Frankel, E.N., Bosanek, C.A., Meyer, A.S., Silliman, K., and Kirk, L.L., Commercial grape juices inhibit
the in vitro oxidation of human low-density lipoproteins. J. Agric. Food Chem., 46, 834–838, 1998.
5. Felice, F., Zambito, Y., Di Colo, G., D’Onofrio, C., Fausto, C., Balbarini, A., and Di Stefano, R., Red
grape skin and seeds polyphenols: Evidence of their protective effects on endothelial progenitor cells
and improvement of their intestinal absorption. Eur. J. Pharm. Biopharm., 80, 176–184, 2012.
6. Bhale, S. and Sonawane, N., Anti-inflammatory potential of alcoholic extract of resveratrol from fruits
of Vitis vinifera Linn. Int. J. Res. Pharm. Sci., 2, 272–275, 2011.
7. Kleinedler, J.J., Pjescic, I., Bullock, K.K., Khaliq, A., Foley, J.D., and Dugas, T.R., Arterial pharma-
cokinetics of red wine polyphenols: Implications for novel endovascular therapies targeting restenosis.
J. Pharm. Sci., 101, 1917–1931, 2012.
8. U.S. Department of Agriculture (USDA), National Nutrient Database for Standard Reference, Release
27, 2014. Published online at: http://ndb.nal.usda.gov (accessed January 19, 2015).
9. Spayd, S.E., Tarara, J.M., Mee, D.L., and Ferguson, J.C., Separation of sunlight and temperature effects
on the composition of Vitis vinifera cv. Merlot berries. Am. J. Enol. Vitic., 53, 171–182, 2002.
10. Tenore, G.C., Manfra, M., Stiuso, P., Coppola, L., Russo, M., Gomez Monterrey, I.M., and Campiglia,
P., Antioxidant profile and in vitro cardiac radical-scavenging versus pro-oxidant effects of commercial
red grape juices (Vitis vinifera L. cv. Aglianico N.). J. Agric. Food Chem., 60, 9680–9687, 2012.
11. Miyagi, Y., Miwa, K., and Inoue, H., Inhibition of human low-density lipoprotein oxidation by flavo-
noids in red wine and grape juice. Am. J. Cardiol., 80, 1627–1631, 1997.
12. Landbo, A.K. and Meyer, A.S., Ascorbic acid improves the antioxidant activity of European grape
juices by improving the juices’ ability to inhibit lipid peroxidation of human LDL in vitro. Int. J. Food
Sci. Technol., 36, 727–735, 2001.
13. Bub, A., Watzl, B., Heeb, D., Rechkemmer, G., and Briviba, K., Malvidin-3-glucoside bioavailabil-
ity in humans after ingestion of red wine, dealcoholized red wine and red grape juice. Eur. J. Nutr.,
40, 113–120, 2001.
14. Monagas, M., Nunez, V., Bartolome, B., and Gomez-Cordoves, C., Anthocyanin-derived pigments
in Graciano, Tempranillo, and Cabernet Sauvignon wines produced in Spain. Am. J. Enol. Vitic.,
54, 163–169, 2003.
Grape Juice 279
15. Hyun, J.W. and Chung, H.S., Cyanidin and malvidin from Oryza sativa cv. Heugjinjubyeo mediate
cytotoxicity against human monocytic leukemia cells by arrest of G(2)/M phase and induction of apop-
tosis. J. Agric. Food Chem., 52, 2213–2217, 2004.
16. Hassimotto, N.M.A. and Lajolo, F.M., Antioxidant status in rats after long-term intake of anthocyanins
and ellagitannins from blackberries. J. Sci. Food Agric., 91, 523–531, 2011.
17. Prior, R.L., Absorption and metabolism of anthocyanins: Potential health effects, in Phytochemicals:
Mechanisms of Action, Meskin, M., Bidlack, W.R., Davies, A.J., Lewis, D.S., and Randolph. R.K., Eds.,
CRC Press / Taylor & Francis Group, Boca Raton, FL, 2004, pp. 1–19.
18. Rastija, V., Srečnik, G., and Medić-Šarić, M., Polyphenolic composition of Croatian wines with differ-
ent geographical origins. Food Chem., 115, 54–60, 2009.
19. Talcott, S.T. and Lee, J.H., Ellagic acid and flavonoid antioxidant content of Muscadine wine and juice.
J. Agric. Food Chem., 50, 3186–3192, 2002.
20. Gawel, R., Red wine astringency: A review. Aust. J. Grape Wine Res., 4, 74–95, 1998.
21. Castillo-Muñoz, N., Gómez-Alonso, S., García-Romero, E., and Hermosín-Gutiérrez, I., Flavonol pro-
files of Vitis vinifera red grapes and their single-cultivar wines. J. Agric. Food Chem., 55, 992–1002,
2007.
22. Boots, A.W., Wilms, L.C., Swennen, E.L., Kleinjans, J.C., Bast, A., and Haenen, G.R., In vitro and
ex vivo anti-inflammatory activity of quercetin in healthy volunteers. Nutrition, 24, 703–710, 2008.
23. Perez-Vizcaino, F., Duarte, J., Jimenez, R., Santos-Buelga, C., and Osuna, A., Antihypertensive effects
of the flavonoid quercetin. Pharmacol. Rep., 61, 67–75, 2009.
24. Shan, B.E., Wang, M.X., and Li, R.Q., Quercetin inhibits human SW480 colon cancer growth in associa-
tion with inhibition of cyclin D1 and survivin expression through Wnt/beta-catenin signaling pathway.
Cancer Invest., 27, 604–612, 2009.
25. Zhang, W. and Zhang, F., Effects of quercetin on proliferation, apoptosis, adhesion and migration, and
invasion of HeLa cells. Eur. J. Gynaecol. Oncol., 30, 60–64, 2009.
26. Stervboa, U., Vanga, O., and Bonnesenb, C., A review of the content of the putative chemopreventive
phytoalexin resveratrol in red wine. Food Chem., 101, 449–457, 2007.
27. Eybl, V., Kotyzova, D., and Koutensky, J., Comparative study of natural antioxidants—curcumin, resve-
ratrol and melatonin—in cadmium-induced oxidative damage in mice. Toxicology, 225, 150–156, 2006.
28. Brito, P.M., Mariano, A., Almeida, L.M., and Dinis, T.C., Resveratrol affords protection against
peroxynitrite-mediated endothelial cell death: A role for intracellular glutathione. Chem. Biol. Interact.,
15, 157–166, 2006.
29. Wang, Y., Romigh, T., He, X., Orloff, M.S., Silverman, R.H., Heston, W.D., and Eng, C., Resveratrol
regulates the PTEN/AKT pathway through androgen receptor-dependent and -independent mechanisms
in prostate cancer cell lines. Hum. Mol. Genet., 19, 4319–4329, 2010.
30. Hsieh, T.C., Uptake of resveratrol and role of resveratrol-targeting protein, quinone reductase 2, in
normally cultured human prostate cells. Asian J. Androl., 11, 653–661, 2009.
31. Leifert, W.R. and Abeywardena, M.Y., Grape seed and red wine polyphenol extracts inhibit cellular
cholesterol uptake, cell proliferation, and 5-lipoxygenase activity. Nutr. Res., 28, 842–850, 2008.
32. Saija, A., Scalese, M., Lanza, M., Marzullo, D., Bonina, F., and Castelli, F., Flavonoids as antioxidant
agents: Importance of their interaction with biomembranes. Free Radic. Biol. Med., 19, 481–486, 1995.
33. O’Byrne, D.J., Devaraj, S., Grundy, S.M., and Jialal, I., Comparison of the antioxidant effects of Concord
grape juice flavonoids and α-tocopherol on markers of oxidative stress in healthy adults. Am. J. Clin.
Nutr., 76, 1367–1374, 2002.
34. Sánchez-Moreno, C., Larrauri, J.A., and Saura-Calixto, F., Free radical scavenging capacity and inhi-
bition of lipid oxidation of wines, grape juices and related polyphenolic constituents. Food Res. Int.,
32, 407–412, 1999.
35. Urquiaga, I. and Leighton, F., Wine and health: Evidence and mechanisms. World. Rev. Nutr. Diet.,
95, 122–139, 2005.
36. Fernandéz-Pachón, M.S., Villaño, D., Troncoso, A.M., and García-Parrilla, M.C., Determination of the
phenolic composition of sherry and table white wines by liquid chromatography and their relation with
antioxidant activity. Anal. Chim. Acta, 563, 101–108, 2006.
37. Das, D.K. and Maulik, N., Protection against free radical injury in the heart and cardiac performance,
in Exercise and Oxygen Toxicity, Sen, C.K., Packer, L., and Hanninen, O., Eds., Elsevier Science,
Amsterdam, the Netherlands, 1995, 359–388.
38. Arroyo, C.M., Kramer, J.H., Dickens, B.F., and Weglicki, W.B., Identification of free radicals in myo-
cardial ischemia/reperfusion by spin trapping with nitrone DMPO. FEBS Lett., 221, 101–104, 1987.
39. Vanden Hoek, T.L., Shao, Z.H., Li, C.Q., Schumacker, P.T., and Becker, L.B., Mitochondrial electron
transport can become a significant source of oxidative injury in cardiomyocytes. J. Mol. Cell. Cardiol.,
29, 2441–2450, 1997.
40. Shao, Z.-H., Becker, L.B., Vanden Hoek, T.L., Schumacker, P.T., Li, C.-Q., Zhao, D., Wojcik, K. et al.,
Grape seed proanthocyanidin extract attenuates oxidant injury in cardiomyocytes. Pharmacol. Res.,
47, 463–469, 2003.
41. Shao, Z.-H., Wojcik, K.R., Dossumbekova, A., Hsu, C., Mehendale, S.R., Li, C.-Q., Qin, Y. et al., Grape
seed proanthocyanidins protect cardiomyocytes from ischemia and reperfusion injury via Akt-NOS
signaling. J. Cell. Biochem., 107, 697–705, 2009.
42. Baydar, N.G., Özkan, G., and Yaşar, S., Evaluation of the antiradical and antioxidant potential of grape
extracts. Food Control, 18, 1131–1136, 2007.
43. Shao, Z.-H., Vanden Hoek, T.L., Xie, J., Wojcik, K., Chan, K.C., Li, C.-Q., Hamann, K. et al., Grape
seed proanthocyanidins induce pro-oxidant toxicity in cardiomyocytes. Cardiovasc. Toxicol., 3,
331–339, 2003.
44. Shao, Z.H., Hsu, C.W., Chang, W.T., Waypa, G.B., Li, J., Li, D., Li, C.Q. et al. Cytotoxicity induced by
grape seed proanthocyanidins: Role of nitric oxide. Cell. Biol. Toxicol., 22, 149–158, 2006.
45. Lluís, L., Muñoz, M., Nogués, M.R., Sánchez-Martos, V., Romeu, M., Giralt, M., Valls, J., and Solà, R.,
Toxicology evaluation of a procyanidin-rich extract from grape skins and seeds. Food Chem. Toxicol.,
49, 1450–1454, 2011.
46. Thornberry, N.A. and Lazebnik, Y., Caspases: Enemies within. Science, 281, 1312–1316, 1998.
47. Picq, M., Dubois, M., Munari-Silem, Y., Prigent, A.F., and Pacheco, H., Flavonoid modulation of protein
kinase C activation. Life Sci., 44, 1563–1571, 1989.
48. Keizer, H.G., Pinedo, H.M., Schuurhuis, G.J., and Joenje, H., Doxorubicin (Adriamycin): A critical
review of free radical-dependent mechanisms of cytotoxicity. Pharmacol. Ther., 47, 219–231, 1990.
49. van Acker, F.A.A., Boven, E., Kramer, K., Haenen, G.R.M.M., Bast, A., and van der Vijgh, W.J.,
Frederine, a new and promising protector against doxorubicin induced cardiotoxicity. Clin. Cancer
Res., 7, 1378–1384, 2001.
50. Chang, W.T., Li, J., Haung, H.H., Liu, H., Han, M., Ramachandran, S., Li, C.-Q. et al., Baicalein protects
against doxorubicin-induced cardiotoxicity by attenuation of mitochondrial oxidant injury and JNK
activation. J. Cell. Biochem., 112, 2873–2881, 2011.
51. Du, Y. and Lou, H., Catechin and proanthocyanidin B4 from grape seeds prevent doxorubicin-induced
toxicity in cardiomyocytes. Eur. J. Pharmacol., 591, 96–101, 2008.
52. Regulation (EC), Regulation No. 258/97 of the European Parliament and of the Council of 27
January 1991 concerning novel foods and novel food ingredients, European Commission, Brussels,
Belgium, 1991.
23
Grapefruit Juice
İncinur Hasbay
CONTENTS
23.1 Introduction.................................................................................................................................... 281
23.2 Nutritional Characteristics............................................................................................................. 282
23.3 Bioactives and Antioxidant Efficacy.............................................................................................. 283
23.4 Health Effects................................................................................................................................. 287
23.5 Novel Products/Formulations and Future Trends...........................................................................291
23.6 Conclusion..................................................................................................................................... 292
References............................................................................................................................................... 292
23.1 Introduction
Commercial processing of grapefruit as a juice started in the late l920s in order to expand the market
outlets for rapidly increasing production in Florida and Texas [1]. Grapefruit and its juice have been
popular worldwide for their refreshing flavor and beneficial health effects. Accordingly, consumption
and production of grapefruit in the world have increased significantly in the past two decades. The world
grapefruit production increased from ~4.5 million tons (MT) in 1987–1989 to ~6 MT in 2012–2013 [2,3].
In recent years, China has become the largest producer of grapefruit in the world, followed by the United
States and South Africa [2].
The highest consumption of grapefruit and its juice is in China and the United States, being commonly
served at breakfast [2,4]. In the United States, grapefruit juice is regularly consumed by about 21% of all
households [4]. It has been reported that about half of the total grapefruit production in the United States
is mainly processed into juice [2].
Grapefruit juice can generally be found in three different colors, red, pink, and white, depending on
the presence of lycopene in the fruit. Pigmented cultivars such as Ray Ruby, Star Ruby, and Rio Red have
become popular and are currently preferred over white grapefruit on the market [5], mainly due to their
high content of bioactive compounds such as vitamin C, limonoids, and carotenoids [6,7] in addition to
their pleasing red or pink color. Maintaining the color of these cultivars during commercial production is
a critical issue, since thermal processing, as well as storage of the juice, leads to the oxidation, isomeriza-
tion, and other chemical changes in lycopene and β-carotene [8]. The USDA has reported the importance
of color as a quality indicator in grapefruit juice. A grade scale from 0 to 100 points was developed to
ascertain the grade of grapefruit juice products such as regular juice, juice from concentrate, frozen
concentrated juice, concentrated juice for manufacturing, and dehydrated juice [9]. This chapter high-
lights the nutrient composition, phytochemical content, and antioxidant potential of grapefruit juice, the
preclinical and clinical evidences for its beneficial health effects, as well as the commercial products and
novel processing techniques.
281

Citrus fruits have long been regarded as one of the most important sources of vitamin C, exceeding the
minimum daily requirement of 60 mg in a serving (about 240 mL) of juice [10]. Grapefruit juice can
also be considered as a good source of folate, potassium [10–12], and vitamin A (particularly for pink
grapefruit juice) [12] (Table 23.1). Reference data for labeling purposes report that one cup (about 247 g)
of pink grapefruit juice provides 156% of the daily value for vitamin C, 22% for vitamin A, 11% for
potassium, and less than 10% for thiamin, folate, magnesium, calcium, niacin, and vitamin B6 [13]. The
nutrient density of pink grapefruit juice has been demonstrated to be the highest among the commonly
consumed fruit juices (apple, grape, pink grapefruit, white grapefruit, orange, pineapple, and prune
juices) [14].
The nutrient composition of grapefruit juice is affected by numerous factors including the raw material
properties such as variety, geographic location, climate, harvest time, maturity, and cultural practices
[6,15–17], as well as the processing factors, packaging, and storage conditions [6,18–20]. The red-col-
ored varieties are richer in vitamin C, as well as limonoids and carotenoids [6]. In addition to genetics,
environmental factors such as climate and harvest time also significantly affect the nutrient content of
grapefruits, especially vitamin C. Two independent research studies have demonstrated that grapefruits
grown in coastal areas of California generally had more vitamin C than those grown in desert areas of
TABLE 23.1
Compositional and Nutrient Characteristics of White and Pink Grapefruit Juices (per 100 g)
Nutrient White Grapefruit Juice Pink Grapefruit Juice
Water g 90.00 90.00
Energy kcal 39 39
Protein g 0.50 0.50
Lipid (fat) g 0.10 0.10
Total sugars g 9.10 na
Total dietary fiber g 0.1 na
Minerals
Calcium mg 9 9
Iron mg 0.20 0.20
Magnesium mg 12 12
Phosphorus mg 15 15
Sodium mg 1 1
Zinc mg 0.05 0.05
Vitamins
Niacin mg 0.200 0.200
Vitamin E (ATE) mg 0.22 na
Source: Adapted from the U.S. Department of Agriculture (USDA), National Nutrient Database for Standard
Abbreviations: DFE, dietary folate equivalents; RAE, retinol activity equivalents; ATE, alpha-tocopherol equiva-
lents; na, not available.
Grapefruit Juice 283
California and Arizona [15]. Grapefruits harvested in early season had higher vitamin C content than
those of their late season ones [16]. Cultural practices and inputs such as fertilizer, soil type, previous
crop, and mineral concentration can also significantly affect the nutritional quality of grapefruits and
hence the juice products. It has been reported in a comparative study that the ascorbic acid concentration
and sugar content were significantly higher in organic grapefruit juice, whereas the opposite was the case
for lycopene [17].
In addition to the aforementioned factors, processing and storage conditions play a critical role on
the nutritional content of grapefruit juice. Thermal processing during juice production may cause irre-
versible losses of nutritional quality and antioxidant activity [18,19]. Degradation of ascorbic acid can
take place by oxidation during processing or by anaerobic degradation during storage [20]. This may
cause quality problems such as browning as well as nutrient loss [20]. Ascorbic acid degradation in citrus
juice concentrates, including grapefruit juice, was shown to increase with increasing storage temperature
during an 8-week storage study at 28°C, 37°C, and 45°C [20]. On the other hand, vitamin C content of
the whole grapefruit was not affected that much during storage [20]. Minimal degradation of vitamin C
after storage at 9°C for 4 weeks was also reported in another study in which carotenoids experienced
significant level of degradation [6].

The major bioactive groups in grapefruit juice include flavonoids, carotenoids, limonoids, furanocouma-
rins, and organic acids [6,7,11] that primarily contribute to the sensory attributes such as flavor and color,
besides the health-promoting properties. White grapefruit juice has been reported to be slightly richer in
flavonoids than the pink and red ones [21], whereas the red-colored varieties have been demonstrated to
be richer in limonoids and carotenoids [6].
The major group of phenolic compounds responsible for the health-promoting properties of grapefruit
juice are flavonoids that act as free radical scavengers, inhibitors of cellular proliferation, promoting dif-
ferentiation, and modulating enzymatic activities such as tyrosine kinases [22,23] and have antibiotic,
antiallergenic, antidiarrhea, antiulcer, and anti-inflammatory activities [22]. It has been proposed that
biological effects of flavonoids are due to their structural similarity to adenosine triphosphate (ATP) that
provides these compounds to compete with ATP for binding at various enzymatic sites [23]. Structures
of flavonoid compounds commonly found in grapefruit juice are given in Figure 23.1.
Flavonoids in grape fruit juice are mainly of flavanone type, including a number of glycosides
of three main aglycones: naringenin (5,7,4-trihydroxyflavanone), hesperetin (4-methoxy-3,5,7-
trihydroxyflavanone), and eriodictyol (5,7,3,4-tetrahydroxyflavanone) [23]. Naringin (naringenin-7-O-
neohesperoside) is the major flavanone glycoside in grapefruit juice [21,24,25], which possesses a wide
range of biological activities such as antioxidant, antiulcer, and anti-inflammatory activities [26]. Despite
its unique health-promoting properties, naringin is the major compound responsible for the bitterness
in grapefruit juice. The acceptability of bitterness of naringin is related to genetic taste markers [27].
As a result of several consumer acceptance studies, it has been reported that naringin was detectable
by humans at concentrations below 19 µg/g [17]. Typical concentrations of naringin in grapefruit juice
(about 40 mg/100 mL) can still be acceptable by consumers in terms of bitterness [28]; however, juice
from early-season grapefruit contains excessive amounts of naringin (about 120 mg/100 mL) as well as
limonin that impart an extremely bitter flavor to the juice [10,28]. Narirutin (naringenin 7-O-rutinoside)
is another glycoside of naringenin that is present at high concentrations in grapefruit juice, although
less than naringin [21,29,30]. The aglycone, naringenin, is found in lower concentrations, although it
has been recognized to be a distinctive component of grapefruit juices [21]. Hesperidin (hesperetin-7-O-
rutinoside) and neohesperidin (hesperetin-7-O-neohesperidoside) are two glycosides of hesperetin with
proven antioxidant properties and potential beneficial health effects. Although their concentrations reach
a maximum of 2% of total flavanones in grapefruit juice, which are not as high as naringin and narirutin
[24], they contribute to the biological activities of the juice due to their anticarcinogenic, antioxidative,
and estrogenic properties [31]. Other flavonoids such as didymin, poncirin, and eriocitrin [7,21], as well
as quercetin [30] and its glycoside rutin [32], have also been detected in some grapefruit juices.
CH3 O OH
HO HO
O OH
O HO O OH
OH
HO O OH
OH O
HO OH OH
OH CH3
Rutinose Neohesperidose
Skeleton Flavanone R
OH Naringenin OH
(Aglycone)
R O
Narirutin Rutinoside
(Naringenin 7-O-rutinoside)
Naringin Neohesperidoside
OH O (Naringenin 7-O-neohesperidoside)
O Hesperetin OH
CH3
(Aglycone)
R O
OH Hesperidin Rutinoside
(Hesperetin-7-O-rutinoside)
Neohesperidin Neohesperidoside
OH O (Hesperetin 7-O-neohesperidoside)
OH Eriodictyol OH
(Aglycone)
R O
OH
Eriocitrin Rutinoside
(Eriodictyol-7-O-rutinoside)
Neoeriocitrin Neohesperidoside
OH O
(Eriodictyol-7-O-neohesperidoside)
O Isosakuranetin OH
CH3 (Aglycone)
R O
Didymin Rutinoside
(Isosakuranetin 7-O-rutinoside)
Poncirin Neohesperidoside
OH O (Isosakuranetin 7-O-neohesperidoside)
OH Quercetin OH
(Aglycone)
HO O
OH Rutin Rutinoside
(Quercetin 3-O-rutinoside)
R
OH O
FIGURE 23.1 Major flavonoid aglycones and their respective glycosides in grapefruit juice.
In addition to phenolic compounds, vitamin C has a considerable contribution to the antioxidant capac-
ity of grapefruit juice. Gardner et al. [33] assessed the relative contribution of vitamin C, carotenoids, and
phenolics to the antioxidant potential of grapefruit juice by determining the ability of each compound
to reduce potassium nitrosodisulfonate (Fremy’s radical), a synthetic free radical species. According
to their findings, the percentage contributions of vitamin C to the total antioxidant capacity were 89%
for white grapefruit juice and 77% for pink grapefruit juice, calculated from the relationship that one
molecule of vitamin C reduces 2.48 Fremy’s radical [33]. Vitamin C is known to be among the bioactive
compounds beneficial to human health due to its antiscorbutic properties as well as its strong antioxidant
activity [6], which may inhibit the development of major clinical conditions, including coronary heart
disease (CHD) and certain cancers [33].
Limonoids are another bioactive group that contributes substantially to the sensory properties as well
as health-promoting effects of grapefruit juice. Citrus limonoids were shown to inhibit the formation of
chemically induced neoplasia in the oral cavity, forestomach, small intestine, colon, lung, and skin of
laboratory animals. However, their antioxidant activities were reported to be low as compared to the fla-
vonoids [22]. Limonin and nomilin (Figure 23.2) are the two major limonoid aglycones in grapefruit juice
that have been demonstrated to have anticarcinogenic properties [6]. In addition to the aglycone forms,
glycosides such as deacetyl nomilinicacid glucoside (DNAG) have also been determined in grapefruit
juice [7]. Limonin is responsible for the so-called delayed bitterness of citrus juices. A tasteless limonin
precursor, released when fruit tissue is damaged, is gradually converted to limonin, which results in bit-
terness [28]. As in the case of naringin, bitterness due to limonoids also involves a major problem for the
consumer acceptance and the citrus industry. A wide variety of patented techniques have been developed
to remove or adsorb excess naringin and limonin from citrus juices [28].
Among carotenoids, lycopene and β-carotene are found in grapefruit juice in varying amounts [6,7].
They contribute to both the flesh color and the health-promoting effects. Lycopene is known to be more
potent antioxidant than other carotenoids and is reported to be a highly efficient quencher of singlet oxy-
gen. In addition to lycopene, the role of β-carotene to provide both color and provitamin A activity has
been reported as an important functionality of some types of grapefruit juices [34].
Coumarins, such as bergamottin, 6′,7′-dihydroxybergamottin, 5-gernyloxy-7-methoxycoumarin
(5-G-7-MC), and paradisin A, have been identified in grapefruit juice [7,11] (Figure 23.3). These com-
pounds are rather known with their adverse effects on individuals using certain types of drugs. In 1991,
it was discovered that grapefruit juice intake is associated with interactions with some commonly used
drugs [35], which was later attributed to the presence of furanocoumarins in grapefruit [36]. Studies on
these compounds have been focused on the mechanism of interaction and adverse effects on individuals
using certain types of drugs. Nevertheless, there are a number of reports on bioactive properties of some
coumarins such as anticarcinogenic and antithrombotic activities [22], as well as inhibition of formation
O
O
O
O CH3
CH3
H3C
O O O O
CH3 CH3 CH3
O O
O O O O
H3C O O O
H3C H3C CH3
Limonin Nomilin
FIGURE 23.2 Major limonoids found in grapefruit juice.
CH3
CH3
O O O
CH3
O
CH3 H3C CH3
O HO O
H3C
HO
O O O O H3C CH3
O O CH3
Bergamottin 5-G-7-MC 6΄,7΄-Dihydroxybergamottin
FIGURE 23.3 Major coumarins found in grapefruit juice.

of biofilms that are associated with over 80% of all microbial infections related to the urinary tract,
catheters, dental plaque, and gingivitis [37]. Furanocoumarins from grapefruit have also been shown
to inhibit the activities of some autoinducers (N-acylhomoserine lactone and a furanosyl borate diester
molecule) that enable bacterial cells to control and coordinate gene expression [37]. Limited data are
available on the antioxidant activities of grapefruit coumarins. The antioxidant activity of bergapten, a
coumarin from grapefruit, was reported to be low as compared to flavonoids that contain a chromanol
ring system [22].
As in most of the citrus fruits, phenolic acids exist in grapefruits either in methanol-soluble ester-
bound form (soluble phenolic esters) or in methanol-soluble glycoside-bound form (soluble phenolic gly-
cosides) as well as in the free forms. Ferulic acid was determined to be the major component, followed
by sinapic, p-coumaric, caffeic, and vanillic acids [38].
Quantitative values of the major bioactives reported in the literature (Table 23.2) show variations
mainly due to methodologies used for sample preparation and analyses [39], as well as to the varia-
tions of these compounds according to the preharvest [6,7,11,17] and postharvest conditions [7,11,40,41].
The preharvest factors such as cultivar, season, and location [11], as well as cultural practices such as
fertilization and use of pesticide [6], have been reported to be the major ones. Research findings on the
effect of organic versus conventional production on the concentration of bioactive compounds in whole
TABLE 23.2
Ranges of Bioactive Compounds in Commercial and Laboratory-Prepared Grapefruit Juices
Reported in the Literature (mg/100 mL)
Compound Commercial Juices Laboratory-Prepared Juices References
Flavanones
Didymin 0.04–1.72 0.43 [7,30]
Eriocitrin na 0.27–0.54 [21]
Hesperidin 0.24–3.12 1.79 [30,32]
Naringin 6.69–63.8 4.46–47.6 [7,23,30,32,38,44,70]
Narirutin 2.25–12.20 7.01–9.40 [7,23,30,38]
Neoeriocitrin na 0.30–0.33 [21]
Neohesperidin 0.04–1.12 0.79–26.52 [7,23,30,38]
Poncirin 0.12–2.36 0.61 [7,23,30]
Flavonols
Rutin na 3.26 [32]
Aglycons
Hesperetin na 0.74 [32]
Naringenin nd nd–1.29 [32,70]
Quercetin 0.20–0.88 0.17–0.21 [21,23,30]
Coumarins
5-G-7-MC na 0.38 [7]
6′,7′-Dihydroxybergamottin 0.022–5.25 0.14–1.03 [7,11,44]
Bergamottin 0.26–3.63 0.09–0.31 [7,11,44]
Phenolic Acids
Caffeic acid 0.023 0.254 [38,72]
Chlorogenic acid 0.085 na [72]
Ferulic acid 0.015 1.113 [38,72]
p-Coumaric acid 0.007 0.260 [38,72]
p-Hydroxybenzoic acid 0.007 0.107 [38,72]
Protocatechuic acid 0.012 0.071 [38,72]
Sinapic acid na 0.388 [38]
Vanillic acid na 0.286 [38]
Abbreviations: 5-G-7-MC, 5-gernyloxy-7-methoxycoumarin; na, not available; nd, not detected.
grapefruit and its juice have demonstrated that the concentrations of vitamin C, nomilin, naringin, and
neohesperidin were higher in organically produced grapefruit juice, whereas the concentrations of lyco-
pene, β-carotene, bergamottin, and 6′,7′-dihydroxybergamottin along with its dimers were significantly
higher in conventional juices [6,17]. The higher content of some bioactive compounds in organic grape-
fruits was explained as the increased level of phytoalexins as a response to biotic stress, which would
lead to the increase in bioactive compounds like limonoids that are known to be antifeedants that are
primarily produced by plants as a response to pests and diseases [6].
Postharvest conditions such as processing, storage, and packaging also affect the quantity of bioac-
tive compounds in grapefruit juice. Significant decreases in narirutin, hesperidin, and total flavonoid
contents were reported in grapefruit juices stored at 4°C for 10 and 15 days [40]. In a comparative study
that involved the effect of postharvest conditions on the level of furanocoumarins in grapefruit juice,
it was reported that hand-squeezed juice contained higher contents of 6′,7′-dihydroxybergamottin and
bergamottin as compared to the processed juice [11]. Moreover, levels of furocoumarins decreased in
the juices with the progress of storage, and the decrease was higher in the ones stored at 24°C than those
stored at 9°C [11]. In the same study, it was found that the amount of furocoumarins were higher in juices
packed in cartoon containers than those in cans and cardboard containers [11]. A survey study conducted
to determine the variations in the content of furanocoumarin in commercial grapefruit juices produced
in the United States indicated that storage temperature affected the amount of furanocoumarins in com-
mercial products [41]. Within the survey, a total of 29 white and 29 red commercial grapefruit juice
samples were collected over two seasons as pasteurized juices or as concentrates to be reconstituted into
juice. All but one of the red juice types were refrigerated products, whereas 11 of the white grapefruit
juices were shelf-stable or nonrefrigerated products. The authors reported that shelf-stable juices had sig-
nificantly lower amounts of 6′,7′-dihydroxybergamottin, but higher amounts of bergamottin [41]. In the
same study, it was also reported that concentration, pasteurization, and squeeze pressure settings did not
have any significant effect on the furanocoumarin content, although extreme evaporator and pasteuriza-
tion temperatures were not tested [41].
Until recently, a number of studies available reported data on the bioavailability of flavonoids after
grapefruit juice intake. Analytical methods allowing the analysis of naringenin and hesperetin from
human plasma and urine were first developed by Erlund et al. [31], who consequently reported that nar-
ingenin and hesperetin were bioavailable from the studied juices. The resulting plasma concentrations
after single ingestion of orange juice or grapefruit juice (8 mL/kg body weight) were demonstrated to be
comparatively high (up to 4 mg/L or 15 μmol/L). The elimination half-lives were between 1.3 and 2.2 h,
which shows a relatively fast clearance from the general circulation. The concentration of flavanones
in plasma after ingestion was relatively high, which suggests that individuals consuming grapefruit or
orange juice could benefit from their considerable health effects [31]. In this latter study, interindividual
variation in bioavailability was remarkable, which has also been reported in a previous research study of
the same group [25]. Such variation has been suggested to be due to physiological (differences in body
weight, body composition, and gastric motility) and molecular factors (differences in the activity or
synthesis of transporters or enzymes involved in biotransformation) [25].
23.4 Health Effects

Grapefruit juice, like all other citrus fruits, has long been valued as a health-promoting drink due to
its high content of bioactive compounds such as vitamin C, folate, dietary fiber, carotenoids, and fla-
vonoids, which have been suggested to be responsible for the prevention of cancer and degenerative
diseases [42].
The major groups of compounds that contribute to the beneficial health effects of grapefruit juice
are flavonoids. A large number of studies exist on the in vitro and in vivo antioxidative, anticarcino-
genic, antiallergic, and anti-inflammatory effects of flavonoids, specifically naringin, naringenin, and
hesperidin [32]. Moreover, epidemiological studies have shown an inverse relationship between dietary
flavonoid intake and cardiovascular disease (CVD) [43]. Accordingly, grapefruit and its juice have been
labeled as “Hearth Healthy” by the American Heart Association (AHA) [4,44].
TABLE 23.3
Selected Preclinical Studies on Possible Health Effects of Grapefruit Juice
Possible Health Effects Experimental Model Possible Mode of Action References
Could be used as a Rats fed cholesterol- Reduced plasma lipids and increased [45]
valuable supplement for containing and cholesterol- antioxidant activity (red grapefruit juice
disease-preventing diets free diets preferable to naringin).
Cholesterol-lowering Mildly hypercholesterolemic Reduced VLDL + LDL cholesterol. [46]
effect rats
Cholesterol-lowering Rabbits with high LDL Reduced serum LDL cholesterol by 32% and [47]
effect cholesterol levels decreased cholesterol excretion by 48%.
Hepatoprotective in low Rats Improved liver histology at low doses of [48]
doses, but hepatotoxic grapefruit juice (200 mg/kg), but poor
in high doses histological profiles and increased liver
oxidative stress at high doses (400 and
600 mg/kg).
Antigenotoxic effect Mice treated with Dau Inhibition of the MNPE induced by Dau. [51]
Antigenotoxic effect Mice treated with BaP Inhibition of the MNPE induced by Dau BaP. [52]
AFB1 Rats given 5 mg/kg AFB1 by Reduced AFB1-induced DNA damage in the [59]
gavage liver.
Abbreviations: AFB1, antigenotoxic effect against aflatoxin B1; LDL, low-density lipoprotein; Dau, daunorubicin; BaP,
benzo(a)pyrene; VLDL, very-low-density lipoprotein; MNPEs, micronucleated polychromatic erythrocytes.
Preclinical studies demonstrating the positive health effects of grapefruit juice mainly focus on the
antigenotoxic, hepatoprotective, and cholesterol-lowering effects (Table 23.3). The in vitro and in vivo
research studies on the functionality of grapefruit juice are more scarce than those on the effects of pure
flavonoid compounds. However, in one of the preclinical studies, it was reported that fresh red grapefruit
juice was preferable to pure naringin in terms of lowering the plasma lipid levels and increasing plasma
antioxidant activity [45]. In this study, 42 male Wistar rats were randomly divided into six groups and fed
with basal diet, basal diet with naringin, grapefruit, cholesterol, cholesterol/naringin, and cholesterol/red
grapefruit juice. After 30 days of feeding, it was determined that diets supplemented with red grapefruit
juice and to a lesser degree with naringin improved the plasma lipid levels mainly in rats fed cholesterol
and increased the plasma antioxidant activity. In conclusion, the researchers suggested that naringin was
a powerful plasma lipid–lowering and plasma antioxidant activity increasing flavanone, but the effects of
fresh red grapefruit juice were higher [45].
In another preclinical study on the cholesterol-lowering effects of dietary citrus juices and their flavo-
noids, it was shown that dietary grapefruit juice tended to reduce very-low-density lipoprotein (VLDL) +
low-density lipoprotein (LDL) cholesterols in mildly hypercholesterolemic rats with chemically induced
breast cancer [46]. The same research group investigated whether orange juice and grapefruit juice
(double strength) could influence cholesterol metabolism in rabbits with high LDL cholesterol level
induced by feeding a semipurified, cholesterol-free, casein diet [47]. At the end of a 3-week intervention
period, it was revealed that replacing drinking water with either orange juice or grapefruit juice reduced
serum LDL cholesterol by 43% and 32%, respectively (P < 0.05) [47]. It was suggested that this was asso-
ciated with total liver cholesterol reduction in the orange juice group (−18%, P < 0.05) and with hepatic
cholesterol ester reduction in both juice groups (−42%, P < 0.05) [47].
As a result of a preclinical study on the hepatoprotective effect of grapefruit juice on rats, it was sug-
gested that high doses of grapefruit juice intake may cause possible hepatic derangement, whereas lower
doses were hepatoprotective [48]. In that study, three groups of Wistar rats were fed with 200, 400, and
600 mg/kg of grapefruit juice per day for 60 days. The control group was given 5 mL/kg daily, orally
of distilled water. The group fed with 200 mg/kg juice showed improved liver histology and a largely
preserved liver oxidative status. However, groups that were treated with both 400 and 600 mg/kg juice
exhibited poor histological profiles and increased evidence of liver oxidative stress [48].
Various grapefruit juice constituents have been shown to inhibit DNA damage induced by xenobiot-
ics. In recent years, the antigenotoxic potential of the juice itself has also been investigated in studies
that reveal the in vivo protection of grapefruit juice against mutagens with different mechanisms of
action [49–52]. The effect of grapefruit juice intake on 2-amino-1-methyl-6-phenylimidazo[4,5-b]
pyridine (PhIP)-induced colon DNA damage was determined by a comet assay in F344 rats given
60 mg/kg of PhIP by gavage [49]. The rats that were allowed free access to grapefruit juice for 5 days
experienced clearly reduced DNA damage in the colon to a 40% level of control rats [49]. The capacity
of grapefruit juice to prevent DNA damage induced by hydrogen peroxide (HP) in human lympho-
cytes was determined in another comet assay [50]. For that purpose, cells were exposed to HP for
5 min, washed, and treated with 0.1%, 0.5%, and 1% (v/v) grapefruit juice for 10–90 min. All three
concentrations of grapefruit juice were reported to decrease the damage caused by HP at different
levels depending on the concentration and the exposure time [50]. When the comet assay was applied
by incorporating formamidopyrimidine DNA glycosylase (an enzyme involved in the initial steps of
the DNA repair), the strongest increase of DNA damage by HP over the control level was experienced,
and the strongest reduction of such damage by grapefruit juice was determined [50]. In an acute assay
in mice, the capacity of grapefruit juice to inhibit the micronucleated polychromatic erythrocytes
(MNPEs) produced by daunorubicin (Dau), a genotoxic agent that produces cytogenetic damage and
DNA breaks in mammalian cells, was evaluated and the antioxidant potential of the juice in mouse
hepatic microsomes was determined [51]. Throughout the examined schedule (from 24 to 96 h), grape-
fruit juice produced no toxic or genotoxic damage, but generated a significant reduction of the MNPE
formed by Dau. With respect to the antioxidant potential of grapefruit juice, a 50% decrease in liver
microsomal lipid peroxidation produced by Dau was determined by quantifying malondialdehyde for-
mation. The researchers suggested that grapefruit juice had an efficient anticlastogenic potential prob-
ably related to its antioxidant capacity or to the alterations of Dau metabolism [51]. In a similar study,
the capacity of grapefruit juice in inhibiting the rate of MNPE was determined in mice treated with
benzo(a)pyrene (BaP), an environmental contaminant that is biotransformed by Cyp1a1 and is a strong
genotoxic agent [52]. As a result, it was suggested that the protective effect of grapefruit juice against
the genotoxicity of BaP may be related to the inhibition of Cyp1a1 enzyme activity [52]. Although the
antigenotoxic effect of grapefruit juice indicates its potential relevance as a chemopreventive agent, its
real protective effect in humans still requires clarification by means of well-planned preclinical and
clinical studies [50].
Citrus flavonoids are known to be effective inhibitors of human breast cancer cell proliferation in vitro,
especially in combination with quercetin [53]. Naringin and naringenin, as well as grapefruit juice and
orange juice concentrates were tested for their ability to inhibit the development of mammary tumors
induced by 7,12-dimethylbenz[a]anthracene (DMBA) in female Sprague–Dawley rats [53]. Two experi-
ments were conducted in which groups of 21 rats were fed a semipurified diet containing 5% corn oil
and were given a 5 mg dose of DMBA intragastrically at approximately 50 days of age while in diestrus.
One week later, in the first experiment, individual groups were given double-strength grapefruit juice or
orange juice or fed naringin or naringenin at levels comparable to those provided by the grapefruit juice.
In the second experiment, the rats were fed a semipurified diet containing 20% corn oil at that time. In
both experiments, tumor development was delayed in the groups given orange juice, rather than grape-
fruit juice. Naringin has also been reported to inhibit mammary carcinogenesis [53].
A number of clinical trials have also been reported on beneficial health effects of the grapefruit juice
or its extracts. The major positive effects reported included decreasing the serum cholesterol levels in
patients with hyperlipidemia and reducing arterial pressure [54–56]. The hypocholesterolemic effect
of grapefruit juice, as well as other citrus juices, in patients with hyperlipidemia has been reported in
a number of studies [54,55]. Jonsson and Ellegard [54] investigated the effect on serum lipid levels in
healthy adults rather than in patients and compared the effect of grapefruit juice with apple juice in the
same individuals. The results showed that the consumption of grapefruit juice decreased LDL choles-
terol by 6%, but with no significant differences compared with the apple juice or washout periods [54].
The antihypertensive effect of grapefruit juice has also been reported on normotensive and hyper-
tensive human [56]. Thus, the arterial pressure was measured at 5-, 10-, 20-, 30-, and 40-min intervals
after the intake of grapefruit juice, and also orange juice, cow milk, and vitamin C–supplemented bev-
erage as control. Grapefruit juice had a significant effect on decreasing the diastolic blood pressure,
systolic blood pressure, and mean arterial pressure compared with the other beverages used as controls.
It has been suggested that the active ingredients associated with this hypertensive effect of grapefruit
juice may be naringin and narirutin [56].
Despite all these potential health benefits of grapefruit juice, its consumption has been limited for
patients taking some groups of drugs since the early 1990s. It has been proven by a large number of
research findings that intake of grapefruit juice is associated with interactions with some commonly
used drugs as it alters oral drug pharmacokinetics by different mechanisms. This interaction was first
reported by Bailey et al. [35], who discovered that absorption of the cardiovascular drug felodipine
was significantly higher in patients who had consumed grapefruit juice. Further studies up to now have
shown the interaction effect of grapefruit juice with other types of drugs like cardiovascular drugs such
as calcium channel blockers (e.g., nifedipine), cholesterol-lowering drugs such as 3-hydroxy-3-methyl-
glutaryl-CoA reductase inhibitors (statins, e.g., simvastatin, lovastatin, and atorvastatin), angiotensin II
type 1 receptor antagonists (e.g., losartan), benzodiazepines (e.g., midazolam and triazolam), antihista-
mines (e.g., terfenadine), and immunosuppressants (e.g., cyclosporine) [57,58].
The major interference of grapefruit consumption with drugs has been reported as the increase in oral
drug bioavailability, for example, the potency of the therapeutic dose, causing overdose effects that result
in severe side effects. It has been reported that a single normal amount (e.g., 200–300 mL) of grapefruit
juice or whole fresh fruit causes irreversible inactivation of cytochrome P450 3A4 (CYP3A4), a cyto-
chrome P450 enzyme that is present in the liver and intestine [57,58]. Several lines of evidence suggest
that the major site of CYP3A4 inhibition by grapefruit juice is the intestine rather than the liver [36], and
the increased drug bioavailability can occur 24 h after juice consumption [58]. Several other mechanisms
of interactions of grapefruit with drugs have also been reported such as interactions with the efflux trans-
porter P-glycoprotein and organic anion transporting polypeptides [57,58].
Since the interaction effect has been observed only with grapefruit juice and not with orange juice,
early studies have suggested the predominant flavonoid naringin and its aglycone, naringenin, as poten-
tial inhibitors of CYP3A4-mediated metabolism [57]. However, further investigations confirmed that
bergamottin and dihydroxybergamottin were more potent inhibitors than naringin and naringenin [36].
Currently, more than 85 drugs have been reported or predicted to interact with grapefruit and its juice,
43 of which being known to possibly generate serious side effects [4]. Scientific data have shown that a
single usual intake (200–250 mL) of grapefruit juice or a whole grapefruit has the potency to cause the
inhibitory effect that may last as long as 24–48 h depending on the concentration of furanocoumarins
in the fruit [4]. Therefore, patients taking drugs susceptible for interaction have been suggested to forgo
grapefruit and grapefruit juice or to be treated with an alternate drug [4,57].
Despite those adverse effects on patients using the aforementioned types of drugs, it has been
reported that inhibition of intestinal CYP3A by grapefruit juice may also have positive outcomes
such as the antimutagenic ability against aflatoxin B1 (AFB1) in vivo, which could be attributed to the
inhibitory effect of grapefruit juice on CYP3A activity [50,52,59]. A comet assay has been applied
on F344 rats given 5 mg/kg AFB1 by gavage to examine the influence of grapefruit juice intake on
AFB1-induced liver DNA damage [59]. Rats treated with grapefruit juice extract (100 mg/kg per
os), as well as those allowed free access to grapefruit juice for 5 days prior to AFB1 administration,
resulted in clearly reduced DNA damage in the liver to 74% and 65% of the level in rats that did not
receive grapefruit juice, respectively. No significant differences in the portal blood and liver con-
centrations of AFB1 were observed between the rats that consumed grapefruit juice and the controls
[59]. In the same study, the capacity for metabolic activation of AFB1 in rat liver microsomes was
evaluated by an Ames assay using Salmonella typhimurium TA98. Accordingly, it was determined
that numbers of revertant colonies were significantly lower with liver microsomes prepared from rats
administered grapefruit juice compared with those of the control rats [59]. Microsomal testosterone
6β-hydroxylation was also lower with rats given grapefruit juice than with control rats. A significant
decrease in hepatic CYP3A content in microsomes of grapefruit juice–treated rats was seen as com-
pared to nontreated rats. In microsomal systems, grapefruit juice extract inhibited AFB1-induced
mutagenesis in the presence of a microsomal activation system from livers of humans as well as rats
[59]. These results suggest that grapefruit juice intake suppresses AFB1-induced liver DNA damage
in rats, which can be attributed to a decrease in the capacity for metabolic activation of AFB1 through
hepatic CYP3A activity [59].

Commercial grapefruit juice products can be found on the market for different types (white, pink, or
red), packages (glass, paper, plastic, or metal), and processed forms (reconstituted from concentrate
or not from concentrate, refrigerated, or shelf stable, with or without pulp) [30,41,60,61]. In addition to
the 100% of grapefruits are juice products, grapefruit is also manufactured as nectars (25%–50% juice
content) and still drinks (up to 25% juice content) [62,63] or blends with other fruit juices [62], mainly to
provide less expensive alternatives on the market [63] and to reduce the bitterness of grapefruit juice [26],
respectively. Although consumers tend to consume natural and 100% juices, nectars and drinks are often
preferred as cheaper alternatives [63]. Nectars, still drinks, and juice blends also have the advantage of
reduced dependency on fruit crops as a source of supply [62].
As mentioned earlier, thermal treatment of commercial grapefruit juice decreases the nutritional qual-
ity and bioactive compounds as well as the sensory properties that are valued by the consumers such
as flavor, color, and phase stability [64]. This has led to the search for new processing technologies that
cause minimum damage to the nutritional and sensory properties, such as high-pressure processing
(HPP), sonication, and the use of microwave energy.
HPP has long been demonstrated to inactivate pathogens and inhibit degradative enzymes, while pre-
venting the degradation of bioactive compounds [7]. This technique has already found applications in
the citrus juice industry in recent years, being more frequently applied for orange juice. The compara-
tive analysis of bioactive compounds in grapefruit juice processed by HPP and thermal processing was
first reported by Uckoo et al. [7]. It has been demonstrated that HPP treatment maintained the levels of
bioactive compounds such as ascorbic acid, flavonoids (naringin, narirutin, neohesperidin, didymin,
and poncirin), limonoids (limonin and DNAG), coumarins (bergamottin, dihydroxybergamottin, and
5-gernyloxy-7-methoxycoumarin), and carotenoids (all trans lycopenes and β-carotene). Accordingly,
the authors suggested that HPP technique could be applied for providing fresh-like grapefruit juice with-
out altering the levels of beneficial health bioactive compounds [7]. Although HPP has been suggested
as a nonthermal alternative to the thermal treatment of juices, it has been reported that some pectin
methylesterase (PME) isozymes that are responsible for the cloud loss are pressure stable and their inac-
tivation can often only be achieved by heating [65]. Accordingly, a combined high-pressure (low/mild)
heat treatment has been suggested, which can eliminate up to 80% of the total PME activity and hence
significantly limits the cloud-loss defect in grapefruit juices [65].
Another potential technique suggested for the prevention of cloud loss is sonication, which is a novel
technique applied for improving the quality of fruit juices, being economical due to reduced processing
time and energy input, as well as environmentally friendly [66]. Scientific results demonstrate that ultra-
sound alone is not sufficient to inactivate microorganisms reliably for food preservation and suggest that
ultrasound may be used in combination with other preservative compounds, electric fields, HHP, heat,
or combination of heat with moderate static pressure [67]. Sonication of the grapefruit juice samples
improves the cloud value, while increasing the total antioxidant capacity, 2,2-diphenyl-1-picrylhydrazyl
(DPPH) free radical scavenging activity, ascorbic acid, total phenolics, flavonoids, and flavonols [66].
The increase has been suggested to be due to the release of bound phenolic compounds as a result of cavi-
tation pressure exerted by sonication or due to the production of hydroxyl groups by sonication, which
can further bind to the aromatic ring of phenolic compounds [66].
Microwave processing has also been recommended as an alternative method for the pasteurization
of grapefruit juice [18,64]. This technique has long been demonstrated for its efficiency on microbial
inactivation. More recently, its effect on nutritional and sensory properties of grapefruit juice has been
investigated. In comparative studies, it has been shown that microwave treatment preserved the thermo-
labile nutrients such as ascorbic and citric acids [18], as well as the physical properties such as color,
particle size distribution, flow behavior, and density [64], while decreasing the PME activity as much as
the conventional treatment [18].
As mentioned previously, the bitterness of grapefruit juice, mainly due to naringin, is a major problem
in terms of sensory quality and consumer acceptance. To address this challenge, a number of processes
such as chemical treatments, physical separation processes, blending with nonbitter citrus juices and
sugars, and enzymatic treatment have been reported and applied until now [26]. Debittering of grapefruit
juice is most frequently carried out in the industry by adsorption onto macroporous resin beads or cross-
linked styrene divinylbenzene resins [68] or cellulose acetate [26]. During the past decade, consumer
demand for the maximum preservation of the health properties, as well as sensory and nutritional quali-
ties of the products, has led to the search for new technologies that cause minimum detrimental effects on
grapefruit juice [26]. Although physical debittering techniques have proven to exert no detrimental effect
on nutritional value [68,69], its effects on sensory quality such as loss of juice acidity, flavor, sweetness,
and turbidity have also been reported [70]. Accordingly, debittering, using the enzyme naringinase that
catalyzes the hydrolysis of naringin to naringenin, glucose, and rhamnose, has gained significance [69].
The industrial citrus fruit juice production results in the accumulation of large amounts of by-products
[29]. During grapefruit juice production, about half of the fruit is obtained as juice with the remainder
being by-product or waste that consists of peels, seeds, and segment membranes [71]. It has been sug-
gested that the peels be used for the production of ethanol and other products after the enzymatic hydro-
lysis [71], as well as production of molasses, pectin, essential oils, and limonene [29]. Most grapefruit
peel waste is dried and sold as a low-value cattle feed known as citrus pulp pellets [71]. In recent years,
citrus by-products have been suggested as major sources of phenolic compounds, since the peels have
been found to contain higher amounts of total phenolics compared to the edible portions. Grapefruit peel
is rich in flavonoids naringin and hesperidin [32]; this led to the development of new processing methods
for the utilization of by-products of grapefruit juice production. As a result of a comparative study on the
distribution of bioactive compounds between the juice and peel, it has been suggested as an important
source of naringin due to the considerable accumulation of this flavonoid in the peel [32]. Recently, more
scientific attention has been focused on the extraction of phenolic compounds from citrus peels with the
aim of being further used as natural antioxidants in foods instead of the synthetic products such as butyl-
ated hydroxyanisole and butylated hydroxytoluene [29].
23.6 Conclusion
Among citrus juices, grapefruit juice has been valued for its high vitamin C content for years. However,
recently, it has gained more attention due to its positive health effects such as antiatherosclerotic actions,
especially after the labeling of “heart healthy food” by AHA. The beneficial health effects of grapefruit
juice appear to be mainly derived from its flavanones, especially naringin, which also imparts a bitter
taste to the fruit. Furanocoumarins such as bergamottin and dihydroxybergamottin also contribute to
the functional properties of grapefruit juice. Preclinical studies on the positive health effects of grape-
fruit juice demonstrate its antigenotoxic, hepatoprotective, and cholesterol-lowering effects. A number
of clinical studies have also reported the beneficial health effects of grapefruit juice or extracts, such
as decreasing the cholesterol levels in patients with hyperlipidemia and reducing the arterial pressure.
In addition to these beneficial health effects of grapefruit juice, research has also focused on its adverse
effects on individuals using several medications after it was discovered about two decades ago that
grapefruit juice intake is associated with interactions with some commonly used drugs. Today, more
than 85 drugs have been reported to interfere with grapefruit juice, some of which can possibly gener-
ate serious side effects. Accordingly, patients taking these drugs have been advised to avoid consuming
grapefruit and its juice or be treated with an alternate drug.
REFERENCES
1. Shear, S.W., Spectacular rise of commercial fruit juice industry significant development in American
food business. Calif. Agric., 15, 2–15, 1961.
2. National Agricultural Marketing Council (NAMC), Beautiful country, beautiful fruit, South African
Fruit Trade Flow, 10. NAMC, Pretoria, South Africa, 2013.
3. Food and Agriculture Organization (FAO), Medium-term prospects for agricultural commodities:
Projections to the year 2010. FAO Commodities and Trade Technical Paper 1, FAO, Rome, Italy, 2003.
4. Agarwal, S.K., Grapefruit: A nutritional fruit fraught with danger of severe drug interactions.
Drug Discov., 3, 43–44, 2013.
5. Jayaprakasha, G.K., Girennavar, B., and Patil, B.S., Radical scavenging activities of Rio Red grapefruits
and Sour orange fruit extracts in different in vitro model systems. Bioresour. Technol., 99, 4484–4494,
2008.
6. Chebrolu, K.K., Jayaprakasha, G.K., Jifon, J., and Patil, B.S., Production system and storage tempera-
ture influence grapefruit vitamin C, limonoids, and carotenoids. J. Agric. Food Chem., 60, 7096–7103,
2012.
7. Uckoo, R.M., Jayaprakasha, G.K., Somerville, J.A., Balasubramaniam, V.M., Pinarte, M., and Patil,
B.S., High pressure processing controls microbial growth and minimally alters the levels of health
promoting compounds in grapefruit (Citrus paradisi Macfad.) juice. Innov. Food Sci. Emerg., 18, 7–14,
2013.
8. Lee, H.S. and Coates, G.A., Thermal pasteurization effects on color of red grapefruit juices. J. Food
Sci., 64, 663–666, 1999.
9. U.S. Department of Agriculture (USDA), United States Standards for Grades of Grapefruit Juice.
Federal Register, Washington, DC, 2012.
10. Bates, R.P., Morris, J.R., and Crandall, P.G., Principles and practices of small- and medium-scale fruit
juice processing. Agricultural Services Bulletin 146, FAO, Rome, Italy, 2001.
11. Girennavar, B., Jayaprakasha, G.K., and Patil, B.S., Influence of pre- and post-harvest factors and
processing on the levels of furocoumarins in grapefruits (Citrus paradisi Macfad.). Food Chem., 111,
387–392, 2008.
26. National Technical Information Service, USDA, Springfield, VA, 2013.
13. SELF nutrition data, grapefruit juice, pink, raw, 2014, published online at: http://nutritiondata.self.com/
facts/fruits-and-fruit-juices/2079/2 (accessed May 13, 2014).
14. Rampersaud, G.C., A comparison of nutrient density scores for 100% fruit juices. J. Food Sci., 72,
S261–S266, 2007.
15. Lee, S.K. and Kader, A.A., Preharvest and postharvest factors influencing vitamin C content of horti-
cultural crops. Postharvest Biol. Technol., 20, 207–220, 2000.
16. Lee, H.S. and Coates, G.A., Vitamin C contents in processed Florida citrus juice products from
1986–1995 survey. J. Agric. Food Chem., 45, 2550–2555, 1997.
17. Lester, G.E., Manthey, J.A., and Buslig, B.S., Organic vs conventionally grown Rio red whole grapefruit
and juice: Comparison of production inputs, market quality, consumer acceptance, and human health-
bioactive compounds. J. Agric. Food Chem., 55, 4474–4480, 2007.
18. Igual, M., García-Martínez, E., Camacho, M.M., and Martínez-Navarrete, N., Effect of thermal treat-
ment and storage on the stability of organic acids and the functional value of grapefruit juice. Food
Chem., 118, 291–299, 2010.
19. Igual, M., García-Martínez, E., Camacho, M.M., and Martínez-Navarrete, N., Changes in flavonoid con-
tent of grapefruit juice caused by thermal treatment and storage. Innov. Food Sci. Emerg., 12, 153–162,
2011.
20. Burdurlu, H.S., Koca, N., and Karadeniz, F., Degradation of vitamin C in citrus juice concentrates dur-
ing storage. J. Food Eng., 74, 211–216, 2006.
21. Gattuso, G., Barreca, D., Gargiulli, C., Leuzzi, U., and Caristi, C., Flavonoid composition of citrus
22. Yu, J., Wang, L., Walzem, R.L., Miller, E.G., Pike, L.M., and Patil, B.S., Antioxidant activity of citrus
limonoids, flavonoids, and coumarins. J. Agric. Food Chem., 53, 2009–2014, 2005.
23. Vanamala, J., Reddivari, L., Yoo, K.S., Pike, L.M., and Patil, B.S., Variation in the content of bioactive
flavonoids in different brands of orange and grapefruit juices. J. Food Compos. Anal., 19, 157–166,
2006.
24. De Lourdes Mata Bilbao, M., Andrés-Lacueva, C., Jáuregui, O., and Lamuela-Raventós, R.M.,
Determination of flavonoids in a citrus fruit extract by LC-DAD and LC-MS. Food Chem., 101,
1742–1747, 2007.
25. Erlund, I., Review of the flavonoids quercetin, hesperetin, and naringenin. Dietary sources, bioactivities,
bioavailability, and epidemiology. Nutr. Res., 24, 851–874, 2004.
26. Cavia-Saiz, M., Muñiz, P., Ortega, N., and Busto, M.D., Effect of enzymatic debittering on antioxidant
capacity and protective role against oxidative stress of grapefruit juice in comparison with adsorption
on exchange resin. Food Chem., 125, 158–163, 2011.
27. Drewnowski, A., Henderson, S.A., and Shore, A.B., Taste responses to naringin, a flavonoid, and the
acceptance of grapefruit juice are related to genetic sensitivity to (6)-n-propylthiouracil. Am. J. Clin.
Nutr., 66, 391–397, 1997.
28. Drewnowski, A. and Gomez-Carneros, C., Bitter taste, phytonutrients, and the consumer: A review.
Am. J. Clin. Nutr., 72, 1424–1435, 2000.
29. Khan, M.K., Zill-E-Huma, and Dangles, O., A comprehensive review on flavanones, the major citrus
polyphenols. J. Food Compos. Anal., 33, 85–104, 2014.
30. Ross, S.A., Ziska, D.S., Zhao, K., and Elsohly, M.A., Variance of common flavonoids by brand of grape-
fruit juice. Fitoterapia, 71, 154–161, 2000.
31. Erlund, I., Meririnne, E., Alfthan, G., and Aro, A., Plasma kinetics and urinary excretion of the flava-
nones naringenin and hesperetin in humans after ingestion of orange juice and grapefruit juice. J. Nutr.,
131, 235–241, 2001.
32. Wu, T., Guan, Y., and Ye, J., Determination of flavonoids and ascorbic acid in grapefruit peel and juice
by capillary electrophoresis with electrochemical detection. Food Chem., 100, 1573–1579, 2007.
33. Gardner, P.T., White, T.A.C., McPhail, D.B., and Duthie, G.G., The relative contributions of v itamin C,
carotenoids and phenolics to the antioxidant potential of fruit juices. Food Chem., 68, 471–474, 2000.
34. Lee, H.S., Characterization of carotenoids in juice of red navel orange (Cara Cara). J. Agric. Food
Chem., 49, 2563–2568, 2001.
35. Bailey, D.G., Spence, J.D., Munoz, C., and Arnold, J.M.O., Interaction of citrus juices with felodipine
and nifedipine. Lancet, 337, 268–269, 1991.
36. Eagling, V.A., Profit, L., and Back, D.J., Inhibition of the CYP3A4-mediated metabolism and
P-glycoprotein-mediated transport of the HIV-1 protease inhibitor saquinavir by grapefruit juice com-
ponents. Br. J. Clin. Pharmacol., 48, 543–552, 1999.
37. Girennavar, B., Cepeda, M.L., Soni, K.A., Vikram, A., Jesudhasan, P., Jayaprakasha, G.K., Pilla, S.D.,
and Patil, B.S., Grapefruit juice and its furocoumarins inhibits autoinducer signaling and biofilm forma-
tion in bacteria. Int. J. Food Microbiol., 125, 204–208, 2008.
38. Xu, G., Liu, D., Chen, J., Ye, X., Ma, Y., and Shi, J., Juice components and antioxidant capacity of citrus
varieties cultivated in China. Food Chem., 106, 545–551, 2008.
39. Bronner, W.E. and Beecher, G.R., Extraction and measurement of prominent flavonoids in orange and
grapefruit juice concentrates. J. Chromatogr. A, 705, 247–256, 1995.
40. Del Caro, A., Piga, A., Vacca, V., and Agabbio, M., Changes of flavonoids, vitamin C and antioxidant
capacity in minimally processed citrus segments and juices during storage. Food Chem., 84, 99–105,
2004.
41. Widmer, W. and Haun, C., Variation in furanocoumarin content and new furanocoumarin dimers in
commercial grapefruit (Citrus paradisi Macf.) juices. J. Food Sci., 70, C307–C312, 2005.
42. Ejaz, S., Ejaz, A., Matsuda, K., and Lim, C.W., Limonoids as cancer chemopreventive agents. J. Sci.
Food Agric., 86, 339–345, 2006.
43. Tripoli, E., La Guardia, M., Giammanco, S., Di Majo, D., and Giammanco, M., Citrus flavonoids:
Molecular structure, biological activity and nutritional properties: A review. Food Chem., 104, 466–479,
2007.
44. De Castro, W.V., Mertens-Talcott, S., Rubner, A., Butterweck, V., and Derendorf, H., Variation of fla-
vonoids and furanocoumarins in grapefruit juices: A potential source of variability in grapefruit juice–
drug interaction studies. J. Agric. Food Chem., 54, 249–255, 2006.
45. Gorinstein, S., Leontowicz, H., Leontowicz, M., Krzeminski, R., Gralak, M., Delgado-Licon, E., Ayala,
A.L.M., Katrich, E., and Trakhtenberg, S., Changes in plasma lipid and antioxidant activity in rats as a
result of naringin and red grapefruit supplementation. J. Agric. Food Chem., 53, 3223–3228, 2005.
46. Kurowska, E.M., Borradaile, N., Meade, M., Spence, J.D., and Carroll, K.K., Cholesterol-lowering
effects of dietary citrus juices and their flavonoids. Studies in rats, mice and rabbits. Atherosclerosis,
134, 330(Abstract), 1997.
47. Kurowska, E.M., Borradaile, N.M., Spence, J.D., and Carroll, K.K., Hypocholesterolemic effects of
dietary citrus juices in rabbits. Nutr. Res., 20, 121–129, 2000.
48. Saalu, L.C., Oyewopo, A.O., Enye, L.A., Ogunlade, B., Akunna, G.G., Bello, J., and Ogunmodede, O.S.,
The hepato-rejuvenative and hepatotoxic capabilities of Citrus paradise Macfad. fruit juice in Rattus
norvegicus. Afr. J. Pharm. Pharmaco., 6, 1056–1063, 2012.
49. Miyata, M., Takano, H., Takahashi, K., Sasaki, Y.F., and Yamazoe, Y., Suppression of 2-amino-1-
methyl-6-phenylimidazo[4,5-b]pyridine-induced DNA damage in rat colon after grapefruit juice intake.
Cancer Lett., 183, 17–22, 2002.
50. Razo-Aguilera, G., Baez-Reyes, R., Álvarez-González, I., Paniagua-Perez, R., and Madrigal-Bujaidar,
E., Inhibitory effect of grapefruit juice on the genotoxicity induced by hydrogen peroxide in human
lymphocytes. Food Chem. Toxicol., 49, 2947–2953, 2011.
51. Álvarez-González, I., Madrigal-Bujaidar, E., Martino-Roaro, L., and Espinosa-Aguirre, J.J.,
Antigenotoxic and antioxidant effect of grapefruit juice in mice treated with daunorubicin. Toxicol.
Lett., 152, 203–211, 2004.
52. Álvarez-González, I., Mojica, R., Madrigal-Bujaidar, E., Camacho-Carranza, R., Escobar-García, D.,
and Espinosa-Aguirre, J.J., The antigenotoxic effects of grapefruit juice on the damage induced by
benzo(a)pyrene and evaluation of its interaction with hepatic and intestinal Cytochrome P450 (Cyp)1a1.
Food Chem. Toxicol., 49, 807–811, 2011.
53. So, F.V., Guthrie, N., Chambers, A.F., Moussa, M., and Carroll, K.K., Inhibition of human breast cancer
cell proliferation and delay of mammary tumorigenesis by flavonoids and citrus juices. Nutr. Cancer,
26, 167–181, 1996.
54. Jonsson, C. and Ellegård, L., Grapefruit juice and serum lipids in healthy adults. Scand. J. Food Nutr.,
50, 118–123, 2006.
55. Gorinstein, S., Caspi, A., Libman, I., Lerner, H.T., Huang, D., Leontowicz, H., Leontowicz, M. et al.,
Red grapefruit positively influences serum triglyceride level in patients suffering from coronary athero-
sclerosis: Studies in vitro and in humans. J. Agric. Food Chem., 54, 1887–1892, 2006.
56. Díaz-Juárez, J.A., Tenorio-López, F.A., Zarco-Olvera, G., del Valle-Mondragón, L., Torres-Narváez,
J.C., and Pastelín-Hernández, G., Effect of Citrus paradisi extract and juice on arterial pressure both
in vitro and in vivo. Phytother. Res., 23, 948–954, 2009.
57. Seden, K., Dickinson, L., Khoo, S., and Back, D., Grapefruit-drug interactions. Drugs, 70, 2373–2407,
2010.
58. Bailey, D.G. and Dresser, G.K., Interactions between grapefruit juice and cardiovascular drugs. Am. J.
Cardiovasc. Drugs, 4, 281–297, 2004.
59. Miyata, M., Takano, H., Guo, L.Q., Nagata, K., and Yamazoe, Y., Grapefruit juice intake does not
enhance but rather protects against aflatoxin B1-induced liver DNA damage through a reduction in
hepatic CYP3A4 activity. Carcinogenesis, 25, 203–209, 2004.
60. Guo, L.-Q., Fukuda, K., Ohta, T., and Yamazoe, Y., Role of furanocoumarin derivatives on grapefruit
juice-mediated inhibition of human CYP3A activity. Drug Metab. Dispos., 28, 766–771, 2000.
61. Sass-Kiss, A. and Sass, M., Distribution of various peptides in citrus fruits (grapefruit, lemon, and
orange). J. Agric. Food Chem., 50, 2117–2120, 2002.
62. Morris, R.A., The U.S. orange and grapefruit juice markets: History, development, growth, and change.
EDIS document FE834, Institute of Food and Agricultural Sciences, University of Florida, Gainesville,
FL, 2010.
63. Tetra Pak, Juice, nectar, still drinks. Tetra Pak Magazine, 97, Tetra Pak International, Lund, Sweden, 2009.
64. Igual, M., Contreras, C., Camacho, M.M., and Martinez-Navarrete, N., Effect of thermal treatment and
storage conditions on the physical and sensory properties of grapefruit juice. Food Bioprocess Technol.,
7, 191–203, 2014.
65. Guiavarc’h, Y., Segovia, O., Hendrickx, M., and Van Loey, A., Purification, characterization, thermal
and high-pressure inactivation of a pectin methylesterase from white grapefruit (Citrus paradisi). Innov.
Food Sci. Emerg. Technol., 6, 363–371, 2005.
66. Aadil, R.M., Zeng, X., Han, Z., and Sun, D., Effects of ultrasound treatments on quality of grapefruit
juice. Food Chem., 141, 3201–3206, 2013.
67. Valero, M., Recrosio, N., Saura, D., Muňoz, N., Martí, N., and Lizama, V., Effects of ultrasonic treat-
ments in orange juice processing. J. Food Eng., 80, 509–516, 2007.
68. Kranz, P., Adler, P., and Kunz, B., Sorption of citrus flavour compounds on XAD-7HP resin during the
debittering of grapefruit juice. Int. J. Food Sci. Technol., 46, 30–36, 2011.
69. Wu, M.H., Zhu, L., Zhou, Z.Z., and Zhang, Y.Q., Coimmobilization of naringinases on silk fibroin
nanoparticles and its application in food packaging. J. Nanopart., 2013, 1–5, 2013.
70. Ribeiro, I.A. and Ribeiro, M.H.L., Naringin and naringenin determination and control in grapefruit
juice by a validated HPLC method. Food Control, 19, 432–438, 2008.
71. Wilkins, M.R., Widmer, W.W., Grohmann, K., and Cameron, R.G., Hydrolysis of grapefruit peel waste
with cellulase and pectinase enzymes. Bioresour. Technol., 98, 1596–1601, 2007.
72. Gruz, J., Novák, O., and Strnad, M., Rapid analysis of phenolic acids in beverages by UPLC-MS/MS.
Food Chem., 111, 789–794, 2008.
24
Guava Juice
İncinur Hasbay and Emine Aytunga Arık Kibar
CONTENTS
24.1 Introduction................................................................................................................................... 297
24.4 Health Effects................................................................................................................................ 302
24.5.1 Novel Products and Processing Technologies.................................................................. 304
24.5.2 Guava Juice Blends and Fortification............................................................................... 306
24.5.3 Guava Juice Processing By-Products............................................................................... 306
24.6 Conclusion..................................................................................................................................... 306
References............................................................................................................................................... 307
24.1 Introduction
Guava is a delicious tropical fruit having a fleshy, yellow ovoid berry about 5 cm in diameter with
an edible pink mesocarp containing numerous small hard white seeds [1]. The name “guava” usually
refers to Psidium guajava L., which is the most common guava species of the genus “Psidium.” Indeed,
there are 150 other guava species, among which “cattley guava” (P. cattleianum Sabine), “pear guava”
(P. pyriferum L.), and “apple guava” (P. pomiferum L.) species are the most important [2]. Guava, which
is native to Mexico, the Caribbean, Central America, and the northern part of South America, grows well
in all tropical and many subtropical regions [1]. India is the leading guava producer country followed by
China. These two countries accounted for about 60% of global market share in 2012 [3].
During the last decade, it has become a necessity for the producers to develop products that have both
nutritional value and health benefits. In this context, guava has a great potential due to its excellent nutri-
tive value, pleasant flavor, good palatability, and availability in abundance at moderate price [4]. Guava
is often considered as a precious fruit due to its rich content of vitamins C and A as well as a high dietary
fiber level. Guava also has adequate levels of minerals, potassium, and magnesium and contains two
major classes of bioactives, namely, carotenoids and polyphenols, which provide it with relatively high
dietary antioxidant value [5].
The ripened guava fruit has a limited shelf life due to its highly perishable nature. Therefore, it is
important to process the fruit into products in order to increase the shelf life. Guava can be processed
into various commercial products including juice, puree, instant guava powder, paste, canned slices in
syrup, and fruit bars, among others. Of these products, guava juice has become economically important
in the market [4].
Guava juice has a delicate color and flavor and is generally produced as a clarified fruit juice that is usu-
ally more acceptable to the consumers [6]. The manufacturing of clear guava juice includes processing
steps similar to those for the other fruits such as mashing, enzyme treatment, pressing, fining, filtration,
and heat treatment. However, manufacturing of clear juice from guava is difficult due to the colloidal
particles that cause turbidity that carries flavor substances and antioxidants [7]. Therefore, conventional
297
processing techniques should be carefully designed or alternative processing technologies developed in

order to maintain unique nutritive content and flavor (taste and aroma) of guava juice.
Guava juice contains numerous bioactive compounds such as polyphenols, carotenoids, minerals, and
vitamins, among others [8]. This chapter highlights the nutritional and phytochemical contents of guava
juice, its effects on human health as well as the novel processing methods, and future trends in the guava
juice processing industry.

Guava is considered to be a highly nutritious fruit mainly due to its high level of vitamin C, which has
been reported to be one of the highest among all fruits [9]. Thus, guava was used in the form of puree by
the allied troops during World War II in order to fortify their rations [10]. Despite the losses during pro-
cessing, guava juice and nectar are still excellent sources of vitamin C, which can reach levels fourfold
higher in guava juice than that of orange juice [11]. The vitamin C content of guava juice is reported
to be at 76.2 mg/100 mL [12] that exceeds the minimum Recommended Dietary Allowances (RDA) of
60 mg/day [13]. Guava and its juice are also good sources of vitamin A as well as some minerals such
as potassium, calcium, manganese, and magnesium [11,14–16] (Table 24.1). Reference data for labeling
purposes report that one cup (about 251 g) of guava nectar provides 82% of the RDA for vitamin C,
2% for vitamin A, 3% for potassium, 3% for calcium, 5% for manganese, and 2% for magnesium [16].
TABLE 24.1
Compositional and Nutritional Characteristics of Guava and Guava Nectar (per 100 g)
Nutrient Unit Guava (Raw) Guava Nectar (Canned) References
Water g 80.80 83.51 [15,16]
Energy kcal 68 63 [15,16]
Protein g 2.55 0.09 [15,16]
Lipid (fat) g 0.95 0.06 [15,16]
Carbohydrate g 14.32 16.25 [15,16]
Total sugars g 8.92 12.95 [15,16]
Total dietary fiber g 5.4–10.61 1.0 [15,16,27]
Minerals
Calcium mg 18 8 [15,16]
Copper mg 0.2 0.017 [15,16]
Iron mg 0.26 0.38 [15,16]
Magnesium mg 22 2 [15,16]
Manganese mg 0.2 0.058 [15,16]
Phosphorus mg 40 2 [15,16]
Potassium mg 417 39 [15,16]
Sodium mg 2.0 6 [15,16]
Zinc mg 0.23 0.03 [15,16]
Vitamins
Folate (DFE) µg 49 3 [15,16]
Niacin mg 1.084 0.170 [15,16]
Riboflavin mg 0.040 0.003 [15,16]
Thiamin mg 0.067 0.003 [15,16]
Vitamin A IU 624 106 [15,16]
Vitamin B6 mg 0.110 0.01 [15,16]
Vitamin C mg 95.4–228 13.8–62.32 [12,15,16,21,22,24]
Vitamin E (ATE) mg 0.73 0.05 [15,16]
Abbreviations: DFE, dietary folate equivalents; ATE, alpha-tocopherol equivalents.
Guava Juice 299
Along with these data, the nutritional value of guava is often included among super fruits [17], and its
juice is considered to be a highly nutritional beverage for human consumption [18], which can be an
alternative to the traditional caffeine-containing beverages [19].
The nutritional composition of guava juice is affected by numerous factors including the raw material
properties such as cultivar, harvest time, maturity, geographical location, and horticultural practices
[12,20,21] as well as the processing factors, packaging, and storage conditions [12]. In a study on changes
in the chemical composition of four guava cultivars during development and ripening, it was determined
that the ascorbic acid content varied among cultivars and significantly increased with fruit growth and
development in all cultivars studied [20]. Processing and storage conditions play a critical role on the
nutrient composition of guava juice [9,12,21,22]. Especially, the vitamin C content decreases to different
extent depending on the processing techniques employed as well as the different steps involved in the
same technique. As temperature, time, oxygen, and light affect the stability of ascorbic acid, the most
critical factor involved is storage temperature [12]. It was also reported that ascorbic acid content in
laboratory-prepared guava nectars from four guava varieties decreased during storage period at ambi-
ent conditions for 150 days, possibly due to oxidation or irreversible conversion of l-ascorbic acid into
dehydroascorbic acid in the presence of ascorbic acid oxidase [21]. Similar findings were reported for the
decrease in ascorbic acid content of guava nectars stored at room temperature for 8 months [23]. It was
shown that guava nectars stored at 4°C retained their l-ascorbic acid content for a longer period than
those stored at 25°C; thus the retention of ascorbic acid during storage was dictated mainly by storage
temperature [9]. Pink guava nectars stored at 10°C for 240 days experienced a progressive decrease in
vitamin C content during the storage period, especially between days 0 and 60 [22].

The major bioactives in guava juice are carotenoids, especially β-carotene and lycopene in the pink vari-
eties [24,25]. Lycopene has been reported to be the principal carotenoid in pink guava, contributing 86%
to the total content [26]. Although most fruits and vegetables have higher carotenoid levels in the peel
than in the pulp, lycopene is concentrated in the pulp of pink-fleshed guava [25,27]. Guava has also been
reported to be a good source of soluble fiber, bioactive flavonoids [28], and carotenoids other than those
mentioned such as rubixanthin, lutein, cryptoflavin, phytofluene [24], zeinoxanthin, and trihydroxy-5,8-
epoxy-β-carotene [29] (Figure 24.1 and Table 24.2).
In addition to carotenoids, guava juice contains phenolic compounds, mostly phenolic acids and cat-
echin (Figure 24.2). Among the phenolic acids in guava juice, ferulic acid is the major component fol-
lowed by vanillic, gallic, and trans-cinnamic acids. Catechin, which is another phenolic compound in
flavan-3-ol group, has been reported as being present at higher amounts than those in other tropical fruit
juices such as bambangan and cocoa pulp [30].
The amount of bioactive compounds in guava juice may change according to the preharvest and post-
harvest conditions of the fruit [20,24,25], as well as the processing and storage conditions [9,24,25].
In a previously mentioned study on changes in the chemical composition of four guava cultivars during
development and ripening, it was shown that the amount of polyphenols varied according to the cultivar
and decreased markedly with fruit growth and development in all tested cultivars [20]. The effect of
processing and storage conditions on lycopene content has been more widely studied. It was reported that
trans-lycopene content of guava nectars decreased by 40% after pasteurization at 87°C for 90 s, whereas
no difference was observed for the ones pasteurized at 82°C for 20 s. However, nectars processed at 87°C
for 90 s did not experience any subsequent loss of trans-lycopene during storage at either 4°C or 25°C,
while the ones processed at 82°C for 20 s exhibited a significant decrease [9]. During the pasteurization
of guava juice for 30 min in boiling water, an insignificant decrease in trans-lycopene and a significant
fivefold increase in cis-lycopene content was reported. The same study showed that both isomers of
lycopene decreased while β-carotene remained unchanged during 10 months of storage at room tem-
perature [31]. In another study on the effect of processing and storage conditions of pink guava nectar,
it was shown that lycopene content decreased during production of nectar from fresh guava. No further
decrease was observed during storage of nectars at 10°C for 240 days [22].
CH3 CH3 CH3 CH3

CH3
H3C
CH3 CH3 CH3 CH3
Lycopene
CH3
H3C CH3 H3C
H3 C
CH3
CH3 CH3 CH3
CH3
β-Carotene
H3C CH3 CH3 CH3

CH3
CH3 CH3 CH3 CH3

HO CH3
Rubixanthin
H 3C OH
H3C CH3 CH3 CH3
CH3 CH3 H3C CH3

HO CH3
Lutein
CH3 CH3 CH3 CH3

CH3
H3C
CH3 CH3 CH3 CH3
Phytofluene
H3C
H3C CH3 CH3 H3C
CH3 H3C H3C CH3

HO CH3
Zeinoxanthin
CH3
O CH3
CH3
H3C CH3
H3C H3C
H3C OH
H3C
CH3
Cryptoflavin
FIGURE 24.1 Major carotenoids found in guava juice.

Guava Juice 301
TABLE 24.2
Ranges of Bioactive Compounds in Guava and Guava Juice
Guava Juice (mg/100 g
Compound Guava (mg/100 g) or mg/100 mL) References
Carotenoids
Lutein 0.27 na [27]
Lycopene 0.77–6.60 0.31–6.0 [10,22,24,27,32,34,35,67]
Trihydroxy-5,8-epoxy-β-carotene 0.21–0.40 na [67]
Zeinoxanthin 0.10–0.19 na [67]
β-carotene 0.1–2.67 0.04–0.37 [27,31,32,67]
Phenolic Acids
Ferulic acid na 1.10 [30]
Gallic acid na 0.51 [30]
Protocatechuic acid na nd [30]
Syringic acid na nd [30]
trans-Cinnamic acid na 0.23 [30]
Vanillic acid na 0.71 [30]
Flavan-3-ols
Catechin na 4.49 [30]
Abbreviations: na, not available; nd, not detected.
HO
OH
HO O
OH
OH
Catechin
H3C
O
HO H3C
O
HO
OH OH
O O
Ferulic acid Vanillic acid
O
HO
OH
HO O
OH HO
Gallic acid trans-Cinnamic acid
FIGURE 24.2 Major phenolic compounds found in guava juice.

The loss of lycopene and β-carotene during processing or storage of guava juice has been attributed to
oxidative degradation as in the case of other processed foods containing highly unsaturated carotenoids.
The oxidation reaction is stimulated by light, heat, metals, enzymes, and peroxides. The degradation is
known to increase with the destruction of the food cellular structure, increase in surface area or poros-
ity, length and severity of the processing conditions, duration and temperature of storage, and use of
packaging material permeable to oxygen and light [32]. Better understanding of the mechanism of this
oxidative degradation is required for avoiding losses during processing and storage. For that purpose,
Rodriguez and Rodriguez-Amaya [25] determined the epoxycarotenoids and apocarotenals formed by
the oxidation of lycopene with atmospheric oxygen in low-moisture and aqueous model systems as well
as in some processed foods. Among the processed foods tested, several oxidation products of lycopene
were determined in guava juice such as three lycopene epoxides, two Z-isomers of lycopene, and two
acid-catalyzed rearrangement products (apo-12′-lycopenal and 2,6-cyclolycopene-1,5-diol). The pres-
ence of these compounds indicates the oxidation of lycopene during processing since epoxidation and
cleavage to apocarotenals are initial steps in the autoxidation of carotenoids in foods [25]. Oxidation and
isomerization of highly unsaturated carotenoids can also take place easily during analysis and/or storage
of samples prior to analysis, which would lead to underestimation of these compounds in the analyzed
samples. Therefore, several measures should be taken to prevent these reactions, such as completion of
the analysis within the shortest possible time, exclusion of oxygen, protection from light, avoiding high
temperature or contact with acid, and use of high purity solvents free from harmful impurities (e.g.,
peroxides) [32].
To the best of our knowledge, studies on the bioavailability of carotenoids and other bioactives of
guava juice following its intake have not been reported until recently. However, it is evident that lyco-
pene is absorbed better from heat-processed foods than unprocessed sources [33], which may imply that
lycopene from pasteurized guava juice may be more bioavailable than that from fresh guava juice. The
reason for this availability improvement after heating foods has been suggested as the consequence of
dissociation of the proteins as well as partial isomerization of lycopene [34]. As mentioned before, pas-
teurization of guava juice leads to the increase in cis isomers of lycopene [31], which were reported to
be more bioavailable than all-trans form due to being more soluble in bile acid micelles and, therefore,
being preferentially incorporated into chylomicrons [33].
24.4 Health Effects

Guava has been widely used in traditional medicine for treatment of several diseases such as gastroin-
testinal and respiratory disturbances, and its leaves are applied on wounds and ulcers, and for rheumatic
pain and also chewed to relieve toothache [1]. However, studies on in vitro and in vivo health effects of
guava and its products are limited, although it has been well documented that the fruit has considerable
amount of bioactive compounds such as vitamin C, lycopene, and β-carotene.
A large number of studies exist on the in vitro and in vivo antioxidative, anticarcinogenic, and
antiatherogenic effects of the major bioactive compounds in guava and guava juice, mainly vitamin
C and lycopene [10,33,35,36]. Epidemiological studies have suggested that a high consumption of
lycopene may protect against cardiovascular disease (CVD) and reduce the risk of several types of
cancer, such as prostate, breast, lung, and digestive tract [35]. The oxidative degradation products in
guava juice and other processed foods have also been reported, and these may have some possible
implications on human health. One example is 2,6-cyclolycopene-1,5-diol, an oxidation product of
lycopene that was determined in processed guava juice [25] and reported to increase the expression
of connexin 43 gene in mouse 10T1/2 cells and in human keratinocytes in organotypic culture to
levels marginally higher than lycopene and consequently indicated a possible stronger preventive
activity against cancer [37].
In addition to research studies on the health effects of bioactives of guava, there are a limited number
of preclinical studies demonstrating the health effects of guava and its juice that focus mainly on their
effects on the metabolic profile and oxidative stress (Table 24.3). In one of those studies conducted to
evaluate the effect of consumption of the pulp and seeds of guava on the metabolic profile of Wistar
Guava Juice 303
TABLE 24.3
Selected Preclinical Studies on Possible Health Effects of Guava Juice
Possible Health Effects Experimental Model Possible Mode of Action References
Control of biochemical variables Wistar rats Significant reduction in glycemia [38]
involved in the incidence of and in the levels of TAG and total
chronic degenerative disorders cholesterol.
Augmented levels of HDL
cholesterol.
Reduced levels of AST and ALT
enzymes.
Hypoglycemic effect Normal and alloxan- A markable hypoglycemic action. [39]
treated diabetic mice
Antidiabetes and/or antiobese Spontaneous non-insulin- Increased amount of initial insulin [40]
effects dependent diabetes secretion in diabetes mellitus rats.
mellitus rats
Effect on oxidative stress, lipid Adult male albino rats Impairment of oxidative damage [41]
peroxidation, and antioxidant better than tomato juice.
substance in plasma and tissues
TAG, triacylglycerols; HLD, high-density lipoprotein; AST, aspartate aminotransferase; ALT, alanine
Abbreviations:
aminotransferase.
rats, the treatment groups were fed with guava pulp juice and guava seeds. After 40 days of feeding,
a significant reduction in glycemia and the levels of triacylglycerols (TAG) and total cholesterol were
observed along with an increase in the levels of high-density lipoprotein (HDL) cholesterol in animals
fed with guava pulp juice and seeds. The levels of aspartate aminotransferase and alanine aminotrans-
ferase enzymes were also reduced. As a result, the researchers suggested that the use of guava pulp
and seeds contributed significantly to the control of biochemical variables involved in the incidence of
chronic degenerative disorders in the population [38].
The hypoglycemic effect of guava juice was examined in normal and diabetic rats, using streptozotocin-
induced diabetes mellitus model. Acute intraperitoneal treatment with 1 g/kg guava juice p roduced a
remarkable hypoglycemic action in normal and alloxan-treated diabetic mice. Accordingly, it was sug-
gested that guava may be employed to improve and/or prevent diabetes mellitus [39]. The antidiabetes
and/or antiobesity effects of long-term ingestion of guava juice was investigated in another preclinical
study, using spontaneous non-insulin-dependent diabetes mellitus Otsuka Long-Evans Tokushima Fatty
rats and its control strain Long-Evans Tokushima Otsuka rats. Thirty rats of each strain were divided into
three groups and fed on glucose, vitamin E, and guava juice. Serum lipid parameters including total cho-
lesterol, TAG, free fatty acids, and HDL cholesterol were measured after ingestion for 23 weeks. Blood
glucose levels and plasma insulin concentrations were measured at the end of the ingestion period of
23 weeks and also 10 weeks after discontinuation of guava juice ingestion. As a result, the blood glucose
level in the guava juice group did not change as compared with the glucose group, but the amount of initial
insulin secretion was significantly increased in diabetes mellitus rats and was restored by discontinuation
of guava juice ingestion. Therefore, the researchers suggested that long-term ingestion of guava juice may
increase plasma insulin concentration in diabetes mellitus rats [40].
Although it has been suggested that the high antioxidant activity of guava may interfere with can-
cers initiated by oxidative and free radical damage to DNA and cell components [10], data on those
health effects of guava and its juice are scarce. A preclinical research study was carried out on the
effect of tomato and guava juice supplementation on oxidative stress, lipid peroxidation, and antioxidant
substances in plasma and tissues of 60 adult male albino rats after strenuous exercise. For that purpose,
xanthine oxidase, glutathione reductase, vitamin C, malondialdehyde (MDA), myeloperoxidase, and
some parameters of liver and kidney functions were measured in groups of rats fed on basal diet supple-
mented with different doses of tomato or guava juice and subjected to strenuous exercise. The researchers
then suggested that guava juice was more effective than tomato juice in impairment of oxidative damage
caused by strenuous exercise leading to significant decrease in plasma and muscle xanthine oxidase,
MDA, and muscle myeloperoxidase after exercise. In addition, the plasma vitamin C levels of the group
fed with guava juice was significantly higher compared with the group fed with tomato juice [41].
A limited number of clinical trials have also been reported on positive health effects of guava or
its extracts. In an early clinical study, guava or its juice was found to lower blood glucose levels in
healthy and maturity-onset diabetic volunteers [39]. In a randomized placebo-controlled trial of guava
juice as a source of ascorbic acid to reduce iron deficiency in Tarahumara indigenous school children
of northern Mexico, 95 boarding school children aged 6–9 years identified as anemic were randomly
allocated to receive 300 mL of natural guava juice containing about 200 mg of ascorbic acid or placebo
(guava-flavored juice free of ascorbic acid) with the main meal. Changes in hemoglobin and plasma fer-
ritin among the iron-deficient subsample at baseline were the main outcomes. As a result of the study,
the researchers suggested that guava juice providing 200 mg ascorbic acid at one meal on each school
day had a marginal effect on hemoglobin and plasma ferritin concentrations in children consuming
high-phytate diets fortified with iron [42]. The scarcity of data on the health effects of guava despite its
superior nutritional and bioactive properties suggests that more clinical and preclinical studies should be
conducted to investigate the unexploited potential of this fruit.

24.5.1 Novel Products and Processing Technologies
Guava juice processing includes unit operations similar to those for the other fruits such as extraction, filtra-
tion, and heat treatment [7]. The design and application of those processing steps are very important since
processing can significantly affect the product quality. Particularly for foods containing high amounts of
bioactive compounds, various physical, chemical, and/or biological changes may occur during processing,
such as a loss of nutrients and color due to the degradation of carotenoids and anthocyanins [43].
Extraction is one of the most important unit operations in fruit juice processing, and conventionally,
it consists of fruit crushing and pressing [44]. After the extraction, untreated guava juice is cloudy due
to colloidal suspensions and needs to be clarified, since a clear guava juice is usually more acceptable
and marketable [7,18,45]. However, the manufacturing of clear guava juice is difficult, since the colloidal
particles carry many flavor substances and antioxidants [7]. The guava juice is usually extracted by cold,
hot, or enzymatic methods [45,46]. Recent studies have shown that that enzymatic treatment significantly
increased juice recovery yields, nutritional value, and clarification performance [6,7,18,45,47]. Cellulase,
hemicellulase, and pectinase were the common enzymes effectively used during extraction to degrade
cell wall and plant tissue of fruits [44]. Several authors have shown that application of 0.12%–0.13%
(w/w) cellulase to guava mash at optimized conditions increased the extraction yield and total phenol
content by approximately 21%–22% and 12%–16%, respectively [18,44]. Pectolytic enzymes are also
effectively used in guava juice processing. For instance, guava mash treatment with 700 ppm pectolytic
enzyme at 50°C for 1.5 h resulted in a 51% reduction in viscosity, 13% increase in ascorbic acid content,
and 18% increase in the yield as well as a significant reduction in the turbidity of the guava juice [6].
A similar study, conducted on the commercial pectinase application to guava pulp, showed that 0.70 mg
enzyme addition to 100 g guava pulp at 45°C for 7 h significantly increased vitamin C retention and
improved the clarity of guava juice [45]. The enhanced clarity of guava juice by enzyme treatment is
generally explained by the breakdown of pectin molecules, which facilitates the formation of pectin–
protein flocs and favors the removal of the colloidal suspension [45,46]. In addition, tannase (tannin acyl
hydrolase) enzyme was successfully applied to guava juice in order to remove the cloudiness and bit-
terness caused by tannins in guava juice. It was noted that remarkable reduction of tannic acid in guava
juice was obtained and organoleptic properties were improved correspondingly [47].
Guava juice, like other fruit juices, is pasteurized to reduce the initial microbial load and to inhibit the
activity of native enzymes. However, thermal processing may cause detrimental effects, such as degra-
dation of heat-sensitive phytochemicals, off-favor formation, and darkening of the product [48]. Several
technologies may be used as alternatives to thermal processing of guava juice, such as ultrasound, micro-
wave, and ultraviolet (UV) radiation [49–51].
Guava Juice 305
The ultrasound processing, also called as sonication, is a nonthermal technology that has been effec-
tive in the inactivation of microorganisms and enzymes related to degradation of fruit juices [52]. Recent
studies have shown that sonication can be used as a preservation technique for guava juice processing
[49,51]. Sonication improves the product quality and allows producing guava juice with higher ascorbic
acid content, lower turbidity, and maintaining color with respect to conventional heat treatment [51].
Cheng et al. [49] found that the ascorbic acid content was higher in guava juice samples treated with
carbonation and/or sonication than in the conventionally treated sample. It is suggested that application
of sonication eliminates the dissolved oxygen that is essential for ascorbic acid degradation during cavi-
tation. This phenomenon is enhanced with carbonation due to the fact that dissolved CO2 could serve as
nuclei sites for cavitation [49]. Microwave technology offers advantages over the conventional process-
ing methods due to fast heating rates that result in shorter processing times [53]. Salazar-González et al.
[53] successfully pasteurized guava nectar by microwave heating (2450 MHz) at 90°C using 500 or
950 W. It was reported that microbial counts remained below the detectable limit, 94% of vitamin C was
retained, and color was better preserved after microwave treatment. Similar to sonication, UV treatment
is performed at low temperatures and classified as a nonthermal disinfection method. In comparison
to conventional pasteurization, application of UV radiation to guava juice provided promising results,
where adequate microbial load reduction was achieved and the taste and color profiles were highly
conserved [50].
Instant guava powder is another important guava product that is used as a common ingredient in
many formulated drinks, baby foods, and confectionary products. It is obtained by dehydration of clear
guava juice with various methods, such as freeze-drying, spray-drying, and tunnel-drying. Several stud-
ies have examined the feasibility of different drying methods to produce instant guava powder with
acceptable quality at reasonable cost [6,54–56]. Chopda and Banrett [6] used a freeze-drying method
to convert guava juice into a powder and reported that oxidative loss of ascorbic acid was considerably
lower in freeze-drying (18.8%) compared to spray-drying (21%) and tunnel drying (32.2%). In addition,
sensory tests showed that freeze-dried samples resulted in a better color and a higher flavor retention.
In a recent study, the effect of freeze-drying and hot-air-drying at 70°C on the bioactives of guava
was compared [43]. Total anthocyanins of fresh red guava were maintained after the drying process
(freeze-dried and hot-air-dried samples) by 45% and 2%, respectively. In addition, total carotenoid
content was higher in freeze-dried guava sample than the fresh fruit, while in hot-air-dried samples,
total carotenoids decreased by ~61%. The higher carotenoid level in freeze-dried guava was explained
by the rupture and the decompression of the tissues, which would facilitate contact with the solvent
and enhance extraction at the time of analysis [43]. Although it is possible to obtain high-quality guava
powder with freeze-drying in terms of nutritional and organoleptic properties, it is known to be the most
expensive method of drying.
Spray-drying is another advantageous drying technique for heat-sensitive materials due to its ability to
meet industrial specifications such as enabling high-capacity, continuous, and automated systems. Guava
juice was spray-dried, and the processing conditions were optimized by different researchers [54–56].
The main outcome of these studies indicated that spray-drying was an adequate technology for guava
powder production with the spray-dried product having a bright color and attractive taste in contrast to
tunnel-dried product. However, it was also noted that concentrated guava juice without additives such
as maltodextrin cannot be satisfactorily spray-dried because of hygroscopic and thermoplastic nature of
the product [54–56].
One of the important technological problems encountered in guava nectar processing is the
nonenzymatic browning, especially, in glass-packaged products [57]. Nonenzymatic browning reactions
involve caramelization, ascorbic acid degradation, and Maillard reaction [58]. In addition to brown pig-
ment formation, loss of nutrients and the formation of undesirable compounds such as furfural and
5-hydroxymethly furfural may also occur during storage period [59]. The common antibrowning agents
used in fruit juice industry are sulfiting compounds. Due to their adverse health effects, alternative
natural compounds are being explored. In this context, green tea extract and l-cysteine were explored
as natural antibrowning agents in guava nectar in a recent study [57]. The results revealed that 400 ppm
l-cysteine or green tea extract was an adequate dosage for inhibiting nonenzymatic browning in glass-
bottled guava nectar [57].
24.5.2 Guava Juice Blends and Fortification

Fruits, which are rich in nutrients but not acceptable due to high acidity, poor taste, and flavors, could
be blended with other fruits to improve their acceptability. In addition, blending of fruit juices helps to
improve the nutritional value of the beverage [60]. Therefore, guava juice has a potential to be used in
fruit juice blends due to its high nutritional and sensory value. Various fruit juices, guava juice blends, and
ready-to-serve (RTS) beverages have been investigated recently [60–63]. For instance, blending of guava
pulp with bael pulp resulted in the development of a beverage rich in vitamins, especially vitamin C and
minerals; and, taste, flavor, and overall acceptability improved remarkably [60]. In another study, 30%
papaya pulp was blended with 70% guava, and results revealed that the carotenoid content and sensory
qualities of the RTS beverage were increased and the shelf life of the RST was extended to 6 months at
room temperature [62]. Selvi et al. [61] evaluated the formulation of a guava/lime/ginger RTS beverage as
a therapeutic drink. They fixed the fruit ratio of guava/lime/ginger as 10:3:2 (v/v/v) and concluded that the
formulated drink was highly acceptable from both technological and economical points of view.
The unique red flower of the roselle (Hibiscus sabdariffa) plant is used to prepare juices and is a good
source of calcium, magnesium, iron, and anthocyanins, such as delphinidin 3-sambubioside, cyanidin
3-sambubioside, delphinidin 3-glucoside, and cyanidin-3-glucoside [63]. However, its high acidity usually
limits its direct consumption; therefore, formulating a guava–roselle blend can be a promising solution to
the palatability problem encountered with roselle flower extract. Mgaya et al. [63] studied the nutritional
value and phytochemical characteristics of the 80:20 (v/v) guava–roselle blend. Values were as follows
as per mg/100 g dry weight: calcium (23.4), magnesium (54), phosphorus (37.8), iron (2.7), sodium (3.5),
zinc (6.4), total monomeric anthocyanin content (118.2 as cyanidin-3-glucoside equivalents), and total
polyphenols content (47.3 as gallic acid equivalents [GAE]). They concluded that the guava–roselle blend
could replace the existing commercially available nonalcoholic beverages on the market.
Whey drinks are light, refreshing, healthful, and nutritious and offer high market potential [64]. The
manufacture of whey-based fruit beverages require the mixing of appropriate fruit juices and minimally
processed whey with the selection of suitable stabilizer. The development and characterization of guava
pulp–whey–based beverage was investigated [64]. It was found that whey–guava beverage pasteurized at
65°C for 25 min was best in terms of sensory quality after 45 days of storage. The pH, acidity, protein,
vitamin C, total sugars, and reducing sugars were higher than that of other samples. In addition, they
noted that low cost as well as high protein and vitamin C contents of this beverage might allow the low-
income consumers to reach such an enriched beverage with reasonable price.
24.5.3 Guava Juice Processing By-Products

Processing of guava juice is accompanied by an important quantity of by-product residues accounting
for 25% of the fruits’ total weight. Three types of by-products are derived from the crushing, refining,
and sieving steps of processing, namely, guava refiner (12%), guava siever (8%), and guava decanter (5%),
respectively [26]. The first fraction, guava refiner, mainly consists of peels and seeds, the latter ones
mostly consist of pulp and pomace having a slightly red color [26]. The by-products are usually discarded
during processing. However, they have a potential as a source of bioactive components. For instance,
lycopene contents of the refiner, siever, and decanter were measured as 7.3, 6.3, and 1.5 mg/100 g,
respectively, and the total phenolic contents were 4434, 2881, and 1529 mg GAE/100 g, respectively
[65,66]. However, further studies are needed to explore extraction techniques in order to obtain maxi-
mum phenolic and flavonoid compounds from the guava juice processing by-products.
24.6 Conclusion
Guava is considered to be a highly nutritious fruit mainly due to its high vitamin C content, which
has been reported to be one of the highest among all fruits. Indeed, guava is often included among
superfruits, and its juice is considered to be an alternative to the commercial soft beverages. The amount
of bioactive compounds in guava juice may change due to variations of these compounds in the fruit
Guava Juice 307
according to the preharvest and postharvest conditions as well as the processing and storage conditions.
The scarcity of data on the health effects of guava juice despite its superior nutritional and bioactive
properties suggests that more preclinical and clinical studies should be conducted to investigate the
unexploited potential of this product.
REFERENCES
1. Gutiérrez, R.M.P., Mitchell, S., and Solis, R.V., Psidium guajava: A review of its traditional uses,
phytochemistry and pharmacology. J. Ethnopharmacol., 117, 1–27, 2008.
2. Gull, J., Sultana, B., Anwar, F., Naseer, R., Ashraf, M., and Ashrafuzzaman, M., Variation in antioxi-
dant attributes at three ripening stages of guava (Psidium guajava L.) fruit from different geographical
regions of Pakistan. Molecules, 17, 3165–3180, 2012.
3. FAO, Word primary crops data. Food and Agriculture Organization of the United Nations Statistics
Division, August 2014. Published online at: http://faostat.fao.org/site/567/default.aspx#ancor (accessed
March 10, 2014).
4. Kadam, D., Kaushik, P., and Kumar, R., Evaluation of guava products quality. Int. J. Food Sci. Nutr.
Eng., 1, 7–11, 2012.
5. Imungi, J., Scheffeldt, P., and Sainthilaire, P., Physicochemical changes during extraction and contrac-
tion of clear guava juice. LWT—Food Sci. Technol., 13, 248–251, 1980.
6. Chopda, C.A. and Barrett, D.M., Optimization of guava juice and powder production. J. Food Process.
Pres., 25, 411–430, 2001.
7. Brasil, I.M., Maia, G.A., and Figueiredo, R.W., Physical-chemical changes during extraction and clari-
fication of guava juice. Food Chem., 54, 383–386, 1995.
8. Flores, G., Wu, S.B., Negrin, A., and Kennelly, E.J., Chemical composition and antioxidant activity of
seven cultivars of guava (Psidium guajava) fruits. Food Chem., 170, 327–335, 2005.
9. Fender, L., Phytochemical, antioxidant, and storage stability of thermally processed guava (Psidium
guajava) and guava juice blends. PhD thesis, University of Florida, Gainesville, FL, 2005.
10. Sato, R., Dang, K.M., McPherson, B.G., and Brown, A.C., Anticancer activity of guava (Psidium gua-
java) extracts. J. Complement. Integr. Med., 7, 43, 2010.
11. Surajbhan, S., Alka, S., Chetan, J., and Lambert, R., Extraction and optimization of guava juice by using
response surface methodology. Am. J. Food Technol., 7, 326–339, 2012.
12. Jawaheer, B., Goburdhun, D., and Ruggoo, A., Effect of processing and storage of guava into jam and
juice on the ascorbic acid content. Plant Food Hum. Nutr., 58, 1–12, 2003.
13. Rampersaud, G.C., A comparison of nutrient density scores for 100% fruit juices. J. Food Sci., 72,
261–266, 2007.
14. Akesowan, A. and Choonhahirun, A., Effect of enzyme treatment on guava juice production using
response surface methodology. J. Anim. Plant Sci., 23, 114–120, 2013.
Release 27. Agricultural Research Service, USDA, Springfield, VA, 2014.
16. Self Nutrition Data, Guava nectar, canned, 2015. Published online at: http://nutritiondata.self.com/facts/
fruit4s-and-fruit-juices/9794/2 (accessed March 2, 2015).
17. Shruthi, S.D., Adhikari, R., Timilsina, S.S., and Sunita, S., A review on the medicinal plant Psidium
guajava Linn. (Myrtaceae). J. Drug Deliv. Ther., 3, 162–168, 2013.
18. Le, T.T., Nguyen, V.P.T., and Le, V.V.M., Application of cellulase preparation to guava mash treatment
in juice processing: Optimization of treatment conditions by RSM. Asian J. Food Agro Ind., 5, 284–291,
2012.
19. Shamsudin, R., Mohamed, I.O., and Yaman, N.K.M., Thermophysical properties of Thai seedless guava
juice as affected by temperature and concentration. J. Food Eng., 66, 395–399, 2005.
20. El Bulk, R.E., Babiker, E.F.E., and El Tinay, A.H., Changes in chemical composition of guava fruits
during development and ripening. Food Chem., 59, 395–399, 1997.
21. Choudhary, M.L., Dikshit, S.N., Shukla, N., and Saxena, R.R., Evaluation of guava (Psidium guajava
L.) varieties and standardization of recipe for nectar preparation. J. Hortic. Sci., 3, 161–163, 2008.
22. Ordóñez-Santos, L.E. and Vázquez-Riascos, A., Effect of processing and storage time on the vitamin
C and lycopene contents of nectar of pink guava (Psidium guajava L.). Arch. Latinoam. Nutr., 60,
280–284, 2010.
23. Bal, L.M., Ahmad, T., Senapati, A.K., and Pandit, P.S., Evaluation of quality attributes during storage
of guava nectar cv. lalit from different pulp and TSS ratio. J. Food Process Technol., 5, 329, 2014.
24. De Brito, C.A.K., Siqueira, P.B., De Souza, J.C., and Bolini, H.M.A., In vitro antioxidant capacity, phe-
nolic, ascorbic acid and lycopene content of guava (Psidium guajava L.) juices and nectars. Bol. Centro
Pesqui. Process. Aliment, 27, 175–182, 2009.
25. Rodriguez, E.B. and Rodriguez-Amaya, D.B., Lycopene epoxides and apo-lycopenals formed by chemi-
cal reactions and autoxidation in model systems and processed foods. J. Food Sci., 74, C674–C682,
2009.
26. Kong, K.W. and Ismail, A., Lycopene content and lipophilic antioxidant capacity of by-products from
Psidium guajava fruits produced during puree production industry. Food Bioprod. Process., 89, 53–61,
2011.
27. Yahia, E.M. and Ornelas-Paz, J.J.O., The contribution of fruit and vegetable consumption to human
health, in Fruit and Vegetable Phytochemicals: Chemistry, Nutritional Value, and Stability, de la Rosa,
L.A., Alvarez-Parrilla, E., and González-Aguilar, G.A., Eds., Wiley-Blackwell, Oxford, UK, 2009,
pp. 177–222.
28. Granato, D., Ribeiro, J.C.B., Castro, I.A., and Masson, M.L., Sensory evaluation and physicochemical
optimisation of soy-based desserts using response surface methodology. Food Chem., 121, 899–906,
2010.
29. Granato, D. and Masson, M.L., Instrumental color and sensory acceptance of soy-based emulsions:
A response surface approach. Ciênc. Tecnol. Aliment., 30, 1090–1096, 2010.
30. Zabidah, A.A., Kong, K.W., and Amin, I., Antioxidant properties of tropical fruit juices and their effects
on in vitro hemoglobin and low density lipoprotein (LDL) oxidations. Int. Food Res. J., 18, 545–552,
2011.
31. Padula, M. and Rodriguez-Amaya, D.B., Changes in individual carotenoids and vitamin C on process-
ing and storage of guava juice. Acta Aliment., 16, 209–216, 1987.
32. Rodriguez-Amaya, D.B., Kimura, M., Godoy, H.T., and Amaya-Farfan, J., Updated Brazilian database
on food carotenoids: Factors affecting carotenoid composition. J. Food Comp. Anal., 21, 445–463,
2008.
33. Bramley, P.M., Is lycopene beneficial to human health? Phytochemistry, 54, 233–236, 2000.
34. Roldán-Gutiérrez, J.M. and de Castro, M.D.L., Lycopene: The need for better methods for characteriza-
tion and determination. Trend Anal. Chem., 26, 163–170, 2006.
35. Omoni, A.O. and Aluko, R.E., The anti-carcinogenic and anti-atherogenic effects of lycopene: A review.
Trends Food Sci. Technol., 16, 344–350, 2005.
36. Naidu, K.A., Vitamin C in human health and disease is still a mystery? An overview. Nutr. J., 2, 7, 2003.
37. King, T.J., Khachik, F., Bortkiewicz, H., Fukushima, L.H., Morioka, S., and Bertram, J.S., Metabolites
of dietary carotenoids as potential cancer preventive agents. Pure Appl. Chem., 69, 2135–2140, 1997.
38. Farinazzi-Machado, F.M.V., Guiguer, E.L., Barbalho, S.M., de Souza, M.dS.S., Bueno, P.C.dS., Mendes,
C.G., Araujo, A.C. et al., Effects of Psidium guajava on the metabolic profile of Wister rats. J. Med.
Plants Res., 6, 3450–3454, 2012.
39. Cheng, J.T. and Yang, R.S., Hypoglycemic effect of guava juice in mice and human subjects. Am. J.
Chin. Med., 11, 74–76, 1983.
40. Sunagawa, M., Shimada, S., Zhang, Z., Oonishi, A., Nakamura, M., and Kosugi, T., Plasma insulin con-
centration was increased by long-term ingestion of guava juice in spontaneous non-insulin-dependent
diabetes mellitus (NIDDM) rats. J. Health Sci., 50, 674–678, 2004.
41. Abd Elmouttaleb, A.T. and Mostafa, U.E.S., Effect of tomato and guava juices on oxidative stress in rats
after strenuous exercise. Jordan J. Biol Sci., 5, 167–174, 2012.
42. Monarrez-Espino, J., Lopez-Alarcon, M., and Greiner, T., Randomized placebo-controlled trial of guava
juice as a source of ascorbic acid to reduce iron deficiency in Tarahumara indigenous schoolchildren of
northern Mexico. J. Am. Coll. Nutr., 30, 191–200, 2011.
43. Nora, C.D., Müller, C.D.R., de Bona, G.S., Rios, A.O., Hertz, P.F., Jablonski, A., de Jong, E.V., and
Flôres, S.H., Effect of processing on the stability of bioactive compounds from red guava (Psidium cat-
tleyanum sabine) and guabiju (Myrcianthes pungens). J. Food Comp. Anal., 34, 18–25, 2014.
44. Nguyen, V.T., Le, T.T., and Le, V.V.M., Application of combined ultrasound and cellulase preparation to
guava (Psidium guajava) mash treatment in juice processing: Optimization of biocatalytic conditions by
response surface methodology. Int. Food Res. J., 20, 377–381, 2013.
Guava Juice 309
45. Kaur, S., Sarkar, B.C., Sharma, H.K., and Singh, C., Response surface optimization of conditions for the
clarification of guava fruit juice using commercial enzyme. J. Food Process Eng., 34, 1298–1318, 2011.
46. Kaur, S., Sarkar, B.C., Sharma, H.K., and Singh, C., Optimization of enzymatic hydrolysis pretreat-
ment conditions for enhanced juice recovery from guava fruit using response surface methodology.
Food Bioprocess Technol., 2, 96–100, 2009.
47. Sharma, N.K., Beniwal, V., Kumar, N., Kumar, S., Pathera, A.K., and Ray, A., Production of tannase
under solid-state fermentation and its application in detannification of guava juice. Prep. Biochem.
Biotechnol., 44, 281–290, 2013.
48. Evrendilek, G.A., Altuntas, J., Sangun, M.K., and Zhang, H.Q., Apricot nectar processing by pulsed
electric fields. Int. J. Food Prop., 16, 216–227, 2011.
49. Cheng, L.H., Soh, C.Y., Liew, S.C., and The, F.F., Effects of sonication and carbonation on guava juice
quality. Food Chem., 104, 1396–1401, 2007.
50. Keyser, M., Müller, I.A., Cilliers, F.P., Nel, W., and Gouws, P.A., Ultraviolet radiation as a non-thermal
treatment for the inactivation of microorganisms in fruit juice. Innov. Food Sci. Emerg. Technol., 9,
348–354, 2008.
51. Saad, S.M., Elaleem, M.A., Foda, F.F.A., Eissa, H.A., Abdelmoniem, G.M., and İbrahim, W.A., Effects
of thermosonication on apple and guava juices quality. J. Appl. Sci. Res., 53, 23–36, 2013.
52. Rawson, A., Patras, A., Tiwari, B., Noci, F., Koutchma, T., and Brunton, N., Effect of thermal and non
thermal processing technologies on the bioactive content of exotic fruits and their products: Review of
recent advances. Food Res. Int., 44, 1875–1887, 2011.
53. Salazar-González, C., San Martín-González, M.F., Vergara-Balderas, F.T., López-Malo, A., and
Sosa-Morales, M.E., Physical-chemical and microbiological stability during refrigerated storage of
m icrowave-pasteurized guava nectar. Focus. Modern Food Ind., 3, 43–51, 2014.
54. Patil, V., Chauhan, A.K., and Singh, R.P., Optimization of the spray-drying process for developing
guava powder using response surface methodology. Powder Technol., 253, 230–236, 2014.
55. Shishir, M.R.I., Taip, F.S., Aziz, N.A., and Talib, R.A., Physical properties of spray-dried pink guava
(Psidium guajava) powder. Agric. Sci. Proc., 2, 74–81, 2014.
56. Mahendran, T., Physico-chemical properties and sensory characteristics of dehydrated guava concen-
trate: Effect of drying method and maltodextrin concentration. Trop. Agric. Res. Ext., 13, 48–54, 2010.
57. Abd El-Hady, M.M., Abd El-Wahab, E.S., and El-Ashwah, F.A., Role of l-cysteine and green tea extracts
for minimizing the non-enzymatic browning in guava nectar. Ann. Agric. Sci., 54, 109–120, 2009.
58. Clegg, K., Non-enzymatic browning of lemon juice. J. Sci. Food Agric., 15, 878–885, 1964.
59. Burdurlu, H., Koca, N., and Karadeniz, F., Degradation of vitamin C in citrus juice concentrates during
storage. J. Food Eng., 74, 211–216, 2006.
60. Bhardwaj, R.L. and Pandey, S., Juice blends—A way of utilization of under-utilized fruits, vegetables,
and spices: A review. Crit. Rev. Food Sci. Nutr., 51, 563–570, 2011.
61. Selvi, J., Banumathi, P., Kanchana, S., and Ilamaran, M., Formulation of therapeutic drink to boon
human health (guava-lime-ginger RTS beverage). Food Sci. Res. J., 4, 141–146, 2013.
62. Tiwari, R.B., Studies on blending of guava and papaya pulp for RTS beverages. Indian Food Pack., 54,
68–72, 2000.
63. Mgaya, K.B., Remberg, S.F., Chove,. B.E., and Wicklund, T., Physio-chemical, mineral composition and
antioxidant properties of roselle (Hibiscus sabdariffa L.) extract blended with tropical fruit juices. Afr.
J. Food Agric. Nutr. Dev., 14, 8963–8978, 2014.
64. Singh, D., Singh, R., and Bhatt, F., Development, quality evaluation and shelf life studies of whey guava
beverage. Int. J. Curr. Eng. Technol., 4, 2171–2175, 2014.
65. Kong, K., Ismail, A., Tan, S., Prasad, K., and Ismail, A., Response surface optimisation for the extraction
of phenolics and flavonoids from a pink guava puree industrial by-product. Int. J. Food Sci. Technol., 45,
1739–1745, 2010.
66. Kong, K., Rajab, N., Prasad, K., Ismail, A., Markom, M., and Tan, C., Lycopene-rich fractions derived
from pink guava by-product and their potential activity towards hydrogen peroxide-induced cellular and
DNA damage. Food Chem., 123, 1142–1148, 2010.
67. Rodriguez-Amaya, D.B., A Guide to Carotenoid Analysis in Foods. ILSI Press, Washington, DC, 2001.
25
Hawthorn Juice
Petras Rimantas Venskutonis
CONTENTS
25.1 Introduction....................................................................................................................................311
25.3.1 Phenolics............................................................................................................................313
25.3.2 Antioxidant Efficacy..........................................................................................................313
25.3.3 Effects of Storage..............................................................................................................316
25.4 Health Effects.................................................................................................................................316
25.4.1 Benefits on the Heart and Vascular System and Hypotensive Effects..............................317
25.4.2 Hypolipidemic Effects.......................................................................................................317
25.4.3 Anti-Inflammatory and Antioxidant Activities.................................................................317
25.4.4 Other Effects.....................................................................................................................317
25.6 Conclusion......................................................................................................................................318
References................................................................................................................................................319
25.1 Introduction
Hawthorns are a large genus of shrubs and trees belonging to the Rosaceae family, Amygdaloideae
subfamily, Maleae tribe, Malinae subtribe Crataegus Tourn. ex L. genus, which are native to temper-
ate regions of the northern Hemisphere in Europe, Asia, and North America. Some botanists in the
past recognized a thousand or more species; however, today, it is estimated that a reasonable number is
200 species [1]. The name “hawthorn” was originally applied to the species native to northern Europe,
especially the common hawthorn (Crataegus monogyna), which is often so used in some countries.
Crataegus spp. are shrubs or small trees, mostly growing to 5–15 m tall, with yellow, orange, dark-red,
or black small pome fruit and (usually) thorny branches. Some Crataegus spp. fruits are edible, but their
flavor has been compared to overripe apples. They are sometimes used to make jelly or homemade wine,
while the leaves are edible and, if picked in the spring when still young, are tender enough to be used
in salads. In Europe, the fruits, leaves, and flowers of the plant were traditionally employed in the treat-
ment of heart problems because of their antispasmodic, cardiotonic, hypotensive, and antiatherosclerotic
effects. Today, the plant is mainly used to treat cardiovascular disease (CVD) [2], and it continues to be
a therapeutic agent for cancer, diabetes, cough, flu, asthma, stomachache, rheumatic pain, nephritis, and
hemorrhoids in Mediterranean traditional medicine.
C. monogyna is commonly cultivated in Mediterranean countries [3], while the Chinese hawthorn
(C. pinnatifida) has long been used in traditional Chinese medicine and European herbal medicine and
is widely consumed as food, in the form of juice, drink, jam, and canned fruit. Among 18 species,
C. pinnatifida Bge. and its “Shanlihong” variety (C. pinnatifida Bge. var. major N.E.Br.) are the most
important, due to their large and delicious fruits with a characteristic acidic taste. Moreover, currently,
only fruits of C. pinnatifida and C. pinnatifida var. major are included in the Chinese Pharmacopoeia.
311
The fruits of C. pinnatifida and C. scabrifolia have traditionally been used as a peptic agent in oriental
medicine and recently in a local soft drink product, in jams, juices, and tinned foods, and as a basic
ingredient for making wines and various sweet foods [4]. This chapter highlights nutritional characteris-
tics, bioactive compounds, antioxidant efficacy, and health effects of hawthorn fruit and juice, as well as
future trends in their processing and wider applications.

Compositional and nutritional characteristics of hawthorn juice depend on the fruits used for juice production.
The fruits contain various nutrients. For instance, 15 Crataegus accessions were sampled in Hatay (Turkey),
and it was reported that fruit weight was from 1.4 to 14.9 g (mean 6.1 g), flesh/seed ratio from 2.56 to 9.23 g
(mean 5.30 g), soluble solids from 6.1% to 23.5% (mean 16.5%), pH from 3.03 to 3.99 (mean 3.19), and acid-
ity from 0.53% to 1.98% (mean 1.49%) [5]. Water is the main quantitatively constituent of fresh fruits and
depending on plant genotype and fruit harvesting time may be in the range of 57%–81% [6]. For instance, the
moisture of C. monogyna fruit reduced during ripening from 76.81% (unripe) to 60% (ripe) [7]. The moisture
and soluble solid contents in C. monogyna and C. azarolus fruits were almost similar, 73.49% and 71.08%,
and 28.50% and 24.80% (sucrose), respectively, while titratable acidity was rather different, 053 and 1.24,
respectively [8]. Consequently, the quality of such hawthorn product as a juice source may be highly variable.
Organic acids and sugars are important constituents of fruit juice both as nutrients and flavor
compounds. Malic, citric, succinic, ascorbic, tartaric, quinic, protocatechuic, 3- and 4-hydroxybenzoic,
salicylic, syringic, caffeic, ellagic, chlorogenic, ursolic, and oleanolic acids were quantified in Crataegus
spp. [9]. Citric, quinic, and malic acids were the most abundant in hawthorn fruit; however, their con-
centration varied considerably. Liu et al. [10] evaluated 22 cultivars/origins of three species of haw-
thorn from China and reported the contents of citric, quinic, malic, and the total acids in the range of
2.0–8.4, 0.5–5.6, 0.3–1.1, and 3.12–11.76 g/100 g dry weight (DW), respectively. The concentration of
various sugars and sugar alcohols was reviewed in 10 Crataegus spp. and was found to be in the ranges
(in mg/g) given: fructose (1.97–134), glucose (2.98–122), sucrose (0.15–152), sorbitol (1.7–140), myo-
inositol (trace-2.2), xylose (0.2–662), hexose (143–511), pentose (190–662), reducing sugars (17–79),
and total sugars (53–170) [9]. Juice content from fully ripened fruits of 14 Crataegus genotypes from
three locations in Tunisia was 2.0%–11.9% (v/w). The physicochemical characteristics of fresh fruit
juice were as follows: pH (3.2–4.2), titratable acidity (0.9–1.9 g malic acid/100 mL), formol index
(2.8–4.4 mL NaOH/100 mL), and total soluble solids (16.3°Brix–21.5°Brix) [11]. The compositional and
nutritional characteristics of hawthorn juice are presented in Table 25.1. Ascorbic acid was reported as
a major vitamin in hawthorn juice, which is also rich in some minerals such as calcium and potassium.

Bioactive constituents and antioxidant efficacy of hawthorn fruits have been extensively studied. The
chemistry of the genus Crataegus [9], the composition of phenolic compounds [12], and the polyphenolic
profiles and biological activity of Chinese hawthorn (C. pinnatifida) [4] fruits were comprehensively
reviewed. Many studies on hawthorn fruits have been conducted with their extracts isolated by using dif-
ferent solvents. Therefore, the concentrations of such substances in the diluted fruit juices or drinks may
be remarkably different. On the other hand, it is important to know the overall potential of the fruits in
order to develop new improved technologies for their processing into juices and other products.
Based on the information published in numerous articles, hawthorn bioactive compounds may be
conditionally grouped into flavonoids, anthocyanins, proanthocyanidins, vitamin C, and other constitu-
ents [9,12]. Some studies reported total phenolic content (TPC), total anthocyanin content (TAC), total
flavonoid content (TFC), and total proanthocyanidin content (TPAC) measured by spectrophotometric
methods and expressed in the equivalents of the selected reference compounds, for example, gallic acid
(GAE), rutin (RE), quercetin (QE), and cyanidin glucoside (CGE), while other studies identified and
Hawthorn Juice 313
TABLE 25.1
Compositional and Nutritional Characteristics Hawthorn Juice (per 100 g)
Nutrient Hawthorn Juice References
Protein g 1.92 [37]
Lipid (fat) g 0.4 [37]
Total sugars g 5.3–17 [11]
Total dietary fiber g 5.3 [37]
Minerals
Calcium mg 286–399 [11]
Magnesium mg 54 [37]
Manganese mg 55 [37]
Potassium mg 139–690 [11,37]
Sodium mg 30.4–43.8 [11]
Zinc mg 24 [37]
Vitamins
Vitamin C mg 31.4 [37]
quantified individual constituents in hawthorn. The examples of the reported results on bioactive con-
stituents of hawthorn and their variations are presented in Table 25.2, whereas the structures of the most
abundant phenolics are given in Figure 25.1.
Ascorbic acid as a water-soluble vitamin C is usually one of the most important bioactive constituent
in many fruit juices. The concentrations of total vitamin C, ascorbic acid, and dehydroascorbic acid in C.
monogyna collected from different sites between 2007 and 2009 were in the following ranges: 16.01–39.40,
0.58–5.01, 15.34–39.01 mg/100 g fresh weight (FW), respectively [6]. However, according to Barros et al.
[7], although the concentration of vitamin C in the ripe fruits of C. monogyna was higher than that in the
unripe ones (220 vs 130 mg/100 g DW), in overripened fruits, it severely decreased to 28.40 mg/100 g DW.
25.3.1 Phenolics
Flavonoids are a big group of bioactive constituents of hawthorn. Edwards et al. [9] grouped them into
apigenin- and quercetin-derived compounds; vitexin (up to 3.06 mg/g) and hyperoside (up to 3.45 mg/g)
being the most abundant. The contents of isoquercitrin and rutin found to be up to 2.3 and 2.2 mg/g,
respectively, while TFC and TPC were up to 147.3 mg/g (in C. monogyna) and 249 mg (in C. pinnati-
fida var. major), respectively [9]. Yang and Liu [12] grouped hawthorn polyphenolics into procyanidins,
flavonols, and C-glycosyl flavones, among others, and their concentrations extended over a wide range
(Table 25.2). For instance, TAC in C. monogyna was 0.0229–0.580 mg/g, TPAC up to 19.29 mg/g, total
procyanidins 0–22.2 mg/g, catechin up to 1.85 mg/g, epicatechin up to 110 mg/g, and B2 dimer up to 71.9
mg/g [9]. Gallic acid (34.44%), chlorogenic acids (5.15%), epicatechin (13.32%), quercetin 3,4-diclucoside
(51.56%), quercetin 3,7, 4-triclucoside (19.20%), and cyanidin 3-galactoside (100%) were reported in C.
monogyna [6]. A traditional C. pinnatifida var. major water extract, which is used in Chinese medicine
formula to treat CVD, contained 0.12% of each of procyanidin B2 and (−)-epicatechin [15].
25.3.2 Antioxidant Efficacy

Various methods have been employed for evaluating antioxidant activity of hawthorn fruits, mainly
extracted by different solvents and methods (Table 25.2). Dietary fiber, concentrated hawthorn juice,
and sugarless polyphenolic fractions were obtained from C. pinnatifida var. major fruits by extraction
with ethanol–water followed by further fractionation with petroleum ether, ethyl acetate, and water.
The juice and sugarless polyphenolics fractions contained 1.63 and 20.73 g/100 g DW TPC, respectively,
TABLE 25.2
Bioactives and Antioxidant Activities of Hawthorn Fruit and Juice
Species, Cultivar, and Bioactives or
Characterization of Antioxidant Activity Content or
the Assayed Samples Assay Unit Antioxidant Activity References
C. pinnatifida var. Procyanidin B2, mg/100 g 374, 349, 78, 21.8, and [20]
major; fruits (−)-epicatechin, 14.9
chlorogenic acid,
hyperoside, and
isoquercitrin
C. pinnatifida var. Procyanidin B2, mg/100 mL 25.6, 21.9, 5.02, 0.10, [20]
major; drink (8 g fresh (−)-epicatechin, and 0.63
fruit extract and 100 chlorogenic acid,
mL water) hyperoside, and
isoquercitrin
C. pinnatifida var. Ideain, chlorogenic acid, μg/mg 13.4, 18.5, 199, 22.1, [33]
major; fruits (peel/flesh epicatechin, hyperoside, 15.7, and 2.3
polyphenolic extracts) isoquercitrin, and
quercitrin
C. monogyna; fruits Phenolic acids mg GAE/100 g FW 242–879 [6]
from 2 sites harvested Flavonols mg RE/100 g FW 60.88–448
between 2007 and 2009 Anthocyanins mg PGE/ 100 g FW 10.66–47.32
TPC mg/100 g FW 347–1374
C. pinnatifida; TPC M/W/E mg GAE/g 499/146/174 [36]
methanolic (M), water TFC M/W/E mg QE/g 472/2.8/87.8
(W), and 95% ethanolic TTRC M/W/E mg ursolic acid/g 1.7/10.2/13.0
(E) extracts
TTC M/W/E %/mg extract 35/13.8/8.3
C. azarolus; fruit peel (−)-Epicatechin mg/100 g FW 12.24 (E) and 18 (U) [13]
(E) and pulp (U) Neochlorogenic acid mg/100 g FW 0.09 (E) and 0.1 (U)
Cryptochlorogenic acid mg/100 g FW 0.08 (E)
Chlorogenic acid mg/100 g FW 0.24 (E) and 0.10 (U)
Hyperoside mg/100 g FW 103 (E) and 6.84 (U)
Isoquercitrin mg/100 g FW 27.21 (E) and 1.36 (U)
Total mg/100 g FW 143 (E) and 26.31 (U)
C. pinnatifida; ethyl ORAC/IC50 mmol/g/mg/mL 28/10.5 (EA) [15]
acetate (EA) extract 375/0.41 (W)
and water (W) extract
C. monogyna; unripe/ TPC mg GAE/g extract 702/274/247 [7]
ripened/overripened TFC mg CE/g extract 436/21.7/22.3
fruits
C. monogyna/C. TPC mg GAE/100 g FW 217/379 [8]
azarolus; extracts with Total carotenoids mg β-carotene/ 1.37/0.58
phosphate buffer 100 g FW
C. monogyna; fruits TPC mg GAE/ 100 g FW 449–1439 [6]
from 2 sites harvested ABTS•+/DPPH• mmol TE/100 g FW 1.68–6.12/0.76–27.03
between 2007 and 2009 FRAP mmol TE/100 g FW 3.28–10.99
C. azarolus (yellow TPC mg GAE/100 g FW 305/190 [14]
azarole); pulp/syrup of TEAC μmol/g FW 55.58/5.27
wild fruits from Tunisia FRAP μmol Fe2+/g FW 33.75/18.07
C. monogyna (red TPC mg GAE/100 g FW 228/263 [14]
azarole); pulp/syrup of TEAC μmol/g FW 10.81/5.52
wild fruits from Tunisia FRAP μmol Fe2+/g FW 29.23/37.3
(Continued )
Hawthorn Juice 315

Bioactives and Antioxidant Activities of Hawthorn Fruit and Juice
Species, Cultivar, and Bioactives or
Characterization of Antioxidant Activity Content or
the Assayed Samples Assay Unit Antioxidant Activity References
C. pinnatifida; HRP-luminol-H2O2 assay, μg/mL 544/762/603 [36]
methanolic/water/95% IC50
ethanol extracts Pyrogallol–luminol assay, μg/mL 329/>1000/1000
IC50
CuSO4–Phen–Vc–H2O2 μg/mL 13.2/540/350
assay,
IC50 μg/mL 1.1/56.9/7.4
Luminol–H2O2 assay, IC50
C. aronia; water extract Inhibition of β-carotene μg/mL 79% at 100 [19]
from dried unripe fruits oxidation
and leaves Inhibition of AAPH- μg/mL 90% at 667
induced plasma oxidation
Inhibition of Fe2+-induced μg/mL 90% at 1000, IC50 43
lipid peroxidation in rat
liver homogenates
O2•−scavenging in NBT μg/mL IC50, 42
reduction assay
C. monogyna; unripe/ DPPH•, IC50, μg/mL extract 20.93/121.3/130.2 [7]
ripened/overripened Reducing power μg/mL extract 17.42/55.8/75.54
β-Carotene bleaching μg/mL extract 6.38/80.50/100.4
TBARS inhibition μg/mL extract 5.42/32.90/49.21
C. monogyna/C. TEAC μM TE/g FW 8.43/4.10 [8]
azarolus extracted with H2O2 scavenging % at 5 mg/mL 86.39/12.90
phosphate buffer OH• scavenging % at 5 mg/mL 81.04/78.61
Abbreviations: AAPH, 2,2′-azobis(2-amidino-propan) dihydrochloride); ABTS, 2,2′-azino-bis(3-ethylbenzothiazoline-
6-sulphonic acid); CE, catechin equivalents; DPPH, 2,2-diphenyl-1-picrylhydrazyl; EC50; IC50, effective
concentration (scavenging 50% of radicals present in the reaction); FRAP, ferric-reducing antioxidant
power; FW, fresh weight; GAE, gallic acid equivalents; HRP, horseradish peroxidase; NBT, nitro blue tet-
razolium, ORAC, oxygen radical absorbance capacity; PGE, pelargonidin-3-glucoside equivalents; QE,
quercetin equivalents; RE, rutin equivalents; TBARS, thiobarbituric acid–reactive substances; TE, trolox
equivalents; TEAC, trolox equivalent antioxidant capacity; TFC, total flavonoid content; TPC, total pheno-
lic content; TTC, total tannin content; TTRC, total triterpenoid content.
whereas concentrated juice demonstrated O2•− and HO • scavenging capacity with IC50 values of 127 and
41 μg/mL, respectively [16]. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity
of ethanolic extracts from 20 genotypes of 8 Crataegus spp. expressed as inhibition concentration 50
(IC50) was in the range of 0.06–0.46 μg/mL. The values of TPC, TFC, and ascorbic acid were in the
range of 52–558 mg GAE/100 g, 3–36 mg QE/100 g, and 27.51–84.15 mg/100 g, respectively [17].
The water extract of C. pinnatifida produced at 100°C and containing 6.9% and 2.2% of flavonoids
and procyanidins, respectively, scavenged DPPH at IC50 of 0.1 mg/mL [18]. Antioxidant properties were
tested in various systems. For instance, hot water extract of C. pinnatifida significantly slowed the relative
electrophoretic mobility of low-density lipoprotein (LDL) in copper-induced LDL-oxidation assay and
reduced thiobarbituric acid reactive substances (TBARS). The extracts completely blocked the sodium
nitroprusside–mediated macrophage-induced LDL oxidation at 0.05 and 0.10 mg/mL, also achieved
by (+)-catechin and (−)-epicatechin at 0.001 mg/mL and 0.01 mg/mL, respectively, in RAW (Abelson
murine leukemia virus transfromed) 264.7 cells in 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium
bromide (MTT) assay (colorimetric assay for assessing cell viability; MTT is a yellow tetrazole) [18].
Water extract from dried unripe fruits of C. aronia caused a concentration-dependent increase in glutathi-
one (GSH) levels and its oxidized form, glutathione disulfide content [19].
OH
OH OH
HO O HO O
OH
OH OH
OH OH
Catechin Epicatechin
OH
OH
HO
OH OH
O OH
HO O HO
OH HO O
O O
OH O HO
OH
OH OH O
Hyperoside Vitexin
OH
HO O OH
OH OH OH
HO OH
O O O HO O
OH O
H3C O OH
HO O O
HO OH
HO OH O
OH OH
Rutin Isoquercitrin
FIGURE 25.1 Most abundant bioactive compounds in hawthorn juice.
25.3.3 Effects of Storage

Low-temperature storage is recommended for maintaining the quality and efficacy of hawthorn fruit and its
preparations as it was demonstrated in the studies of polyphenolics, which were stable at 4°C and relatively
unstable at 23°C and 40°C with varied extents of degradation [20]. TPC of hawthorn during 12 months
of storage decreased from 891 to 681 mg GAE/100 g FW, TFC from 537 to 420 mg catechin equivalents
(CE)/100 g FW, and TAC from 16 to 9 mg CGE/100 g FW. During storage, the ferric-reducing antioxi-
dant power (FRAP) values of hawthorn ranged between 4.3 (at months 2) and 6.4 mmol Fe2+ /100 g FW
(at months 3), while a 6% loss in reducing power was detected at the end of storage (5.9 mmol Fe2+ /100 g FW)
in comparison to the initial value (6.3 mmol Fe2+ /100 g FW) [21]. In another study, the same authors showed
that storage at 4°C did not adversely influence the content of hawthorn phytochemicals, except for TAC,
which was 15.1% lower at the end of storage. TFC and TAC decreased from 537 to 444 mg CE/100 FW
and from 15.47 to 7.08 mg CGE/100 g FW, respectively, after 14 days of storage at 25°C. The loss in trolox
equivalents (TE) antioxidant capacity in 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS)
assay was 22.9% of the initial value of 7.27 mmol TE/100 g FW after 1 year of frozen storage [22].
25.4 Health Effects

Hawthorn fruits accumulate various bioactive constituents, mainly belonging to several classes of poly-
phenolics. Health effects of Crataegus spp. are mainly attributed to their phenolic compounds [12]. Many
of these compounds are also found in other fruits and have been tested by various in vitro and in vivo
Hawthorn Juice 317
assays demonstrating their activities, which may be promising in terms of health benefits. The reports
on direct health effects of hawthorn juice or drinks are rather scarce. The majority of studies have been
performed with extracts, tinctures, or other preparations obtained from various Crataegus spp. fruits by
different procedures and solvents. Such extracts usually contain higher concentrations of bioactive com-
pounds than the juice. However, most are also present in the juice, and, therefore, it is reasonable to briefly
review the relevant studies on health benefits of preparations obtained from hawthorn fruits. In general,
the health benefits of hawthorn may be grouped into effects on the heart and vascular system, hypotensive
and hypolipidemic effects, antioxidant, radical scavenging, and anti-inflammatory activities [12].
25.4.1 Benefits on the Heart and Vascular System and Hypotensive Effects
In general, hawthorn extract is used as an adjuvant therapy for patients with congestive heart failure [12].
Benefits of Crataegus preparations in symptom control and physiologic outcomes [23] as well as their
significant potential in the treatment of CVD [24] have been demonstrated in numerous published
studies. However, these studies provide rather controversial results that are both positive [2,25–27] or
with no significant effects [28]. It has also been reported that overdose can cause cardiac arrhythmia and
dangerously low blood pressure, whereas milder side effects include nausea and sedation [29].
25.4.2 Hypolipidemic Effects

Lipid-lowering effects of hawthorn extract and powder have been demonstrated in animals fed high-fat
diets. The ability of extracts in lowering the level of total cholesterol, LDL, and triacylglycerols (TAG)
and increasing the level of high-density lipoprotein (HDL) was reported in several studies and explained
by several possible mechanisms [12]. The mice fed on hawthorn fruit diet developed significantly
decreased atherosclerotic lesions, serum lipids, and very-low-density lipoprotein (VLDL) and LDL [30].
25.4.3 Anti-Inflammatory and Antioxidant Activities

Anti-inflammatory, antioxidant, and radical scavenging activities of the main bioactive compounds, which
are present in hawthorn fruits, are reported in many studies. In vitro antioxidant properties, mainly mea-
sured as radical scavenging capacity in various assays, were presented in Table 25.2. Such properties
were also studied in assays, which were more related to the biological systems [12]. For instance, it was
suggested that therapeutic benefits of C. aronia can be, at least, partially attributed to the efficient scav-
enging of O2•− and possible increase in GSH biosynthesis. Water-soluble extracts also inhibited oxidation
of β-carotene, 2,2′-azobis(2-amidinopropane) dihydrochloride–induced plasma oxidation, and Fe2+-
induced lipid peroxidation in rat liver homogenates [19]. In vitro assays showed that the flavonoid extract
of C. pinnatifida decreased the production of prostaglandin E2 and nitric oxide (NO) as induced by lipo-
polysaccharide (LPS) in macrophage cells, whereas in in vivo (rats), it significantly attenuated the increase
in activities of hepatic enzymes, reduced the incidence of liver lesions, decreased the hepatic expression of
inducible NO synthase (iNOS), and cyclooxygenase 2, all induced by LPS [31]. Increased NO production
is involved in the process of inflammation; therefore, inhibitors of iNOS might protect from inflammation
and hepatic damage–induced toxins [12]. Hawthorn phenolics may play a crucial role in antioxidant and
anti-inflammatory processes. Hydroethanolic extract of C. pinnatifida fruit composed of 19.86% procyan-
idin B2, 15.27% epicatechin, 3.10% chlorogenic acid, 2.91% hyperoside, and 1.34% isoquercitrin improved
antioxidant indicators in the serum, liver, and brain of senescence-accelerated mice. The activities of
superoxide dismutase (SOD), catalase, and glutathione peroxidase (GSH-Px) increased, meanwhile the
malondialdehyde content declined [32]. The mRNA expression levels of the SOD1, SOD2, and GSH-Px3
were higher in the livers of mice on the hawthorn fruit diet compared with those in the control mice [30].
25.4.4 Other Effects

Hawthorn fruit extract was tested as a natural chemopreventive agent. Peel and flesh polyphenolic extracts
of hawthorn inhibited Michigan Cancer Foundation-7 (MCF-7) cell growth in a dose-dependent man-
ner with the IC50 of 88.6 and 175.5 μg/mL, respectively. The extracts mediated the cell cycle arrest at the
S-phase, which led to apoptosis of MCF-7 cells via the mitochondrial pathway, as evidenced by the activa-
tion of caspase-3 and caspase-9 and the elevation of intracellular reactive oxygen species (ROS) production
[33]. The aqueous extract of C. aronia at 100–500 mg/kg significantly altered the bleeding time and the
closure time of male albino Wistar rats, as determined by the platelet function analyzer-100 and thrombox-
ane B2 levels, suggesting significant platelet function inhibition [34]. Supplementation with water extract of
C. monogyna proved useful against reproductive toxicity during cyclosporine (immunosuppressant drug)
treatment in a rat model [35]. Water, methanolic, and ethanolic extracts of C. pinnatifida fruit at 0.5–5 μg/
mL protected PC12 neuronal cells against H2O2-induced cell death and showed their potential to serve
as novel neuroprotective agents in nutraceutical products for preventing oxidative-related disorders [36].

Hawthorn fruits have mainly been used for medicinal preparations. For food purposes, hawthorn berries
have traditionally been used for producing jams, compotes, and pressing juice. However, juice yield is
rather low. Dried fruit can also be ground into flour and mixed into baked goods as a special flavor-
ing and healthy ingredient. Commercial hawthorn juice, concentrated juice, and powdered products
are mainly produced in China [37]; some production is also made in Europe, especially in Poland [38].
Various technologies are applied in the processing of hawthorn fruits, including filtration, ultrahigh
treatment sterilization, vacuum evaporation, and spray- and freeze-drying. Juice and its concentrate can
be used for drinks, health-care products, baby foods, puffed foods, baking foods, ice creams, oatmeal,
candy fillings, desserts, breakfast cereals, and yogurt flavorings and in other application where a fresh
fruit flavor is desired. Hawthorn juice and its concentrate are particularly suitable for jellies and sauces
where a boost of flavor without the addition of liquid is necessary [37].
The relevant data on the global production of hawthorn juice and other products are not available.
However, considering fast developments in the area of functional foods, it may be expected that process-
ing of hawthorn may increase by producing both traditional and innovative products. In addition to the
traditional production of juice by pressing, the following trends can be observed: (1) optimization of juice
processing technologies in terms of better extraction of bioactive compounds and improved flavor proper-
ties, (2) development of bioactive compound–enriched fruit fractions, (3) development of hawthorn juice
blends with other fruits, and (4) development of specialty ingredients for various applications in foods. Some
examples of such innovations include the production of flavonoid-rich juice from C. pinnatifida containing
1.83 g RE/100 mL that was established by cellulose hydrolysis and membrane filtration [39]. Five different
blend combinations made of juice from one cultivar of muscadine grape and varying levels of juice from one
cultivar of mayhaw (Crataegus opaca) “Texas star” fruit were tested both for individual juice quality and
for juice-blend compatibility. A consumer preference test showed that juices from 60/40, 30/70, and 40/60
mayhaw/muscadine had the best flavor and overall acceptability [40]. A mixed beverage was also devel-
oped from mushroom juice (36.4%), hawthorn juice (45.4%), and apple juice (18.2%) [41]. The ultrasound-
assisted extraction of health-promoting flavonoids from hawthorn seed, an important by-product of the
hawthorn industry, was achieved. Under optimum conditions, the extraction rate was up to 91.7%, yielding
16.45 mg/g flavonoid that was 1.32-fold the yield of conventional extraction under reflux conditions [42].
Hawthorn juice is generally a safe product. However, contamination with various hazardous toxicants
may occur. For instance, 1 out of 13 analyzed samples from Chinese market was contaminated with
patulin, a toxic secondary metabolite produced by several fungal species of Penicillium, Aspergillus, and
Byssochlamys, at a level of 12.26 μg/kg [43].
25.6 Conclusion
Hawthorn contains various bioactive phytochemicals and has nutritional and functional potential, pro-
viding chemical compounds with biological properties. However, it still remains as an underutilized wild
fruit requiring further research in different directions for better valorization of its applications in the
production of juice and functional ingredients for foods and nutraceuticals.
Hawthorn Juice 319
REFERENCES
1. Phipps, J.B., O’Kennon, R.J., and Lance, R.W., Hawthorns and Medlars, Royal Horticultural Society,
Cambridge, UK, 2003.
2. Fong, H. and Bauman, J., Alternative medicines for cardiovascular diseases: Hawthorn. J. Cardiovasc.
Nurs., 16, 1–8, 2002.
3. Caliskan, O., Mediterranean hawthorn fruit (Crataegus) species and potential usage, in The
Mediterranean Diet: An Evidence-Based Approach, Preedy, V.R. and Watson, R.T, Eds., Academic
Publisher, London, UK, 2015, pp. 621–628.
4. Jurikova, T., Sochor, J., Rop, O., Mlcek, J., Balla, S., Szekeres, L., Adam, V., and Kizek, R., Polyphenolic
profile and biological activity of Chinese hawthorn (Crataegus pinnatifida Bunge) fruits. Molecules, 17,
14490–14509, 2012.
5. Serçe, S., Şimşek, Ö., Toplu, C., Kamiloğlu, Ö., Çalişkan, O., Gündüz, K., Özgen, M., and Kaçar, Y.A.,
Relationships among Crataegus accessions sampled from Hatay, Turkey, as assessed by fruit character-
istics and RAPD. Gen. Res. Crop Evol., 58, 933–942, 2011.
6. Ruiz-Rodríguez, B.M., de Ancos, B., Sanchez-Moreno, C., Fernández-Ruiz, V., Sánchez-Mata, M.D.,
Cámara, M., and Tardío, J., Wild blackthorn (Prunus spinosa L.) and hawthorn (Crataegus monogyna
Jacq.) fruits as valuable sources of antioxidants. Fruits, 69, 61–73, 2014.
7. Barros, L., Carvalho, A.M., and Ferreira, I.C.F.R., Comparing the composition and bioactivity of
Crataegus monogyna flowers and fruits used in folk medicine. Phytochem. Anal., 22, 181–188, 2011.
8. Egea, I., Sánchez-Bel, P., Romojaro, F., and Pretel, M.T., Six edible wild fruits as potential antioxidant
additives or nutritional supplements. Plant. Food Hum. Nutr., 65, 121–129, 2010.
9. Edwards, J.E., Brown, P.N., Talent, N., Dickinson, T.A., and Shipley, P.R., A review of the chemistry of
the genus Crataegus. Phytochemistry, 79, 5–26, 2012.
10. Liu, P.Z., Kallio, H., Lu, D.G., Zhou, C.S., Ou, S.Y., and Yang, B.R., Acids, sugars, and sugar alcohols
in Chinese hawthorn (Crataegus spp.) fruits. J. Agric. Food Chem., 58, 1012–1019, 2010.
11. Bahri-Sahloul, R., Ammar, S., Grec, S., and Harzallah-Skhiri, F., Chemical characterisation of Crataegus
azarolus L. fruit from 14 genotypes found in Tunisia. J. Hortic. Sci. Biotechnol., 84, 23–28, 2009.
12. Yang, B. and Liu, L., Composition and health effects of phenolic compounds in hawthorn (Crataegus
spp.) of different origins. J. Sci. Food. Agric., 92, 1578–1590, 2012.
13. Belkhir, M., Rebai, O., Dhaouadi, K., Sioud, B., Amri, M., and Fattouch, S., Antioxidant and antimi-
crobial activities of Tunisian azarole (Crataegus azarolus L.) leaves and fruit pulp/peel polyphenolic
extracts. Int. J. Food Prop., 16, 1380–1393, 2013.
14. Belkhir, M., Rebai, O., Dhaouadi, K., Congiu, F., Tuberoso, C.I.G., Amri, M., and Fattouch, S.,
Comparative analysis of Tunisian wild Crataegus azarolus (yellow azarole) and Crataegus monogyna
(red azarole) leaf, fruit, and traditionally derived syrup: Phenolic profiles and antioxidant and antimi-
crobial activities of the aqueous-acetone extracts. J. Agric. Food Chem., 61, 9594–9601, 2013.
15. Chen, C.-Y., Li, H., Yuan, Y.-N., Dai, H.-Q., and Yang, B., Antioxidant activity and components of a
traditional Chinese medicine formula consisting of Crataegus pinnatifida and Salvia miltiorrhiza. BMC
Complement. Altern. Med., 13, 99, 2013.
16. Cui, T., Nakamura, K., Tian, S., Kayahara, H., and Tian, Y.-L., Polyphenolic content and physiological
activities of Chinese hawthorn extracts. Biosci. Biotechnol. Biochem., 70, 2948–2956, 2006.
17. Garcia-Mateos, R., Ibarra-Estrada, E., and Nieto-Angel, R., Antioxidant compounds in hawthorn fruits
(Crataegus spp.) of Mexico. Revista Mexicana de Biodiversidad, 84, 1298–1304, 2013.
18. Chu, C.Y., Lee, M.J., Liao, C.L., Lin, W.L., Yin, Y.F., and Tseng T.H., Inhibitory effect of hot-water
extract from dried fruit of Crataegus pinnatifida on low-density lipoprotein (LDL) oxidation in cell and
cell-free systems. J. Agric. Food Chem., 51, 7583–7588, 2003.
19. Ljubuncic, P., Portnaya, I., Cogan, U., Azaizeh, H., and Bomzon, A., Antioxidant activity of Crataegus
aronia aqueous extract used in traditional Arab medicine in Israel. J. Ethnopharmacol., 101, 153–161,
2005.
20. Chang, Q., Zuo, Z., Chow, M.S.S., and Ho, W.K.K., Effect of storage temperature on phenolics stability
in hawthorn (Crataegus pinnatifida var. major) fruits and a hawthorn drink. Food Chem., 98, 426–430,
2006.
21. Šamec, D. and Piljac-Žegarac, J., Fluctuations in the levels of antioxidant compounds and antioxidant
capacity of ten small fruits during one year of frozen storage. Int. J. Food Prop., 18, 21–32, 2015.
22. Šamec, D. and Piljac-Žegarac, J., Postharvest stability of antioxidant compounds in hawthorn and cor-
nelian cherries at room and refrigerator temperatures—Comparison with blackberries, white and red
grapes. Sci. Hortic., 131, 15–21, 2011.
23. Pittler, M.H., Guo, R., and Ernst, E., Hawthorn extract for treating chronic heart failure. Cochrane
Database Syst. Rev., CD005312, 2008.
24. Tassell, M., Kingston, R., Gilroy, D., Lehane, M., and Furey, A., Hawthorn (Crataegus spp.) in the treat-
ment of cardiovascular disease. Pharmacogn. Rev., 4, 32–41, 2010.
25. Zhang, Z., Chang, Q., Zhu, M., Huang, Y., Ho, W.K., and Chen, Z., Characterization of antioxidants
present in hawthorn fruits. J. Nutr. Biochem., 12, 144–152, 2001.
26. Degenring, F.H., Suter, A., Weber, M., and Saller, R., A randomised double blind placebo controlled
clinical trial of a standardised extract of fresh Crataegus berries (Crataegisan) in the treatment of
patients with congestive heart failure NYHA II. Phytomedicine, 10, 363–369, 2003.
27. Rigelsky, J.M. and Sweet, B.V., Hawthorn: Pharmacology and therapeutic uses. Am. J. Health Syst.
Pharm., 59, 417–422, 2002.
28. Zick, S.M., Vautaw, B.M., Gillespie, B., and Aaronson, K.D., Hawthorn extract randomized blinded
chronic heart failure (HERB CHF) trial. Eur. J. Heart Fail., 11, 990–999, 2009.
29. Dasgupta, A., Kidd, L., Poindexter, B.J., and Bick, R.J., Interference of hawthorn on serum digoxin
measurements by immunoassays and pharmacodynamic interaction with digoxin. Arch. Pathol. Lab.
Med., 134, 1188–1192, 2010.
30. Zhang, Y., Zhang, L., Geng, Y., and Geng, Y., Hawthorn fruit attenuates atherosclerosis by improving
the hypolipidemic and antioxidant activities in apolipoprotein E-deficient mice. J. Atheroscler. Thromb.,
21, 119–128, 2014.
31. Kao, E.-S., Wang, C.-J., Lin, W.-L., Yin, Y.-F., Wang, C.-P., and Tseng, T.-H., Anti-inflammatory poten-
tial of flavonoid contents from dried fruit of Crataegus pinnatifida in vitro and in vivo. J. Agric. Food
Chem., 53, 430–436, 2005.
32. Wang, H., Zhang, Z., Guo, Y., Sun, P., Lv, X., and Zuo, Y., Hawthorn fruit increases the antioxidant
capacity and reduces lipid peroxidation in senescence-accelerated mice. Eur. Food Res. Technol., 232,
743–751, 2011.
33. Li, T., Zhu, J., Guo, L., Shi, X., Liu, Y., and Yang, X., Differential effects of polyphenols-enriched
extracts from hawthorn fruit peels and fleshes on cell cycle and apoptosis in human MCF-7 breast car-
cinoma cells. Food Chem., 141, 1008–1018, 2013.
34. Shatoor, A.S., Soliman, H., Al-Hashem, F., El- Gamal, B., Othman, A., and El-Menshawy, N., Effect of
hawthorn (Crataegus aronia syn. Azarolus (L)) on platelet function in Albino Wistar rats. Thromb. Res.,
130, 75–80, 2012.
35. Armand, Z., Najafi, G., Farokhi, F., and Jalali, A.S., Attenuation of cyclosporine-induced sperm impair-
ment and embryotoxicity by Crataegus monogyna fruit aqueous extract. Cell J., 15, 198–205, 2013.
36. Chang, C.L., Chen, H.S., Shen, Y.C., Lai, G.H., Lin, P.K., and Wang, C.M., Phytochemical composi-
tion, antioxidant activity and neuroprotective effect of Crataegus pinnatifida fruit. S. Afr. J. Bot., 88,
432–437, 2013.
37. Alibaba, Published online at: http://www.alibaba.com/showroom/hawthorn-juice_2.html (accessed
April 13, 2015).
38. Eko Medica, Published online at: http://zdrowie.bioieko.pl/soki/eko-medica-sok-glog-500.html
(accessed February 20, 2015).
39. Sun, J.L., Fan, Y.F., Li, G., and Zhao, R.X., Preparation of flavonoid-rich hawthorn juice with cellulose
hydrolysis and membrane filtration, in Proceedings of the Second Conference on Horticulture Science
and Technology (CHST 2010), Zhang, Y., Ed., Science Publications, London, UK, 2010, pp. 41–44.
40. Trappey, A.F. and Johnson, C.E., Consumer acceptance of mayhaw (Crataegus opaca Hook. and Arn.) fruit
juice blended with muscadine (Vitis rotundifolia Michx.) grape juice. Hortic. Sci., 40, 1068–1068, 2005.
41. Hou, X.J., Zhang, N., Xiong, S.Y., Li, S.G., and Yang, B.Q., Extraction of BaChu mushroom polysac-
charides and preparation of a compound beverage. Carbohydr. Polym., 73, 289–294, 2008.
42. Pan, G., Yu, G., Zhu, C., and Qiao, J., Optimization of ultrasound-assisted extraction (UAE) of flavo-
noids compounds (FC) from hawthorn seed (HS). Ultrason. Sonochem., 19, 486–490, 2012.
43. Zhou, Y.C., Kong, W.J., Li, Y., Logrieco, A.F., Xu, J., and Yang, M.H., A new solid-phase extraction and
HPLC method for determination of patulin in apple products and hawthorn juice in China. J. Sep. Sci.,
35, 641–649, 2012.
26
Indian Gooseberry (Amla) Juice
Neelima Garg and Pushpa Chethan Kumar
CONTENTS
26.1 Introduction....................................................................................................................................321
26.4 Health Effects................................................................................................................................ 325
26.6 Conclusion..................................................................................................................................... 327
References............................................................................................................................................... 327
26.1 Introduction
Indian gooseberry, also known as amla, has been considered as a miracle fruit in Indian medicine
(Ayurveda). According to two main classic texts on Ayurveda, Charak Samhita and Sushrut Samhita,
amla is regarded as “the best among rejuvenative herbs,” “useful in relieving cough and skin disease,”
and “the best among the sour fruits” [1]. It plays a major role in herbal medicine to treat or cure many
diseases because of its nutraceutical properties. India is the largest producer in the world of amla. It is
commercially cultivated in the Uttar Pradesh, Maharashtra, Gujarat, and Rajasthan states of India. Many
of the traditional products prepared from amla include squash, preserves, syrups, jams, candy, shreds,
sauces, and juice. Except for amla juice, other products produced from amla are rich in calories. With
rising health consciousness among consumers, amla juice is gaining importance.
Unlike other fruit juices, amla juice is not used as conventional fruit drink. Consumers do not relish
this fruit juice in a large quantity because of its highly acidic and astringent nature and is consequently
used for medicinal purposes only. In India, a number of large companies as well as small-scale industries
involved in amla processing are manufacturing amla juice for national and international markets. This
chapter highlights the nutraceutical properties of amla juice and associated novel products.

Amla juice is obtained by pulping and pressing amla fruits (Figure 26.1). It is a rich source of ascorbic
acid (300–1095 mg/100 mL) (Table 26.1) and polyphenols (2.9–3.6 g/100 mL) (Table 26.2). The fresh
amla juice contains 20 times more vitamin C as compared with orange juice. A single amla fruit is
equivalent to one or two oranges in vitamin C content. The daily recommended allowance for v itamin C
is about 45–90 mg/day (aged 9–70), whereas for lactating mothers, it is 120 mg/day (aged 19–50).
Consumption of a fresh amla fruit or a glass of amla juice will fulfill the daily vitamin C requirement.
Amla is not only an exceptional fruit among others due to its ascorbic acid content, but it also contains
substances that partially protect the ascorbic acid from destruction upon heating or drying due to high
acidity [2].
321
Selection of fruits
Washing
Direct extraction of juice

Shredding
Extraction of juice
Filter
Pasteurization
Filling in bottle and sealing
Store
FIGURE 26.1 Flowchart for the preparation of amla juice.
TABLE 26.1
Compositional and Nutritional Characteristics of Amla Juice
Nutrient Unit Amla Juice References
Moisture % 97 [9]
TSS °Brix 4.6 [9]
Acidity as citric acid % 1.82 [9]
Reducing sugar % 0.6 [9]
Ascorbic acid mg/100 mL 300–1095 [3,16]
Phosphorous g/100 g 0.08 [9]
Calcium mg/kg 17.7 [9]
Iron mg/kg 4.8 [9]
Abbreviation: TSS, total soluble solid.
TABLE 26.2
Polyphenols in Amla Juice (per 100 mL)
Polyphenols Unit Amla Juice References
Polyphenols g 2.9–3.6 [9,16]
Gallic acid mg 0.15 a
Kaempferol mg 1.57 a
Caffeic acid mg 0.04 a
a Unpublished data.
Indian Gooseberry (Amla) Juice 323
Jain and Khurdiya [4] standardized a procedure for the extraction of juice from amla fruit. Blanching
of fruits prior to juice extraction significantly improved the juice recovery, increased the density and
tannin content of the juice, but reduced the vitamin C content by 12%. Blending of amla juice with other
fruit juices for the preparation of ready-to-drink beverages was demonstrated to enhance the nutritional
quality and sensory acceptability of the amla drink [5–8].
Garg et al. [9] monitored the nutritive constituents of amla juice decreased during storage. The loss
of ascorbic acid content during processing and storage was highly significant in all products including
amla juice. Rathnam and Srinivasam [10] suggested vacuum processing technique for the preparation
of stable vitamin C concentrate from amla. To offset losses during storage, enrichment of the fruit pulp
with ascorbic acid is recommended. A considerable loss in ascorbic acid during heating, possibly due
to its thermal destruction, was reported [5]. However, the loss of the ascorbic acid content was high by
processing at 80°C for 20 min as compared to 90°C for 1 min.
Bhosale et al. [11] carried out a study to detect the changes in color and quality attributes of amla juice
during storage following pasteurization at different temperatures. After extracting juice from amla, it was
pasteurized at five different temperatures and preserved with 500 ppm SO2 in polyethylene terephthalate
(PET) bottles under ambient conditions. Juice was periodically analyzed for color and chemical param-
eters up to 9 months of storage. Though the contents of ascorbic acid and polyphenols decreased in juice
with increasing storage period, the effect of pasteurization temperature was not significant. High degree
of browning was observed in juice heated at higher temperatures (90°C and 95°C) as compared to lower
temperatures (75°C and 80°C) throughout the storage period as indicated by increase in nonenzymatic
browning values. The degree of browning was further confirmed by higher negative numerical values of
whiteness index in Hunter’s scale for intensity of color.
Damame et al. [12] reported that amla juice was the most acceptable preserved product with respect to
its maximum retention potential of vitamin C content (298 mg/100 g) at the end of storage period. The
high water content in juice probably permitted better solubility of vitamin C and hence its maximum
retention in amla juice. Amla juice could be served as the best medicinal ready-to-serve (RTS) drink due
to high retention potential of vitamin C. Mobaserri [13] found that amla juice contained 20-fold higher
vitamin C than orange juice. Physiconutritional qualities of fruits such as apple, lime, pomegranate,
Perlette grape, and Pusa Navrang grape were analyzed and compared with amla juice and found that the
latter contained the highest content of vitamin C (479 mg/100 mL). Hence, when amla juice was blended
with other fruit juices for the preparation of RTS beverages, it boosted their nutritional quality in terms
of vitamin C content [14].

Amla juice has been studied for its antioxidant activity. Ascorbic acid and polyphenols are the major anti-
oxidants present in amla juice. Antioxidants are involved in the prevention of cellular damage, which is
the common pathway for cancer, aging, and a variety of other diseases. Bhattacherjee et al. [15] reported
the presence of polyphenols, including gallic acid, kaempferol, and caffeic acid in high quantities in amla
juice (Figure 26.2). Other phenolic compounds such as chlorogenic acid, (+)-catechin, (–)-epicatechin,
p-coumaric acid, and p-hydroxybenzaldehyde were also identified in amla juice. A study reported that
total polyphenol content remained unchanged during storage, but gallic acid content increased sharply
at 40°C [16]. High-performance liquid chromatography (HPLC) data indicated that the content of gal-
lic acid in juice decreased initially but increased sharply as the storage period prolonged [11]. After
9 months of storage, a higher amount of gallic acid was detected in amla juice pasteurized at 95°C than in
that heated at 75°C. The contents of kaempferol and caffeic acid decreased throughout the storage period
irrespective of pasteurization temperature. Scartezzini et al. [1] indicated that the Ayurvedic traditional
method of processing the fruit (svaras bhavana), which involved repeated treatment of the dried fruit
with its fresh juice up to 21 times, increased its beneficial characteristics and the amount of ascorbic
acid, polyphenols, and antioxidant activity than in dried fruit. Amla juice has shown very high reducing
power. Both aqueous and alcoholic extracts of amla showed protective activity against tert-butyl hydro-
peroxide (t-BOOH)-induced cytotoxicity and reactive oxygen species (ROS) generation in cultured C6
O OH
HO OH
OH
Gallic acid
OH
HO O
OH
OH O
Kaempferol
OH
HO
OH
Caffec acid
OH
O OH
O
O O C
OH
OH
O C
O O O OH
O C C O
OH
HO OH
HO OH HO OH
Emblicanin A
OH
O OH
O
O O C OH
OH
O C
O O O OH
O C C O
OH
HO OH
HO OH HO OH
Emblicanin B
FIGURE 26.2 Chemical structures of major tannoids present in amla juice. (Continued)
OH
O OH
OH
O O C
OH
OH
O C
O O O OH
O C C O
OH
HO OH
HO OHHO OH
Pedunculagin
HO2C
OH
H OH OH
O
HO H
H O C
H OH OH
OH
OH2C C
O OH
OH
Punigluconin
FIGURE 26.2 (Continued) Chemical structures of major tannoids present in amla juice.
glioma cells at a dose of 50 mg/mL [17]. The ethyl acetate fraction of phenolics present in amla included
geraniin, quercetin 3-β-d-glucopyranoside, kaempferol 3-β-d-glucopyranoside, isocorilagin, quercetin,
and kaempferol, which showed strong antioxidant and radical scavenging activities. The antioxidant
activity of flavonoids (quercetin 3-β-d-glucopyranoside, kaempferol 3-β-d-glucopyranoside, quercetin,
and kaempferol) was moderate to weak in 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, and flavonoids
belonging to the quercetin class (quercetin and quercetin 3-β-d-glucopyranoside) had higher antioxidant
activities due to their possessing of an O-dihydroxy B-ring structure, which conferred higher stability in
the radical form and participated in electron delocalization [18].
26.4 Health Effects

The potential of amla juice as a medicine has been demonstrated in many epidemiological studies. Herbal
medicine is still favored and used by about 75%–80% of the world’s population, mainly in developing
countries for primary health care due to better cultural acceptability, better compatibility with the human
body, and lesser side effects. Studies have proven that amla can be used as an herbal medicine, and the
significant nutritive value of amla showed that it can be used as a dietary supplement [19]. The juice of
amla has been used in Ayurveda (Indian medicine) to cure many disorders. Some of the medicine indi-
cated that consumption of amla juice with honey is useful in the treatment of conjunctivitis and glaucoma.
Regular use of a tablespoon of amla juice with honey can promote vigor of the body and its consumption
when mixed with a cup of fresh bitter gourd juice can stimulate the islets of Langerhans thus reducing the
blood sugar level in diabetic patients. The juice from fresh amla fruit is given as tonic, diuretic, and anti-
bilious remedy. It also helps to overcome burning sensation, thirst, dyspepsia, and other digestive prob-
lems. Weakness of the body, heart, and mind is dispelled by taking fresh amla juice between meals [20].
Administration of amla juice reduced the age-related dyslipidemia by inhibiting the protein expres-
sion involved in oxidative stress during the aging process when fed to the aged rats, and thus it may be
beneficial in preventing age-related atherosclerotic cardiovascular disease (CVD) [21]. Several reports
have shown that hyperlipidemia diminishes the antioxidant defense systems, decreasing the activities of
catalase and superoxide dismutase (SOD), and thereby elevating the level of lipid peroxides, resulting
in the production of toxic intermediates. High fat induces decrease in normal activities of glutathione
peroxidase and glutathione reductase enzymes and the content of glutathione in the tissues. The treat-
ment with flavonoids extract from amla at a dose of 10 mg/kg body weight/day significantly elevated the
activities of free radical scavenging enzymes and significantly decreased the content of lipid peroxides
in flavonoid-treated hypercholesterolemic rats [22]. Ngamkitidechakul et al. [23] observed that the juice
of amla, which contains tannins (43%), uronic acid (11%), and gallic acid (21%), inhibited the growth
of A549 (lung), HepG2 (liver), HeLa (cervical), MDA-MB-231 (breast), SK-OV3 (ovarian), and SW620
(colorectal) cells in vitro. However, at the same concentration, the aqueous extract did not cause similar
level of cytotoxicity in the MRC5, normal lung fibroblast, suggesting it to be safe for normal cells.
Amla juice is also rich in emblicanin A and B and tannins including punigluconin and pedunculagin
[24] (Figure 26.2). Emblicanin A may be transformed into emblicanin B, which in turn also attacks free
radicals and is transformed into emblicanin oligomers. This provides amla juice with one of the best free
radical scavenging properties. An emblicanin A (37%)- and emblicanin B (33%)-enriched fraction of
fresh juice of Emblica fruits (EOT) was investigated for antioxidant activity against ischemia–reperfusion
injury (IRI)-induced oxidative stress in rat heart [25]. Vitamin E was used as the standard antioxidant
agent. The IRI-induced effects were prevented by the administration of EOT (50 and 100 mg/kg body
weight) and vitamin E (200 mg/kg body weight) given orally twice daily for 14 days prior to the sacrifice
of the animals and initiation of the perfusion experiments. The study confirmed the antioxidant effect
of Emblica officinalis juice and indicated that it may have a cardioprotective effect [25]. Amla juice was
also effective in reducing cyclophosphamide-induced suppression of humoral immunity and to restore
the levels of glutathione and the antioxidant enzymes in the kidneys and liver of mice [26].
Tannoids (combination of tannins and alkaloids) of amla juice are found to be potent inhibitors of
aldose reductase (AR), which is involved in the development of secondary complications of diabetes,
including cataract. The isolated tannoids prevented not only the AR activation in rat lens organ culture
but also sugar-induced osmotic changes [27]. The elevation in serum glucose levels by high-fructose
diet indicates the progression of insulin resistance. The amla fruit would probably play a protective
role against the abnormal metabolism of carbohydrate induced by a high-fructose diet. It significantly
attenuated the increase in liver weight by the high-fructose diet and significantly decreased the weight of
epididymal fat pads increased by the diet in a rat model [28].
The phenolics present in amla such as geraniin, quercetin 3-β-d-glucopyranoside, kaempferol 3-β-d-
glucopyranoside, isocorilagin, quercetin, and kaempferol were studied for their immunomodulatory and
anticancer activities on mouse splenocytes and human breast cancer cells (MCF-7) and human embry-
onic lung fibroblast cells (HELF). The assay of anticancer activities suggested that geraniin and isocori-
lagin had higher cytotoxicities against MCF-7 and HELF and significant stimulatory effect on splenocyte
proliferation [29]. Preclinical studies have shown that amla juice causes a c oncentration-dependent cyto-
toxic effect on L929 cells in vitro and that the IC50 was 16.5 μg/mL. The extract also caused apoptosis
in Dalton’s lymphoma ascites and CeHa cell lines [30,31]. A study conducted to evaluate the effect of
aqueous extract of E. officinalis, Phyllanthus amarus, and Picrorhiza kurroa on N-nitrosodimethylamine
(NDEA) induced hepatocarcinogenesis and showed a significant inhibition of hepatocarcinogenesis
induced by NDEA in a dose-dependent manner. Male Wistar rats treated with 250 mg of aqueous extract
of E. officinalis did not develop any tumors at 20 weeks, and the animals treated with 125 and 50 mg
showed a reduction in tumor incidence of 60% and 30%, respectively [32].

Ram et al. [33] used amla and bael (Aegle marmelos) fruits for the preparation of blended RTS bever-
ages. The blended RTS beverage prepared with 25% amla and 75% bael pulp with 15°Brix total soluble
solid (TSS) and 0.25% titratable acidity was similar to the beverage prepared with only bael pulp and
was better than pure amla beverage. They found that increased concentration of amla pulp decreased the
acceptability of mixed beverages. Chandan et al. [34] prepared amla RTS beverage with drained amla
syrup. They obtained the drained syrup from blanched slices of amla steeped in salt for 2 h, followed by
steeping in 70°Brix syrup for 24 h, and adjusted it to 20°Brix containing 2% lime juice +1% ginger juice,
which was found to be acceptable with good organoleptic scores.
Bhattacherjee et al. [35] reported that 100 g spray-dried amla powder contained 3282 mg ascor-
bic acid and 22 g polyphenols. Polyphenols identified were gallic acid, caffeic acid, (+)-catechin, and
(–)-epicatechin [16]. Spray-dried powder is one of the most value-added products from amla, which can
be used as RTS health drinks as well as in encapsulated medicine and has tremendous future market
potential and export value.
Fermentation is a cheap and energy-efficient means of preserving perishable raw materials. It requires
very little sophisticated equipment, to carry out either the fermentation or for subsequent storage of the
fermented product. Cider is considered to be a beverage made “wholly or partly from the fermented juice
of apples,” though there are also reports of cider preparation from pear, peach, and raspberry, among
others. Both polyphenols and acids are required in moderate amounts for cider preparation. Since amla
is rich in polyphenols and ascorbic acid, it has been used for cider preparation. Amla cider is a fermented
drink having unique properties of amla along with globally acceptable taste and was developed at the
Central Institute for Subtropical Horticulture, Lucknow, India. Amla juice was fermented with yeast
(Saccharomyces cerevisiae) to afford a sweet fermented drink having 10°Brix TSS, 4% alcohol, and
0.4% polyphenols. Amla cider is a novel health drink with nutritional and medicinal benefits and global
acceptance potential.
26.6 Conclusion
Amla is known for its therapeutic properties from the ancient times in India and is considered as a won-
der fruit for health-conscious people. Unfortunately, despite being a rich source of ascorbic acid, it is
used more for medicinal purposes rather than as a part of a daily diet. The fruits are less utilized for fresh
consumption and generally used after processing because of high astringency and acidity. However, the
loss of nutritional value in processed products, including juice, is very high due to thermal treatment‒
based processing. However, a demand for amla juice is expected to increase manifold in the future
because of the awareness of international community about its health attributes. To benefit the global
consumer, there is a need to develop juice-based products using nonthermal processing techniques for
the best preserved properties of amla along with a globally acceptable taste.
REFERENCES
1. Scartezzini, P., Antognoni, F., Raggi, M.A., Poli, F., and Sabbioni, C., Vitamin C content and antioxidant
activity of the fruit and of the Ayurvedic preparation of Emblica officinalis Gaertn. J. Ethnopharmacol.,
104, 113–118, 2006.
2. Gopalan, C., Rama Sastri, B.V., and Balasubramanian, S.C., Nutritive Value of Indian Foods, Indian
Council of Medical Research, Hyderabad, India, 2011.
3. Parmar, C. and Kaushal, M.K., Wild Fruits, Kalyani Publishers, New Delhi, India, 1982.
4. Jain, S.K. and Khurdiya, D.S., Physico-chemical characteristics and post-harvest technology of aonla
(Phyllanthus emblica L.)—A resume. Indian Food Packer, 47, 46–49, 2002.
5. Prasad, P.S.R.K., Surya Prakash Rao, P.V., Nageshwara Rao, G., and Girdhar, N., Some preliminary
studies on utilization of amla (Phyllanthus emblica Linn. L.) N. Indian Food Packer, 22, 8–11, 1968.
6. Singh, I.S. and Kumar, S., Studies on processing of aonla fruits: II Aonla products. Prog. Hortic., 27,
39–47, 1995.
7. Nath, V., Delicacies of Aonla. Indian J. Hortic., 44, 15–17, 1999.
8. Deka, B.C., Sethi, V., Prasad, R., and Baba, P.K., Application of mixture methodology for beverages
from mix fruit juice/pulp. J. Food Sci. Technol., 38, 615–618, 2001.
9. Garg, N., Sonkar, P., and Bhriguvanshi, S.R., Nutritional and microbial quality evaluation of commer-
cial samples of amla chyavanprash, amla preserve and amla juice. J. Food Sci. Technol., 45, 193–195,
2008.
10. Rathnam, C. and Srinivasam, M., Behavior of ascorbic acid in Indian gooseberry to heat treatment.
J. Sci. Ind. Res., 18, 132–33, 1959.
11. Bhosale, V.I., Kute, L.S., and Kadam, S.S., Studies on preparation of RTS beverage from amla. Beverage
Food World, 27, 24–27, 2000.
12. Damame, S.V., Gaikwad, R.S., Patil, S.R., and Masalkar, S.D., Vitamin C content of various amla
products during storage. Orissa J. Hortic., 30, 19–22, 2002.
13. Mobaserri, R., Ayurveda. Published online at: http://www.chakarpaniayurveda.com/news/July2004.pdf
(accessed November 8, 2013).
14. Jain, S.K. and Khurdiya, D.S., Vitamin C enrichment of fruit juice based ready-to-serve beverages
through blending of Indian gooseberry (Emblica officinalis Gaertn.) juice. Plant Food Human Nutr., 59,
63–66, 2004.
15. Bhattacherjee, A.K., Dikshit, A., and Tandon, D.K., High-performance liquid chromatographic deter-
mination of ascorbic acid and polyphenols in aonla juice and spray-dried powder, in Proceedings of the
International Symposium on Minor Fruits and Medicinal Plants for Health and Ecological Security,
Ghosh, S.N., Ed., ISMF & MP, Kalyani, West Bengal, India, 2012, pp. 264–270.
16. Bhattacherjee, A.K., Tandon, D.K., Dikshit, A., and Kumar, S., Effect of pasteurization temperature on
quality of amla juice during storage. J. Food Sci. Technol., 48, 269–273, 2011.
17. Shukla, V., Vashistha, M., and Singh, S.N., Evaluation of antioxidant profile and activity of amalaki
(Emblica officinalis), spirulina and wheat grass. Indian J. Clin. Biochem., 24, 70–75, 2009.
18. Liu, X., Cui, C., Zhao, M., Wang, J., Luo, W., Yang, B., and Jiang, Y., Identification of phenolics in the
fruit of emblica (Phyllanthus emblica L.) and their antioxidant activities. Food Chem., 109, 909–915,
2008.
19. Rajashree, R., Divya, G., Sushma, P., and Kanchan, I., Amla, ashwagandha and shatavari formulations
as herbal medicines and nutraceuticals. Res. J. Pharmaceutical Sci., 1, 10–15, 2012.
20. Sampath Kumar, K.P., Bhowmik, D., Dutta, A., Yadav, P.A., Paswan, S., Srivastava, S., and Deb, L.,
Recent trends in potential traditional Indian herbs Emblica Officinalis and its medicinal importance.
J. Pharmacog. Phytochem. 1, 24–32, 2012.
21. Yokozawa, T., Kim, H.Y., Kim, H.J., Okubo, T., Djoing-Chi, C., and Juneja, R.L., Amla (Emblica
officinalis Gaertn.) prevents dyslipidaemia and oxidative stress in the ageing process. Br. J. Nutr., 97,
1187–1195, 2007.
22. Anila, L. and Vijayalakshmi, N.R., Antioxidant action of flavonoids from Mangifera indica and Emblica
officinalis in hypercholesterolemic rats. Food Chem., 83, 569–574, 2003.
23. Ngamkitidechakul, C., Jaijoy, K., Hansakul, P., Soonthornchareonnon, N., and Sireeratawong, S.,
Antitumour effects of Phyllanthus emblica L: Induction of cancer cell apoptosis and inhibition of in vivo
tumour promotion and in vitro invasion of human cancer cells. Phytother. Res., 24, 1405–1413, 2010.
24. Ghosal, S., Tripathi, V.K., and Chauhan, S., Active constituent of Emblica officinalis: Part 1st—The
chemistry and antioxidant effects of two new hydrolysable tannins, emblicanin A and B. Indian
J. Chem., 35, 941–948, 1996.
25. Bhattacharya, A., Chatterjee, A., Ghosal, S., and Bhattacharya, S.K., Antioxidant activity of active tan-
noid principles of Emblica officinalis (amla). Indian J. Exp. Biol., 37, 676–680, 1999.
26. Haque, R., Bin-Hafeez, B., Ahmad, I., Parvez, S., Panday, S., and Raisuddin, S., Protective effects of
Emblica officinalis Gaertn. in cyclophosphamide-treated mice. Hum. Exp. Toxicol., 20, 643–650, 2001.
27. Suryanarayana, P., Anil Kumar, P., Saraswat, M., Mark Petrash, J., and Reddy, G.B., Inhibition of aldose
reductase by tannoid principles of Emblica officinalis: Implications for the prevention of sugar cataract.
Mol. Vision, 10, 148–154, 2004.
28. Kim, H.Y., Okubo, T., Juneja, L.R., and Yokozawa, T., The protective role of amla (Emblica officinalis
Gaertn.) against fructose-induced metabolic syndrome in a rat model. Br. J. Nutr., 103, 502–512, 2010.
29. Liu, X., Zhao, M., Wu, K., Chai, X., Yu, H., Tao, Z., and Wang, J., Immunomodulatory and anticancer
activities of phenolics from emblica fruit (Phyllanthus emblica L.). Food Chem., 131, 685–690, 2012.
30. Jose, J.K., Kuttan, G., and Kuttan, R., Antitumour activity of Emblica officinalis. J. Ethnopharma., 75,
65–69, 2001.
31. Rajeshkumar, N.V., Pillai, M.R., and Kuttan, R., Induction of apoptosis in mouse and human carcinoma
cell lines by Emblica officinalis polyphenols and its effect on chemicals carcinogenesis. J. Exp. Clin.
Cancer Res., 22, 201–212, 2003.
32. Jeena, K.J., Joy, K.L., and Kuttan, R., Effect of Emblica officinalis, Phyllanthus amarus and Picrorrhiza
kurroa on N-nitrosodiethylamine induced hepatocarcinogenesis. Cancer Lett., 136, 1, 11–16, 1999.
33. Ram, R.B., Meena, M.L., Sonkar, P., Lata, R., and Upadhyay, A.K., Standardization and evaluation of
blended amla (Emblica officinalis Gaertn.) and bael (Aegle marmelos Correa) RTS beverages. Plant
Arch. 11, 205–208, 2011.
34. Chandan, K., Prashanth, S.J., Nataraj, S.K., Indudhara, S.M., and Rokhade, A.K., Preparation of dehy-
drated slices and RTS beverages from amla (Emblica officinalis Gaertn.) fruits. Int. J. Agric. Sci., 6,
300–304, 2010.
35. Bhattacherjee, A.K., Chaurasia R., and Tandon, D.K., Quality evaluation of aonla powder obtained by
different drying techniques. Prog. Hortic., 4, 110–113, 2012.
27
Kiwifruit Juice
Asim K. Duttaroy
CONTENTS
27.1 Introduction.....................................................................................................................................331
27.2 Nutritional Characteristics..............................................................................................................332
27.3 Bioactives and Antioxidant Efficacy...............................................................................................332
27.4 Health Effects................................................................................................................................. 334
27.5 Novel Products/Formulations and Future Trends...........................................................................335
27.6 Conclusion......................................................................................................................................335
References................................................................................................................................................335
27.1 Introduction
Kiwifruit is the edible berry of the woody vine Actinidia. Among the cultivars of Actinidia, the most
popular are Actinidia deliciosa “Hayward” (green kiwifruit) and Actinidia chinensis “Hort 16A,”
ZESPRI® (gold kiwifruit) [1]. The A. deliciosa has a soft texture and a unique flavor [2,3] and com-
mercially grown in several countries such as in Italy, New Zealand, Brazil, the United States, and Chile.
While the total world production production of kiwifruit has increased by over 50% during the last
decade, the kiwifruit remains a niche fruit, taking up an estimated 0.22% of the global fruit bowl, which
is dominated by apples, oranges, and bananas.
A growing body of scientific evidence supports kiwifruit’s health benefits, including their effects
on metabolic health, iron nutrition, digestion, antioxidant activity, and immune function [4]. Despite
kiwifruit having several health beneficial effects, its juice is not widely consumed for several reasons.
The production of high-quality kiwifruit juice or extract is affected by a number of factors including
excessive browning, formation of haze/precipitate, and flavor changes. To a large extent, this market
does not exist now owing to the difficulty in producing products that retain the desirable aroma, green
color, and bioactive compounds in kiwifruit juice. Phenolic compounds in kiwifruits are important
contributors to the health effects, color, flavor, and aging characteristics of fruit products such as juice
or extract [5]. Kiwifruit generally has a low pH (3.0–3.5) and suffers from browning upon exposure of
its juice to air [5]. Kiwifruit juice contains oxalic acid; however, the levels depend on the preparation
process [6]. A small number of people appear to be allergic to the fruit [7]. In order to avoid all these
problems, a specific aqueous extract of kiwifruit with bioactivity and free from inactive materials with
much longer shelf life was developed. As a further modification to the extraction process, a juice or
pulp fraction was boiled, centrifuged, filtered, delipidated, and desugarized. The especially prepared
aqueous extract of kiwifruit exhibited the ability to inhibit platelet aggregation and reduce plasma
angiotensin-converting enzyme (ACE) activity [8]. This chapter provides insight into kiwifruit juice
or extract composition, phytochemical profiles, potential health benefits, and future perspectives of
this industry.
331
TABLE 27.1
Physicochemical, Nutritional, and Functional Characteristics of Kiwifruit Juices
Component Unit Nonclarified Kiwifruit Juice Clarified Kiwifruit Juice
Total soluble solids °Brix 14.40 14.26
Titratable acidity % 1.24 1.20
pH 3.43 3.53
Reducing sugars % 8.34 8.17
Nonreducing sugars % 2.25 1.97
Total sugars % 10.59 10.14
Vitamin C mg/100 mL 196 154
Pectin % calcium pectate 0.92 0.12
Total phenolics μg GAE/L 389 240
Total flavonoids μg GAE/L 55 45
Source: Adapted from Vaiyda, D. et al., Nat. Prod. Rad., 8, 388, 2009. With permission.
Abbreviation: GAE, gallic acid equivalents.

Kiwifruit is a nutrient-dense fruit [9]. It is particularly high in vitamin C, fat-soluble vitamins, folic
acid, carotenoids, potassium, and fiber and contains a range of phytochemicals [1,9]. Green kiwifruit
has a higher total dietary and insoluble fiber content than other commonly consumed fruits. Kiwifruit
contains significant levels of fat-soluble vitamins such as vitamins E and K [9]. Table 27.1 shows
the physicochemical, nutritional, and functional characteristics of nonclarified and clarified kiwifruit
juices [10]. Nonclarified and clarified kiwi fruit juices contain 10.59 and 10.14 mg/100 g of vitamin
C, respectively. The consumption of kiwifruit resulted in increased plasma vitamin E concentrations
[11,12]. Kiwifruit is an excellent source of carotenoids, including β-carotene, lutein, and zeaxanthin.
The carotenoids contribute to the color of the kiwifruit, but the unique green color of green kiwifruit
is attributed to the retention of chlorophylls during ripening [13,14]. Kiwifruit also contains a range of
other phytochemicals/polyphenols, although many of the phenolics, including flavonoids, are yet to be
identified [15].

The bioactivity of kiwifruit is largely attributed to the presence of nutrients and antioxidants such
as vitamins C and E, caffeic acid, naringenin, quercetin, and epicatechin [16,17]. The major pheno-
lic compounds in kiwifruit juice have been identified as coumaric and caffeic acid derivatives includ-
ing chlorogenic acid, protocatechuic acid, and a derivative of 3,4-dihydroxybenzoic acid (Figure 27.1).
It also contains epicatechin, catechin, and procyanidins in small quantities. Flavonols are present as
the glycosides of quercetin (glucoside, rhamnoside, and rutinoside) and kaempferol (rhamnoside and
r utinoside) [5]. Kiwifruit’s nutritional components and desirable bioactives, including polyphenols, vita-
mins, and water-soluble polysaccharides, may be advantageous for functional food applications, increas-
ing the range of kiwifruit products. Phenolic compounds are present, at the levels of <1–7 mg/L, in
clarified juice [5]. Green kiwifruit is lower in sugars and higher in organic acids and calcium oxalate
compared with gold kiwifruit [18].
Gold kiwifruit contained the highest level of polyphenols (1.04 mg/mL), followed by green kiwifruit
(0.85 mg/mL), navel orange, mandarin orange, grapefruit, and apple in this order. The clarified kiwifruit
juice has the same levels of polyphenols. Boiling of kiwifruit juice at 100°C for 60 min did not change
its polyphenol concentrations [19]. Nonclarified and clarified kiwi fruit juices contain 389 and 240 mg
gallic acid equivalents (GAE)/L of total phenolics and total flavonoids, respectively (Table 27.1) [10].
Kiwifruit Juice 333
O
OH
O O
OH OH
HO
HO OH
Chlorogenic acid
O OH
O
HO
OH
OH
OH HO
Protocatechuic acid 3,4-Dihydroxybenzoic acid
FIGURE 27.1 Chemical structures of common phenolic acids found in kiwifruit juice.
In order to investigate the effect of kiwifruit on platelet aggregation and ACE activity, kiwifruit
juice was prepared [8]. Kiwifruit juice inhibited both adenosine diphosphate (ADP)-induced plate-
let aggregation and plasma ACE inhibitory activity in a dose-dependent manner. The in vitro plate-
let aggregation experiments suggest that green and gold kiwifruit extracts inhibited both ADP- and
collagen-induced whole blood platelet aggregations [20]. The fruit extract may contain a wide variety
of different types of compounds that have antiplatelet activity and that affect different mechanisms of
activation and aggregation. Incubation with kiwifruit juice inhibited ADP-induced platelet aggrega-
tion in a dose-dependent manner in platelet-rich plasma. It also inhibited collagen-induced platelet
aggregation; however, the levels of inhibition were lower compared with those observed with ADP-
induced platelet aggregation [20]. Arachidonic acid–induced platelet aggregation was also inhibited
by kiwifruit juice, which inhibited the platelet aggregation and ACE activity in a dose-dependent man-
ner. Kiwifruit juice at 30 μL (0.006 mg catechin equivalents [CE]/mL) inhibited 80% of plasma ACE
activity (Table 27.2). Among all fruits tested for their in vitro antiplatelet activity, tomato and kiwifruit
had the highest activity followed by grapefruit, melon, and strawberry, whereas pear and apple had
little or no activity [20,21].
The compounds responsible for the observed activity in kiwifruit juice were isolated as a mixture,
and inactive soluble sugars were removed. These compounds in kiwifruit were water soluble and heat
stable, and their molecular mass was less than 1000 Da [20,22]. Delipidation followed by ultrafiltra-
tion of the kiwifruit extract indicated that the active factors in kiwifruit were water soluble and heat
stable, with a molecular mass of >1000 Da. Kiwifruit extract had average glucose (8.85 mg/mL), fructose
TABLE 27.2
Effects of Freshly Prepared Kiwifruit Juice (100%) on Platelet Aggregation and Serum ACE
Activity
Inhibition of Platelet Inhibition of ACE
Aggregation (%) Activity (%)
Control 0.0 0.0
Kiwifruit juice (10 μL) (0.002 mg CE/mL PRP) 23 ± 11a 61.43 ± 3.41a
Source: Adapted from Dizdarevic, L.L. et al., Platelets, 25, 567, 2014. With permission.
a P < 0.05.
Abbreviations: ACE, angiotensin-converting enzyme; CE, catechin equivalents; PRP, platelet-rich plasma.
TABLE 27.3
Effects of Sugar-Free Kiwifruit Extract and Captopril on Plasma ACE Activity
Kiwifruit Extract ACE Activity Captopril ACE Activity
(mg/mL) μg CE/mL (%) (μg/mL) (%)
0 0 100 ± 0 0 100 ± 0
0.12 2.25 90 ± 12 1.2 40 ± 5a
0.34 6.12 68 ± 8a 2.5 19 ± 3a
0.63 11.34 57 ± 7a 5.0 11 ± 3a
1.25 22.50 38 ± 8a 7.5 5 ± 2a
2.06 37.08 20 ± 6a 10.0 4 ± 2a
Source: Adapted from Dizdarevic, L.L. et al., Platelets, 25, 567, 2014. With permission.
a P < 0.001.
Abbreviations: ACE, angiotensin-converting enzyme; CE, catechin equivalents.
(9.86 mg/mL), and sucrose (2.31 mg/mL). Soluble sugars were removed by using solid-phase extraction
column chromatography [8]. Typically, 100 g of kiwifruit produced an average of 66.34 mg (1.20 mg CE)
of sugar-free kiwifruit extract containing both antiplatelet and anti-ACE activities [8].
Sugar-free kiwifruit extract inhibited platelet aggregation induced by different aggregating agents,
ADP, collagen, and arachidonic acid [5]. The IC50 for ADP-induced platelet aggregation was 1.52 mg/mL
of kiwifruit extract (0.026 mg CE/mL), and for collagen-induced aggregation, this average value was
1.83 mg/mL (0.032 mg CE/mL). Kiwifruit extract inhibited the release of PF4 and TxB2, but without alter-
ing cyclic AMP levels in platelets. Its extract inhibited human serum ACE activity in a dose-dependent
manner (Table 27.3). The IC50 of kiwifruit extract for serum ACE was 0.60 mg/mL (11 μg CE/mL),
whereas the IC50 of captopril was a thousand-fold less (0.56 μg/mL) [8]. Boiling of kiwifruit juice at
90°C for 20 min did not destroy antiplatelet or anti-ACE activities. Rather boiling of the kiwifruit juice
imparted stability to the inhibitory activity of the juice, as the nonboiled juice lost 75% of its activity
within a week of preparation, whereas the antiplatelet activity of the boiled juice remained the same till
the day 18. At present, there is no detailed information on the nature of compounds present in the sugar-
free kiwifruit extract.
27.4 Health Effects

Several in vitro and in vitro studies using kiwifruit extract/juice or whole kiwifruit demonstrated
antioxidant activities and other beneficial effects on lowering markers for cardiovascular disease
(CVD) and cancer as well as anti-inflammatory effects [3,8,11,12,16,17,19,20,22–26]. Another impor-
tant aspect of the health benefits of kiwifruit has been the kiwifruit proteolytic enzyme, actinidin
[27]. This has proved intriguing for consumers and provides a platform for messages about digestion
and gut health. The health benefits of consuming kiwifruit are well documented, but there are insuf-
ficient data available on human supplementation studies with kiwifruit juice or extract. Studies are
required to examine whether various bioactive components in kiwifruit are available when kiwifruit
juice is consumed.
In a volunteer study, supplementation with 500 mg kiwifruit extract resulted in a significant decrease
in the platelet aggregation response at 2 h, whereas the control supplement resulted in no change. Platelet
aggregation responses to ADP were significantly lower than baseline values by 12.9% (8.1%–15.4%)
n = 9, compared with the control P < 0.05 [8]. Kiwifruit juice was shown to regulate the expression of
genes related to adaptive immune pathways in C57BL/6J mice [28]. As mentioned earlier, some people
are allergic to the kiwifruit. IgE-mediated kiwifruit allergy is often associated with birch and grass
pollinosis as well as with latex allergy. Eleven green kiwifruit allergens recognized to date are termed
as Act d 1 through Act d 11. Bet v 1 homologue (Act d 8) and profilin (Act d 9) are important allergens
in polysensitized subjects, whereas actinidin (Act d 1) is important in kiwifruit monosensitized sub-
jects [29]. The largest group of subjects with kiwifruit allergy is patients with a pollen–fruit syndrome.
Kiwifruit Juice 335
Differences in allergenicity have been found among different cultivars [29]. Limited information is
available in the literature on the prevalence of kiwifruit allergy.

Kiwifruit is mainly eaten as whole fruit, because of technical issues such as nutrient and color loss
encountered in processing and storage of its juice as mentioned earlier. The few examples where kiwi-
fruit has been processed into products include frozen desserts and blended juices [30], and more recently,
a few natural kiwifruit drinks such as Kiwi CrushTM (Vital Food Processors Ltd., Manukau City,
Auckland, New Zealand) has been introduced. ENZAFOODS New Zealand Ltd. processes kiwifruit
into high-quality kiwifruit juice concentrate. In some cases, kiwifruit juice is supplied frozen and can be
used in a range of potential products for the hospitals, catering, and food-processing industries as well as
household consumers. It is a light green frozen product with a fine crystal texture. At −18°C, the product
retains a consistency such that it can be scooped. Kiwifruit juice concentrate may be thawed by dilution
with water (2 parts water to 1 part frozen kiwifruit juice concentrate) to produce a ready chilled drink.
On reconstitution, the fine kiwifruit particles are evenly dispersed and suspended in the drink resulting
in a natural appearance of kiwifruit juice.
The specially prepared sugar-free aqueous kiwifruit extract, as described earlier, exhibits an abil-
ity to inhibit both platelet aggregation and serum ACE activity in vitro and in vivo. The water-soluble,
colorless components (molecular weight <1000 Da) in kiwifruit extract can be mixed with any juice for
consumption. The active components were found to be primarily associated with, or extractable from,
the juice, the flesh surrounding the pips of the kiwifruit. Kiwifruit extract can be exploited for develop-
ment of cardioprotective functional juice. These claims still require further studies to elucidate their
effects in vivo, including bioavailability, stability, and metabolism of these compounds. As the IC50 value
of kiwifruit extract was higher than those of the prescribed drugs for hypertension, it could be used as
preventative juice against hypertension. In order to use specially purified kiwifruit juice as a constituent
of a health drink, human intervention trials using the extract and studies on toxicity, bioavailability, and
structure–activity relationships are required.
27.6 Conclusion
Although it is evident that kiwifruit has many health benefits, its juice has several drawbacks such as
browning, instability, and discoloration, as well as possible presence of some allergens. Specially pre-
pared sugar-free aqueous kiwifruit extracts exhibited an ability to inhibit both platelet aggregation and
serum ACE activity. As the IC50 value of components present in kiwifruit extract was 1000-fold higher
than that of the prescribed drugs for hypertension (e.g., captopril), kiwifruit extract may be used as a
cardioprotective agent and can be exploited for the development of cardioprotective functional juice.
Modulation of platelet reactivity and plasma ACE activity by kiwifruit extract could be of potentially
prophylactic and therapeutic benefit in preventing and halting pathologic processes that lead to CVD.
The active components were found to be primarily associated with, or extractable from, the juice, the
flesh surrounding the pips of the kiwifruit. These claims still require further studies to elucidate their
effects in vivo, including bioavailability, stability, and metabolism of these compounds.
REFERENCES
1. Ferguson, A.R. and Ferguson, L.R., Are kiwifruit really good for you? in Proceedings of the Fifth
International Symposium on Kiwifruit, Hongwen, H., Ed., International Society for Horticultural
Science, Leuven, Belgium, 2003, pp. 131–138.
2. Nishiyama, I., Yamashita, Y., Yamanaka, M., Shimohashi, A., Fukuda, T., and Oota, T., Varietal dif-
ference in vitamin C content in the fruit of kiwifruit and other Actinidia species. J. Agric. Food Chem.,
52, 5472–5475, 2004.
3. Stonehouse, W., Gammon, C.S., Beck, K.L., Conlon, C.A., von Hurst, P.R., and Kruger, R., Kiwifruit:
Our daily prescription for health. Can. J. Physiol. Pharm., 91, 442–447, 2013.
4. Singletary, K., Kiwifruit: Overview of potential health benefits. Nutr. Today, 47, 133–147, 2012.
5. Dawes, H.M. and Keene, J.B., Phenolic composition of kiwifruit juice. J. Agric. Food Chem., 47,
2398–2403, 1999.
6. Nguyen, H.V.H. and Savage, G., Oxalate contents of kiwifruit (Actinidia deliciosa) juice extracted by
pressing or enzyme extraction. J. Food Agric. Environ., 11, 228–230, 2013.
7. Le, T.M., Bublin, M., Breiteneder, H., Fernandez-Rivas, M., Asero, R., Ballmer-Weber, B., Barreales, L.
et al., Kiwifruit allergy across Europe: Clinical manifestation and IgE recognition patterns to kiwifruit
allergens. J. Allergy Clin. Immunol., 131, 164–171, 2013.
8. Dizdarevic, L.L., Biswas, D., Uddin, M.M., Jorgenesen, A., Falch, E., Bastani, N.E., and Duttaroy, A.K.,
Inhibitory effects of kiwifruit extract on human platelet aggregation and plasma angiotensin-converting
enzyme activity. Platelets, 25, 567–575, 2014.
9. Drummond, L., The composition and nutritional value of kiwifruit. Adv. Food Nutr. Res., 68, 33–57,
2013.
10. Vaiyda, D., Vaidya, M., Sharma, S., and Ghanshayam, Enzymatic treatment for juice extraction and
preparation and preliminary evaluation of kiwifruits wine. Nat. Prod. Rad., 8, 388–391, 2009.
11. Chang, W.H. and Liu, J.F., Effects of kiwifruit consumption on serum lipid profiles and antioxidative
status in hyperlipidemic subjects. Int. J. Food Sci. Nutr., 60, 709–716, 2009.
12. Hunter, D.C., Skinner, M.A., Wolber, F.M., Booth, C.L., Loh, J.M., Wohlers, M., Stevenson, L.M., and
Kruger, M.C., Consumption of gold kiwifruit reduces severity and duration of selected upper respiratory
tract infection symptoms and increases plasma vitamin C concentration in healthy older adults. Br. J.
Nutr., 108, 1235–1245, 2012.
13. McGhie, T.K. and Ainge, G.D., Color in fruit of the genus actinidia: Carotenoid and chlorophyll com-
positions. J. Agric. Food Chem., 50, 117–121, 2002.
14. Nishiyama, I., Fukuda, T., and Oota, T., Cultivar difference in chlorophyll, lutein and beta-carotene
content in the fruit of kiwifruit and other Actinidia species, in Proceedings of the Sixth International
Symposium on Kiwifruit, Martin, R.A., Ed., International Society for Horticultural Science, Leuven,
Belgium, 2007, pp. 473–478.
15. Tarascou, I., Souquet, J.M., Mazauric, J.P., Carrillo, S., Coq, S., Canon, F., Fulcrand, H., and Cheynier,
V., The hidden face of food phenolic composition. Arch. Biochem. Biophys., 501, 16–22, 2010.
16. Sun-Waterhouse, D., Chen, J., Chuah, C., Wibisono, R., Melton, L.D., Laing, W., Ferguson, L.R., and
Skinner, M.A., Kiwifruit-based polyphenols and related antioxidants for functional foods: Kiwifruit
extract-enhanced gluten-free bread. Int. J. Food Sci. Nutr. 60(Suppl. 7), 251S–264S, 2009.
17. Collins, B.H., Horska, A., Hotten, P.M., Riddoch, C., and Collins, A.R., Kiwifruit protects against oxi-
dative DNA damage in human cells and in vitro. Nutr. Cancer, 39, 148–153, 2001.
18. Perera, C.O., Hallett, I.C., Nguyen, T.T., and Charles, J.C., Calcium-oxalate crystals—The irritant factor
in kiwifruit. J. Food Sci., 55, 1066–1069, 1990.
19. Iwasawa, H., Morita, E., Yui, S., and Yamazaki, M., Anti-oxidant effects of kiwi fruit in vitro and
in vivo. Biol. Pharm. Bull., 34, 128–134, 2011.
20. Duttaroy, A.K. and Jorgensen, A., Effects of kiwi fruit consumption on platelet aggregation and plasma
lipids in healthy human volunteers. Platelets, 15, 287–292, 2004.
21. Duttaroy, A.K., Crosbie, L., and Gordon, M.J., Effects of tomato extract on human platelet aggregation
in vitro. Platelets, 12, 218–227, 2001.
22. Duttaroy, A.K., Cardioprotective properties of kiwifruit. Adv. Food Nutr. Res., 68, 273–282, 2013.
23. Collins, A.R., Kiwifruit as a modulator of DNA damage and DNA repair. Adv. Food Nutr. Res., 68,
283–299, 2013.
24. Gaziano, J.M., Manson, J.E., Branch, L.G., Colditz, G.A., Willett, W.C., and Buring, J.E., A prospective
study of consumption of carotenoids in fruits and vegetables and decreased cardiovascular mortality in
the elderly. Ann. Epidemiol., 5, 255–260, 1995.
25. Brevik, A., Gaivao, I., Medin, T., Jorgenesen, A., Piasek, A., Elilasson, J., Karlsen, A. et al.,
Supplementation of a western diet with golden kiwifruits (Actinidia chinensis var.’Hort 16A’:) effects on
biomarkers of oxidation damage and antioxidant protection. Nutr. J., 10, 54, 2011.
Kiwifruit Juice 337
26. Karlsen, A., Svendsen, M., Seljeflot, I., Laake, P., Duttaroy, A.K., Drevon, C.A., Arnesen, H., Tonstad, S.,
and Blomhoff, R., Kiwifruit decreases blood pressure and whole-blood platelet aggregation in male
smokers. J. Hum. Hypertn., 27, 126–130, 2013.
27. Boland, M., Kiwifruit proteins and enzymes: Actinidin and other significant proteins. Adv. Food Nutr.
Res., 68, 59–80, 2013.
28. Edmunds, S.J., Roy, N.C., Davy, M., Cooney, J.M., Barnett, M.P., Zhu, S., Park, Z., Love, D.R., and
Laing, W.A., Effects of kiwifruit extracts on colonic gene and protein expression levels in IL-10
gene-deficient mice. Br. J. Nutr., 108, 113–129, 2012.
29. Bublin, M., Kiwifruit allergies. Adv. Food Nutr. Res., 68, 321–340, 2013.
30. Cassano, A., Figoli, A., Tagarelli, A., Sindona, G., and Drioli, E., Integrated membrane process for the
production of highly nutritional kiwifruit juice. Desalination, 189, 21–30, 2006.
28
Lemon Juice
Lucia Maria Jaeger de Carvalho, Lara de Azevedo Sarmet Moreira Smiderle,

Ediane Maria Gomes Ribeiro, and José Luiz Viana de Carvalho
CONTENTS
28.1 Introduction................................................................................................................................... 339
28.4 Health Effects................................................................................................................................ 343
28.6 Conclusion..................................................................................................................................... 344
References............................................................................................................................................... 344
28.1 Introduction
The main citrus-producing countries in the world are China, Brazil, and the United States, followed
by Mexico, Spain, and Italy. However, Brazil is the first world producer of oranges and lemons by
conventional cultivation, followed by the United States, while China stands out in the production of
tangerines [1]. Although a large number of varieties or species of citrus fruits exist, lemon (Citrus limon
L. Burm. f.) is the most widespread, used citrus fruit for juice processing.
The lemon tree is sensitive to cold, but it can be cultivated cold regions since its acidity and level of
juice are reached at the time of harvest. The Lisbon, Cosmopolitan, Bernia, Spain, and Interdonato are
the main varieties of the winter, while Villafranca, Cosmopolitan, Femminello, and Monachello (Italy)
cannot grow in the winter season. Currently, there are about 70 varieties of lemons. The citrus flavor
and aroma are pleasant, and in its composition, there are essential nutrients and micronutrients, such as
vitamins, minerals, and fiber [2,3]. Phenolic compounds present in lemon and lemon juices neutralize
free radicals, thereby rendering the antioxidant activity [4,5].
Lemon juice can be found commercially in the regular strength, concentrate, and the dried form or
added in the formulation of soft drinks and new beverages as sport drinks. Lemon juice is known and con-
sumed as a refreshing drink rich in vitamin C. Recently, it has been shown that lemon juice has other bioac-
tive compounds such as polyphenols. Among polyphenols, flavonoids have been widely studied due to their
antioxidants, anticarcinogenic, and anti-inflammatory properties and as inhibitors of lipid peroxidation [6].
Several authors have previously reported that citrus fruits are important sources of ascorbic acid and
flavonoids [6–11] and are still frequently used in industry for the extraction of flavanones [10,12]. This con-
tribution highlights the nutritional characteristics, bioactive compounds, antioxidant efficacy, and health
effects of lemon juice. Furthermore, the novel products, formulations, and future trends are also discussed.

Lemon juice normally has a pH of 2.78–2.81, citric acid content of 5.98–6.05 g/100 g, soluble solids
of 7.63–8.42oBrix, fructose content of 0.66–0.98 g/100 g, glucose at 0.54–0.89 g/100 g, and sucrose at
0.07–0.12 g/100 g. It also has vitamin C (22.80–22.86 mg/mL) and some minerals, of which the most
339
TABLE 28.1
Proximate Compositions of Lime and Lemon Juices (per 100 g)
Component Unit Lime Juice [13] Lemon Juice [14]
Water g 92.52 92.31
Energy kcal 21 22
Protein g 0.25 0.35
Lipid (fat) g 0.23 0.24
TABLE 28.2
Nutritional Characteristics of Lemon Juice (per 100 g)
Nutrient Unit Concentration
Minerals
Calcium mg 6
Copper mg 0.016
Iron mg 0.08
Magnesium mg 6
Manganese mg 0.012
Phosphorus mg 8
Potassium mg 103
Selenium μg 0.1
Sodium mg 1
Zinc mg 0.05
Vitamins
Choline mg 5.1
Folate (DFE) μg 20
Niacin mg 0.091
Riboflavin mg 0.015
Thiamin mg 0.024
Vitamin A IU 6
Vitamin B6 mg 0.046
Vitamin C mg 38.7
Carotenoids
β-Carotene μg 1
β-Cryptoxanthin μg 4
Lutein + zeaxanthin μg 15
Reference, Release 26, Statistical Report 09152, Lemon Juice, Raw, USDA, Beltsville, MD, 2013.
Abbreviations: DFE, dietary folate equivalents; IU, international unit; ATE, alpha-tocopherol equivalents.
abundant are potassium (376.79 μg/g) and calcium (23.24 μg/g) [13,14]. The proximate compositions of
lime and lemon juices are given in Table 28.1. In addition, Table 28.2 shows the mineral, vitamin, and
carotenoid content of lemon juice. The content of vitamin C was 38.7 mg/100 g [14]. The major carotenoids
in 100 g lemon juice are lutein + zeaxanthin (15 μg), β-cryptoxanthin (4 μg), and β-carotene (1 μg) [14].
Additionally, Agócs et al. [15] reported different carotenoids in lemon juice, namely, lutein (5.7%), α- and
β-cryptoxanthin (36% and 8.7%, respectively), and α- and β-carotenes (1.2% and 11.1%, respectively).
Lemon Juice 341

The bioactive compounds are important in citrus species because of their antioxidant capacity pro-
moted by vitamin C and some phenolic compounds (particularly flavonoids), as well as minerals, dietary
fiber, essential oils, and carotenoids. Additionally, lemon is the third most important citrus species as an
important health-promoting fruit.
Nearly 50 years ago, Vandercook and Stephenson [16] identified the most abundant phenolic a glycones
and glucosides in the fraction of single-strength lemon juice. They quantified eriodictyol (20 mg/100 mL),
hesperidin (1.4 mg/100 mL), quercetin (2.2 mg/100 mL), phloroglucinol (2 mg/100 mL), and umbel-
liferone (0.2 mg/100 mL). Eriocitrin had stronger antioxidant activity than the other citrus flavonoid
compounds, and it was abundantly present in lemon and its juice [17]. The chemical structures of impor-
tant phenolic compounds (naringin, hesperidin, diosmin, luteolin-7-O-rutinoside, and eriocitrin) found
in lemon juice are presented in Figure 28.1.
Wang et al. [18] determined the bioactive compounds in edible portions of eight varieties of citrus
fruit juices. Diosmin was the major flavones in kumquat (0.699 mg/g) and lemon juice (0.323 mg/g).
OH
HO
HO
OH H3C O
OH OH
HO O O OCH3
HO O O
HO O
O O O
HO
H3C O OH
HO OH O
OH
OH OH O
Naringin Hesperidin
OH OH
HO O O
OH
OH O
HO
HO OH O OH O
O
O O O HO
O O OH O
HO OH HO CH3
OH OH O OH
Diosmin Luteolin-7- Ο-rutinoside
HO
OH
O O OH
HO
O
OH
O OH O
OH
O
H 3C OH
OH
Eriocitrin
FIGURE 28.1 Chemical structures of flavonoids found in lemon juice.

TABLE 28.3
Flavonoid Content of Different Varieties of Lemon Juices Processed at Different
Extraction Systems (mg/L)
Extraction Eriocitrin Hesperidin Diosmin Luteolin-7-O-Rutinoside
Fino
ES-1 81 104 10 22
ES-2 203 240 41 65
ES-3 205 253 51 64
Verna
ES-1 111 224 13 15
ES-2 193 395 41 42
ES-3 391 410 28 28
Source: Adapted from Marín, F.R. et al., Food Chem., 78, 319, 2002. With permission.
Abbreviation: ES, extraction system.
Additionally, chlorogenic acid was the major phenolic acid in wendun and lemon, containing 103 and
92.6 μg/g, respectively.
The flavonoids in different varieties of lemon juice were identified by Marín et al. [6] (Table 28.3).
Eriocitrin content varied from 81 to 205 and from 111 to 391 mg/L, hesperidin from 104 to 253 and
from 224 to 410 mg/L, diosmin from 10 to 51 and from 13 to 41 mg/L, and luteolin-7-O-rutinoside from
22 to 64 and from 15 to 42 mg/L in Fino and Verna varieties, respectively.
The main flavanones in lemon juice were eriodictyol (4.9 mg/100 g), hesperetin (14.5 mg/g), narin-
genin (1.4 mg/100 g), and quercetin (0.4 mg/100 g) [14,19]. In addition, some flavonoids in lemon juice
of the Sicilian cultivar, determined by high-performance liquid chromatography–diode array detector
(HPLC-DAD) coupled with electrospray ionization–mass spectrometry (ESI-MS), were evaluated [20].
They found significant amount of an unusual compound (6,8-di-C-glucopyranosyldiosmetin) after puri-
fication, together with eriocitrin, hesperidin, and diosmin.
According to Lorente et al. [21], flavanones are the main flavonoids in the genus Citrus. Hesperidin
and eriocitrin dominated the flavanone profile of lemons and sweet oranges [22]. The concentrations
of hesperidin in direct and reconstituted juices were 365 and 367 mg/L, respectively [22]. Previously,
some authors had reported lower hesperidin levels in juices [23–28], probably due to differences in the
extraction method used. In this study [22], lemon juice was obtained by industrial processing, while in
all previous studies, the juices were manually prepared.
The flavonoids of citrus juice were also studied by Gattuso et al. [29], and the main compounds
found were flavanone-O-glycosides and flavone-O- or -C-glycosides. Similarly, Abeysinghe et al.
[30] evaluated the phenolic compounds and total antioxidant capacity by ferric-reducing antioxidant
power (FRAP) assay in different edible tissues of citrus fruits, namely, juice sacs, segment mem-
brane, and segment, of C. unshiu, C. reticulata, C. sinensis, and C. changshanensis. The highest
total phenolics, total flavonoids, naringin, and total antioxidant capacity were found in the segment
membrane of C. changshanensis, while the highest carotenoid content was found in juice sacs of
C. reticulata. They recommended consuming citrus fruit with all edible tissues rather than juice or
juice sacs alone.
Several authors reported bioactive compounds and antioxidant activity as well as the methods to evalu-
ate and identify these compounds in citrus juices [23,28–32]. The method normally used to determine the
total phenolic content is the Folin–Ciocalteu assay. For the antioxidant capacity, there are some methods
that can be used, such as the 1,1-diphenyl-2picrylhydrazyl (DPPH), peroxyl radical scavenging assay,
trolox equivalent antioxidant capacity (TEAC), FRAP, 2,2-azino-bis (3-ethyl-benzothiazoline-6-sulfonic
acid) (ABTS), oxygen radical absorbance capacity (ORAC), and β-carotene/linoleate model system
[23,30,33–37]. For the quantification of products formed during lipid peroxidation, 2-thiobarbituric acid
reactive substances (TBARS) and low-density lipoprotein (LDL) oxidation are commonly used [38–40].
Lemon Juice 343
28.4 Health Effects

Many health effects of lemon juices are reported in the literature. In Turkey, lemon juice is one of the
most common alternative therapies used in the treatment of acute blood pressure elevation. However,
there is lack of clinical studies proving this effect [41]. Furthermore, in 1850, some doctors had used
lemon juice in hospitals to treat acute articular rheumatism [42]. Lemon juice was also used as a tra-
ditional intravaginal contraceptive throughout the Mediterranean region for hundreds of years. Since
it was well documented that sperm was sensitive to low pH, the spermicidal properties of lemon juice
was possibly due to the high concentration of citric acid [43]. In addition, some authors have suggested
that lemon juice can be an alternative to potassium citrate in the treatment of urinary calcium stones in
patients with hypocitraturia [44]. D’Aquino and Teves [45] also showed that lemon juice could be used
as an affordable disinfectant capable of destroying Vibrio cholerae in drinking water in areas lacking
water treatment plants.
Generally, benefits of citrus are related to the bioactive compounds present in the fruits, such as carot-
enoids, minerals, essential oils, organic acids, dietary fiber, phenolic compounds (mainly flavonoids),
and vitamins, especially ascorbic acid (vitamin C). There is scientific evidence that these substances are
involved in the prevention and treatment of diseases, such as obesity, diabetes, hypercholesterolemia,
cardiovascular disease (CVD), and some cancers.
Miyake et al. [46] evaluated the crude flavonoids, eriocitrin, and hesperidin effects on streptozotocin-
induced diabetic rats. After 28 days, the concentration of the TBARS in the serum, liver, and kidney
of diabetic rats administered with crude flavonoids, eriocitrin, and hesperidin significantly decreased as
compared with that of the diabetic group. Crude flavonoids, eriocitrin, and hesperidin suppressed the
oxidative stress in diabetic rats. These results demonstrate that dietary lemon flavonoids of eriocitrin and
hesperidin play a role as antioxidants in vivo. Eriocitrin had stronger antioxidant activity than the other cit-
rus flavonoids and was abundantly present in lemon fruit/juice showing that dietary flavonoids are impor-
tant antioxidants in vivo [46]. Additionally, lime may have an immune modulatory effect in humans, and
lemon might even ease hangover symptoms [47]. Essential oils from C. aurantium and the monoterpene
limonene have been used medicinally throughout the world to treat gastritis and gastric disorders [48].
In recent years, citrus has gained importance due to its ability to provide a multitude of health ben-
efits not only from vitamin C but also from other bioactive compounds. Until recently, citrus’ health-
promoting properties were always associated with the content of vitamin C. Only in the last decade,
studies have focused on several bioactive compounds, specifically limonoids and flavonoids, which play
a major role in preventing chronic diseases [49]. Despite being considered a flavoring ingredient, the
limonene has an antitumoral, antibiotic, and antiprotozoal activity against Leishmania braziliensis,
L. major, L. chagasi, L. amazonensis, and L. promastigotes both in vitro and in vivo. Arruda et al. [50]
reported that the treatment of L. amazonensis-infected macrophages with 300 mM limonene resulted
in 78% reduction in infection rates. L. amazonensis-infected mice treated topically or intrarectally with
limonene had significant reduction of lesion sizes. The intrarectal treatment was also highly effective in
decreasing the parasite burden, healing established lesions, and suppressing the dissemination of ulcers.
Limonene presents low toxicity in humans and has been shown to be effective as an agent for enhancing
the percutaneous permeation of drugs.
The potential benefits of bioactive compounds such as curcumin, flavonols, and isoflavones in pancreatic
cancer prevention have been reported by many authors [51–54]. In addition to the antioxidant activity, hesper-
idin is known to act as an anticancer agent through prostaglandin and chemical carcinogens inhibition [55].
The other major flavonoid in lemon fruits and juices is rutin, also known as quercetin-3-O-rutinoside.

Beverages are largely consumed by different types of consumers, especially in the form of soft drinks and
juices. These days, the constant demand for new products, including fruit juices, which also provide health
benefits, is a reality. Therefore, new products with functional properties have become essential. The use
of technologies for juice processing to preserve their nutritional value, as well as bioactive compounds
with high antioxidant activity that could be present in the original product or added in the final process,
should be considered.
A new approach in the formulation is production of blends and soft drinks with clarified lemon juice
obtained by microfiltration and ultrafiltration as well as the ones concentrated by reverse osmosis and osmotic
distillation alternative processes carried out at room temperature. Cassano et al. [56] elaborated a clarified and
concentrate lemon juice, among others (orange and carrot), using an integrated membrane system (ultrafiltra-
tion, reverse osmosis, and osmotic distillation). The concentrated lemon juice had a high antioxidant activity.
High-pressure process is another alternative technology for fruit juice sterilization [57]. Lemon juice
that was high-pressure (300 MPa) processed demonstrated good shelf life with minor changes in its
constituents and physicochemical properties. No fungi were detected in pressure-treated lemon juices,
whereas the control samples were spoiled by yeasts and fungi after 10 days [58].
New drinks have been searched for potential market launch, if bioactive compounds are added. One
example of this is the drink based on lemon juice with aronia (Aronia melanocarpa) concentrate that has
high antioxidant activity [24]. According to Silva et al. [59], an additional application for citrus juices,
including lemon juice, is the modification of the flavonoids by enzymatic deglycosylation (hesperidinase,
naringinase, and β-D-glucosidase) to produce functional beverages with higher antioxidant activity.
González-Molina et al. [25] designed a new beverage rich in polyphenols. In this, lemon and pome-
granate juices were mixed in different proportions (25:75, v/v). They suggested that the combination of
75% pomegranate and 25% lemon juices had high antioxidant capacity and desirable color and served as
a promising new functional beverage.
Lemon is processed into juice or concentrate in order to fulfill demands of the consumers in any season
with affordable cost and to meet recommended daily intake values [60]. Lemon juice is an interesting food
matrix for designing new beverages, as well as being a suitable source of value-added products. Lemonade
is a noncarbonated drink that has attracted much attention and has been in focus for consumers. The
demand for healthy and convenient functional foods affects consumer trends through innovative products.
For this reason, lemonade could be enriched with different natural herbal extracts such as ginger, linden,
and mint. Gironés-Vilaplana et al. [28] elaborated new beverages using lemon juice and maqui (Aristotelia
chilensis), rich in flavonoids and vitamin C. They reported that beverages produced had high in vitro anti-
oxidant capacity, mainly due to the maqui polyphenols, with a strong stability throughout the study period.
A new mixture composed by a blend of black chokeberry and lemon juices was proposed by Gironés-
Vilaplana et al. [28]. It promoted inhibitory effects of the cholinesterase and high antioxidant activ-
ity. The cholinesterase is of interest with regard to neurodegenerative disorders such as Alzheimer’s
and Parkinson’s diseases and senile dementia. Chornomaz et al. [61] obtained clarified lemon juice
using microfiltration membranes composed by polyvinylidene fluoride and polyvinylpyrrolidone that
presented similar nutraceutical quality (hesperidin) to the unprocessed juice.
28.6 Conclusion
Various lemon juices present many components with different functions that can prevent diseases and
promote human health among other benefits. The bioactive compounds in lemon juice, rich in phenolic
compounds and high antioxidant activity, are used to improve the nutritional quality of food, and new
beverage formulations can be introduced to the market. However, the processes applied for the preven-
tion of postharvest physical injuries are essential and must be effective to minimize damage in order to
maintain the nutritional quality and integrity of bioactive compounds of lemon juice.
REFERENCES
1. Food and Agriculture Organization (FAO), FAOSTAT, Statistics Database, Agriculture, FAO, Rome,
Italy, 2008.
2. Morton, L.W., Cacettaa, R.A., Puddey, I.B., and Croft, K.D., Chemistry and biological effects of dietary phe-
nolic compounds: Relevance to cardiovascular disease. Clin. Exp. Pharmacol. Physiol., 27, 152–159, 2000.
Lemon Juice 345
3. Pellegrini, N., Serafini, M., Colombi, B., and Del Rio, D., Total antioxidant capacity of plant foods bev-
erages and oils consumed in Italy assessed by three different in vitro assays. J. Nutr., 133, 2812–2819,
2003.
4. Harborne, J.B. and Williams, C.A., Advances in flavonoid research since 1992. Phytochemistry, 52,
481–504, 2000.
5. Dhuique-Mayer, C., Caris-Veyrat, C., Ollitrault, P., Curk, F., and Amiot, M.J., Varietal and interspecific
influence on micronutrient contents in citrus from the Mediterranean area. J. Agric. Food Chem., 53,
2140–2145, 2005.
6. Marín, F.R., Martinez, M., Uribesalgo, T., Castillo, T.S., and Frutos, M.J., Changes in nutraceutical
composition of lemon juices according to different industrial extraction systems. Food Chem., 78,
319–324, 2002.
7. Benavente-García, O., Castillo, J., and Del Río, J.A., Changes in neodiosmin levels during development
of Citrus aurantium leaves and fruits. Postulation of neodiosmin biosynthetic pathway. J. Agric. Food
Chem., 41, 1916–1919, 1993.
8. Castillo, J., Benavente-García, O., and Del Río, J.A., Hesperetin 7-O-glucoside and prunin in Citrus
species (C. aurantium and C. paradisi). A study of their quantitative distribution in immature fruits
and as immediate precursors of neohesperidin and naringin in C. aurantium. J. Agric. Food Chem.,
41, 1920–1924, 1993.
9. Del Río, J.A., Fuster, M.D., Sabater, F., Porras, I., GarcíaLidón, A., and Ortuño, A., Selection of citrus
varieties highly productive for the neohesperidin-dihydrochalcone precursor. Food Chem., 59, 433–437,
1997.
10. Del Río, J.A., Arcas, M.C., Benavente, O., Sabater, F., and Ortuño, A., Changes of polymethoxylated
flavones levels during development of Citrus aurantium (cv. Sevillano) fruits. Planta Med., 64, 575–576,
1998.
11. Ortuño, A., Arcas, M.C., Benavente-García, O., and Del Río, J.A., Evolution of polymethoxy flavones
during development of tangelo Nova fruits. Food Chem., 66, 217–220, 1999.
12. Baer, A., Borrego, F., Benavente-García, O., Castillo, J., and Del Río, J.A., Neohesperidin-
dihydrochalcone: Properties and applications. LWT—Food Sci. Technol., 23, 371–376, 1990.
13. Rangel, C.N., Carvalho, L.M.J., Fonseca, R.B.F., Soares, A.G., and Jesus, E.O., Nutritional value of
organic acid in lime juice (Citrus latifolia T.), cv. Tahiti. Food Sci. Technol., Campinas, 31, 918–922,
2011.
Release 26, Statistical Report 09152, Lemon Juice, Raw. USDA, Beltsville, MD, 2013.
15. Agócs, A., Nagy, V., Szabó, Z., Márk, L., Ohmacht, R., and Deli, J., Comparative study on carotenoid
composition of the peel and pulp of different citrus species. Innov. Food Sci. Emerg. Technol., 8,
390–394, 2007.
16. Vandercook, K.E. and Stephenson, R.G., Lemon juice composition. Identification of the major phenolic
compounds and estimation by paper chromatography. J. Agric. Food Chem., 14, 5450–5454, 1966.
17. Miyake, Y., Yamamoto, K., Tsujihara, N., and Osawa, T., Protective effects of lemon flavonoids on
oxidative stress in diabetic rats. Lipids, 33, 689–695, 2010.
18. Wang, Y.C., Chuang, Y.C., and Ku, Y.H., Quantitation of bioactive compounds in citrus fruits cultivated
in Taiwan. Food Chem., 102, 1163–1171, 2007.
19. González-Molina, E., Domínguez-Perles, R., Moreno, D.A., and García-Viguera, C., Natural bioactive
compounds of Citrus lemon for food and health. J. Pharm. Biomed. Anal., 51, 327–345, 2010.
20. Caristi, C., Bellocco, E., Panzera, V., Toscano, G., Vadala, R., and Leuzzi, U., Flavonoids detection by
HPLC-DAD-MS-MS in lemon juices from Sicilian cultivars. J. Agric. Food Chem., 51, 3528–3534.
2003.
21. Lorente, J., Vegara, S., Martí, N., Ibarz, A., Coll, L., Hernández, J., Valero, M., and Saura, D.,
Chemical guide parameters for Spanish lemon (Citrus limon (L.) Burn.) juices. Food Chem., 162,
186–191, 2014.
22. Peterson, J.J., Beecher, G.R., Bhagwat, S.A., Dwyer, J.T., Gebhardt, E., Haytowitz, D.B., and Holden,
J.M., Flavanones in grapefruit, lemons, and limes: A compilation and review of the data from the
analytical literature. J. Food Comp. Anal., 19, S74–S80, 2006.
24. González-Molina, E., Moreno, D.A., and García-Viguera, C., Genotype and harvest time influence the
phytochemical quality of Fino lemon juice (Citrus limon [L.] Burn. F.) for industrial use. J. Agric. Food
Chem., 56, 1669–1675, 2008.
25. González-Molina, E., Moreno, D.A., and García-Viguera, C., A new drink rich in healthy bioactives
combining lemon and pomegranate juices. Food Chem., 115, 1364–1372, 2009.
26. González-Molina, E., Moreno, D.A., and García-Viguera, C., Comparison of “Verna” lemon juice
quality for new ingredients and food products. Sci. Hortic., 120, 353–359, 2009.
27. Barreca, D., Bellocco, E., Caristi, C., Leuzzi, U., and Gattuso, G., Flavonoid profile and radical-scaveng-
ing activity of Mediterranean sweet lemon (Citrus limetta) juice. Food Chem., 129, 417–422, 2011.
28. Gironés-Vilaplana, A., Mena, P., García-Viguera, C., and Moreno, D.A., A novel beverage rich in anti-
oxidant phenolics: Maqui berry (Aristotelia chilensis) and lemon juice. LWT- Food Sci. Technol., 47,
279–286, 2012.
30. Abeysinghe, D.C., Li, X., Sun, C., Zhang, W., Zhou, C., and Chen, K., Bioactive compounds and anti-
oxidant capacities in different edible tissues of citrus fruit of four species. Food Chem., 104, 1338–1344,
2007.
31. Ramful, D., Tarnus, E., Aruoma, O.I., Bourdon, E., and Bahorun, T., Polyphenol composition, vitamin
C content and antioxidant capacity of Mauritian citrus fruit pulps. Food Res. Int., 44, 2088–2099, 2011.
32. Beh, L.K., Zakaria, Z., Beh, B.K., Ho, W.Y., Yeap, S.K., Banu, N., and Alitheen, M., Comparison of total
phenolic content and antioxidant activities of freeze-dried commercial and fresh fruit juices. J. Med.
Plants Res., 6, 5857–5862, 2012.
33. Marco, G., A rapid method for evaluation of antioxidants. J. Am. Oil Chem. Soc., 45, 594–598, 1968.
34. Miller, H.E., A simplified method for the evaluation of antioxidant. J. Am. Oil Chem. Soc., 48, 91, 1971.
35. Pellegrini, N., Proteggnente, A., Pannala, Y.M., and Rice-Evans, C.A., Antioxidant activity applying an
improved ABTS radical cation decolorization assay. Free Radical Bio. Med., 26, 1231–1237, 1999.
36. Molyneux, P., The use of the stable free radical diphenylpicryl-hydrazyl (DPPH), for estimating antioxi-
dant activity. Songklanakarin J. Sci. Technol., 26, 213–219, 2004.
37. Scherer, R. and Godoy, H.T., Antioxidant activity index (AAI) by the 2,2-diphenyl-1-picrylhydrazyl
method. Food Chem., 112, 654–665, 2009.
38. Frankel, E.N. and Meyer, A.S., The problem of using one dimensional method to evaluate multifunc-
tional food and biological antioxidants. J. Sci. Food Agric., 80, 1925–1941, 2000.
39. Sánchez-Moreno, C., Review: Methods used to evaluate the free radical scavenging activity in foods and
biological systems. Food Sci. Technol. Int., 8, 121–137, 2002.
40. Aruoma, O. I., Methodological characterizations for characterizing potential antioxidant actions of
bioactive components in plant foods. Mutat Res., 523–524, 9–20, 2003.
41. Adibelli, Z., Dilek, M., and Akpolat, T., Lemon juice as an alternative therapy in hypertension in Turkey.
Int. J. Cardiol., 135, e58–e59, 2009.
42. Rees, O., Guy’s hospital: Acute articular rheumatism treated with lemon juice. Lancet, 56, 551, 1850.
43. Clarke, G.N., McCoombe, S.G., and Short, R.V., Sperm immobilizing properties of lemon juice. Fertil.
Steril., 85, 1529–1530, 2006.
44. Aras, B., Kalfazade, N., Kemahli, E., Polat, H., Ozbay, B., and Tasci, A.I., Can lemon juice be an alter-
native to potassium citrate in the treatment of urinary calcium stones in patients with hypocitraturia?
(Prospective randomised study). Eur. Urol. Suppl., 7, 152, 2008.
45. D’Aquino, M. and Teves, S.A., Lemon juice as a natural biocide for disinfecting drinking water.
B. PAHO, 28, 324–330, 1994.
46. Miyake, Y., Yamamoto, K., Morimitsu, Y., and Osawa, T., Characteristics of antioxidative flavonoid
glycosides in lemon fruit. Food Sci. Technol. Int. Tokyo, 4, 48–53, 1998.
47. Ivarez-Ariasa, B.A. and Ramón-Lacab, L., Pharmacological properties of citrus and their ancient and
medieval uses in the Mediterranean region. J. Ethnopharmacol., 97, 89–95, 2005.
48. Moraes, T.M., Kushima, H., Moleiro, F.C., Santos, R.C., Rocha, L.R.M., Marques, M.O., Vilegas, W.,
and Hiruma-Lima, C.A., Effects of limonene and essential oil from Citrus aurantiumon gastric mucosa:
Role of prostaglandins and gastric mucus secretion. Chem. Biol. Interact., 180, 499–505, 2009.
49. Tian, Q., Miller, E.G., Ahmad, H., Tang, L., and Patil, B.S., Differential inhibition of human cancer cell
proliferation by citrus limonoids. Nutr. Cancer, 40, 180–184, 2001.
Lemon Juice 347
50. Arruda, D.C., Miguel, D.C., Jenicer, K.U., Yokoyama-Yasunaka, K.U., Katzin, A.M., and Uliana,
S.R.B., Inhibitory activity of limonene against Leishmania parasites in vitro and in vivo. Biomed.
Pharmacother., 63, 643–649, 2009.
51. Aggarwal, B. and Sung, B., Pharmacological basis for the role of curcumin in chronic diseases: An age-
old spice with modern targets. Trends Pharmacol. Sci., 30, 2, 85–94, 2009.
52. Zhang, Y., Li, M., Wang, H., Fisher, W.E., Peter, H.L., Yao, Q., and Chen, C., Profiling of 95 microRNAs
in pancreatic cancer cell lines and surgical specimens by real-time PCR analysis. World J. Surg., 33,
698–709, 2008.
53. Awale, S., Li, F., Onozuka, H., Esumi, H., Tekusa, Y., and Kadota, S., Constituents of Brazilian red
propolis and their preferential cytotoxic activity against human pancreatic PANC–1 cancer cell line in
nutrient-deprived condition. Bioorgan. Med. Chem., 16, 181–189, 2008.
54. Patil, J.R., Studies on isolation and characterization of bioactive compounds in lime [Citrus aurantifolia
(Christm) Swingle], their antioxidant and anticancer properties, PhD thesis, University of Agricultural
Sciences, Dharwad, India, 2009.
55. Kupfer, D. and Bulger, W.H., Metabolic activation of pesticides with progestrogenic activity. Fed. Proc.,
46, 1864–1869, 1987.
56. Cassano, A., Drioli, E., Galaverna, G., Marchelli, R., DiSilvestro, G., and Cagnasso, P., Clarification and
concentration of citrus and carrot juices by integrated membrane processes. J. Food Eng., 57, 153–163,
2003.
57. Rastiogi, N.K., Recent Developments in High Pressure Processing Foods, Springer, Berlin, Germany,
2013.
58. Donsi, G., Ferrari, G., Matteo, M.D., and Bruno, M.C., High pressure stabilization of lemon juice. Ital.
Food Bever. Technol., 14, 14–16, 1998.
59. Silva, C.M.G., Contesini, F.J., Sawaya, A.C.H.F., Cabral, E.C., Cunha, I.B.S., Eberlin, M.N., and
Carvalho, P.O., Enhancement of the antioxidant activity of orange and lime juices by flavonoid enzy-
matic de-glycosylation. Food Res. Int., 52, 308–314, 2013.
60. Yekeler, F.Z., Ozyurek, H., and Tamer, C.E. A Functional beverage: Lemonade. Int. J. Agri. Biosci.
Eng., 7, 249–252, 2013.
61. Chornomaz, P.M., Pagliero, C., Marchese, J., and Ochoa, N.A., Impact of structural and textural
membrane properties on lemon juice clarification. Food Bioprod. Process., 91, 67–73, 2013.
29
Lime Juice
Lucia Maria Jaeger de Carvalho, Gisela Maria Dellamora-Ortiz,

and José Luiz Viana de Carvalho
CONTENTS
29.1 Introduction................................................................................................................................... 349
29.4 Health Effects.................................................................................................................................352
29.6 Conclusion......................................................................................................................................355
References................................................................................................................................................355
29.1 Introduction
Citrus fruits (Rutaceae) are grown worldwide in regions with mild climate in winter and in
appropriate soils. Although different varieties or species of citrus fruits exist, lemons (Citrus limon
L. Burm. f.) and limes (C. latifolia and C. aurantifolia) are the most widespread ones used for juice
processing [1–4].
Brazil is the world’s largest producer of citrus crops. Recent studies have shown that irrigated areas
of citrus in the country have been increasing significantly during the last few years [5]. Citriculture was
established in the Brazilian scenery as one of the most important agricultural activities. Today, Brazil
is the world’s largest orange producer, and the state of São Paulo is responsible for 85% of the country’s
citrus fruit production, mainly destined for the concentrated orange juice industry [6]. It does not differ
with the “Tahiti” lime, and the state of São Paulo dominates this internal market, with about 70% of the
entire country’s production. In this context, cultivation of the “Tahiti” lime has drawn the growers’ atten-
tion because of its high profitability and expansion possibilities, as the application of new techniques
allows for a secondary crop during the off-season, when fruit prices are more attractive [7].
According to FAO [8], the main citrus-producing countries include China, Brazil, and the United
States, followed by Mexico, Spain, and Italy. The citrus flavor and aroma are pleasant, and there are
essential nutrients and micronutrients in its composition, such as minerals, vitamins, and bioactives [1,9].
Vitamin C and phenolic compounds present in lime possess antioxidant activity due to their ability to
scavenge free radicals [10]. There is considerable evidence that citrus fruits have antioxidant and antimu-
tagenic properties and positive associations with bone, cardiovascular, and immune system health [11,12].
Fruits and vegetables are the main source of natural antioxidants in the human diet and are associ-
ated with a lower risk of cardiovascular disease (CVD), diabetes, and immune system deficiency, among
others [13]. Polyphenols, present in different foods and drinks, are a heterogeneous group composed
of several classes of substances with antioxidant properties. Typical compounds that possess anti-
oxidant activity include simple phenols, phenolic acids and their derivatives, carotenoids, flavonoids
(anthocyanins and anthoxanthins), tocopherols, phospholipids, amino acids, phytic acid, ascorbic acid,
349
and sterols [14]. Marín et al. [15] and others [16–18] have reported that citrus fruits are important sources
of ascorbic acid and flavonoids, frequently used in the industry for the extraction of flavanones [19,20].
The large, green, and seedless lime found in the supermarket is the “Persian” or “Tahiti” lime, a
hybrid developed in the early twentieth century. They are picked slightly immature while they are still
green in color (they turn yellow when fully ripe and might be confused with lemon) [21]. This chapter
highlights the nutritional characteristics, bioactive compounds, antioxidant efficacy, and health effects of
lime juice. Novel products, formulations, and future trends are also discussed.

Lime juice is rich in micro- and macronutrients, with low caloric value and high antioxidant activity due
to the presence of carotenoids such as β-carotene [22]. Vitamin C and potassium are the major constitu-
ents. The compositional and nutritional characteristics of lime juice (canned or bottled and unsweetened)
are given in Table 29.1 [21].
Rangel et al. [23] compared some parameters of conventional (C. latifolia Tanaka) and organic biody-
namic lime juices from the same species. They found the following results: pH (2.81 and 2.78), acidity in
citric acid (6.05 and 5.98 g/100 g), soluble solids (8.42 and 7.63°Brix), fructose (0.98 and 0.66 g/100 g),
glucose (0.89 and 0.54 g/100 g), sucrose (0.07 and 0.12 g/100 g), vitamin C (22.86 and 22.80 mg/mL),
TABLE 29.1
Compositional and Nutritional Characteristics of Lime Juice (per 100 g)
Nutrient Unit Canned or Bottled, Unsweetened
Water g 92.52
Energy kcal 21
Protein g 0.25
Lipid (fat) g 0.23
Carbohydrate g 6.69
Total sugars g 1.37
Minerals
Calcium mg 12
Iron mg 0.23
Magnesium mg 7.0
Phosphorus mg 10
Potassium mg 75
Sodium mg 16
Zinc mg 0.06
Vitamins
Folate (DFE) µg 8.0
Niacin mg 0.163
Riboflavin mg 0.003
Thiamin mg 0.033
Vitamin A (RAE) µg 1.0
Vitamin B6 mg 0.027
Vitamin C mg 6.4
Vitamin K µg 0.5
Reference Release 27, 2014, Published online at: http://ndb.nal.usda.gov/ndb/foods (accessed April 20, 2015).
Lime Juice 351
and minerals. Potassium (37.7 mg/100 g) and calcium (2.32 mg/100 g) were the most abundant minerals
in both juices.
The physical and chemical characteristics for lime and lemon juices are provided by the Brazilian
legislation standards [24]. Compared to lemon, lime contains less vitamin C but still provides 35% of
the Recommended Dietary Allowances (RDA) value per 100 g serving. Lime is a good source of dietary
fiber and contains numerous other nutrients in small quantities.
Among other compounds, volatiles are responsible for some bioactivities of citrus juices, especially
lemon and lime. Fonseca [25] found that d-limonene is the major component in lime juice (20.8 µg/g).
The peel oils of lime and lemon are rich in limonene that is the most abundant essential oil component
(62% in lime essential oil) in lime juice [26]. Additionally, lemon and lime are the third most important
Citrus species as important health-promoting fruits rich in phenolic compounds. Essential oils from
limes reduced anxiety and prolonged ether sleeping time in mice [27].

The bioactive compounds are important in Citrus species because of the antioxidant capacity ascribed
to vitamin C and some phenolic compounds (flavonoids), as well as minerals, dietary fiber, essential oils,
and carotenoids. Additionally, lemon and lime are the third most important Citrus species as important
health-promoting fruits rich in phenolic compounds. Lime flesh and peel contain diverse phytochemicals,
including polyphenols and terpenes, many of which are under basic research for their potential properties
in humans [2]. On the other hand, Gorgus et al. [27] evaluated the phototoxic and photogenotoxic prop-
erties in combination with ultraviolet radiation of furocoumarins in citrus-containing beverages. The
coumarin derivative limettin was mainly found in lime products. The main bioactive compounds found
in the Mexican lime (C. aurantifolia Swingle) juice were rutin, neohesperidin, hesperidin, quercetin,
and hesperetin (Figure 29.1). In addition, the limonoids’ limonexic acid, isolimonexic acid, limonin, and
nomilin (bitter) were also found [28].
Barros et al. [29] evaluated the chemical composition, antioxidant activity, and total polyphenols of
the peels and pulps of four Citrus species (C. sinensis cvs. Pera and Lima, C. latifolia Tanaka cv. Tahiti,
C. limettioides Tanaka cv. Sweet lime, and C. reticulata cv. Ponkan) grown in Brazil. Peels presented
significantly higher contents of all bioactive compounds than the pulps (0.05%), with the exception of
OH O
OH
HO OH
HO O
O O
OH
O
OH O OH
O OCH3
OH O HO OO OH
OH O OH O
O CH3 O HO
O O O
O
OH OH CH3
OH OH OCH3
OH OH OH
OH OH OH OH OH
(a) (b) (c)
OH OH
OH OH
HO O HO O
OH OH
OH O OH O
(d) (e)
FIGURE 29.1 Some bioactive compounds found in lime juice: (a) rutin, (b) hesperidin, (c) neohesperidin, (d) quercetin,
and (e) naringenin.
TABLE 29.2
Content of Some Bioactive Compounds in Lime Juice (per 100 g)
Compound Unit Lime Juice References
Carotenoids
β-Carotene μg 30 [21]
Flavonoids
Eriodictyol mg 2.2 [21]
Hesperetin mg 9.0 [21]
Naringenin mg 0.4 [21]
Quercetin mg 0.5 [21]
Limonoids
Limonin and nomilin mg 150–300 [28]
the Pera orange pulp that presented the highest ascorbic acid content (68 mg/100 mL), while Tahiti lime
peel presented the lowest content (8 mg/100 g) of vitamin C. Peels presented higher contents of miner-
als and phenolics than pulps. The antioxidant activity of citrus was correlated with both vitamin C and
phenolics. Aside from citrus pulps, the peels are also good sources of bioactive compounds and minerals
and can be explored for their health-promoting potential in food products.
Lime contains a number of bioactive compounds that are important contributors to the antioxidant
power of vitamin C, the most abundant of which are limonoids in the peels. Limonene is a monoterpene
that helps to trigger detoxification in the liver and small bowel, increasing the number of enzymes, stimu-
lating glutathione S-transferases, and thereby helping to eliminate carcinogens. Animal studies indicate
that limonene is a chemopreventive and anticancer agent.
Lime juice contains a number of flavonoids [21]. The most abundant flavanone is hesperetin
(9 mg/100 g), followed by eriodictyol (2.2 mg/100 g) and naringenin (0.4 mg/100 g), and quercetin
(0.5 mg/100 g) is the flavonol present in the edible portion and lime juice. In addition, β-carotene is the
unique carotene present in lime juice (30 µg/100 g). In addition, limonoids (limonin and nomilin) were
reported to be between 150 to 300 mg/100 g in lime juice (Table 29.2). Several authors have studied
bioactive compounds (phenolics) and antioxidant activity as well as the methods to evaluate and identify
these compounds in citrus juices [30–34].
According to Beh et al. [33], fruits contain various natural antioxidants that play an important role in
human health. They evaluated the total phenolic content and antioxidant activity of various freeze-dried
fresh fruit blends or fruit juices including lime. The commercial lime juice had the lowest antioxidant
activity and did not show any meaningful EC50 (effective concentration [scavenging 50% in the reaction
present radicals]) even up to 10 mg/mL. The highest antioxidant activity and EC50 were found in organic
lime juice compared to the conventional lime juice [35].
On the other hand, studies on antioxidant activities of lime peel powders (Persian and Mexican)
revealed that lime fiber had the highest polyphenol contents, and polyphenols associated with dietary
fiber in both lime peel varieties showed a good antioxidant activity [32]. Xu et al. [32] evaluated the
antioxidant activity of citrus fruits. Tangerine showed 29.67% and lime 24.50% inhibition in the
2,2-diphenyl-1-picrylhydrazyl (DPPH) method. These data are similar to previously reported results for
citrus fruits by Yoo et al. [36]. The flavonoids of citrus juices were studied, and the main compounds
found were flavanone-O-glycosides and flavone-O- or -C-glycosides [31].
29.4 Health Effects

Citrus fruits, especially lemon, lime, and orange, are similar in their health benefits and are excellent
sources of vitamin C, potassium, folate, flavonoids, and the outstanding phytochemical limonene. Lime
and lemon are rich in phytochemicals with high antioxidant activity. They render beneficial effects on
human health such as anticarcinogenic, antibiotic, and antimicrobial effects. Limonene has anticancer
effects and helps increase the level of enzymes that detoxify carcinogens.
Lime Juice 353
Rutin promotes the inhibition of platelet aggregation, thus improving circulation, has anti-inflammatory
activity, and inhibits aldose reductase that helps change glucose into sorbitol; it also prevents hyperglyce-
mia and assists in preventing sorbitol accumulation [37].
Naringin and its aglycone naringenin are flavonoids that are found to display strong anti-inflammatory
and antioxidant activities. Naringin supplementation has been shown to be beneficial for the treatment of
obesity, diabetes, hypertension, and metabolic syndrome. A number of molecular mechanisms underly-
ing its beneficial activities have been elucidated [38].
Dietary antioxidants may offer some protection against the early stage of diabetes mellitus and the
development of associated complications. Jung et al. [39] investigated the effect of citrus flavonoids on
blood glucose level, hepatic glucose-regulating enzymes activities, hepatic glycogen concentration, and
plasma insulin levels and assessed the relations between plasma leptin, body weight, blood glucose,
and plasma insulin in mice. Hesperidin and naringin supplementation significantly reduced blood glu-
cose compared with the control group. Hepatic glucokinase activity and glycogen concentration were
both significantly elevated in the hesperidin- and the naringin-supplemented groups compared with the
control group. Naringin also markedly lowered the activity of hepatic glucose-6-phosphatase and phos-
phoenolpyruvate carboxykinase compared with the control group. Plasma insulin, C-peptide, and leptin
levels in the db/db mice from two bioflavonoid-supplemented groups were significantly higher than those
of the control group. The results suggested that hesperidin and naringin both play important roles in
preventing the progression of hyperglycemia, partly by increasing hepatic glycolysis and glycogen con-
centration and/or by lowering hepatic gluconeogenesis [39].
Hesperetin, naringin, and naringenin are flavonoid glycosides commonly found in citrus fruits.
Naringenin is found to have bioactive effect on human health as antioxidant, free radical scavenger, anti-
inflammatory, and immune system modulator. This substance has also been shown to reduce oxidant
injury to DNA in the cells in in vitro studies.
In order to develop novel prevention strategies to reduce human immunodeficiency virus (HIV) trans-
mission, Fletcher et al. [40] evaluated lime juice as a topical microbicide. They found a good effect in
HIV. However, the adverse effects on genital mucosa of women must be taken into account when consid-
ering this as a prevention approach.
The inactivation of Escherichia coli O157:H7 was evaluated in lime and lemon juices. A greater than
5-log reduction of stationary-phase cells was achieved in both lemon and lime juices after 72 h of incu-
bation at 22°C [41]. Storage at room temperature can, thus, be an alternative to heat treatment for juices.
Generally, health benefits of citrus are related to bioactive compounds present in the fruits, such as
carotenoids, essential oils, organic acids, dietary fiber, phenolic compounds (mainly flavonoids), miner-
als, and vitamins (mainly vitamin C). There is scientific evidence that these substances are involved in
the prevention and treatment of diseases, such as obesity, diabetes, hypercholesterolemia, CVD, and
certain types of cancers (Table 29.3).
Kummer et al. [26] investigated the anti-inflammatory activity in C. latifolia essential oil and limonene
effects using cell viability and in vitro assays in mouse models. In cells, 90 μg/mL doses promoted low
cytotoxicity, and regarding the zymosan-induced peritonitis, limonene (500 mg/kg) decreased the infil-
tration of peritoneal exudate leukocytes and the number of polymorphonuclear leukocytes. Limonene
isolated from C. latifolia essential oil had potential anti-inflammatory effects, inhibiting proinflamma-
tory mediators present in inflammatory exudate and leukocyte chemotaxis.
The potential benefits of bioactive compounds (curcumin, flavonols, and isoflavones) in pancreatic can-
cer prevention have been reported [42–45]. Patil et al. [28] confirmed the involvement of apoptosis in induc-
tion of cytotoxicity in pancreatic cells by expression of Bax, Bcl-2, caspase-3, and p53. These results clearly
indicated that antioxidant activity is proportional to the content of flavonoids and that proliferation inhibi-
tion ability is proportional to the content of both flavonoids and limonoids present in Mexican lime juices.
An impressive array of research on lime juice from the National Library of Medicine indicates that it could
either cure or greatly accelerate healing time from a variety of life-threatening illnesses including sickle cell
anemia and malaria [46], bacterial agents in food (Vibrio parahaemolyticus and Salmonella enterica) [47],
disinfecting water enhancing (E. coli) [48], killing pancreatic cancer, and stopping smoking [49].
On the other hand, toxic effects can be observed in lime rind and juice/pulp. Nigg et al. [50] evalu-
ated the presence of coumarins in the rind and pulp of Persian and Key limes. In the rind of Persian
TABLE 29.3
Health Benefits Attributed to Lime and Lime Juice
Diseases Benefits/Effects References
Scurvy Vitamin C reduces the deficiency. [27]
Digestion Aids primary digestion. [55]
Constipation High acidity (citric acid). [23,55]
Diabetes High levels of soluble fiber, ideal dietary aid to help regulate the body’s absorption of [12]
sugar into the bloodstream. Low GI.
Heart disease Soluble fiber can also lower blood pressure and eliminate the presence of LDL [12,54]
cholesterol. Can cut down on inflammation of the blood vessels, which is a known
preventative measure against CHD.
Peptic ulcer Flavonoids, limonoids, and limonin glucoside, which have antioxidant, [55]
anticarcinogenic, antibiotic, and detoxifying properties, stimulating the healing
process of peptic and oral ulcers.
Respiratory Flavonoid-rich oil of lime used in anticongestive medicines: balms, vaporizers, and [55]
disorders inhalers (kaempferol).
Arthritis Citric acid is a solvent in which uric acid can dissolve, increasing the amounts that are [41]
eliminated in the urine.
Eye care Vitamin C (antioxidant) protects eyes from aging and macular degeneration. [27]
Flavonoids protect the eyes from infections.
Fever Vitamin C fever-reducing qualities. [27]
Gout Lime can help prevent despite its source of antioxidants and detoxifiers (vitamin C and [27]
flavonoids), reducing the free radicals.
Lime gums Vitamin C (scurvy, which gives bleeding and spongy gums) and microbial growth. [55]
Flavonoids inhibit microbial growth and potassium. Flavonoids help heal ulcers and
wounds.
Piles Helps heal ulcers and wounds in the digestive system and excretory system, while [12]
providing relief from constipation. It eradicates all the root causes of piles.
Cholera Lime juice, when added to potentially infected water, is a very effective disinfectant, [48]
and even when it was consumed regularly after someone had been exposed to
cholera-infected water, fatalities were reduced.
Weight loss Citric acid, present in lime and lime juice, is an excellent fat burner. [12,23]
Urinary The high potassium content of lime is very effective in removal of the toxic substances [12]
disorders and the precipitates, which get deposited in kidneys and the urinary bladder.
Other benefits Can help to cure rheumatism, prostate and colon cancers, arteriosclerosis, and fatigue. [12,37]
Abbreviations: CHD, coronary heart disease; GI, glycemic index; LDL, low-density lipoprotein.
lime, coumarin concentrations were in the decreasing order of limettin > bergapten > isopimpinellin >
xanthotoxin > psoralen. In the rind of Key lime, psoralen and xanthotoxin were analytically absent;
limettin was 10-fold more concentrated than either bergapten or isopimpinellin, which were equal in
concentration. Coumarin content in Persian lime pulp was in the order of isopimpinellin > limettin >
bergapten > xanthotoxin > psoralen. For Key lime pulp, the concentrations of limettin, isopimpinellin,
and bergapten were equal; psoralen and xanthotoxin were not detected. Coumarins in lime pulp were
13–182-fold less concentrated than those in the peel. Based on the amounts and types of coumarins,
Persian lime appears to be potentially more phototoxic than Key lime. Although bergapten may be the
main component of both limes responsible for phytophotodermatitis, dermatological interaction assays,
further research using psoralen, bergapten, xanthotoxin, and limettin should be conducted.

The yield of essential oil obtained from lime peel extraction is about 1 kg of oil to 2500 fruits. The oil is rich
in limonene, citronellal, and citral, which are responsible for the citrus fruit aroma and possess antioxidant
activity, thus being used by the pharmaceutical and food industries in a wide range of applications [51].
Lime Juice 355
Processing of fruit juice has been carried out using membrane technology, for clarification using
ultrafiltration and microfiltration, as well as for concentration by reverse osmosis. These processes are
preferred in comparison to others due to the high efficiency and low temperature. Filtration rate and
consequently product quality can be affected by product preparation, membrane selection, and operat-
ing parameters. Development of new membranes, improvements in process engineering, and a better
understanding of fruit beverage constituents have increased the range of membrane separation processes.
Electrodialysis and pervaporation are among the new technologies that increase the possibilities for
applications in processing of lime juices and beverages [52,53].
The development of functional drinks and foods has opened a new market for consumers that want to
be healthy. The lime juice combined with many functional raw materials provides a new option. Bhuiyan
et al. [54] developed a new beverage with black cumin seed (containing crystalline nigellone), 15 amino
acids, carbohydrates, fatty acids, volatile oils, alkaloids, dietary fiber, and minerals. Honey, garlic paste,
and lime juice are effective in reducing cholesterol. Better retention of the chemical and sensory proper-
ties was ascertained by storage at refrigerator temperature than at room temperature.
29.6 Conclusion
Various lime juices (C. latifolia, specially) contain several components with different functions and
health benefits. Lime displays the same health benefits as lemon, depending on the part of the fruit
that is used. Antioxidant activity, low caloric value, dietary fiber, and essential oils, among others, are
important to prevent many diseases and promote human health as well as other benefits. Lime is a very
versatile fruit for food industries for the formulation of new products including functional beverages or
as an additive. The combination of lime juice with many functional raw materials is a new option.
REFERENCES
1. Morton, L.W., Cacetta, R.A., Puddey, I.B., and Croft, K.D., Chemistry and biological effects of dietary
phenolic compounds: Relevance to cardiovascular disease. Clin. Exper. Pharmacol. Physiol., 27,
152–159, 2000.
2. Lime, Lime fruit, 2015. Published online at: http://en.wikipedia.org/wiki/Lime_(fruit)#Plants_known_
as_.22lime.22 (accessed March 5, 2015).
3. Lime, Lime fruit data: Yield, sugar, acidity, tannin, 2011. Published online at: http://www.brsquared.
org/wine/CalcInfo/FruitDat.htm (accessed March 5, 2015).
4. Morton, J.F., Tahiti lime, in Fruits of Warm Climates, Morton, J.F., Ed., Echo Point Books & Media,
LLC, Miami, FL, 1987, pp. 172–175.
5. Barboza Júnior, C.R.A., Escola Superior de Agricultura Luiz de Queiroz (in Portuguese), MSc thesis,
São Paulo State University, Piracicaba, Brazil, 2007.
6. Mattos, D., Jr. De Negri, J.D., Figueiredo, J.O., and Pompeu, J. Jr., Citros: Principais Informações
e Recomendações de Cultivo. Published online at: www.iac.sp.gov.br (accessed February 28, 2015).
7. Stuchi, E.S., Martins, A.B.G., Lemo, R.R., and Cantuarias-Avilés, T., Fruit quality of ‘’Tahiti’’ lime
(Citrus latifolia Tanaka) grafted on twelve different rootstocks. Ver. Bras. Frutic., 31, 454–460, 2009.
8. FAO, FAOSTAT Statistics Database, Food Agriculture Organization, Rome, Italy, 2008.
9. Pellegrini, N., Serafini, M., Colombi, B., and Del Rio, D., Total antioxidant capacity of plant foods bever-
ages and oils consumed en Italy assessed by three different in vitro assays. J. Nutr., 133, 2812–2819, 2003.
10. Dhuique-Mayer, C., Caris-Veyrat, C., Ollitrault, P., Curk, F., and Amiot, M.J., Varietal and interspe-
cific influence on micronutrient contents in citrus from the Mediterranean area. J. Agric. Food Chem.,
53, 2140–2145, 2005.
11. Codoñer-Franch, P. and Valls-Bellés, V., Citrus as functional foods. Curr. Trends Nutraceut. Res.,
8, 173–183, 2010.
12. Turner, T. and Burri, B.J., Potential nutritional benefits of current citrus consumption: Review.
Agriculture, 3, 170–187, 2013.
13. Harborne, J.B. and Williams, C.A., Advances in flavonoid research since 1992. Phytochemistry,
55, 481–504, 2000.
14. Roesler, R., Catharino, R.R., Malta, L.G., Eberlin, M.N., and Pastore, G., Antioxidant activity of
Caryocar brasiliense (pequi) and characterization of components by electrospray ionization mass spec-
trometry. Food Chem., 110, 711–717, 2008.
15. Marín, F.R., Martinez, M., Uribesalgo-Castillo, T.S., and Frutos, M.J., Changes in nutraceutical com-
position of lemon juices according to different industrial extraction systems. Food Chem., 78, 319–324,
2002.
16. Benavente-García, O., Castillo, J., and Del Río, J.A., Changes in neodiosmin levels during development
of Citrus aurantium leaves and fruits. Postulation of neodiosmin biosynthetic pathway. J. Agric. Food
Chem., 41, 1916–1919, 1993.
17. Del Río, J.A., Arcas, M.C., Benavente, O., Sabater, F., and Ortuño, A., Changes of polymethoxylated
flavones levels during development of Citrus aurantium (cv. Sevillano) fruits. Planta Med., 64, 575–576,
1998.
18. Ortuño, A., Arcas, M.C., Benavente-García, O., and Del Río, J.A., Evolution of polymethoxy flavones
during development of tangelo Nova fruits. Food Chem., 66, 217–220, 1999.
19. Baer, A., Borrego, F., Benavente-García, O., Castillo, J., and Del Río, J.A., Neohesperidin-
dihydrochalcone: Properties and applications. LWT—Food Sci. Technol., 23, 371–376, 1990.
20. Del Río, J.A., Fuster, M.D., Sabater, F., Porras, I., García Lidón, A., and Ortuño, A., Selection of citrus
varieties highly productive for the neohesperidin dihydrochalcone precursor. Food Chem., 59, 433–437,
1997.
21. U.S. Department of Agriculture (USDA), USDA National Nutrient Database for Standard Reference
Release 27, 2014. Published online at: http://ndb.nal.usda.gov/ndb/foods (accessed April 20, 2015).
22. Key Limes, 2015. Published online at: http://www.foodreference.com/html/artkeylimes.html (accessed
March 5, 2015).
23. Rangel, C.N., Carvalho, L.M.J., Fonseca, R.B.F., Soares, A.G., and Jesus, E.O., Nutritional value of
organic acid lime juice (Citrus latifolia T.), cv. Tahiti. Food Sci. Technol. Campinas, 31, 918–922, 2012.
24. Ministry of Agriculture, Livestock, and Supply of Brazil, Identity and Quality Standards for Acid Lime
Juice, Official Gazette (January 10th), Rio de Janeiro, Brazil, 2000.
25. Fonseca, R.B.F., Identification of volatile compounds in acid lime (Citrus latifolia, Tanaka), cv. Tahiti,
from conventional and biodynamic cultivations, MSc thesis, Federal University of Rio de Janeiro, Rio
de Janeiro, Brazil, 2007.
26. Kummer, R., Fachini-Queiroz, F.C., Silva, C.F.E., Grespan, R., Silva, E.L., Bersani-Amado, C.A.,
and Cuman, R.K.N., Evaluation of anti-inflammatory activity of Citrus latifolia Tanaka essential
oil and limonene in experimental mouse models. eCAM., Article ID 859083, 8pages http://dx.doi.
org/10.1155/2013/859083, 2013.
27. Gorgus, E., Lohr, C., Raquet, N., Guth, S., and Schrenk, D., Limettin and furocoumarins in beverages
containing citrus juices or extracts. Food Chem. Toxicol., 48, 93–98, 2010.
28. Patil, J.R., Murthy, K.N.C., Jayaprakasha, G.K., Chetti, M.B., and Patil, B.S., Bioactive compounds
from Mexican lime (Citrus aurantifolia) juice induce apoptosis in human pancreatic cells. Aliment
Pharmacol. Ther., 1521, 435–444, 2005.
29. Barros, H.R.M.F., Castro, T.A.P., and Genovese, M.I., Antioxidant capacity and mineral content of pulp
and peel from commercial cultivars of citrus from Brazil. Food Chem., 134, 1892–1898, 2012.
30. Abeysinghe, D.C., Xian, L., Chong, D.S., Wang, S.Z., Chun, H.Z., and Kun, S.C., Bioactive compounds
and antioxidant capacities in different edible tissues of citrus fruit of four species. Food Chem., 104,
1338–1344, 2007.
33. Beh, L.K., Zakaria, Z., Beh, B.K., Ho, W.Y., Yeap, S.K., Banu, N., and Alitheen, M., Comparison of total
Plants Res., 6, 5857–5862, 2012.
34. Viana, D.S., Acid lime (Citrus latifolia Tanaka), cv. Tahiti, from biodynamic organic cultivations:
Evaluation of the antioxidant capacity of fresh juices and clarified by microfiltration membranes, MSc
thesis, Rio de Janeiro Federal University, Rio de Janeiro, Brazil, 2010.
Lime Juice 357
35. Ubando-Rivera, A.J., Navarro-Ocaña, V., and Lopez, M.A., Mexican lime peel: Comparative study on
contents of dietary fibre and associated antioxidant activity. Food Chem., 89, 57–61, 2005.
36. Yoo, K.M., Lee, K.W., Park, J.B., Lee, H.J., and Hwang, I.K., Variation in major antioxidants and total
antioxidant activity of yuzu (Citrus junus Sieb ex Tanaka) during maturation and between cultivars.
J. Agric. Food Chem., 52, 5907–5913, 2004.
37. Sahelian, R., Rutin supplement health benefits, 2012. Published online at: www.rayshelian.com
38. Alam, M.A., Subhan, N., Rahman, M.M., Uddin, S.J., Reza, H.M., and Sarker, S.D., Effect of citrus
flavonoids, naringin and naringenin, on metabolic syndrome and their mechanisms of action. Adv. Nutr.,
5, 404–417, 2014.
39. Jung, U.J., Lee, M.K., Jeong, K.S., and Choi, M.S., The hypoglycemic effects of hesperidin and naringin
are partly mediated by hepatic glucose-regulating enzymes in C57BL/KsJ-db/db mice. J. Nutr., 134,
2499–2503, 2004.
40. Fletcher, P.S., Harman, S.J., Boothe, A.R., Doncel, G.F., and Shattock, R.J., Preclinical evaluation of
lime juice as a topical microbicide candidate. Retrovirology, 5, 3, 2008.
41. Enache, E., Chen, Y., and Elliott, P.H., Inactivation of Escherichia coli O157:H7 in single-strength
lemon and lime juices. J Food Prot. 72, 235–240, 2009.
42. Aggarwal, B. and Sung, B., Pharmacological basis for the role of curcumin in chronic diseases: An
age-old spice with modern targets. Trends Pharmacol. Sci., 30, 85–94, 2009.
43. Zhang, Y., Li, M., Wang, H., Fisher, W.E., Peter, H.L., Yao, Q., and Chen, C., Profiling of 95 microRNAs
in pancreatic cancer cell lines and surgical specimens by real-time PCR analysis. World J. Surg., 33,
698–709, 2008.
44. Awale, S., Li, F., Onozuka, H., Esumi, H., Tekusa, Y., and Kadota, S., Constituents of Brazilian red
propolis and their prefential cytotoxic activity against human pancreatic PANC–1 cancer cell line in
nutrient-deprived condition. Bioorgan. Med. Chem., 16, 181–189, 2008.
45. Patil, B.S., Jayaprakasha, G.K., Murthy, K.N.C., and Vikram, A., Bioactive compounds: Historical per-
spectives, opportunities, and challenges, J. Agric. Food Chem., 57, 8142–8160, 2009.
46. Adegoke, S.A., Oyelami, O.A., Olatunya, O.S., and Adeyemi, L.A., Effects of lime juice on malaria
parasite clearance. Phytother. Res., 25, 1547–1550, 2011.
47. Mathurand, P. and Schaffner, D.W., Effect of lime juice on Vibrio parahaemolyticus and Salmonella
enterica inactivation during the preparation of the raw fish dish ceviche. J. Food Prot., 76, 1027–1030,
2013.
48. Rodrigues, A., Sandström, A., Cá, T., Steinsland, H., Jensen, H., and Aaby, P., Protection from cholera
by adding lime juice to food—Results from community and laboratory studies in Guinea-Bissau, West
Africa. Trop. Med. Int. Health., 5, 418–22, 2000.
49. Rungruanghiranya, S., Ekpanyaskul, C., Sakulisariyaporn, C., Watcharanat, P., and Akkalakulawas, K.,
Efficacy of fresh lime for smoking cessation. J. Med. Assoc. Thai., 95(Suppl.), S76–S82, 2012.
50. Nigg, H.N., Nordby, H.E., Beier, R.C., Dillman, A., Macias, C., and Hansen, R.C., Phototoxic couma-
rins in limes. Food Chem. Toxicol., 31, 331–335, 1993.
51. Mendonça, L.M.V.L., Conceição, A., Piedade, J., Carvalho, V.D., and Theodoro, V.C.A., Characterization
of the chemical composition and the yielding of industrial residues from thaiti lime (Citrus latifolia
Tanaka). Food Sci. Technol., Campinas, 26, 870–874, 2006.
52. Girard, B. and Fukumoto, L.R., Membrane processing of fruit juices and beverages: A review. Crit. Rev.
Food Sci. Nutr., 40, 91–157, 2000.
53. Ilame, S.A. and Singh, S.V., Application of membrane separation in fruit and vegetable juice processing:
A review. Crit. Rev. Food Sci. Nutr., 55, 964–987, 2015.
54. Bhuiyan, M.H.R., Shams-Ud-Din, M., and Islam, M.N., Development of functional beverage based on
taste preference. J. Environ. Sci. Nat. Resour., 5, 83–87, 2012.
55. Lime, Health benefits of lime, 2015. Published online at: www.organicfacts.net/health-benefits/fruit/
health-benefits-of-lime.html (accessed April 20, 2015).
30
Mango Juice
Sui Kiat Chang and Amin Ismail
CONTENTS
30.1 Introduction................................................................................................................................... 359
30.3 Bioactives and Antioxidant Capacity............................................................................................ 362
30.3.1 Commercial Mango Juice................................................................................................. 362
30.3.2 Juice Produced from Mangifera pajang (Bambangan).................................................... 363
30.3.3 Polyphenol Content of Mango and Its Juice..................................................................... 364
30.4 Health Effects................................................................................................................................ 367
30.4.1 Bioavailability of Bioactives from Mango Juice in Relation to Its Intake....................... 367
30.4.2 Results from In Vitro, Animal, and Human Intervention Studies................................... 367
30.6 Conclusion..................................................................................................................................... 370
References............................................................................................................................................... 370
30.1 Introduction
Mango (Mangifera indica L.) is one of the most important tropical fruit crops in the world and is ranked
fifth in production among major fruit crops worldwide [1,2]. The Food and Agricultural Organization (FAO)
estimated that the worldwide production of mango in 2014 was 82.28 million tons. Sixty-nine percent of the
total production is from Asia and the Pacifics (India, China, Pakistan, the Philippines, and Thailand) [3].
The genus Mangifera contains various species cultivated in different countries. Most of the fruits that
are commonly known belong to the species of M. indica L. [1,2]. There are also underutilized mango
species, for example, M. pajang that is found in Malaysia, Brunei, and Indonesia (Borneo Island), with
their size being three times the size of commercial mangoes [4]. The mango is one of the few fruits that
can be utilized in all stages of maturity. The fruit is used as a dessert and a table fruit between meals
and is also processed for various food products, including pulp, juice, squash, nectar, pickles, chutney,
preserves, jams, canned slices, dried powder, and mango toffee [2]. Furthermore, mango has high nutri-
tive value with its high vitamins, carotenoids, minerals, and polyphenols [1].
Fruit juices have been used as a convenient substitute for fresh fruits. Fruit juices contain the
physicochemical and nutritional characteristics of fruits from which they are produced. Hence, fruit
juice consumption should also contribute to the maintenance of human health, making fruit juices
as functional beverages. Health benefits of fruit juices are usually attributed to various bioactive
compounds present [5–7]. The quality of mango juice, such as its bioactive compounds and health
benefits, has been scarcely reviewed in the recent past. Hence, this chapter highlights the nutritional
characteristics, phenolic contents, antioxidant capacity, health effects, novel formulations, and future
trends of mango juice.
359

Proximate composition and nutritional characteristics of mango juice found in the U.S. market are given
in Table 30.1 [8]. Mango juice and nectar are important beverages prepared on a commercial scale. Mango
juice and nectar contain 12% and 20% of pulp, respectively [2]. Table 30.1 shows that all B-vitamins,
vitamin E, vitamin K, and some minerals, such as magnesium, phosphorus, and zinc, are not detected
in commercial juice compared to mango nectar and fresh mango pulp [8]. Mango juice is a good source
of vitamin A, as its content in commercial mango juice is almost two- and threefold higher than that
of mango pulp and mango nectar, respectively. However, mango juice has the lowest vitamin C content
(2.5 mg/100 g) compared to fresh mango pulp (36.4 mg/100 g) and mango nectar (15.4 mg/100 g). Huge
amounts of vitamin C are lost during the processing of mango juice from its pulp. A study by Mahdavi
et al. [9] demonstrated that vitamin C content of natural fresh mango juice (14.65 mg/100 mL) and
commercial mango juice (12.57 mg/100 mL) from Iran do not differ significantly. Commercial orange,
TABLE 30.1
Compositional and Nutritional Characteristics of Mango Juice, Mango Nectar, and Raw Mango Pulp
(Mangifera indica L.)
Mango Juice, Smoothie Mango Nectar, Canned Raw Mango Pulp
Nutrient Unit (per 100 g) (per 100 g) (per 100 g)
Water g 83.46 86.63 83.46
Protein g 0.42 0.11 0.82
Lipid (fat) g 0.00 0.06 0.38
Total sugars g 12.50 12.45 13.66
Total dietary g 0.0 0.3 1.6
fiber
Minerals
Calcium mg 8 17 11
Iron mg 0.15 0.36 0.16
Magnesium mg — 3 10
Phosphorus mg — 2 14
Sodium mg 4 5 1
Zinc mg — 0.02 0.09
Vitamins
Folate (DFE) μg — 7 43
Niacin mg — 0.08 0.669
Riboflavin mg — 0.003 0.038
Thiamin mg — 0.003 0.028
Vitamin A IU 2083 692 1082
Vitamin B12 μg — 0.00 0.00
Vitamin B6 mg — 0.015 0.119
Vitamin C mg 2.5 15.2 36.4
Vitamin E (ATE) mg — 0.21 0.9
Vitamin K μg — 0.8 4.2
Abbreviations: DFE, dietary folate equivalents; IU, International Unit; ATE, alpha-tocopherol equivalents.
Mango Juice 361
sour cherry, and pomegranate juice has 15.86, 16.44, and 17.34 mg/100 mL of vitamin C, respectively;
the value is higher than that of mango juice [9].
Although mango pulp has 1.6 g dietary fiber/100 g, no fiber is present in mango juice [8]. Mango
juice contains low protein content. The energy value of the mango juice (63 kcal/100 g) is consistent
with that of the mango pulp (60 kcal/100 g) (Table 30.1). The total sugar content of mango juice is 12.5
g/100 g, which is also consistent with the study conducted on the Thai mango beverage (12.08 g/100 g).
This value is lower compared to tamarind (15.83 g/100 g) and pineapple juices (15.6 g/100 g) in
Thailand [10]. Comparatively, the energy value of mango juice (48.37 kcal/100 g) is lower compared
to tamarind (63.3 kcal/100 g) and pineapple juice (62.4 kcal/100 g) [10]. The sugars of mango juice
from Tommy Atkins cultivar were predominantly sucrose and fructose, with a lower amount of glu-
cose. The proportions of sucrose, fructose, and glucose in mango juice were 40.8%, 34.6%, and 24.6%,
respectively [11]. Mango juice is a good source of potassium (142 mg/100 g). In addition, it also has
low amount of sodium (4 mg/100 g). The iron content of mango juice is almost similar with the mango
pulp (Table 30.1).
The proximate composition and nutritional characteristics of juice produced from an underutilized
mango species; M. pajang Kostermans (Bambangan) found in the indigenous area of Borneo Island are
shown in Table 30.2. The pulp of bambangan has a high fiber content (5.26 g/100 g) compared to its
juice powder (0.80 g/100 g). Comparatively, the pulp of bambangan contains higher amount of insoluble
fiber, while its juice powder contains higher soluble fiber content. Bambangan juice powder is rich in
total carbohydrate (76.09 g/100 g) compared to its pulp (21.02 g/100 g). Furthermore, bambangan juice
powder has also higher protein and ash contents (3.78 and 3.3 g/100 g) compared to its pulp (1.13 and
0.43 g/100 g). The vitamin C content of bambangan juice powder is also higher (132.14 mg/100 g) com-
pared to its pulp (46.31 mg/100 g) (Table 30.2). However, the β-carotene content of bambangan juice
powder (35.6 mg/100 g) is lower than that of its pulp (42.21 mg/100 g). Interestingly, the bambangan
juice powder demonstrated lower energy value (335 kcal/100 g) compared to its pulp (429 kcal/100 g)
TABLE 30.2
Chemical Composition, Total Phenolic Content, and Antioxidant Capacities of
Mangifera pajang (Bambangan) Pulp and Its Juice Powder (per 100 g)
Unit Pulp Juice Powder
Chemical Composition
Moisture g 86.84 10.01
Protein g 1.13 3.78
Lipid (fat) g 1.98 1.75
Ash g 0.43 3.3
Insoluble fiber g 4.84 0.12
Soluble fiber g 0.42 0.68
Total fiber g 5.26 0.80
Energy kcal 429 335
Antioxidant Parameters
Ascorbic acid mg 46.31 132.14
β-carotene mg 42.21 35.59
Total phenolic content mg GAE 26.09 19.30
FRAP nM 26.50 36.58
DPPH % 43.25 52.61
Source: Adapted from Ibrahim, M. et al., Afr. J. Biotechnol., 9, 4392, 2010.
Abbreviations: FRAP, ferric-reducing antioxidant power; DPPH, 2,2-diphenyl-1-picrylhydrazyl;
GAE, gallic acid equivalents.
despite its higher total carbohydrate and fat contents. The reasons underlying this observation remain
to be elucidated.
By considering the nutritional compositions of mango (M. indica L.) and bambangan (M. pajang
Kostermans) juice powder, bambangan juice has the advantages of higher vitamin C, total dietary fiber,
and protein content compared to the juice of M. indica L. Thus, mango juice provides sufficient amount
of vitamin A and some minerals. Furthermore, it also has a lower energy value compared to other fruit
juices. This makes mango juice a functional beverage. There are some knowledge gaps on the mineral
and vitamin compositions of bambangan juice powder. Moreover, the nutritional compositions of the
mango juice produced from different mango cultivars need to be further studied.
30.3 Bioactives and Antioxidant Capacity

30.3.1 Commercial Mango Juice
Due to the presence of high amount of both carotenoids and polyphenols, mango is a rich source of
antioxidants [1,2,12]. Due to the improved economy, commercial fruit juices have been chosen as the
fresh fruit juice substitutes due to their convenience [7]. Commercial mango juice still has high level
of phenolic content and antioxidant capacity as compared to fresh mango (Table 30.3). Commercial
mango juice in Malaysian market has a total phenolic content (TPC) of 12.56 mg gallic acid equivalents
(GAE)/L compared to fresh mango juice (9.26 mg GAE/L). Commercial mango juice demonstrated com-
parable antioxidant capacity to the fresh fruit juice as determined by the 2,2-diphenyl-1-picrylhydrazyl
(DPPH) radical scavenging assay. This result is also in agreement with the TPC and antioxidant capacity
of commercial and fresh orange juices [7]. However, the TPC of the fresh (56.72 mg GAE/100 mL) and
commercial (28.57 mg GAE/100 mL) mango juice in the Iranian market differ significantly [9]. A recent
study also reported that there was no difference between the TPC and total flavonoid content (TFC) of
fresh (54.22 mg GAE/mL and 4.32 mg rutin equivalents [RE]/mL) and commercial (51.04 mg GAE/mL
and 4.36 mg RE/mL) mango juice produced from cultivar Banganapalli [13]. However, the antioxidant
capacities of the fresh (61.49% and 0.232 mM/g) and commercial (63.79% and 0.243 mM/g) mango
juice do not differ as determined using ferric-reducing antioxidant power (FRAP) and DPPH assay
(Table 30.3). Moreover, the antioxidant capacities of the fresh and commercial mango juice correlated
well with its TPC and TFC, indicating the contribution of phenolic compounds from mango juice to the
observed antioxidant capacities [13].
TABLE 30.3
Reported Total Phenolic Content and Antioxidant Capacities of Commercial and Fresh Mango Juice
Types of Mango
Juices TPC DPPH PCL FRAP References
Commercial 12.56 mg GAE/L IC50: 0.27 mg/mL — — [7]
mango juice
Commercial 10.0 mg GAE/100 mL 18.9 μmol 14.1 μmol — [10]
mango juice TE/100 mL TE/100 mL
17.8 μmol 51.1 μmol
AAE/100 mL AAE/100 mL
IC50: 11.2% (v/v)
Commercial 51.04 mg GAE/100 mL 63.79% — 0.243 mM/g [13]
mango juice
Fresh mango juice 9.26 mg GAE/L IC50: 0.22 mg/mL — — [7]
Fresh mango juice 54.22 mg GAE/100 mL 61.49% — 0.232 mM/g [13]
Abbreviations: TPC, total phenolic content; DPPH, 2,2-diphenyl-1-picrylhydrazyl; PCL, photochemiluminescence;
FRAP, ferric-reducing antioxidant power; GAE, gallic acid equivalents; TE, trolox equivalents; AAE,
ascorbic acid equivalents.
Mango Juice 363
The TPC of the commercial mango juice reported by Lek et al. [7] and Mahdavi et al. [9] is higher than
the mango juice found in the Thai market [10] (Table 30.3). The antioxidant capacity of the mango juice
from Thai market demonstrates 18.9 μmol trolox equivalents (TE)/100 mL and 17.8 μmol ascorbic acid
equivalents (AAE)/100 mL determined using DPPH radical scavenging assay, 14.1 μmol TE/100 mL
and 51.1 μmol AAE/100 mL determined using the photochemiluminescence assay. Another study [14]
reported that antioxidant capacities determined using oxygen radical absorbance capacity (ORAC) and
DPPH radical scavenging assay are 3.41 μmol TE/L and 605.1 AAE/L, respectively. Antioxidant capaci-
ties determined using ORAC and DPPH assays are significantly correlated with its TPC, showing that
phenolic compounds may contribute to the observed antioxidant capacities [14]. A Latino mango nectar
beverage found in the United States had the antioxidant capacity of 0.2 mmol TE/100 g or 0.44 mmol TE/
serving, as determined by using the FRAP assay [15].
The variations between the TPC and antioxidant capacities of commercial and fresh mango juices
reported by different studies [7,9,10] could be due to different varieties of fruit, the percentage of pure
juice in the final product, processing, and manufacturing techniques [9]. Falguera et al. [16] demonstrated
that the activity of polyphenol oxidase (PPO) in freshly made mango juice was low (0.016 unit/mL of
PPO activity) compared to fresh mangosteen juice (0.1435 unit/mL of PPO activity). This indicates that
the phenolic compounds of fresh mango juice will not be easily oxidized by external factors, such as
oxygen and high temperature, and hence, this will not affect its antioxidant capacity [16].
Mango juice is sold as 100% juice or nectar, or blended with other juices, such as orange, peach, and
pineapple. Blended fruit juices often marketed as skim milk mixture are gaining popularity among
consumers [17]. It has been reported that a mixture of fruit juice containing mango and skim milk
contains good TPC and antioxidant capacity [17]. These juices, orange and mango; orange, mango,
pineapple, and lemon; mango and peach; and mango and pineapple, have 75.7, 56.3, 49.2, and 26.5 mg
GAE/100 mL of TPC, respectively, with corresponding antioxidant capacities of 2.99, 3.41, 2.35, and
1.48 mmol TE/L [17]. However, there were no correlations between the TPC of these four fruit juice
mixtures with their observed antioxidant capacities. This may be due to the presence of skim milk,
which affects the concentrations of phenolic compounds, vitamins C, and A, which in turn influences
the observed antioxidant capacities. Storage and processing conditions also play important roles in
affecting their TPC and antioxidant capacities [17]. By comparing TPC of commercial and fresh mango
juice shown in Table 30.3, mixtures of fruit juices were found to contain more TPC and, hence, high
antioxidant capacities. For example, juice containing a mixture of orange, mango, pineapple, and lemon
had a lower TPC (56.3 mg GAE/100 mL) than juice containing a mixture of orange and mango (75.5 mg
GAE/100 mL). Thus, mixtures of fruit juices with complement of mango juice should be recommended
instead of mango juice alone.
30.3.2 Juice Produced from Mangifera pajang (Bambangan)

Ibrahim et al. [18] determined the TPC and antioxidant capacities of bambangan juice where its TPC
was 19.30 mg GAE/100 g. In addition, its ascorbic acid and β-carotene contents were 132.14 and
35.6 mg/100 g, respectively (Table 30.2). In this study, the antioxidant capacities of bambangan juice
were 52.61% (DPPH radical scavenging assay) and 36.58 mM/100 g (FRAP assay). In another study,
Zabidah et al. [19] evaluated the TPC and antioxidant capacities of bambangan juice where they found
that bambangan juice with a low TPC (10.01 mg GAE/100 mL) exhibited the highest antioxidant capac-
ity as determined by using DPPH radical scavenging activity (IC50: 0.03 mg GAE/mL) and β-carotene
bleaching activity (76%). The low TPC in bambangan juice can be attributed to the fact that 78% of its
TPC occurs in the kernel, while 17% occurs in the peel [20]. TPC value in this study was significantly
lower than that obtained by Ibrahim et al. [18]. In comparison, the gallic acid content of bambangan juice
was the highest compared to guava (0.51 mg/100 mL) and cocoa (0.22 mg/100 mL) [19].
In addition, bambangan juice demonstrated the highest inhibition of malondialdehyde (MDA) forma-
tion (0.70 μM MDA) in in vitro hemoglobin oxidation compared to guava (0.74 μM MDA) and cocoa
(0.736 μM MDA) [19]. For low-density lipoprotein (LDL) oxidation test, guava juice (0.27 μM MDA) dem-
onstrated a slightly higher inhibition of LDL oxidation than the bambangan juice (0.30 μM MDA) [19].
The gallic acid content of bambangan juice is comparable to the gallic acid content of bayberry juice
(0.6–2.4 mg/100 mL) [21]. However, the TPC value of bambangan juice is negatively correlated with its
antioxidant capacity [19]. Hence, it can be postulated that compounds other than phenolic compounds
may contribute to their antioxidant capacities.
A significant correlation also existed between the TPC value and inhibition activity against hemo-
globin oxidation of bambangan juice. At the same time, a strong positive correlation is found between
the antioxidant capacities and inhibition effects on LDL oxidation. This shows that polyphenols may
contribute to the inhibitory effects of bambangan juice on hemoglobin and LDL oxidation [19]. This
observation is in consistent with the findings of Kulisic-Bilusic et al. [22], showing that a strong correla-
tion existed between the antioxidant capacities of fruit (red grape, strawberry, cherry, and sour cherry)
juices and their inhibition on LDL oxidation. Bambangan juice, produced from an underutilized mango
species of M. pajang, could be regarded as a potential new functional beverage due to its possible
health-promoting properties.
30.3.3 Polyphenol Content of Mango and Its Juice

The polyphenol content of many foods has been documented in nutrient databanks such as the Phenol-
Explorer and USDA flavonoid and proanthocyanidin databases [23]. However, no data exist on the poly-
phenol contents of mango juice in both databases, except for mango pulp. Data on the content and
composition of polyphenols from Phenol-Explorer [24,25] and USDA flavonoids database [23] are also
inadequate to cover the polyphenol contents of mango from different varieties or cultivars. The major
polyphenol classes in mango pulp are anthocyanidins, flavan-3-ols, flavones, and flavonols as found in the
USDA flavonoids database [26–29]. Some 5–20 mg/100 g or 100 mL of flavanols in mango (M. indica L.)
pulp have been reported [23]. Other researchers have reported 1.72 mg/100 g fresh weight of flavanols
([+]-catechin) in mango (M. indica L.) pulp [25,29].
Gu et al. [30] reported that the total proanthocyanidins content of mango pulp was 12.8 mg/100 g. The
type of proanthocyanidins found in mango pulp was procyanidins with the amount of monomers, dimers,
trimers, and 4–6mers being 2.3, 1.8, 1.4, and 7.2 mg/100 g, respectively. Variations between the reported
flavanols content of mango pulp [23,25,29] might be due to the existing differences in determination meth-
ods, extraction conditions, and the conditions used during the identification and quantification of phenolic
compounds using high-performance liquid chromatography (HPLC) as well as mass spectrometry (MS)
[23,26]. Future research should be carried out to identify and quantify the phenolic compounds of mango
pulp from different cultivars to fill in the knowledge gap in the Phenol-Explorer and USDA databases.
The main phenolic compounds identified in commercial mango (M. indica L.) puree concentrate were
gallic acid, mangiferin, seven different quercetin and five kaempferol glycosides, and many unchar-
acterized hydrolyzable tannins, called gallotannins [31]. In most mango varieties, free gallic acid
(3,4,5-trihydroxybenzoic acid) was the predominant compound present and found to possess a high anti-
oxidant capacity [1,31]. A recent study by Naresh et al. [13] reported the phenolic compounds in mango
(M. indica L.) juice produced from the cultivar Banganapalli. The phenolic compounds of the mango
juice comprise mostly of phenolic acids, followed by flavan-3-ols and flavonols (Figures 30.1 and 30.2
and Table 30.4). Gallic acid, p-coumaric acid, m-coumaric acid, and protocatechuic acid are the four
predominant phenolic acids found in mango juice, with the rest being chlorogenic acid, syringic acid,
vanillic acid, ellagic acid, caffeic acid, synapic acid, ferulic acid, and p-OH-benzoic acid. (+)-Catechin,
quercetin, and rutin were the type of flavan-3-ols and flavonols reported in mango juice [13].
Abu Bakar et al. [32] reported that phenolic groups of the pulp of bambangan consist of flavanones
(naringin and hesperidin), flavonols (quercetin, kaempferol, and rutin), and phenolic acids (p-coumaric
acid, caffeic acid, and chlorogenic acid). In comparison, Zabidah et al. [19] reported the phenolics
profile of bambangan juice as gallic acid (1.69 mg/100 mL), vanillic acid (0.49 mg/100 mL), and trans-
cinnamic acid (0.20 mg/100 mL). Only seven phenolic standards were used for identification in this study
conducted by Zabidah et al. [19]. Hence, more research should be conducted to identify the complete
profile of phenolic compounds of bambangan juice in order to compare any similarities or differences
between juices from M. indica L. and M. pajang. However, mangiferin, being the main phenolic com-
pound of mango pulp [31], was absent in bambangan juice [19] and mango juice produced from cultivar
Banganapalli [13].
Mango Juice 365
O
O
R OH
HO
O
R
Gallic acid, R H;
Syringic acid, R OCH3
OH
HO
p-Coumaric acid
O
O
HO
OH HO
OH
R O
Protocatechuic acid, R H m-Coumaric acid
Vanilic acid, R OCH3
O
O O
O OH
OH
HO
HO O
Ferulic acid Sinapic acid
O
O
HO
O R
OH
HO
Caffeic acid, R H; trans-Cinnamic acid
Chlorogenic acid, R 5-quinony1
O
OH
O
HO
OH
O
HO
O
Ellagic acid
FIGURE 30.1 Chemical structures of major phenolic acids reported in mango juice.
OH
HO O
OH
OH
OH
(+)-Catechin
OH
HO O
OH
OH
OH O
Quercetin
OH
HO O
OH
OH
OH
O O OH
OH O O
H3C O
HO
HO
OH
Rutin
FIGURE 30.2 Chemical structures of major flavonoids (flavan-3-ols and flavonols) reported in mango juice.
TABLE 30.4
Reported Polyphenol Content of Mango Juice
Types of Polyphenols Mango Juice (μg/mL) [13] Bambangan Juice (mg/100 mL) [19]
Phenolic Acids
Gallic acid 175.94 1.69
Protocatechuic acid 77.02 —
p-OH-benzoic acid 19.81 —
Vanillic acid 13.83 0.49
Chlorogenic acid 20.07 —
Syringic acid 3.58 —
Caffeic acid 35.77 —
p-Coumaric acid 120.68 —
m-Coumaric acid 82.05 —
Ferulic acid 45.37 —
Sinapic acid 37.84 —
Ellagic acid 8.94 —
trans-Cinnamic acid 0.20
Flavan-3-ols
(+)-Catechin 16.07 —
Flavonols
Quercetin 13.43 —
Rutin 15.02 —
Mango Juice 367
30.4 Health Effects

30.4.1 Bioavailability of Bioactives from Mango Juice in Relation to Its Intake
Mango and its products are popular due to their high nutraceutical and pharmaceutical values [1,2].
However, no information is available on the absorption, distribution, metabolism, and excretion of poly-
phenols in human, especially about the bioaccessibility and bioavailability of antioxidant polyphenols
from mango juice in humans [24–26,33]. Bub et al. [34] have determined the effect of daily consumption
of two fruit juices, A and B (mixture of orange, mango, and apple juice), for 2 weeks on the appearance
of polyphenols in plasma and excretion in the urine. The consumption of both fruit juices A (anthocyanin
rich) and B (flavanol rich) did not result in increased plasma polyphenols in subjects. Consumption of juice
A resulted in a significant increase in total polyphenol excretion in urine. Consumption of juice B also
increased total polyphenol excretion, but there was no significant difference when compared to the end of
the washout period. This shows that the polyphenols were already eliminated from the circulating blood
[34]. This result indicates that polyphenols and/or their metabolites from mango juices were absorbed and
excreted in part by the kidneys. However, the metabolites of polyphenols from juices A and B after uri-
nary excretion were not determined by Bub et al. [34]. Hence, further research on this matter is warranted.
The TPC of both fresh and commercial mango juice was low (Table 30.3), since the daily phenolic com-
pounds intake from the diet is estimated to range between 150 mg and 1 g/day [23,33,35]. For example,
the mean total phenolic intake of Spanish PREDIMED (Prevención con Dieta Mediterránea) cohort was
820 mg/day (443 mg/day of flavonoids and 304 mg/day of phenolic acids), where fruits were the major
source [35]. The mean total phenolic intake of SUVIMAX (Suppléments en Vitamines et Minéraux
Antioxydants—Antioxidant Vitamin and Mineral Supplements) French cohort was 206 mg/day, where
fruits were the second largest phenolic compounds provider after coffee (520 mg/day) [36]. All these results
indicate that the mean daily total phenolic intake could reach more than 1 g [36]. Practically, the daily
intake of 100 mL of commercial mango juice (with 5% or 5 g of fruit concentrate) only contributes a maxi-
mum of 65 mg of phenolic compounds (13 mg of phenolic compound in 1 g of mango juice). This result
reflects that relying on either fresh fruit or commercial fruit juice alone does not contribute sufficiently to
the daily intake of phenolic compounds [7]. However, fresh fruit juice consumption would still result in an
important contribution to total polyphenol intake.
30.4.2 Results from In Vitro, Animal, and Human Intervention Studies

Interestingly, human or animal studies that examine the effect of mango (flesh or juice) on health param-
eters are scarce. Studies determining the effects of the consumption of mango flesh or juice on nutrient
intake, diet quality, and health biomarkers in humans are also lacking [12]. Percival et al. [14] demon-
strated that the juice produced from M. indica L. cultivar Palmer had anticancer and antioxidant effects.
Fresh mango juice inhibited the growth cycle of an immortal cancer cell line (HL-60) in vitro in a dose-
dependent manner. In addition, whole mango juice (0.01% and 0.1%) reduced the number of transformed
foci at 50% and 70%, respectively, in the neoplastic transformation assay using BALB/3T3 cells. In fact,
carcinogenesis has three stages: initiation, promotion, and progression. Percival et al. [14] showed that
the ability of mango juice to inhibit foci formation was not during initiation stage, but rather at a later
stage of carcinogenesis, which was during the promotion stage of carcinogenesis, of which the progres-
sion of cancer was irreversible (antipromotion activity). However, whether these anticancer properties are
maintained after digestion, absorption, and metabolism is unknown. Anyway, the observed anticancer
activity was found to be a result of other functions and not mainly due to the antioxidant capacity of the
mango juice [14].
A recent study reported an excellent in vitro radioprotective effect of mango juice (cultivar
Banganapalli) against gamma ray (γ)-irradiation-induced DNA (pUC19 plasmid) damage [13]. There
was complete degradation of plasmid DNA when exposed to γ-irradiation (1, 3, and 5 kGy) dose depend-
ently, whereas in the presence of mango juice, the degradation was prevented significantly [13]. In the
same study, mango juice was also demonstrated to possess nitric oxide (NO) scavenging activity in vitro
(62.16%). NO, a free radical produced during infection and inflammation, has serious implications for
the pathogenesis of cancer, hypertension, and heart disease [13]. The presence of phenolic compounds
in mango juice has been postulated to contribute to the radioprotective and NO scavenging activity [13].
Ibrahim et al. [37] examined the effects of bambangan (M. pajang) juice powder (BJP) drink on plasma
vitamins and antioxidant enzyme levels, liver, and kidney function in healthy subjects for 4 weeks. This BJP
drink (250 mL with 50 g of mango pulp powder) contained 66 mg of ascorbic acid and 18 mg of β-carotene.
As a result, plasma total antioxidant status increased significantly (18%) after BJP consumption compared to
baseline value. At the same time, plasma ascorbic acid (72.10 vs. 92.43 μM) and β-carotene (1.48 vs. 2.30 μM)
also increased significantly by 28% and 45%, respectively, compared to the baseline values. However, there
were no changes in plasma antioxidant enzymes (glutathione peroxidase and superoxide dismutase) or non-
enzymatic antioxidants for both the treatment and placebo group after 4 weeks of intervention [37].
In addition, BJP drink consumption also did not affect liver function parameters (plasma gamma-
glutamyl transferase, alkaline phosphatase, alanine transaminase, and aspartate transaminase) as well
as kidney function parameters (plasma total protein, albumin, creatinine, and urea concentrations) [37].
Hence, no adverse cytotoxic effects were observed due to the BJP drink consumption, as demonstrated
by increasing in liver enzymes in the placebo groups at the end of the study [37]. In other words, BJP
drink consumption does protect against liver disease.
Another study conducted by Bub et al. [34] demonstrated that the consumption of two types of fruit
juice mixtures (juices A and B) containing apple, mango, and orange juice (330 mL/day) for 2 weeks
modulated and enhanced antioxidant status and immune functions; it also reduced level of oxidative
DNA damage in 27 nonsmoking German male subjects. These two juices contained 236 mg (A) and
226 mg (B) of polyphenols with cyanidin glycosides (A) and epigallocatechin gallate (B) as the major
polyphenolic compounds. Plasma total antioxidant status increased lightly after consumption of juice
B, whereas consumption of juice A did not result in any changes on plasma antioxidant status. Plasma
MDA decreased with time during juice interventions, thus indicating physiological inhibition of lipid
peroxidation [34]. However, plasma LDL oxidation did not change after consumption of these two juices.
Interleukin-2 (IL-2) secretion by activated lymphocytes and the lytic activity of natural killer cells were
significantly increased by both juices, indicating the stimulated immune functions in human body.
However, intervention of juices A and B had no effect on single DNA strand breaks but significantly
reduced oxidative DNA damage in lymphocytes [34]. The understanding of how fruit juice, such as
mango juice, benefit human health requires future studies and clarification. Thus, more research should
be carried out to determine the health-promoting effects of mango juice, validated with the study of its
bioavailability in animals or humans.

Mango juice is rich in phenolic compounds and antioxidant capacity. Hence, it is commonly produced as
a functional juice. A variety of novel products produced from mango juice have been developed to sup-
ply essential nutrients for humans. Mango yogurt is a good substitute for the conventional high-calorie
dessert pudding, which contain sufficient amount of curd with cream, providing daily recommended
amounts of essential fats [2]. Mango shake and lassi, a popular and nutritious drink during summer
months commonly found in India, is prepared from mango powder with curd. These mango shake and
lassi have been rated well in terms of their color, flavor, and texture and hence overall acceptability [2].
Mango juice has also been fermented to produce mango wine rich in phenolic compounds with good
antioxidant capacities [38]. Meanwhile, wine produced from mango was comparable to the commercial
grape wine in terms of flavor (taste and aroma) and acceptability [39].
Recently, a mango tablet was prepared from ripe and unripe mango, where the mango tablets can
be used as an effective vitamin C supplement. The tablets can easily be consumed by chewing or by
dissolving in water [40]. Mango juice has also been used as a vehicle to supplement prebiotics, for
example, fructooligosaccharide (FOS). It is a type of prebiotic with similar sweetness of sucrose, which
is commonly used in the production of fruit and vegetable juices. The overall quality of the mango juice
fortified with FOS for 4 months of storage at room temperature was acceptable, as indicated by sensory
Mango Juice 369
analysis (color, consistency, flavor, and overall quality) [41]. Future research should be carried out on the
changes of the nutritional quality of mango juice after fortified with FOS.
Various processing methods have been reported to increase the content of vitamin C, carotenoids, and
polyphenols in relation to their antioxidant capacities in mango juice. These methods are thermal pas-
teurization, ultrasonication, and ultraviolet (UV) light treatment [42–44]. Ultrasound sonication (40 kHz
frequency with 1.36–1.44 W/cm3 of acoustic energy density) and UV-C treatment (3.525 J/m2 of radiation
dose) of freshly made Chokanan mango (M. indica L.) juice for 15 and 30 min demonstrated signifi-
cant improvement in carotenoids (4%–9%), phenolic compounds (30%–35%), flavonoids (3%–6%), and
antioxidant capacities (determined using reducing power and DPPH radical scavenging activity), when
compared to freshly squeezed mango juice treated with heat pasteurization [42,44].
Chokanan mango juice treated with combined ultrasound sonication and UV-C for 15 and 30 min
exhibited significant improvements in clarity, antioxidant capacity, and extractability of carotenoids
(15%), phenolic compounds (37%), and flavonoids (35%), when compared to freshly squeezed juice [43].
In addition, a combined action of ultrasound sonication and UV-C treatment demonstrated complete
inactivation of coliforms and aerobic bacteria, together with significant reduction in yeast and mold
count. The combine treatment, therefore, is a promising method for better retention of mango juice
quality and a feasible alternative to the conventional thermal pasteurization [44]. In line with increasing
consumer demand for healthy beverages from fruits and vegetables and, at the same time, ensuring food
safety and quality, more research should be carried out to determine novel preservation technology that
retains a fresh-like quality in fruit juices [41].
Mango juice treated with the combination of Panax ginseng (2% v/v), malic acid (0.5% v/v), and potas-
sium sorbate (0.05% v/v) also demonstrated the highest antimicrobial effectiveness against Salmonella
enterica ser. Saintpaul and Escherichia coli O157:H7, compared to P. ginseng, malic acid, or potassium
sorbate alone, in addition to a higher microbiological inhibition during storage for 21 days [45]. Sensory
attributes such as flavor and color of mango juice have also been enhanced. This indicates that natural
antimicrobials from plant sources, such as P. ginseng, could be an effective alternative in the food indus-
try for ensuring the microbial safety and quality in mango juice, compared to chemical food additives,
such as malic acid and potassium sorbate [45]. More research should be carried out to determine the
effect of other potential plant sources on the microbial safety, quality, and nutritional compositions in
fruit juices other than mango juice.
A recent work [13] reported that mango juice treated with γ-irradiation doses (1, 3, and 5 kGy) resulted
in significant increases in TPC, TFC, and antioxidant capacities determined using DPPH (6.2%), FRAP
(20.3%), and NO scavenging activity (13.5%). However, the γ-irradiation resulted in a significant decrease
in ascorbic acid content (70.75%). Gamma-irradiation also affected the profile of phenolic compounds in
the said mango juice [13]. Specifically, the contents of gallic, syringic, and chlorogenic acids increased
by 3.2-, 2.5-, and 2.3-fold, respectively, whereas the contents of ferulic and sinapic acids decreased by
4.5- and 2.7-fold, respectively, in the irradiated mango juice samples. The amount of other polyphenolic
compounds, such as protocatechuic acid, p-OH-benzoic acid, ellagic acid, caffeic acid, p-coumaric acid,
and rutin also increased significantly with increasing irradiation dose (1, 3, and 5 kGy) in the mango
juice samples [13]. The γ-irradiation also inhibited the growth of aerobic bacteria, yeast, and molds in
mango juice completely at irradiation dose of 5 kGy. The overall quality of the mango juice irradiated
with gamma rays was acceptable as indicated by sensory analysis (color, consistency, flavor, and overall
quality). This shows that γ-irradiation treatment could be an effective method for microbial decontami-
nation, improving quality and for enhancing sensory attributes of mango juice once the safety aspect of
this radiation treatment has been ascertain in the future [13].
Gad et al. [46] utilized whey in the production of mango beverage with the use of mango pulp powder
with the incorporation of flaxseed oil. As a result, this whey–mango beverage is a new functional bever-
age with little sedimentation and low viscosity, exhibiting a pleasant mango flavor. This beverage is an
excellent source of nutrients, such as minerals (calcium, potassium, and phosphorus), vitamins (vitamins
C and E), essential amino acids, omega-3 fatty acids, and digestible carbohydrate (lactose) [46]. Whey
protein is an excellent source of essential amino acids, such as cysteine, methionine, leucine, isoleucine,
and valine. This beverage is also an excellent source of antioxidants, including phenolic compounds,
vitamins C, and carotenoids, which demonstrates good antioxidant capacities (determined using DPPH,
FRAP, and ORAC assays). It can be stored best at refrigeration temperature with minimum effects on
their physicochemical and sensory quality. The presence of various components such as, phenolic com-
pounds, carotenoids, and vitamins in the “whey–mango beverage” is all acting together synergistically
to improve consumer health [46].
A functional beverage formulated from a mixture of fruits, such as orange, lemon, apple, mango, and
plum, which can help to reduce hangover symptoms, such as dizziness and fatigue, has been patented
in Korea (2007-08-08) [5]. There have been enormous opportunities in the formulation of fruit-origin
functional beverages evolving and growing at different rates both within and across various countries.
The future of fruit-origin functional beverages depends on their efficacy in promoting health [5]. The sci-
entific community is expecting more exciting results on functional fruit beverages in the coming years.
30.6 Conclusion
Mango juice is a rich source of vitamin A, minerals, and antioxidant phenolic compounds (flavonols,
flavanols, and phenolic acids). Furthermore, mango juice has also good aroma and sensory qualities,
which receives good ratings among consumers. More novel mango juice as functional beverages should
be developed using novel preservation technologies. More clinical trials and human intervention studies
should also be carried out to ascertain the beneficial effects of mango juice in disease risk reduction in
relation to its stability, bioaccessibility, and bioavailability after extensive metabolism in human body.
REFERENCES
1. Masibo, M. and He, Q., Mango bioactive compounds and related nutraceutical properties—A review.
Food Rev. Int., 25, 346–370, 2009.
2. Ravani, A. and Joshi, D.C., Mango and it’s by product utilization—A review. Trends Post Harvest
Technol., 74, 55–67, 2013.
3. Food and Agriculture Organization of the United Nations Statistics Division (FAOSTAT), Food and
Agricultural Commodities Production, 2013. Published online at http://faostat.fao.org/site (accessed
September 26, 2014).
4. Hassan, F.A., Ismail, A., Abdul Hamid, A., and Azlan, A., Identification and quantification of phenolic
compounds in Bambangan (Mangifera pajang Kort.) peels and their free radical scavenging activity.
J. Agric. Food Chem., 59, 9102–9111, 2011.
5. Sun-Waterhouse, D., The development of fruit-based functional foods targeting the health and wellness
market: A review. Int. J. Food Sci. Technol., 46, 899–920, 2011.
6. Corbo, M.R., Bevilacqua, A., Petruzzi, L., Casanova, F.P., and Sinigaglia, M., Functional beverages:
The emerging side of functional foods. Compr. Rev. Food Sci. F., 13, 1192–1206, 2014.
7. Lek, K.B., Zuraini, Z., Boon, K.B., Wan, Y.H., Swee, K.Y., and Noorjahan, B.M.A. Comparison of total
Plants Res., 6, 5857–5862, 2012.
9. Mahdavi, R., Nikniaz, Z., Rafraf, M., and Jouyban, A., Determination and comparison of total poly-
phenol and vitamin C contents of natural fresh and commercial fruit juices. Pak. J. Nutr., 9, 968–972,
2010.
10. Abdullakasim, P., Songchitsomboon, S., Techagumpuch, M., Balee, N., Swatsitang, P., and Sungpuag,
P., Antioxidant capacity, total phenolics and sugar content of selected Thai health beverages. Int. J. Food
Sci. Nutr., 58, 77–85, 2007.
11. Gil, A.M., Duarte, I.F., Delgadillo, I., Colquhoun, I.J., Casuscelli, F., Humpfer, E., and Spraul, M., Study
of the compositional changes of mango during ripening by use of nuclear magnetic resonance spectros-
copy. J. Agric. Food Chem., 48, 1524–1536, 2000.
12. O’Neil, C.E., Nicklas, T.A., and Fulgoni, V.L. III., Mangoes are associated with better nutrient intake,
diet quality, and levels of some cardiovascular risk factors: National Health and Nutrition Examination
Survey. J. Nutr. Food Sci., 3, 185–192, 2013.
Mango Juice 371
13. Naresh, K., Varakumar, S., Variyar, P.S., Sharma, A., and Reddy, O.V.S., Enhancing antioxidant activ-
ity, microbial and sensory quality of mango (Mangifera indica L.) juice by γ-irradiation and its in vitro
radioprotective potential. J. Food Sci. Technol., 1–12, 2014.
14. Percival, S.S., Talcott, S.T., Chin, S.T., Mallak, A.C., Lounds-Singleton, A., and Pettit-Moore, J.,
Neoplastic transformation of BALB/3T3 cells and cell cycle of HL-60 cells are inhibited by mango
(Mangifera indica L.) juice and mango juice extracts. J. Nutr., 136, 1300–1304, 2006.
15. Halvorsen, B.L., Carlsen, M.H., Phillips, K.M., Bøhn, S.K., Holte, K., Jacobs, D.R., and Blomhoff, R.,
Content of redox-active compounds (antioxidants) in foods consumed in the United States. Am. J. Clin.
Nutr., 84, 95–135, 2006.
16. Falguera, V., Sánchez-Riaño, A.M., Quintero-Cerón, J.P., Rivera-Barrero, C.A., Méndez-Arteaga, J.J.,
and Ibarz, A., Characterization of polyphenol oxidase activity in juices from 12 underutilized tropical
fruits with high agroindustrial potential. Food Bioprocess Technol., 5, 2921–2927, 2012.
17. Zulueta, A., Esteve, M.J., Frasquet, I., and Frígola, A., Vitamin C, vitamin A, phenolic compounds and
total antioxidant capacity of new fruit juice and skim milk mixture beverages marketed in Spain. Food
Chem., 103, 1365–1374, 2007.
18. Ibrahim, M., Prasad, K.N., Ismail, A., Azlan, A., and Abd. Hamid, A., Physiochemical composition and
antioxidant activities of underutilized Mangifera pajang fruit. Afr. J. Biotechnol., 9, 4392–4397, 2010.
19. Zabidah, A.A., Kong, K.W., and Amin, I., Antioxidant properties of tropical juices and their effects
on in vitro hemoglobin and low density lipoprotein (LDL) oxidation. Int. Food Res. J., 18, 549–556,
2011.
20. Abu Bakar, M.F., Mohamed, M., Rahmat, A., and Fry, J., Phytochemicals and antioxidant activity of
different parts of Bambangan (Mangifera pajang) and Tarap (Artocarpus odoratissimus). Food Chem.,
113, 479–483, 2009.
21. Fang, Z., Zhang, Y., Lü, Y., Mab, G., Chen, J., Liu, D., and Ye, X., Phenolic compounds and antioxidant
capacities of bayberry juices. Food Chem., 113, 884–888, 2009.
22. Kulisic-Bilusic, T., Schnäbele, K., Schmöller, I., Dragovic- Uzelac, V., Krisko, A., Dejanovic, B., Milos,
M., and Pifat, G., Antioxidant activity versus cytotoxic and nuclear factor kappa B regulatory activities
on HT-29 cells by natural fruit juices. Eur. Food Res. Technol., 228, 417–424, 2009.
23. Pérez-Jiménez, J., Neveu, V., Vos, F., and Scalbert, A., Systematic analysis of the content of 502 poly-
phenols in 452 foods and beverages: An application of the Phenol-Explorer Database. J. Agric. Food
Chem., 58, 4959–4969, 2010.
metabolism and pharmacokinetics in humans and experimental animals. Database, doi: 10.1093/data-
base/bas031, 2012.
25. Neveu, V., Perez-Jiménez, J., Vos, F., Crespy, V., du Chaffaut, L., Mennen, L., Knox, C. et al., Phenol-
Explorer: An online comprehensive database on polyphenol contents in foods. Database, doi: 10.1093/
database/bap024, 2010.
26. Bhagwat, S., Haytowitz, D.B., and Holden, J.M., USDA Database for the Flavonoid Content of Selected
Foods, Release 3.1, 2014. Published online at: http://www.ars.usda.gov/nutrientdata/flav (accessed
November 2, 2014).
27. Arts, I.C.W., van de Putte, B., and Hollman, P.C.H., Catechin content of foods commonly consumed
in the Netherlands. 1. Fruits, vegetables, staple foods and processed foods. J. Agric. Food Chem., 48,
1746–1751, 2000.
28. Franke, A.A., Custer, L.J., Arakaki, C., and Murphy, S.P., Vitamin C and flavonoid levels of fruits and
vegetables consumed in Hawaii. J. Food Comp. Anal., 17, 1–35, 2004.
29. Lako, J., Trenerry, V.C., Wahlqvist, M., Wattanapenpaiboon, N., Sotheeswaran, S., and Premier, R.,
Phytochemical flavonols, carotenoids and the antioxidant properties of a wide selection of Fijian fruit,
vegetables and other readily available foods. Food Chem., 101, 1727–1741, 2007.
30. Gu, L., Kelm, M.A., Hammerstone, J.F., Beecher, G., Holden, J., Haytowitz, D., Gebhardt, S., and Prior,
R.L., Concentrations of proanthocyanidins in common foods and estimations of normal consumption.
J. Nutr., 134, 613–617, 2004.
31. Schieber, A., Ullrich, W., and Carle, R., Characterization of polyphenols in mango puree concentrate
by HPLC with diode array and mass spectrometric detection. Innov. Food Sci. Emerg. Technol., 1,
161–166, 2000.
32. Abu Bakar, M.F., Mohamed, M., Rahmat, A., Burr, S.A., and Fry, J.R., Cytotoxicity and polyphenol
diversity in selected parts of Mangifera pajang and Artocarpus odoratissimus fruits. J. Nutr. Food Sci.,
40, 29–38, 2010.
33. Manach, C.M., Williamson, G., Morand, C., Scalbert, A., and Rémésy, C., Bioavailability and bioef-
ficacy of polyphenols in humans. I. Review of 97 bioavailability studies. Am. J. Clin. Nutr., 81(Suppl.),
230S–242S, 2005.
34. Bub, A., Watzl, B., Blockhaus, M., Briviba, K., Liegibel, U., Müller, H., Pool-Zobel, B.L., and
Rechkemmer, G., Fruit juice consumption modulates antioxidative status, immune status and DNA
damage. J. Nutr. Biochem., 14, 90–98, 2003.
35. Tresserra-Rimbau, A., Medina-Remón, A., Pérez-Jiménez, J., Martínez-González, M.A., Covas, M.I.,
Corella, D., Salas-Salvadó, J. et al., Dietary intake and major food sources of polyphenols in a Spanish
population at high cardiovascular risk: The PREDIMED study. Nutr. Metab. Cardiovasc. Dis., 23,
953–959, 2013.
36. Pérez-Jiménez, J., Fezeu, L., Touvier, M., Arnault, N., Manach, C., Hercberg, S., Galan, P., and Scalbert,
A., Dietary intake of 337 polyphenols in French adults. Am. J. Clin. Nutr., 93, 1220–1228, 2011.
37. Ibrahim, M., Ismail, A., Al-Sheraji, S.H., Azlan, A., and Abdul Hamid, A., Effects of Mangifera pajang
Kostermans juice on plasma antioxidant status and liver and kidney function in normocholesterolemic
subjects. J. Funct. Foods, 5, 1900–1908, 2013.
38. Kondapalli, N., Sadineni, V., Variyar, P.S., Sharma, A., and Obulam, V.S.R., Impact of γ-irradiation on
antioxidant capacity of mango (Mangifera indica L.) wine from eight Indian cultivars and the protection
of mango wine against DNA damage caused by irradiation. Process Biochem., 49, 1819–1830, 2014.
39. Reddy, L.V.A. and Reddy, O.V.S., Production and characterization of wine from mango fruit (Mangifera
indica L.). World J. Microbiol. Biotechnol., 21, 1345–1350, 2005.
40. Ong, M.Y., Yusof, Y.A., Aziz, M.G., Chin, N.L., and Amin, N.A., Characterisation of fast dispersible
fruit tablets made from green and ripe mango fruit powders. J. Food Eng., 125, 17–23, 2014.
41. Renuka, B., Kulkarni, S.G., Vijayanand, P., and Prapulla, S.G., Fructooligosaccharide fortification
of selected fruit juice beverages: Effect on the quality characteristics. LWT—Food Sci. Technol., 42,
1031–1033, 2009.
42. Santhirasegaram, V., Razali, Z., George, D.S., and Somasundram, C., Comparison of UV-C treatment
and thermal pasteurization on quality of Chokanan mango (Mangifera indica L.) juice. Food Bioprod.
Process., 94, 313–321, 2015.
43. Santhirasegaram, V., Razali, Z., and Somasundram, C., Effects of sonication and ultraviolet-C treatment
as a hurdle concept on quality attributes of Chokanan mango (Mangifera indica L.) juice. Food Sci.
Technol. Int., 21, 232–241, 2014.
44. Santhirasegaram, V., Razali, Z., and Somasundram, C., Effects of thermal treatment and sonication on
quality attributes of Chokanan mango (Mangifera indica L.) juice. Ultrason. Sonochem., 20, 1276–1282,
2013.
45. Raybaudi-Massilia, R., Zambrano-Durán, A., Mosqueda-Melgar, J., and Calderón-Gabaldón, M.I.,
Improving the safety and shelf-life of orange and mango juices using Panax ginseng, malic acid and
potassium sorbate. J. Verbrauch. Lebensm., 7, 273–282, 2012.
46. Gad, A.S., Emam, W.H., Mohamed, G.F., and Sayd, A.F., Ulitization whey in production of functional
health beverage ‘whey-mango beverages’. Am. J. Food Technol., 8, 133–148, 2013.
31
Mangosteen Juice
Mark L. Failla, Fabiola Gutierrez-Orozco,

Chureeporn Chitchumroonchokchai, and Florian Diekmann
CONTENTS
31.1 Introduction................................................................................................................................... 373
31.4 Health Effects................................................................................................................................ 377
31.6 Conclusion......................................................................................................................................381
References................................................................................................................................................381
31.1 Introduction
Tropical fruits, such as açai, acerola, camu camu, dragon fruit, goji, lychee, mangosteen, passion fruit,
and pomegranate, as well as products containing these fruits, are currently marketed as “exotic” or
“super” fruits because of their relatively high antioxidant capacity and proposed health-promoting activi-
ties. Mangosteen juice and related products are primarily sold in North America and Europe using direct
sales and a multilevel marketing approach rather than a standard retail model. Sales of mangosteen bev-
erages in 2007 were estimated at $147 million and increased to $210 million in 2010 in the United States
alone [1,2]. The juice generally is prepared from ripened fruit that weighs 75–125 g and is composed
of the edible aril or pulp that accounts for approximately one-third of the total weight, a thick pericarp
or rind, and the skin (Figure 31.1). Standard processing of mangosteen fruit into juice is outlined in
Figure 31.2 [3,4] Generally, a puree is prepared from the aril and mixed with an extract of the pericarp
to produce the “juice.”
This contribution highlights the composition of several of the mangosteen juices with the largest
market in the North American market and the bioactive compounds in mangosteen juice products with
particular attention to xanthones and their health-promoting activities. Recent reports regarding the
absorption and metabolism of mangosteen xanthones from juice and the effects of chronic consumption
of commercial mangosteen juices on inflammatory markers are also discussed.

The USDA National Nutrient Database for Standard References [5] and the Nutrition Division of the
Thai Department of Health [6] have provided data on the nutrient composition of canned mangosteen
aril but not mangosteen juice. The aril has limited nutritional value as there are relatively low quantities
of sugar, protein, fat, ascorbic acid, potassium, and calcium. However, analysis of the fibrous pericarp
has revealed the presence of bioactive xanthones, phenolics, anthocyanins, and proanthocyanidins [7–9].
The composition of mangosteen juice products varies markedly because many of the commercial for-
mulations are blends that include juices and concentrates of other fruits, as well as supplements such as
373
Pericarp
Aril
FIGURE 31.1 Ripened mangosteen fruit (upper left) and mangosteen aril and pericarp (lower left), and mangosteen juice
products.
Mangosteen fruit
Aril and pericarp separated
White pulp/aril Pericarp
Mash/press Xanthones extracted and

dried to xanthone-rich powder
Puree stored cold Blending of aril puree

with xanthone-rich powder
Packed and sterilized Packed and Packed and

sterilized sterilized
Mangosteen juices, supplements, confectionaries, and health and skin products
FIGURE 31.2 Processing of mangosteen fruit to produce juice and other mangosteen-containing products. (Modified
from Office of Agricultural Economics [OAE], Indicators of Agricultural Economics of Thailand 2009, 2010a, Published
online at: http://www.oae.go.th [accessed May 23, 2014]; Office of Agriculture Regulation [OAR], Public Access Database
2011, Published online at: http://m.doa.go.th/ard/ [accessed May 23, 2014].)
aloe vera leaves and green tea. Information about the quantities of nutrients and health-promoting bioac-
tives on product labels and company websites generally is quite limited. Moreover, variability among
lots of mangosteen juices is likely because of the existing differences in the sources of mangosteen and
other fruit juices or concentrates added to the formulation, the effects of preharvest abiotic and biotic
factors, the addition of proprietary mixtures of minerals and vitamins, and methods of postharvest pro-
cessing that may affect the composition of the commercial product. The available information on the
Mangosteen Juice 375
TABLE 31.1
Composition of Mangosteen Juice Products Prepared from Whole Fruits (per 30 mL Serving)
Mangosteen Mangosteen Juice with
Nutrient Unit Blended Juice [10] Nutrient Unit Supplementsa [65]
Energy kcal 13.0 Energy kcal 17.5
Carbohydrate g 3.2 Carbohydrate g 4
Total sugars g 2.7 Vitamins
A as β-carotene IU 5000
Ascorbic acid mg 150
Biotin μg 150
Cholecalciferol (D3) IU 500
Cyanocobalamin (B12) μg 7.5
Folic acid μg 400
Niacin amide mg 10
Pantothenate, calcium salt μg 5
Pyridoxine HCl (B6) mg 2.5
Riboflavin mg 0.85
Thiamin mg 0.75
α-Tocopheryl acetate (E) IU 30
Others
Selenium (as amino acid μg 70
chelate)
Mangosteen, aloe vera, and g 12.6
green tea blend
Mangostins mg 47.1
Catechins (ECGC) mg 1.8
Proprietary plant mineral blend mg 478
Source: Xango, Wikipedia Encyclopedia, Published online at: https://en.wkipedia.org/wiki/Xango (accessed October 6, 2015).
a Modified from information communicated by manufacturer.
Abbreviations: ECGC, epigallocatechin gallate; IU, international unit.
composition of two of the major selling mangosteen juices in western markets is provided in Table 31.1.
According to the respective producers, these juices are prepared with an unspecified quantity of puree
from whole mangosteen fruit. The mangosteen juice from at least one company is supplemented with
vitamins at levels that meet or exceed the recommended daily intake. The proprietary plant mineral mix-
ture increases the content of essential minerals in the juice, but the actual concentrations of the essential
inorganic elements are not provided.

Mangosteen beverages and supplements are primarily marketed on the basis of the proposed health-
promoting activities of several classes of secondary metabolites in the fruit rather than for their nutri-
tional value. Thus, it is surprising that the specific types and amounts of the more abundant compounds
from these classes, and especially the xanthones in the juices, are not provided. More than 60 xanthones
have been identified in mangosteen fruit, and these include α-mangostin (α-MG), β-mangostin (β-MG),
γ-mangostin (γ-MG), garcinones, and gartanin [7,8]. The structures of these xanthones are shown in
Figure 31.3. The compounds consist of a tricyclic aromatic ring system with a C6-C3-C6 skeleton referred
to as xanthene-9-one. The A and B benzyl rings are fused to the central heterocyclic ring containing
oxygen. The various xanthones are differentiated by the number and location of isoprene, hydroxyl, and
methoxy groups. Antioxidant activity is determined by the number of hydroxyl and in particular vicinal
hydroxyl groups on the A and B benzyl rings. Thus, the antioxidant activity of γ-MG is greater than that
of α-MG that exceeds the activity of β-MG [7,9].
OH
O OH O OH
MeO MeO
HO O OH HO O OH
α-Mangostin Garcinone D
O OH
HO
O OH
MeO HO O OH
HO O OMe
β-Mangostin Garcinone E
OH O OH
O OH
O OH
HO
OH
HO O OH
γ-Mangostin Gartanin
FIGURE 31.3 Chemical structures of major xanthone compounds found in mangosteen fruit and juice.
The profile of the more abundant xanthones in the complex matrix of several mangosteen juices is
presented in Table 31.2. The α- and γ-MGs are the most abundant xanthones in the analyzed juices as
in mangosteen whole fruit and its pericarp. The differences in the concentrations of the xanthones in
the analyzed juices likely reflect the relative amounts of mangosteen aril puree and pericarp extract in
the juice product. Wittenaurer et al. [11] reported that the xanthone content of Xango juice (per 100 g
fresh weight) was only 1.1% and 17.6% of that in an equivalent amount of mangosteen pericarp and
aril, respectively. This marked difference in the concentration of the xanthones is due to the low per-
centage (<5%) of mangosteen puree in the final product. These investigators estimated that 90 mL of
the tested juice (e.g., three daily servings) provided a similar dose of mangosteen xanthones as the aril
from a single fruit. It is important to note that the content of xanthones in aril is less than 10% that in
the pericarp.
Phenolic acids are present in all parts of the fruit although their concentrations in the pericarp and
the peel greatly exceed those in the aril [12]. Ester and glycoside conjugates of the phenolic acids
account for the majority of this class of compounds with derivatives of hydroxybenzoic acid such as
protocatechuic, p-hydroxybenzoic, and vanillic acids being most prevalent. The highest concentra-
tions of anthocyanins were present in skin and outer pericarp but absent in aril [13]. Cyanidin-3-O-
sporoside was the most abundant anthocyanin in mangosteen fruit, and there were minor amounts of
cyanidin-3-O-glucoside [14]. These pigments are responsible for the characteristic red color of freshly
prepared mangosteen juice. The concentration of anthocyanins in fresh mangosteen juice mixed with
rosella and grape juices (2:1:2, v/v/v) was reported as 3.7 mg cyanidin-3-O-glucoside equivalents
(C3GE) per 100 mL [15]. Due to the blended nature of the juice, the contribution of the mangosteen
juice to the total anthocyanin content is unknown. Mangosteen pericarp also contains oligomeric pro-
anthocyanidins rich in epicatechin monomers that likely contribute to the peroxyl radical scavenging
activity of the juice [16].
TABLE 31.2
Concentrations of Xanthones in Mangosteen Juices
Mangosteen Juice (mg/30 mL Serving)
Xanthone Product A [11] Product A[a] Product B [24] Product C [a] Product D [a]
α-Mangostin 3.71 7.06 39.18 4.19 13.37
β-Mangostin 0.14 1.53 0.16 0.49
γ-Mangostin 0.90 1.25 4.23 0.41 1.09
Garcinone D 0.05 6.66 0.63 1.23
Garcinone E 0.25 0.33 3.33 0.34 0.86
Gartanin 0.26 0.22 1.86 0.16 0.34
8-Deoxygartanin 0.20 0.26 2.01 0.19 0.44
9-Hydroxycalabaxanthone 0.13 2.34 0.15 0.37
1,7,Dihydroxy-3-methoxy-2- 0.14 — — — —
(3-methylbut-2-enyl)xanthone
1,3,7-Trihydroxy-2,8- 0.16 — — — —
di-(3-methylbut-2-enyl)xanthone
Total xanthones 5.64 9.49 64.74 6.23 18.18
a Data from Chitchumroonchokchai, C. and Failla, M.L., Xanthone content in commercial mangosteen juices. Human
Nutrition Program, The Ohio State University, OH, 2014.
Note: Values are means of 6 replicates.
31.4 Health Effects

Traditional medicine has been practiced in China and other Asian countries for thousands of years.
Information about the identification and preparation of plants and their use for treating various medical
problems has been transferred orally from generation to generation [17]. Mangosteen pericarp has been
used in Southeast Asia, China, and India to treat wounds, diarrhea, and skin diseases. For example,
freshly ground or dried mangosteen pericarp has been boiled in water to prepare a disinfectant for clean-
ing wounds [18]. Alternatively, dried fruit is rubbed against stone and suspended in saturated calcium
hydroxide solution before pouring over the wound [19]. Roasted pericarp powder suspended in water
(4 g/100 mL) is consumed every 2 h to treat diarrhea [18]. Pericarp can also be powdered and dissolved
in rice water or boiled water to treat dysentery. A decoction prepared with dried pericarp is also used to
treat cystitis, gonorrhea, and gleets [20]. We are unaware that the chemical composition of these medici-
nal extracts has been investigated.
Numerous bioactivities of xanthones have previously been discussed in several reviews [7–9,21–23],
as summarized in Table 31.3. Because α-MG is the most abundant xanthone in mangosteen fruit, it
has received the greatest attention in studies with cultured cells and rodent models. The most investi-
gated activities of mangosteen xanthones have addressed their antioxidant activity and the inhibition
of proliferation and induction of apoptosis in transformed cell lines of diverse tissue origins, including
colon, bone marrow, lung, breast, and prostate [24–28]. α-MG and extracts of mangosteen pericarp also
suppress tumor growth in murine xenograft models of glioblastoma, breast, prostate, and colon cancer
[24–26,29–31]. An addition of a xanthone-rich extract containing 76% α-MG and 16% γ-MG extract pre-
pared from mangosteen pericarp to the diet also inhibited chemically induced colon cancer in rats [32].
Anti-inflammatory activities of mangosteen xanthones have been reported in cell and animal models.
For example, α- and γ-MGs attenuated the lipopolysaccharide (LPS)-induced inflammatory response
in primary cultures of human adipocytes and transformed human cell lines [42–44], as well as trans-
formed murine cell lines [45–47]. In contrast to the anti-inflammatory activity of α-MG in cultures of
transformed cells treated with LPS, we have found that pretreatment of human monocyte-derived mac-
rophage-like cells derived from healthy adults with 10 µM α-MG increased both basal and LPS-induced
secretion of the proinflammatory cytokine tumor necrosis factor-alpha [44].
TABLE 31.3
Bioactivity of Mangosteen Xanthones
Bioactivity Model References
Anticancer and antimetastasis Rodents and cultured human cells [24,26,29,31]
Antiangiogenic Biochemical assays [33]
Anti-inflammatory Rodents, transformed, and normal human and rodent cells [43–47]
Antioxidant Biochemical assays, cultured cells, and rats [7,9,34,53]
Antibacterial, antifungal, and antiviral Biochemical assays, bacterial and fungal cultures, and [35,36,57–59]
virus-infected cells in culture
Central nervous system depressant Rodents [48]
Inhibitor of histamine release Rodent cells [37]
Myocardial stimulation Dogs and frogs [48]
Antiobesogenic Biochemical assays and cultured cells [38,42,69]
Aromatase inhibition Biochemical assay [39]
Analgesic Mice [40]
Antihyperglycemic Rat [41]
Inhibition of cytochrome P450 Biochemical assay [71]
An early in vivo study found that intraperitoneal and oral administration of α-MG, 1-isomangostin,
or mangostin triacetate reduced inflammation in rats in response to exposure to chemical irritants [48].
The α- and γ-MGs also attenuated carrageenan-induced paw edema in mice and rats, respectively
[45,49], and inhibited recruitment of inflammatory cells into the airway in a mouse model of allergic
asthma [50].
Evidence of anti-inflammatory activities of mangosteen juice products in humans is limited to two
reports and suggests the possibility that high doses of the juice may be proinflammatory. Healthy vol-
unteers consumed 2 ounces (59 mL) of a blended juice product with mangosteen containing unspecified
quantity of mangosteen puree and 94 mg undefined “mangostins” along with aloe vera leaves, green tea,
vitamins, and selenium before breakfast for 1 month. Plasma C-reactive protein (CRP) was significantly
decreased in the treatment group [51]. However, proinflammatory markers including interleukin-1α
(IL-1α), interleukin-1β (IL-1β), complement C3, and complement C4 were greater in plasma of adults
chronically ingesting the mangosteen juice compared to the placebo. Udani et al. [52] reported that
daily consumption of 18 ounces (540 mL), but not 6 ounces (180 mL) or 9 ounces (270 mL), of another
mangosteen-containing blended juice for 8 weeks by obese subjects significantly decreased plasma CRP
compared to that in obese subjects drinking a placebo. Serum interleukin-12p70 was also significantly
decreased compared to the placebo group after daily consumption of 6, 12, or 18 ounces (180–540 mL)
of the mangosteen juice blend for 8 weeks. However, there were no significant differences (P > 0.05) in
the amounts of several other proinflammatory cytokines (e.g., interferon-inducible protein-10, platelet-
derived growth factor-10, normal T-cell expressed and secreted protein, and macrophage inflamma-
tory protein-1β [MIP-1β]). However, plasma levels of proinflammatory markers including interleukin-1
(IL-1), complement C3 and C4, and MIP-1β increased significantly in subjects consuming the mango-
steen juice product. Because the tested mangosteen juices contained either supplements such as green
tea and minerals or concentrates from numerous other fruits, the observed outcomes cannot be attributed
solely to the mangosteen component.
Mangosteen extracts and xanthones possess free radical scavenging activity [7,9]. γ-MG was found
be more effective at scavenging hydroxyl radicals than ascorbic acid [7], and α-MG inhibited copper-
induced oxidation of human low-density lipoprotein (LDL) [53]. Extracts of mangosteen pericarp also
protected cultured neuroblastoma and neuronal cells against oxidative damage [54,55]. Furthermore,
α-MG decreased lipid peroxidation during isoproterenol-stimulated myocardial infarction in rats [56].
Mangosteen extracts and pure xanthones exhibit antibacterial, antifungal, and antiviral activities.
For example, α- and γ-MGs, garcinone D, and demethyl calabaxanthone all inhibited Mycobacterium
tuberculosis [57]. α-MG inhibited methicillin-resistant Staphylococcus aureus and vancomycin-resistant
enterococci [58]. Other examples of antimicrobial activities of xanthones have been summarized
elsewhere [8,9]. Both α- and γ-MGs also inhibited HIV-1 protease, an enzyme required for viral replica-
tion [59]. The relatively nonspecific antimicrobial activity of xanthones [8,9] and the likely presence of
high concentrations of these compounds in the colonic lumen [24] suggest that chronic consumption of
products containing mangosteen pericarp may affect the gut microbiota. In light of the influence of the
gut microbiota on health [60], this possibility merits critical examination.
Although limited to biochemical, cellular, and animal models, mangosteen xanthones and xanthone-
rich extracts have also been reported to have antiangiogenic, neuromodulatory, myocardial antiobeso-
genic, antihyperglycemic, and analgesic activities (Table 31.3). Whether consumption of mangosteen
juice affects such activities in humans is unknown.
The delivery of ingested xanthones or their metabolites to target tissues is required to exert their
bioactivities on peripheral organs. Reports on the absorption, metabolism, tissue distribution, and
excretion of xanthones using absorptive epithelial cells, rodents, and human participants have recently
appeared. We first reported that α- and γ-MGs in mangosteen aril and pericarp were stable during
simulated gastric and small intestinal digestion [61]. These xanthones were efficiently transferred to
the aqueous fraction of chyme in a bile salt-dependent manner suggesting incorporation into mixed
micelles. The uptake of xanthones in mixed micelles by Caco-2 human intestinal cells was propor-
tional to their extracellular concentration, and α- and γ-MGs were partially metabolized to phase
II metabolites. Both free and phase II conjugates effluxed across the apical and basolateral mem-
branes. Secretion of unconjugated xanthones across the basolateral membrane was dependent in part
on assembly and secretion of chylomicrons. Similarly, we and other several groups of investigators
have reported that a small percentage of orally administered α-MG was absorbed by mice and rats,
and that both the free compound and several phase II metabolites appeared in plasma [24,62–64].
Other xanthones have also been identified in plasma, liver, and colonic tumor tissue in athymic nude
mice chronically fed diet containing α-MG (0.09 g per kg diet; α-MG = 89% total xanthones) [24].
The concentrations of conjugated xanthones in plasma from fasted mice exceeded free α-MG 100-fold.
Other xanthones, including β- and γ-MGs, 9-hydroxycalabaxanthone, garcinones, and gartanin, were
also detected in plasma, and γ-MG, gartanin, 8-deoxygartinin, and 9-hydroxycalabaxanthone were
present in liver and the xenograph tumor. There was also extensive elimination of free and conjugated
α-MG and other xanthones in feces, suggesting exposure of colonic epithelium to high concentrations
of dietary xanthones and their metabolites.
These observations provide the background for the investigation of the bioavailability of xanthones
from mangosteen juices in humans. Kondo et al. [65] administered 2 ounces (59 mL) of a mangosteen
juice product (Table 31.1) to healthy adults (10 males and 10 females) before breakfast. As described
earlier, this product is a blended juice containing mangosteen puree combined with pericarp extract,
whole leaf aloe vera, and green tea, and 94 mg of undefined xanthones “mangostins,” according to the
label. Peak plasma concentration of α-MG was 3.1 ng/mL at 1 h, and approximately one-third of this
concentration was in plasma after 6 h. Information regarding the possible presence of other xanthones
and their metabolites in plasma was not provided, thus likely underestimating the extent of absorption of
xanthones from the juice blend.
We administered 2 ounces (59 mL) of a 100% mangosteen juice product with a western-style breakfast
to healthy adults (5 males and 5 females) [66]. The quantity of analyzed xanthones in the administered
mangosteen juice was 129 mg. Both free and conjugated forms of α-MG were present in plasma of all
subjects from 1 to 24 h postingestion of the juice. There was considerable variation in the maximum
concentration of α-MG in plasma (42–450 nmoL/L), as well as the time after consumption to peak con-
centration (range of 2–8 h with mean of 3.7 h), among participants. γ-MG, garcinone D, 8-deoxygartanin,
and gartanin were also detected in the serum. The profile of xanthones in 24 h urine was similar to that
in plasma and suggested a minimum absorption of 2% of xanthones from the juice.
Collectively, the results from preclinical and two human trials indicate that mangosteen xanthones
are absorbed to a limited extent, subjected to first pass metabolism, possibly biotransformed to other
xanthone compounds, and eliminated relatively slowly from the body in comparison with many other
dietary polyphenols.
The limited absorption of xanthones in mangosteen juice by healthy adults raises the question
of how such low concentrations in plasma and tissues can modulate physiological processes. An
expanding literature shows that α- and γ-MGs attenuate the activation of various signal transduction
pathways, including the nuclear factor kappa B (NF-κB) pathway [49,67], the AMP-activated protein
kinase (AMPK) pathway [30], the extracellular signal-regulated kinase 1 and 2 (ERK1/2) pathway
[28], and the Wnt-signaling pathway [68]. For example, Shen et al. [69] developed stably transfected
3T3–1 fibroblast cell lines with NF-κB and nuclear factor-erythroid-2-related factor 2 (Nrf-2) response
elements in promoters linked to fluorescent reporter genes. Relatively low concentrations of α- and
γ-MGs exhibited anti-obesogenic and anti-inflammatory activities. α-MG mediated this effect by inhib-
iting Nrf-2 promoter activity that in turn decreased the expression of peroxisomal proliferator-activated
receptor-gamma (PPAR-γ), a key inducer of adipogenesis and inflammation in mature adipocytes.
Both α- and γ-MGs also inhibited activation of the NF-κB-driven reporter gene construct resulting in
decreased expression of monocyte chemoattractant protein-1 (MCP-1). These results provide the impe-
tus to determine if the bioavailability of the xanthones in mangosteen juice is sufficient to modulate
such processes in humans.
Another important consideration regarding mangosteen juice is whether chronic consumption may
adversely affect health. There is a single report of possible overt toxicity associated with daily con-
sumption of mangosteen juice (source not specified) containing 25 mg α-MG per ounce (30 mL) for
12 months for the purpose of losing weight [70]. The actual amount of the juice product consumed daily
was unclear. The adult male presented with severe lactic acidosis. The patient had a history of hyper-
tension, metabolic syndrome, pulmonary sarcoidosis, and renal impairment. The patient’s condition
improved during hospitalization when mangosteen juice was not available. The authors suggested that
numerous medical disorders of the patient may have predisposed the individual to the reported inhibi-
tory activity of xanthones on mitochondrial respiration. This clinical report suggests the need for further
examination of the potential toxicological properties of mangosteen juice in human subjects with various
diseases. We are not aware of any controlled studies of potentially adverse interactions of mangosteen
juice with therapeutic drugs, other supplements, and herbals. The need for such investigations is sup-
ported by the report that a xanthone-rich extract of mangosteen pericarp inhibited multiple cytochrome
P450 enzymes and particularly the cytochrome P450 2C8 (CYP2C8) and 2C9 (CYP2C9) isozymes [71].
The investigators indicated that the observed inhibition occurred at concentrations of the xanthones
predicted to be achieved with doses present in some mangosteen-containing products including juices.
Potentially, adverse interactions of mangosteen juices with therapeutic drugs and other supplements and
herbals require consideration.

The successful marketing of juice containing mangosteen has provided an impetus for continued produc-
tion of new formulations and functional beverages, foods, and confectionaries containing whole fruit,
aril, and extracts of pericarp. An area requiring attention is how processing affects the bioactive com-
pounds in mangosteen fruit. Chaovanalikit et al. [15] presented preliminary results on the effect of sev-
eral processing styles on the phenolic and anthocyanin content of mangosteen powder and mangosteen
concentrate. Mangosteen juice was prepared from blanched juices of the fruit and aril before clarifying
by centrifugation and freezing. Upon thawing, mangosteen juice was mixed with rosella and grape juices
(2:1:2, v/v/v) and dried either by heating at 55°C in a vacuum oven or spray drying. Approximately 75%
of total phenolics in the fresh juice were lost during both drying procedures, whereas the loss of antho-
cyanins during spray drying (33% of total) was significantly less than that during vacuum drying (76%).
Mangosteen concentrate was prepared from the juice by either vacuum evaporation (40°C) or direct heat
(60°C). Aliquots of juice for each process were also incubated with pectinase for 2 h. The amounts of
total phenolics and anthocyanins in the concentrate prepared by vacuum evaporation were greater than
those in the concentrate prepared at the higher temperature under atmospheric pressure. Treatment with
pectinase prior to concentration increased the content of total phenolics and preserved several organo-
leptic properties.
31.6 Conclusion
Although there have been numerous testimonials from individuals supporting the efficacy of mango-
steen juice and other mangosteen-containing products, there is only one randomized, double blind, and
placebo-controlled human study testing the bioactivity of chronic consumption of mangosteen juice in
the peer-reviewed literature at this time [51]. This situation has led several health-care professionals to
conclude that claims that mangosteen juice promotes health are overstated [72–74]. The Food and Drug
Administration (FDA) issued a warning letter that the labeling claims for a mangosteen juice suggested
that the product should be classified as a drug [75]. Because required data demonstrating that the product
was both safe and effective had not been submitted and approved by the FDA, the company was directed
to prevent such further promotion of products or be in violation of the Federal Food, Drug, and Cosmetic
Act. Nevertheless, the diverse health-promoting activities of mangosteen xanthones when used as pure
compounds, extracts, and milled pericarp in preclinical models have been impressive and support the
need for further investigation in humans. Properly designed intervention trials using chemically char-
acterized mangosteen juice without other fruit juices and concentrates are needed to determine whether
the reported preclinical health-promoting activities of xanthones and other bioactive compounds in man-
gosteen fruit exhibit preventive and therapeutic activities for healthy and diseased humans, respectively.
It is also essential that the safety of chronic consumption of mangosteen juice and other mangosteen-
containing products is assessed. Financial support from the nutraceutical industry and government agen-
cies is needed to support such inquiries.
REFERENCES
1. Mohamed, M.F. and Frye, R., Effects of herb supplements on drug glucuronidation, review of clinical,
animal, and in vitro studies. Planta Med., 77, 311–321, 2011.
2. Sloan, E.W., Getting ahead of the curve: Phytochemicals. Nutraceutical World, 13, 16–17, 2010.
3. Office of Agricultural Economics (OAE), Indicators of Agricultural Economics of Thailand 2009,
2010a. Published online at: http://www.oae.go.th (accessed May 23, 2014).
4. Office of Agriculture Regulation (OAR), Public Access Database 2011. Published online at:
http://m.doa.go.th/ard/ (accessed May 23, 2014).
Release 26, 2013. Published online at: http://www.ars.usda.gov/services/docs.htm?docid=8964 (accessed
September 20, 2013).
6. Sinwat, S., Nutritive Values of Thai Foods, Nutrition Division, Department of Health, Ministry of Public
Health, Nonthaburi, Thailand, 2001.
7. Chin, Y. and Kinghorn, A.D., Structural characterization, biological effects, and synthetic studies on
xanthones from mangosteen (Garcinia mangostana), a popular botanical dietary supplement. Mini-Rev.
Org. Chem., 5, 355–364, 2008.
8. Obolskiy, D., Pischel, I., Siriwatanametanon, N., and Heinrich, M., Garcinia mangostana L.: A phyto-
chemical and pharmacological review. Phytother. Res., 23, 1047–1065, 2009.
9. Pedraza-Chaverri, J., Cárdenas-Rodríguez, N., Orozco-Ibarra, M., and Pérez-Rojas, J.M., Medicinal
properties of mangosteen (Garcinia mangostana). Food Chem. Toxicol., 46, 3227–3239, 2008.
10. Xango, Wikipedia Encyclopedia. Published online at: https://en.wkipedia.org/wiki/Xango (accessed
October 6, 2015).
11. Wittenauer, J., Falk, S., Schweiggert-Weisz, U., and Carle, R., Characterization and quantification of
xanthones from the aril and pericarp of magosteen (Garcina mangostana L.) and a mangosteen contain-
ing functional beverage by HPLC-DAD-MS. Food Chem., 134, 445–452, 2012.
12. Zadernowski, R., Czaplicki, S., and Naczk, M., Phenolic acid profiles of mangosteen fruits (Garcinia
mangostana). Food Chem., 112, 685–689, 2009.
13. Tulyatan, V., Subhimaros, S., Sukcharoen O., and Dulyapirunhasilp, S., Extraction of anthocyanins
from mangosteen rind. Food, 19, 25–35, 1989.
14. Du, C.T. and Francis, F.J., Anthocyanins of mangosteen, Garcina mangostana. J. Food Sci., 42,
1667–1668, 1977.
15. Chaovanalikit, A., Mingmuang, A., Kitbunluewit, T., Choldumringkool, N., Sondee, J., and Chupratum, S.,
Anthocyanin and total phenolics content of mangosteen and effect of processing on the quality of
mangosteen products. Int. Food Res. J., 19, 1047–1053, 2012.
16. Fu, C., Loo, A.E.K., Chia, P.P., and Huang, D., Oligomeric proanthocyanidins from mangosteen peri-
carps. J. Agric. Food Chem., 55, 7689–7694, 2007.
17. World Health Organization (WHO), General Guidelines for Methodologies on Research and Evaluation of
Traditional Medicine, WHO/EDM/TRM/2000.1, World Health Organization, Geneva, Switzerland, 2000.
18. Trisongti, P. and Termsirikoon, R., Mangosteen in Traditional Herb Medicine, Mahidol University,
Bangkok, Thailand, 1986.
19. Yapwattanaphun, C., Subhadrabandhu, S., Sugiura, A., Yonemori, K., and Utsunomiya, N., Utilization
of some Garcinia species in Thailand. Acta Hortic., 575, 563–570, 2002.
20. Yaacob, O. and Tindall, H.D., Mangosteen Cultivation, FAO Plant Production and Protection Paper
No. 129, Food and Agriculture Organization, Rome, Italy, 1995.
21. Gutierrez-Orozco, F. and Failla, M.L., Biological activities and bioavailability of mangosteen xanthones:
A critical review of the current evidence. Nutrients, 5, 3163–3183, 2013.
22. Pinto, M., Sousa, M., and Nascimento, M.S., Xanthone derivatives: New insights in biological activities.
Curr. Med. Chem., 12, 2517–2538, 2005.
23. Shan, T., Ma, Q., Guo, K., Liu, J., Li, W., Wang, F., and Wu, E., Xanthones from mangosteen extracts as
natural chemopreventive agents: Potential anticancer drugs. Curr. Mol. Med., 11, 666–677, 2011.
24. Chitchumroonchokchai, C., Thomas-Ahner, J.M., Li, J., Riedl, K.M., Nontakham, J., Suksumrarn, S.,
Clinton, S.K., Douglas A.K., and Failla, M., Anti-tumorigenicity of dietary α-mangostin in an HT-29
colon cell xenograft model and the tissue distribution of xanthones and their phase II metabolites. Mol.
Nutr. Food Res., 57, 203–211, 2013.
25. Doi, H., Shibata, M., Shibata, E., Morimoto, J., Akao, Y., Inuma, M., Tanigawa, N., and Otsuki, Y.,
Panaxanthone isolated from pericarp of Garcinia mangostana L. suppresses tumor growth and metas-
tasis of a mouse model of mammary cancer. Anticancer Res., 29, 2485–2495, 2009.
26. Johnson, J., Petiwala, S., Syed, D., Rasmussen, J.T., Adhami, V.M., Siddiqui, I.A., Kohl, A.M., and
Mukhtar, H., α-Mangostin, a xanthone from mangosteen fruit, promotes cell cycle arrest in prostate
cancer and decreases xenograft tumor growth. Carcinogenesis, 33, 413–419, 2012.
27. Matsumoto, K., Akao, Y., Yi, H., Ohguchi, K., Ito, T., Tanaka, T., Kobayashi, E., Iinuma, M., and
Nozawa, Y., Preferential target is mitochondria in α-mangostin-induced apoptosis in human leukemia
HL60 cells. Bioorg. Med. Chem., 12, 5799–5806, 2004.
28. Lee, Y., Ko, K., Shi, M.D., Liao, Y.C., Chiang, T.A., Wu, P.F., Shih, Y.X., and Shin, Y.W., α-Mangostin,
a novel dietary xanthone, suppresses TPA-mediated MMP-2 and MMP-9 expressions through the ERK
signaling pathway in MCF-7 human breast adenocarcinoma cells. J. Food Sci., 75, H13–H23, 2010.
29. Aisha, A., Abu-Salah, K., Ismail, Z., and Majid, A.M., In vitro and in vivo anti-colon cancer effects
of Garcinia mangostana xanthones extract. BMC Complement. Altern. Med., 12, 104, 2012. DOI:
10.1186/1472-6882-12-104.
30. Chao, A.C., Hsu, Y.L., Liu, C.K., and Kuo, P.L., α-Mangostin, a dietary xanthone, induces autophagic
cell death by activating the AMP-activated protein kinase pathway in glioblastoma cells. J. Agric. Food
Chem., 59, 2086–2096, 2011.
31. Shibata, M.A.,, Iinuma, M., Morimoto, J., Kurose, H., Akamatsu, K., Okuno, Y., Akao, Y., and Otsuki, Y.,
α-Mangostin extracted from the pericarp of the mangosteen (Garcinia mangostana Linn) reduces tumor
growth and lymph node metastasis in an immunocompetent xenograft model of metastatic mammary
cancer carrying a p53 mutation. BMC Med., 9, 69, 2011. DOI: 10.1186/1741-7015-9-69.
32. Nabandith, V., Suzui, M., Morioka, T., Kaneshiro, T., Kinjo, T., Matsumoto, K., Akao, Y., Iinuma,
M., and Yoshimi, N., Inhibitory effects of crude α-mangostin, a xanthone derivative, on two different
categories of colon preneoplastic lesions induced by 1, 2-dimethylhydrazine in the rat. Asian Pac. J.
Cancer Prev., 5, 433–438, 2004.
33. Shiozaki, T., Fukai, M., Hermawati, E., Juliawaty, L.D., Syah, Y.M., Hakim, E.H., Puthongking, P. et al.,
Anti-angiogenic effect of α-mangostin. J. Nat. Med., 67, 202–206, 2013.
34. Márquez-Valadez, B., Lugo-Huitrón, R., Valdivia-Cerda, V., Miranda-Ramírez, L.R., Pérez-De La Cruz,
V., González-Cuahutencos, O., Rivero-Cruz, I., Mata, R., Santamaría, A., and Pedraza-Chaverrí, J.,
The natural xanthone α-mangostin reduces oxidative damage in rat brain tissue. Nutr. Neurosci., 12,
35–42, 2009.
35. Gopalakrishnan, G., Banumathi, B., and Suresh, G., Evaluation of the antifungal activity of natural
xanthones from Garcinia mangostana and their synthetic derivatives. J. Nat. Prod., 60, 519–524, 1997.
36. Riscoe, M., Kelly, J., and Winter, R. Xanthones as antimalarial agents: Discovery, mode of action, and
optimization. Curr. Med. Chem., 12, 2539–2549, 2005.
37. Itoh, T., Ohguchi, K., Iinuma, M., Nozawa, Y., and Akao, Y., Inhibitory effect of xanthones isolated
from the pericarp of Garcinia mangostana L. on rat basophilic leukemia RBL-2H3 cell degranulation.
Bioorg. Med. Chem., 16, 4500–4508, 2008.
38. Jiang, H.Z., Quan, X.F., Tian, W.X., Hu, J.M., Wang, P.C., Huang, S.Z., Cheng, Z.Q. et al., Fatty acid
synthase inhibitors of phenolic constituents isolated from Garcinia mangostana. Biooorg. Med. Chem.
Lett., 20, 6045–6047, 2005.
39. Balunas, M.J., Su, B., Brueggemeier, R.W., and Kinghorn, A.D., Xanthones from the botanical dietary
supplement mangosteen (Garcinia mangostana) with aromatase inhibitory activity. J. Nat. Prod., 71,
1161–1166, 2008.
40. Cui, J., Hu, W., Cai, Z., Liu, Y., Li, S., Tao, W., and Xiang, H., New medicinal properties of mangostins:
Analgesic activity and pharmacological characterization of active ingredients from the fruit hull of
Garcinia mangostana L. Pharmacol. Biochem. Behav., 95, 166–172, 2010.
41. Ryu, H.W., Cho, J.K., Curtis-Long, J.C., Yuk, H.J., Kim, Y.S., Jung, S., Kim, Y.S., Byong Won Lee,
B.W., and Park, K.H., α-Glucosidase inhibition and antihyperglycemic activity of prenylated xanthones
from Garcinia mangostana. Phytochemistry, 72, 2148–2154, 2011.
42. Bumrungpert, A., Kalpravidh, R.W., Chitchumroonchokchai, C., Chuang, C.C., West, T., Kennedy, A.,
and McIntosh, M., Xanthones from mangosteen prevent lipopolysaccharide-mediated inflammation and
insulin resistance in primary cultures of human adipocytes. J. Nutr., 139, 1185–1191, 2009.
43. Bumrungpert, A., Kalpravidh, R.W., Chuang, C.C., Overman, A., Martinez, K., Kennedy, A., and
McIntosh, M., Xanthones from mangosteen inhibit inflammation in human macrophages and in human
adipocytes exposed to macrophage-conditioned media. J. Nutr., 140 842–847, 2010.
44. Gutierrez-Orozco, F., Chitchumroonchokchai, C., Lesinski, G., Suksamrarn, S., and Failla, M.L.,
α-Mangostin: Anti-inflammatory activity and metabolism by human cells. J. Agric. Food Chem., 61,
3891–3900, 2013.
45. Chen, L.G., Yang, L.L., and Wang, C.C., Anti-inflammatory activity of mangostins from Garcinia man-
gostana. Food Chem. Toxicol., 46, 688–693, 2008.
46. Nakatani, K., Nakahata, N., Arakawa, T., Yasuda, H., and Ohizumi, Y., Inhibition of cyclooxygenase
and prostaglandin E2 synthesis by γ-mangostin, a xanthone derivative in mangosteen, in C6 rat glioma
cells. Biochem. Pharmacol., 63, 73–79, 2002.
47. Tewtrakul, S., Wattanapiromsakul, C., and Mahabusarakam, W., Effects of compounds from Garcinia
mangostana on inflammatory mediators in RAW264.7 macrophage cells. J. Ethnopharmacol., 121,
379–382, 2009.
48. Shankaranarayan, D., Gopalakrishnan, C., and Kameswaran, L., Pharmacological profile of mangostin
and its derivatives. Arch. Int. Pharmacodyn. Ther., 239, 257–269, 1979.
49. Nguemfo, E.L., Dimo, T., Dongmo, A.B., Azebaze, A.G., Alaoui, K., Asongalem, W.E., Cherrah, Y.,
and Kamtchouing, P., Antioxidative and anti-inflammatory activities of some isolated constituents from
the stem bark of Allanblackia monticola Staner L.C. (Guttifererae). Inflammapharmacology, 17, 37–41,
2009.
50. Jang, H.Y., Kwon, O.K., Oh, S.R., Lee, H.K., Ahn, K.S., and Chin, Y.W., Mangosteen xanthones miti-
gate ovalbumin-induced airway inflammation in a mouse model of asthma. Food Chem. Toxicol., 50,
4042–4050, 2012.
51. Tang, Y.P., Li, P.G., Kondo, M., Ji, H.P., Kou, Y., and Ou, B., Effect of a mangosteen dietary supplement
on human immune function: A randomized, double-blind, placebo-controlled trial. J. Med. Food., 12,
755–763, 2009.
52. Udani, J.K., Singh, B.B., Barrett, M.L., and Singh, V.J., Evaluation of mangosteen juice blend on bio-
markers of inflammation in obese subjects: A pilot, dose finding study. Nutr. J., 8, 48–54, 2009
53. Williams, P., Ongsakul, M., Proudfoot, J., Croft, K., and Beilin, L., Mangostin inhibits the oxidative
modification of human low density lipoprotein. Free Radic. Res., 23, 175–184, 1995.
54. Moongkarndi, P., Srisawat, C., Saetun, P., Jantaravinid, J., Peerapittayamongkol, C., Soi-ampornkul, R.,
Junnu, S. et al., Protective effect of mangosteen extract against β-amyloid-induced cytotoxicity, oxida-
tive stress and altered proteome in SK-N-SH cells. J. Proteome Res., 9, 2076–2086, 2010.
55. Weecharangsan, W., Opanasopit, P., Sukma, M., Ngawhirunpat, T., Sotanaphun, U., and Siripong, P.,
Antioxidative and neuroprotective activities of extracts from the fruit hull of mangosteen (Garcinia
mangostana Linn.). Med. Princ. Pract., 15, 281–287, 2006.
56. Devi Sampath, P. and Vijayaraghavan, K., Cardioprotective effect of α-mangostin, a xanthone derivative
from mangosteen on tissue defense system against isoproterenol-induced myocardial infarction in rats.
J. Biochem. Mol. Toxicol., 21, 336–339, 2007.
57. Suksamrarn, S., Suwannapoch, N., Phakhodee, W., Thanuhiranlert, J., Ratananukul, P., Chimnoi, N.,
and Suksamrarn, A., Antimycobacterial activity of prenylated xanthones from the fruits of Garcinia
mangostana. Chem. Pharm. Bull., 51, 857–859, 2003.
58. Sakagami, Y., Iinuma, M., Piyasena, K., and Dharmaratne, H.R., Antibacterial activity of α-mangostin
against vancomycin resistant Enterococci (VRE) and synergism with antibiotics. Phytomedicine, 12,
203–208, 2005.
59. Chen, S., Wan, M., and Loh, B.N., Active constituents against HIV-1 protease from Garcinia
mangostana. Planta Med., 62, 381–382, 1996.
60. Lozupone, C.A., Stombaugh, J.I., Gordon, J.I., Jansson, J.K., and Knight, R., Diversity, stability and
resilience of the human gut microbiota. Nature, 489, 220–230, 2012.
61. Bumrungpert, A., Kalpravidh, R.W., Suksamrarn, S., Chaivisuthangkura, A., Chitchumroonchokchai,
C., and Failla, M.L., Bioaccessibility, biotransformation, and transport of α-mangostin from Garcinia
mangostana (mangosteen) using simulated digestion and Caco-2 human intestinal cells. Mol. Nutr.
Food Res., 53(Suppl. 1), S54–S61, 2009.
62. Li, L., Brunner, I., Han, A.R., Kinghorn, A.D., Frye, R., and Butterweck V., Pharmacokinetics of
α-mangostin in rats after intravenous and oral application. Mol. Nutr. Food Res., 55 (Suppl. 1), S67–S74,
2011.
63. Ramaiya, A., Li, G., Petiwala, S.M., and Johnson, J.J., Single dose oral pharmacokinetic profile of
α-mangostin in mice. Curr. Drug Targets, 13, 1698–1704, 2012.
64. Faizatun, S. and Rahayu, L., HPLC analysis of mangostin after orally administration in rats. Asian J.
Chem., 22, 6729–6733, 2010.
65. Kondo, M., Zhang, L., Ji, H., Kou, Y., and Ou, B., Bioavailability and antioxidant effects of a xanthone-
rich mangosteen (Garcinia mangostana) product in humans. J. Agric. Food Chem., 57, 8788–8792, 2009.
66. Chitchumroonchokchai, C., Riedl, K.M., Suksumrarn, S., Clinton, S.K., Kinghorn, A.D., and Failla,
M.L., Xanthones in mangosteen juice are absorbed and partially conjugated by healthy adults. J. Nutr.,
142, 675–680, 2012.
67. Shih, Y., Chien, S., Chen, P., Lee, J., Wu, S., and Yin, L., α-Mangostin suppresses phorbol 12-myristate
13-acetate-induced MMP-2/MMP-9 expressions via alphavbeta3integrin/FAK/ ERK and NF-kappaB
signaling pathway in human lung adenocarcinoma A549 cells. Cell Biochem. Biophys., 58, 31–44, 2010.
68. Yoo, J., Kang, K., Jho, E.H., Chin, Y., Kim, J., and Nho, C.W., α- and γ-Mangostin inhibit the prolif-
eration of colon cancer cells via β-catenin gene regulation in Wnt/cGMP signaling. Food Chem., 129,
1559–1566, 2011.
69. Shen, Q., Chitchumroonchokchai, C., Thomas, J.L., Gushchina, L.V., Disilvestro, D., Failla M.L., and
Ziouzenkova, O., Adipocyte reporter assays: Application for identification of anti-inflammatory and
antioxidant properties of mangosteen xanthones. Mol. Nutr. Food Res., 58, 239–247, 2014.
70. Wong, L.P. and Klemmer, P.J., Severe lactic acidosis associated with juice of the mangosteen fruit
Garcinia mangostana. Am. J. Kidney Dis., 51, 829–833, 2008.
71. Foti, R.S., Pearson, J.T., Rock, D.A., Wahlstrom, J.L., and Wienkers, L.C., In vitro inhibition of multiple
cytochrome P450 isoforms by xanthone derivatives from mangosteen extract. Drug Metab. Dispos., 37,
1848–1855, 2009.
72. Lobb, A.L., Science in liquid dietary supplement promotion: The misleading case of mangosteen juice.
Hawaii J. Med. Public Health, 71, 46–48, 2012.
73. Marcason, W., What are the facts and myths, about mangosteen? J. Am. Diet. Assoc., 106, 986, 2006.
74. Yeung, S., Mangosteen for the cancer patient: Facts and myths. J. Soc. Integr. Oncol., 4, 130–134, 2006.
75. Collins, B.B., Warning Letter, Food and Drug Administration (FDA) 2006. Published online at: http://
www.casewatch.org/prod/2006/xango.shtml (accessed May 12, 2014).
76. Chitchumroonchokchai, C. and Failla, M.L., Xanthone content in commercial mangosteen juices.
Human Nutrition Program, The Ohio State University, OH, 2014.
32
Melon Juice
Ayse Karadag and Banu Bayram
CONTENTS
32.1 Introduction................................................................................................................................... 385
32.3 Bioactives and Antioxidant Efficiency.......................................................................................... 388
32.4 Health Effects................................................................................................................................ 390
32.6 Conclusion..................................................................................................................................... 394
References............................................................................................................................................... 395
32.1 Introduction
Melons are members of the Cucurbitaceae (cucurbit) family and produced on vining plants with tendrils.
They are a multicolored family with flesh that can be orange, green, or white, and vary in size and shape
from huge to small and from round to cylindrical [1]. Melons are widely consumed all over the world.
Over 90 genera and 750 species of melons are known, and there are four basic categories of melons
related to four different genera in the Cucurbitaceae family.
Although melons are very popular and widely consumed fruits throughout the world, they are highly
perishable during short periods of storage due to their mild acidity (pH 5.2–6.7) and high water activ-
ity (~0.98), for example, sliced/cut melons are perishable food products. The production of melon juice
needs more sophisticated nonthermal alternative pasteurization steps since the mild, distinctive flavor
of melon juice is very sensitive to conventional thermal treatment. Therefore, commercially available
packaged melon juice is virtually unknown compared to the other fruit juices such as apple, orange,
and mango, and there are only a few studies considering the possible production of melon juice or con-
centrates and defining process effects on nutritional composition and bioactive compounds [2]. Some
important bioactive compounds found in melons are provided in Figure 32.1.
Melon has a high portion of edible part (68%–98%) and the water content of edible part is around
90%–96% [3–5]. This chapter highlights the differences in the composition of nutrients and bioactive
compounds, antioxidant efficacy, and health effects of melons (where possible melon juices) as well as
the future trends and novelties for juice production. However, it should be noted that beyond fresh con-
sumption of fresh fruits, the choices for other processed products including juices and concentrates from
melons are practically nonexistent. Since there are very limited studies specifically about nutritional
characteristics and bioactives of melon juice, in some cases the data in literature are provided with the
edible portion of melon, including its flesh and juice.

Table 32.1 shows the compositional and nutritional characteristics of the edible portion of four
d ifferent types of melons. Due to high water content, they are not generally ranked nutritious on a
fresh weight (FW) basis. However, melons contain large amounts of vitamins A and C in addition to
385
CH3
H3C CH3 H3C H3C
CH3
CH3 CH3 CH3
CH3
β-Carotene
H OH
HO H
H OH
H3C H
O
OH O
O H
H OH
O H
H H OH H3 C
O O
O H
H OH HO
OH
O OH
CH3 O
Hesperidin Vanillic acid
OH
O
O OH O H
HO O
H
O HO O
O O
HO
HO OH
OH
Charantin
FIGURE 32.1 Some important bioactive compounds found in melons.
small amounts of minerals such as potassium, calcium, iron, and magnesium [8,9]. Laur and Tian
[10] reported that β-carotene that exhibits provitamin A activity was present at detectable levels
in both cantaloupe and honeydew melons. It was also found that in honeydew melon, the major
pigments are chlorophylls a and b, while cantaloupe fruit does not contain chlorophylls and accu-
mulates up to 60-fold more β-carotene than honeydew melon. Consuming 1–2 cups of cantaloupe
(about 150–170 g/cup) would meet the Recommended Dietary Allowances (RDA) of vitamins A
and C for healthy adults; in addition, it is a good source of vitamin B6 [10]. Wolbang et al. [11]
studied the β-carotene contents of 10 cultivars of Cucumis melo with orange, light, and green flesh
and their in vitro bioaccessibility and antioxidant activity. The results suggested that β-carotene
present in the melons was highly bioaccessible due to their thin and nonfibrous cell walls with
globular chromoplast structure and the bioaccessibility was not correlated to the amount [11,12].
Some of the data related to nutritional composition of melon juice could only be retrieved from the
few studies about the extraction and processing of melon juice reported in the literature. However, it
should be noted that much of the work has been on experimental basis since commercial processing
of melon juice or juice products has not been reported in the literature. Total soluble solids of fresh
Melon Juice 387
TABLE 32.1
Compositional and Nutritional Characteristics of Various Melons (per 100 g)
Nutrient Unit Winter Melon [4,31] Cantaloupe [6] Honeydew [5] Bitter Melon [7]
Water g 96 90 89.92 83.2–94.03
Energy kcal 13 34 36 17
Protein g 0.4 0.84 0.54 1.0–2.1
Fat g 0.2 0.19 0.19 0.17
Carbohydrate g 3.0 8.16 9.09 3.7
Ash g 0.3 0.65 0.41 1.1
Total dietary fiber g 2.9 0.9 0.8 2.8
Total sugars g na 7.86 8.12 na
Sucrose g na 4.33 2.48 na
Glucose g na 1.54 2.68 na
Fructose g na 1.87 2.96 na
Minerals
Calcium mg 5–23 9 6 19–23
Copper mg 0.02 0.04 0.02 0.03–0.19
Iron mg 0.2–0.5 0.21 0.17 0.43
Magnesium mg 10 12 10 17
Manganese mg 0.06 0.04 0.03 0.09
Phosphorus mg 19 15 11 31–38
Potassium mg 77–131 267 228 171–296
Sodium mg 0.14–6 16 18 2.4–5
Zinc mg 0.61 0.18 0.09 0.46–0.80
Vitamins
Niacin mg 0.2–0.46 0.734 0.41 0.4
Pantothenic acid mg na 0.105 0.15 0.21
Riboflavin mg 0.03–0.11 0.019 0.01 0.04
Thiamin mg 0.02–0.04 0.041 0.03 0.04
Total folate (DFE) µg 5 21 19 72
Vitamin A IU na 3382 50 471
Vitamin B6 mg 0.04 0.07 0.09 0.04
Vitamin C mg 13–68 36.7 18 84–96
Vitamin E (ATE) µg na 50 20
Carotenoids
β-Carotene µg na 2020 30 126–190
α-Carotene µg na 16 na 185
Lutein + zeaxanthin µg na 26 27 170
Abbreviations: DFE, dietary folate equivalents; na, not available; ATE, alpha tocopherol equivalents.
squeezed melon juice were reported to be between 85 and 102 g/L [2,13]. The concentrations of three
main sugars, namely, fructose, glucose, and sucrose were in the range of 13.4–16.1, 11.3–17.3, and
29.9–45.7 g/L, respectively. The vitamin C content of melon juice ranged from 106 to 190 mg/L [2,13].
The content of β-carotene of fresh melon juice was 40.7 mg/L, and all carotenoids were lost during clar-
ification through membrane, possibly due to strong association of β-carotene with membrane and wall
structures of the cell fragments (e.g., the pulp) in both studies [2,13]. When Chen et al. [14] produced
unclarified melon juice by different pasteurization methods, they found that dense-phase carbon dioxide
method nearly completely protected all β-carotene (8.95 mg/kg juice), but 57% of its loss occurred in
heat-treated melon juice.
32.3 Bioactives and Antioxidant Efficiency

The antioxidant efficacy of melon bioactive compounds has been mainly studied in aqueous, hydroalco-
holic, or acetone extracts from the edible portion of melon, depending on the antioxidant assay employed.
Results from these studies provide further insight into the antioxidant activity of melon juice. Table 32.2
shows the antioxidant activities and total phenolics of various melon types.
Vinson et al. [21] stated that among the 20 fruits consumed in the United States, cantaloupe and
honeydew melon had the lowest total phenol antioxidant index similar to the study of Morales-Soto
et al. [22]. On the other hand, cantaloupe melon ranked sixth in terms of contribution to daily phenol
intake depending on the consumption rate and serving size. Wolfe et al. [23] found that cantaloupe
melon along with banana and avocado had the lowest cellular antioxidant activity among the several
tested fruits such as berries, pomegranates, apples, oranges, and peaches. In addition, oxygen radical
absorbance capacity (ORAC) values and total phenolics of cantaloupe and honeydew melons were the
lowest among the tested fruits.
Rodríguez-Pérez et al. [24] characterized the predominant group of phenolic antioxidants in melons as
phenolic acids, which may be further classified into hydroxybenzoic acid derivatives (isomers of gentisic acid
hexoside and hydroxybenzoic acid hexoside), hydroxycinnamic acid derivatives (ferulic acid and p-coumaric
acid hexoside), and other phenolic acid derivatives (vanillic acid dihexoside, vanillic acid hexoside, and
coelovirins A and B). Among the tested melon varieties, cantaloupe was the richest source of phenolic acid
derivatives. As flavonoids, primarily eriodictyol rutinoside and hesperidin were identified [24].
TABLE 32.2
Antioxidant Activities and Total Phenolics of Various Melons
Winter
Unit Melon Cantaloupe Honeydew Bitter Melon References
Antioxidant Activities
TEAC–ABTS mmol TE/kg FW na 1.20 0.65–2.99 na [15,34]
mmol TE/kg DW na 9.44–26.6 9.8–11.7 na [22]
FRAP µM TE/kg FW na 351.3–493.8 na [25]
mmol eq FeSO4/100 g DW na 1.30–3.34 1.78–2.17 na [22]
mmol Fe2+ /kg FW na 5.73 2.27 na [34]
mM TE/g extract 26.71 na na na [16]
mg TE/g extract na na na 25.24–28.19 [27]
DPPH mg TE/g extract na na 11.89–13.12 [27]
Scavenging percentage (%) 74.57 na na na [17]
per 150 mg FW
EC50: µg/mL 195 na na na [16]
IC50: µg/mL na na na 129.4–156.7 [18]
ORAC mmol TE/kg FW na 0.23–0.95 0.97–2.74 na [23,34,60]
mmol TE/kg DW na 4.23–4.70 1.90–4.20 na [8,22]
Total mg GAE/100 g FW 169 16.11 11.52–25.7 26.6 [15–17,19,
Phenolics 23,25]
mg GAE/g DW na na na 5.39–8.90 [26]
mg CE/100 g FW na 33.56–56.44 26 na [20,21]
mg GAE/g extract 185 na na 22.73–68.8 [16,18,27]
Total mg CE/100 g FW na 3.15–6.29 2.25 na [19,20]
Flavonoids
ABTS, 2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid); CE, catechin equivalents; DPPH,
Abbreviations:
2,2-diphenyl-picrylhydrazyl; DW, dry weight; EC50, effective concentration 50; FeSO4, ferrous sulfate;
FRAP, ferric reducing antioxidant power; FW, fresh weight; GAE, gallic acid equivalents; IC50, inhibitory
concentration 50; na, not available; ORAC, oxygen radical absorbance capacity;TE, trolox equivalents;
TEAC, trolox equivalent antioxidant capacity.
Melon Juice 389
Kolayli et al. [25] found total polyphenol content of three types of melons at around 100 mg gallic
acid equivalents (GAE)/100 g FW, and the amount of phenolic acids in aqueous extracts of the melons
varied widely from 0.5 to 30 mg/100 g FW, mainly benzoic acid (5.55–30.06 mg/100 g FW), abscisic
acid (8.35–15.39 mg/100 g FW), vanillic acid (6.36–7.83 mg/100 g FW), and trans-cinnamic acid (2.99–
6.85 mg/100 g FW), p-coumaric acid (3.24–4.16 mg/100 g FW), and ferulic acid (2.91–3.69 mg/100 g
FW); all the samples had high amounts as major phenolic components [25]. Gallic acid, gentisic acid,
catechin, chlorogenic acid, and epicatechin were the phenolics typically abundant in the bitter melon,
and their concentrations were in the range of 8.04–39.76, 16.99–32.39, 23.06–82.45, 4.55–15.83, and
16.14–44.28 mg/100 g dry weight (DW), respectively [26]. Quinic acid, syringic acid, caffeic acid,
and 4-coumaric acid were also analyzed in bitter melon extracts at concentrations of 16.25–145.29,
24.61–56.34, 32.97–175.5, and 6.90–56 μg/g extract, respectively [27]. The total phenolic content of
bitter melon obtained by subcritical water extraction was 48.18 mg GAE/g DW, with gallic acid being
the main phenolic acid (0.65 mg/g DW) [28]. Additionally, the pulp around the seeds of the ripe bitter
melon is a good source of lycopene. The bitter component of fruit has been characterized as having
cucurbitane-type triterpene glycosides. Polypeptide-P (a peptide mimicking insulin that lowers blood
sugar levels), charantin (steroidal saponin acting as a hypoglycemic agent), momordin Ic, oleanolic
acid-3-O-monodesmoside, and oleanolic acid 3-O-glucuronide have also been identified in the fruit [28].
Three main phenolic compounds, astilbin, catechin, and naringenin, were identified in the winter
melon [29]. The fruit also contains many other bioactive compounds, such as isovitexin, 1-sinapoylglucose,
benzylalcolcohol-O-α-l-arabinopyranosyl-(1–6)-β-d-glucopyranoside, isomultiflorenyl acetate, 5-gluten-
3-β-ylacetate, triterpenes (alnusenol, multiflorenol, and isomultiflorenol), and sterols (lupeol, lupeol
acetate, β-sitosterol, and stigmasterol), among others. Among the active triterpenes and sterols, two
triterpenes, alnusenol and multiflorenol, have the ability to strongly inhibit the release of histamine, an
inflammatory mediator [30,31].
In the study of Lester and Hodges [8], nonnetted orange-fleshed honeydew fruit cultivars “Orange
Delight” and “Orange Dew” showed high activities of ascorbate peroxidase, catalase, and superoxide
dismutase (SOD) and least increase in malondialdehyde (MDA) (e.g., lipid peroxidation) during stor-
age. SOD activity was found to vary significantly among the genotypes of melons. For example,
casaba-type melons had average SOD activities that were approximately 16-fold greater than those
of honeydew type and approximately 9-fold greater than those of cantaloupe type [8]. These data
indicated the existence of useful genetic diversity among commercial melon varieties and in exotic
genotypes that could be used to develop C. melo as a functional food with enhanced SOD content.
Cantaloupe melon provides a new class of antioxidant nutraceutical products, called Extramel®,
GliSODin®, and PromutaseTM 200, all containing high level of SOD. Extramel is the freeze-dried
cantaloupe melon juice concentrate that was threefold higher in SOD compared to a classic melon that
contains 14 IU SOD/mg powder. Similarly, Promutase 200 is also a freeze-dried cantaloupe melon
juice concentrate containing 2.6 IU of SOD/mg. GliSODin is another extract covered by polymeric
films of wheat matrix gliadin. In vivo and in vitro studies are available in the literature with these prod-
ucts showing high antioxidant activity that is closely related to their high SOD activity. However, the
antioxidant activity of melon extract was significantly decreased after the heat inactivation (lacking its
SOD activity), which confirmed the inhibitory effect of SOD on radical formation [32].
Vouldoukis et al. [32] evaluated antioxidant and anti-inflammatory properties of cantaloupe melon
extract with high SOD activity (an average of 100 IU/mg of dry extract), but also having catalase activity
(10 IU/mg), residual glutathione peroxidase (GPx) activity, and number of natural antioxidant quench-
ing molecules. Fruit extract inhibited the production of superoxide anion (O2•−) with a maximal effect
at 100 µg/mL of fruit extract. In addition, melon extract inhibited the production of peroxynitrite, thus
strengthening its antioxidant properties. Carillon et al. [33] measured the antioxidant activity and radi-
cal scavenging capacities of Extramel for hydroxyl (HO•), O2•− anion, and hydrogen peroxide (H2O2)
by total antioxidant capacity (TAC), 2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS),
2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) assays. The TAC of
SOD-rich melon concentrate was comparable to that of blackberry [34], where blackberry was ranked as
the fruit having the highest TAC value among 30 fruits. The scavenging ability of SOD-rich melon concen-
trate was higher against O2•− (IC50=0.80 mg/mL) than against HO• and H2O2 (IC50 up to 10 mg/mL) [33].
The antioxidant activities of methanolic extracts from four varieties of the bitter melon ranged from
81.7% to 86.5% inhibition at a level of 500 ppm extract in methyl linoleate. They have moderate to good
inhibition effects on oxidation in comparison with some other plant extracts [26]. Shetty et al. [35]
studied the antioxidant activity of winter melon extract by determining the healing of indomethacin-
induced ulcers in rats. The fruit extract caused significant increase in SOD in red blood cells and antral
homogenate and vitamin C levels in rat plasma. There was an apparent decrease in ulcer index in animals
treated with extract. The authors postulated that the fruit extract probably inhibits gastric mucosal injury
by scavenging the free radicals.
32.4 Health Effects

Most of the studies regarding the health effects of melon juices are mainly focused on bitter melon.
In addition to a vast number of animal studies, in vitro studies with cell culture provide useful data to
determine the effects of juices on different diseases. Melon juices, mainly bitter melon juice, exhibit a
number of biologic effects including antioxidant, antihypertensive, antidiabetic, antiobesity, anticancer,
cytoprotective, neuroprotective, cardioprotective, hypoglycemic, and hypolipidemic effects, among oth-
ers [32,36–39]. Studies regarding the health effects of melon juices with proposed mechanisms of action
are summarized in Table 32.3.
Antidiabetic effect of bitter melon has been well documented through lowering plasma lipids, cho-
lesterol, glucose, insulin resistance, and antioxidant mechanisms. Currently, available drugs, including
well-known metformin, have side effects and fail to control diabetes completely. Therefore, there has
been a growing interest in discovering bioactive compounds from natural sources that may show thera-
peutic potential against diabetes. Alkaloids, polypeptide-P, and charantin appear to be responsible for
the hypoglycemic action of bitter melon. Tan et al. [42] isolated four glycosides (momordicosides Q, R, S,
and T) and karaviloside XI from bitter melon that have beneficial effects on obesity and diabetes work-
ing through adenosine 5′-monophosphate-activated protein kinase (AMPK), a key pathway mediating
glucose uptake and fatty acid oxidation, in L6 myotubes and 3T3-L1 adipocytes. Only a few studies are
available in the literature about cantaloupe and winter melons. Cantaloupe melon has high antioxidant
power related to its high SOD activity. Furthermore, it shows hypoglycemic and cardioprotective effects
as well as ameliorating diet-induced obesity.
The glucose, insulin, and lipid metabolisms are strictly related to diabetes and obesity. High choles-
terol, high glucose, increased oxidative stress, insulin resistance, and impaired leptin/adiponectin
levels are common risk factors for diabetes. There are in vivo and in vitro studies performed with melon
juices showing the antidiabetic and thus antiobesity effect. Singh et al. [43] incubated rat muscle cells
(L6 myotubes) either with insulin (100 nM) or with different concentrations (1–10 µg/mL) of the lyoph-
ilized bitter melon juice. Time-dependent increase in glucose (3H-deoxy-D-glucose) and amino acid
(14C-Me-AIB) uptake with maximum concentration of 5 µg/mL was observed. This effect was similar to
that of 100 nM insulin, demonstrating the presence of insulin-like active ingredients in bitter melon juice
acting as a hypoglycemic agent. The glucose uptake stimulating effect was also reported in Streptozotocin
(STZ)-induced Wistar rats fed bitter melon juice (10 mL/kg body weight) for 10 weeks [44]. In brush
border membrane vesicles isolated from jejunum of rats, glucose uptake was lower in bitter melon juice
group as compared to STZ-induced rats. Similarly, hypoglycemic effect was observed in L6 myotubes
isolated from skeletal muscles, but only when they were applied at a physiological dose similar to that of
insulin. The inhibitory effect was reported to be through the same signal transduction pathway since the
response was blocked by wortmannin, an inhibitor phosphatidylinositol 3-kinase that is associated with
the transport of glucose into muscle cells [44].
Another proposed mechanism of action for the hypoglycemic effect of bitter melon is the protection
of islets where insulin is synthesized. The number of insulin-positive islets was higher in bitter melon–
treated group demonstrating the repair and regeneration activity on the islets of Langerhans [44].
Sitasawad et al. [45] found that the viability of islet cells isolated from Balb/c mice supplemented
with 0.2 mL (equivalent to 10 mL/kg body weight) of 10%, 50%, or 100% bitter melon juice for
5 days was dose-dependently increased. Additionally, MDA levels as a marker of lipid peroxidation
Melon Juice
TABLE 32.3
Summary of In Vivo and In Vitro Studies for Health Effects of Melon Juices
Model Organisms/
Treatment Subjects Experimental Design Mechanisms of Action Effect References
SOD-rich cantaloupe db/db mice Diet + 0.08% extract Urinary albumin ↓, urine and kidney 8-OHdG ↓, diabetes- Antioxidant [36]
extract powder induced oxidative stress ↓, and renal cell injury ↓.
Freeze-dried BM juice C57BL/6 mice HF + 1.5% BM juice Plasma glucose, TAG, and CHL ↓, hepatic TAG ↓, ALT ↓, Hypolipidemic and [37]
and AST↓. hypoglycemic
BM juice Human primary 0.5%, 1%, and 2% BM Lipolysis ↑, lipid content ↓, mRNA level of PPARγ, SREB, Hypolipidemic [38]
preadipocytes juice and perilipin and resistin ↓.
BM juice Pancreatic carcinoma cells; 2%–5% (v/v) Cell viability ↓ and induction of apoptosis, activation of Anticacinogenic [39]
BxPC-3, MiaPa Ca-2, AMPK, and caspases.
AsPC-1, and Capan-2
Lyophilized BM juice Athymic nude mice 5 mg BM juice/100 μL/ Inhibition of MiaPaCa-2 tumor growth by 60%, induction Anticacinogenic [39]
mouse/day of apoptosis, and activation of AMPK.
BM juice Breast cancer cells; MCF-7 1%, 2%, and 5% BM Cell proliferation ↓, apoptotic cell death ↓, and activation Anticarcinogenic [51]
and MDA-MB-231 juice of caspase expression of apoptosis regulatory proteins ↑.
Freeze-dried BM juice Sprague Dawley rats LF diet + 0.75% BM and TAG and CHL in skeletal muscle ↓, plasma glucose↓, Antiobesity and [40]
HF diet + 0.75/1.5% BM serum FFA ↑, and hepatic TAG↓. hypolipidemic
BM juice Wistar STZ-induced 10 mL BM juice/kg body Hepatic GSH concentration ↑ GST activity↓, blood glucose Antioxidant and [41]
diabetic rats weight ↓, and normalization of CYP enzymes. hypoglycemic
Abbreviations:
8-OHdG, 8-hydroxydeoxyguanosine; ALT, alanine transaminase; AMPK, adenosine 5′-monophosphate-activated protein kinase; AST, aspartate aminotransferase; BM, bitter
melon; CHL, cholesterol; CYP, cytocrome-P-450; FFA, free fatty acids; GSH, glutathione; GST, glutathione-S-transferase; HF, high fat; LF, low fat; mRNA, messenger RNA;
PPARγ, peroxisome proliferator-activated receptor gamma; SOD, superoxide dismutase; SREB, sterol regulatory element binding protein; STZ, streptozotocin; TAG,
triacylglycerols.
391
significantly decreased in pancreas, isolated islets, and RIN cells with 0.1%–0.5% juice treatment, and
also antiapoptotic effect of melon juice was reported in RIN cells (islet-derived tumor cells), showing
a cytoprotective action. However, high concentrations of melon juice caused detrimental effects due to
increased oxidative stress in islets [45].
Ahmed et al. [46] studied the effects of bitter melon juice on plasma and liver lipid profiles in normal
and STZ-induced type I diabetic rats after treatment with 10 mL of 100% fruit extract for 10 weeks. Bitter
melon juice decreased total cholesterol; however, no effect was observed in nonesterified cholesterol,
phospholipid, and triacylglycerols (TAG) levels that were significantly increased in STZ-induced rats.
However, treatment did not result a decrease in tissue TAG levels (kidney, brain, and testis) in STZ-
induced rats, except in the liver. The levels of MDA in plasma and kidney were normalized with bitter
melon in diabetic rats [46].
Hypolipidemic and antioxidant effect of melon juice was demonstrated in several studies. Décordé
et al. [47] fed male golden Syrian hamsters for 12 weeks with a high-fat (HF) diet and water or an
aqueous solution of Extramel at a concentration of 0.7, 2.8, and 5.6 mg/day corresponding to 10,
40, and 80 IU SOD/day, respectively. Body fat accumulation and abdominal adipose tissue weight
of hamsters increased with HF diet. Extramel reduced these effects at concentrations of 2.8 and
5.6 mg/day. Increased plasma glucose, TAG, and insulin levels were observed with HF diet. Extramel
decreased TAG and plasma insulin levels; however, no effect was found on glucose. Extramel showed
an antiobesity effect in HF hamsters by decreasing body weight, abdominal fat, TAG, insulin, insulin
resistance, and liver lipids, as well as increasing the adiponectin level in plasma, which is associated
with decreased oxidative stress, hepatic fat content, and improved insulin sensitivity. Furthermore, it
decreased oxidative stress in the liver by decreasing MDA and protein oxidation products. Chan et al.
[48] fed male Sprague Dawley rats with HF diets to induce obesity, and the effect of bitter melon
juice powder was assessed at doses of 0.75%, 1.0%, and 1.25% added to the HF diet. Bitter melon
supplementation in the HF diet–fed rats lowered hepatic and muscle TAG concentrations, normalized
plasma glucose, and improved insulin sensitivity compared with the unsupplemented HF diet–fed
rats. In addition, the two mitochondrial enzymes crucial to lipid oxidation were elevated, namely,
carnitine palmitoyltransferase-I (CPT-I) and acyl-CoA dehydrogenase (AD) in the liver and skeletal
muscles of rats. At the level of 0.75% bitter melon, the increase in CPT-I and AD activities ranged
from 36% to 57% and from 25% to 31%, respectively, compared with the unsupplemented rats. The
reduction in lipid content of liver and gastrocnemius muscle among the bitter melon supplemented
groups was ~40%. Data suggest that bitter melon enhanced mitochondrial transport of long-chain
fatty acids and the decreased organ lipid content was associated with the enhancement in activity of
AD, a key oxidative enzyme [48].
The underlying mechanism of decrease in TAG may be the inhibition of synthesis and secretion.
This mechanism of action was demonstrated [49], with bitter melon juice applied at 0.5% and 1%
applied to human hepatoma cell line, HepG2 cells, as compared with untreated control cells. It was
suggested that bitter melon juice inhibits the synthesis and secretion of TAG in the form of apolipo-
protein B (apoB), a risk factor of hyperlipidemia and coronary heart disease (CHD), as well decreases
mRNA expression of microsomal transfer protein, playing a important role in the assembly and secre-
tion of apoB-containing lipoproteins and lipid metabolism. The results of in vitro studies further
confirmed in vivo experiments showing that this juice lowers plasma apoB-100 and B-48 in C57BL/6
mice fed HF diets [37].
A few studies have also been conducted to investigate the proposed anticancer properties of bitter
melon. In human prostate cancer cells (PC3 and LNCaP), treatment with bitter melon extract at 2% (v/v)
resulted in an induced apoptosis and inhibited cell proliferation through decreasing the expression of
cyclin D1 and cyclin E (critical cell-cycle regulatory proteins) and inhibiting their normal regulation of
cell-cycle progression [50]. This effect was also confirmed by in vivo study conducted with transgenic
adenocarcinoma of mouse prostate mice that fed 100 mL bitter melon juice. This experiment also resulted
in an inhibition of prostate cancer progression by interfering cell cycle progression and proliferation [50].
Together with caspase activation, these mechanisms were shown in other studies performed with human
pancreatic carcinoma cells (BxPC-3, MiaPaCa-2, AsPC-1, and Capan-2) [39] and breast cancer cells
(MCF-7 and MDA-MB-231) [51], suggesting the anticarcinogenic effects of bitter melon (Table 32.3).
Melon Juice 393
Some studies have shown antihypertensive, anti-inflammatory, and antiatherosclerotic effect of

melon juices or melon juice products providing evidence that these may contribute to cardiovascular
health. In a study with golden Syrian hamsters that fed HF diet and water or aqueous solution of
Extramel, plasma total cholesterol decreased by 48%; however, there was no change in plasma TAG,
high-density lipoprotein (HDL), and liver lipid levels at the end of 12 weeks of treatment. Importantly,
aortic cholesterol, aortic fatty streak accumulation, and cardiac hepatic superoxide radical anion
production decreased [52]. In another study, Extramel inhibited angiotensin I converting enzyme,
which increases blood pressure by causing blood vessels to constrict by 99.3% at a concentration of
10 mg/mL. The IC50 value of Extramel was determined as 2.4 mg/mL [33]. Nitric oxide (NO) is the
most important relaxing factor released from endothelial cells lowering blood pressure. Winter melon
condensed juice decreased blood pressure by 11 and 91 mmHg in male Wistar rats when applied at
intravenous injection doses of 0.8 and 1.6 mL/kg, respectively [53]. In the same study, treatment with
3, 10, 30, and 100 µL winter melon juice induced relaxation of isolated aortic rings with endothe-
lium isolated from rats in a concentration-dependent manner. In cultured porcine aorta endothelial
cells, addition of 200 µL winter melon juice to 10 mL of medium significantly increased NO produc-
tion, demonstrating the blood pressure lowering effect of winter melon juice through endothelium-
dependent vasodilation [53].
Lin and Tang [54] investigated the anti-inflammatory effect of lyophilized bitter melon juices at
concentrations of 10–500 μg/mL in primary peritoneal macrophages isolated from mice. Bitter melon
juice (10–500 μg/mL) concurrent administration with lipopolysaccharide (LPS) stimulation to mac-
rophages did not significantly change proinflammatory cytokines interleukin-1β (IL-1β), IL-6, and
tumor necrosis factor-alpha (TNF-α) secretions. However, high dose (500 μg/mL) of bitter melon juice
administration significantly increased the secretion of anti-inflammatory cytokine IL-10 in a dose-
dependent manner. When bitter melon was applied after LPS stimulation, interestingly, an increase in
proinflammatory cytokines was observed. The results suggested that bitter melon juice administration
could not fully inhibit the inflammation in macrophages stimulated by LPS under this experimental
model. In another study conducted with a cantaloupe melon product [55], the levels of proinflamma-
tory cytokines IL-6 and TNF-α decreased in golden Syrian hamsters that received a cafeteria diet
composed of HF, high-sugar, and high-salt products after 1-month supplementation with Extramel
(10 U SOD/day).
The improvement of antioxidant defenses and prevention of oxidative stress has been reported in some
studies performed with melon juice products. In a study conducted by Carillon et al. [55], golden Syrian
hamsters received either a standard diet or a cafeteria diet composed of HF, high-sugar, and h igh-salt
products for 15 weeks. After 1-month supplementation with Extramel (10 U of SOD/day), body weight
and insulin resistance induced by the cafeteria diet—reduced and hepatic oxidative stress was corrected
corresponding with the increase in the expression of the liver antioxidant defense enzymes (Mn-SOD,
Cu/Zn SOD, catalase, and GPx). In addition to inflammatory markers, levels of transcription factor
nuclear factor-kappa B, hepatic F(8)-isoprostane levels, and total superoxide anion production were
decreased. The increase in low-density lipoprotein (LDL) cholesterol and glucose levels was corrected
by 55% and 35%, respectively, in the treatment group as compared to cafeteria diet–fed group. Milesi
et al. [56] examined the antioxidant activity of Extramel in 70 volunteers aged between 30 and 55 who
feel daily stress and fatigue. A dose of 10 mg of Extramel corresponding to 140 IU SOD was given for
4 weeks, and at the end of the period difficulty of contact, concentration weariness, symptoms of stress,
and fatigue were improved in volunteers. It was concluded that Extramel shows antioxidant activity on
cellular level and fights against oxidative stress. An increase in plasma SOD level was observed in pigs
fed the diets supplemented with prediluted Promutase at concentration of 1 and 4 g/kg food, thus pro-
viding 12.5 and 50 IU SOD activity, respectively, at day 14 of the treatment. Supplementation resulted
in a decrease in heat shock proteins (HSP-27, HSP-70, and HSP-90) and neuronal nitric oxide synthase
(nNOS) in the gastrointestinal system of the pigs. The HSP-27, HSP-70, and HSP-90 are highly con-
served and ubiquitous proteins that limit aggregation of nonnative proteins and favor their refolding
and intracellular transport. Melon juice concentrate lowered the levels of all stress proteins as a result
of decreased oxidative stress in the stomach, whereas it decreased HSP-27 (nNOS in the mid–small
intestine) and HSP-70 (nNOS in the colon) [57].

Since conventional thermal processing techniques result in off-flavor formation, color, vitamin, and
aromatic compound degradation, nonthermal alternative pasteurization techniques may be applied
to have commercialized products for the processing of melon to fruit juices and concentrates.
In the application of ultrasound processing [58] and high-pressure processing [59], the use of cross
flow microfiltration and osmotic evaporation [13] seemed very promising ways to preserve some
quality attributes, including the color, browning degree, and most of the main aroma compounds.
Compared to fresh melons, processed melon juices showed decrease in vitamin C and FRAP values
as well as phenolic degradation (15%–35%). The processed melon juices might also show increase
in β-carotene contents in case of pressure application thought to be a result of enhanced extraction
by denaturation of the carotenoprotein complex [59] or improved cloud stability and uniformity
in case of sonication application, presumably by decreasing the size of suspended particles [58].
Although the use of gamma irradiation can improve the microbiological safety and inactivate the
enzymes to some extent, the development of off odor may preclude its use in the process of melon
juice production [60].
Other possible use of melon juice could be the production of fermented melon juice that may
not need special pasteurization step at the end. In the study of Fonteles et al. [61], cantaloupe juice
was used as a substrate for probiotic (Lactobacillus casei) fermentation, since the natural pH of
juice and its composition (natural sugars and amino acids) provide an enabling environment for
the development of probiotic microorganism. In addition, the fermentation process was able to
preserve the juice for 42 days under cold storage without any additive addition or the use of further
preservation method. Herein, it may be considered that melon juice is a suitable vehicle for lactic
acid bacteria (LAB), producing an alternative low-calorie nondairy probiotic product for vegans
as well as for consumers having lactose intolerance or milk protein allergies. Recently, it has been
patented that fermented cantaloupe juice provides an environment for folate producing probiotic
LAB strains (Lactobacillus reuteri or L. plantarum). Increased folate production was achieved dur-
ing microbial fermentation of cantaloupe melon juice with the use of p-aminobenzoic acid [62].
This natural folate fortified product serves as an alternative drink for consumers having vitamin
deficiencies.
Another approach to provide commercialized product types could be the production of melon juice
powder. There are three powdered commercial products, Extramel, Promutase 200, and GliSODin, which
are produced from cantaloupe possessing high antioxidant activity. In addition to these products, involve-
ment of encapsulation technologies may result in value-added food products. By using maltodextrin as a
carrier agent, spray-dried melon juice powder was successfully produced, which might be a good source
of β-carotene and vitamin C in a convenient form [63].
32.6 Conclusion
Melons are very popular and widely consumed fruits throughout the world and provide numerous bio-
active compounds that may contribute to human health, such as vitamins, minerals, and phenolic com-
pounds. However, their commercial products such as melon juice and concentrate are virtually unknown
and it is warranted to find novel processing techniques for the juice production as melon is highly per-
ishable due to its high water activity, mild acidity, and sensitivity to processing conditions. Moreover,
studies are scarce in terms of the effect of process conditions on bioactive compounds of melon and need
to be investigated. Accordingly, the studies dealing with the health effects of melon juice are limited
and the components that are responsible for the proposed mechanisms of action are not fully defined.
Furthermore, placebo-controlled, randomized, and double-blind human clinical trials are warranted in
addition to a vast number of animal or cell culture studies available in order to obtain more reliable results
to confirm health benefits.
Melon Juice 395
REFERENCES
1. Perkins-Veazie, P., Beaulieu, J.C., and Siddiq, M., Watermelon, cantaloupe and honeydew, in Tropical and
Subtropical Fruits: Postharvest Physiology, Processing and Packaging, Siddiq, M., Ed., Wiley-Blackwell,
Oxford, UK, 2012, pp. 549–568.
2. Galeb, A.D.S., Wrolstad, R.E., and McDaniel, M.R., Composition and quality of clarified cantaloupe
juice concentrate. J. Food Process. Preserv., 26, 39–56, 2002.
3. Lester, G.E., Antioxidant, sugar, mineral, and phytonutrient concentrations across edible fruit tis-
sues of orange-fleshed honeydew melon (Cucumis melo Var Reticulatus). J. Agric. Food Chem., 56,
3694–3698, 2008.
4. Lim, T.K., Benincasa hispida (Wax Gourd), in Edible Medicinal and Non-Medicinal Plants, Vol. 2,
Lim, T.K., Ed., Springer, Berlin, Germany, 2012, pp. 167–178.
5. Lim, T.K., Cucumis melo (Inodorus Group), in Edible Medicinal and Non-Medicinal Plants, Vol. 2, Lim,
T.K., Ed., Springer, Berlin, Germany, 2012, pp. 210–218.
6. Schaffer, A.A. and Paris H.S., Melons, squashes and gourds, in Encyclopedia of Food Sciences and
Nutrition, 2nd edn., Caballero, B., Ed., Academic Press, Oxford, UK, 2003, pp. 3817–3826.
7. Behera, T.K., Behera, S., Bharathi, L.K., John, K.J., Simon, P.W., and Staub, J.E., Bitter gourd: Botany,
horticulture, breeding, in Horticultural Reviews, vol. 37, Janick, J., Ed., John Wiley & Sons, Hoboken,
NJ, 2010, pp. 101–141.
8. Lester, G.E. and Hodges, D.M., Antioxidants associated with fruit senescence and human health:
Novel orange-fleshed non-netted honey dew melon genotype comparisons following different seasonal
productions and cold storage durations. Postharvest Biol. Technol., 48, 347–354, 2008.
9. Grover, J.K. and Yadav, S.P., Pharmacological actions and potential uses of Momordica charantia:
A review. J. Ethnopharmacol., 93, 123–132, 2004.
10. Laur, L.M. and Tian, L., Provitamin A and vitamin C contents in selected California-grown cantaloupe
and honeydew melons and imported melons. J. Food Comp. Anal., 24, 194–201, 2011.
11. Wolbang, C.M., Singh, D.P., Sykes, S.R., McInerney, J.K., Bird, A.R., and Treeby, M.T., Influence of
pre- and postharvest factors on β-Carotene content, its in vitro bioaccessibility, and antioxidant capacity
in melons. J. Agric. Food Chem., 58, 1732–1740, 2010.
12. Fleshman, M.K., Lester, G.E., Riedl, K.M., Kopec, R.E., Narayanasamy, S., Curley, R.W., Schwartz,
S.J., and Harrison, E.H., Carotene and novel apocarotenoid concentrations in orange-fleshed Cucumis
melo melons: Determinations of β-Carotene bioaccessibility and bioavailability. J. Agric. Food Chem.,
59, 4448–4454, 2011.
13. Vaillant, F., Cisse, M., Chaverri, M., Perez, A., Dornier, M., Viquez, F., and Dhuique-Mayer, C.,
Clarification and concentration of melon juice using membrane processes. Innov. Food Sci. Emerg.
Technol., 6, 213–220, 2005.
14. Chen, J., Zhang, J., Feng, Z., Song, L., Wu, J., and Hu, X., Influence of thermal and dense-phase carbon
dioxide pasteurization on physicochemical properties and flavor compounds in hami melon juice.
J. Agric. Food Chem., 57, 5805–5808, 2009.
15. Selale, H., Sıgva, H.O., Celik, I., Doganlar, S., and Frary, A., Water soluble antioxidant potential of
melon lines grown in Turkey. Int. J. Food Prop., 15, 145–156, 2012.
16. Abdullah, N., Syida, W.S., Kamarudin, W., Samicho, Z., Zulkifli, K.S., and Aziman, N., Study on anti-
oxidant capacity and phenolic content of various parts of wax gourd (Benincasa hispida). World Appl.
Sci. J., 191, 1051–1056, 2012.
17. Huang, H.Y., Huang, J.J., Tso, T.K., Tsai, Y.C., and Chang, C.K., Antioxidant and angiotensin-
converting enzyme inhibition capacities of various parts of Benincasa hispida (Wax Gourd). Food/
Nahrung, 48, 230–233, 2004.
18. Wu, S.J. and Ng, L.T., Antioxidant and free radical scavenging activities of wild bitter melon
(Momordica charantia Cucumis melo Var Reticulatus abbreviata Cucumis melo Var Reticulatus) in
Taiwan. LWT—Food Sci. Technol., 41, 323–330, 2008.
19. Nattaporn, W. and Pranee, A., Effect of pectinase on volatile and functional bioactive compounds in the
flesh and placenta of “Sunlady” cantaloupe. Int. Food Res. J., 18, 819–827, 2011.
20. Maietti, A., Tedeschi, P., Stagno, C., Bordiga, M., Travaglia, F., Locatelli, M., Arlorio, M., and
Brandolini, V., Analytical traceability of melon (Cucumis melo var. reticulatus): Proximate composi-
tion, bioactive compounds, and antioxidant capacity in relation to cultivar, plant physiology state, and
seasonal variability. J. Food Sci., 77, C646–C652, 2012.
21. Vinson, J.A., Su, X., Zubik, L., and Bose, P., Phenol antioxidant quantity and quality in foods: Fruits.
J. Agric. Food Chem., 49, 5315–5321, 2001.
22. Morales-Soto, A., García-Salas, P., Rodríguez-Pérez, C., Jiménez-Sánchez, C., de la Luz Cádiz-Gurrea, M.,
Segura-Carretero, A., and Fernández-Gutiérrez, A., Antioxidant capacity of 44 cultivars of fruits and
vegetables grown in Andalusia (Spain). Food Res. Int., 58, 35–46, 2014.
23. Wolfe, K.L., Kang, X., He, X., Dong, M., Zhang, Q., and Liu, R.H., Cellular antioxidant activity of
common fruits. J. Agric. Food Chem., 56, 8418–8426, 2008.
24. Rodríguez-Pérez, C., Quirantes-Piné, R., Fernández-Gutiérrez, A., and Segura-Carretero, A.,
Comparative characterization of phenolic and other polar compounds in Spanish melon cultivars by
using high-performance liquid chromatography coupled to electrospray ionization quadrupole-time of
flight mass spectrometry. Food Res. Int., 54, 1519–1527, 2013.
25. Kolayli, S., Kara, M., Tezcan, F., Erim, F.B., Sahin, H., Ulusoy, E., and Aliyazicioglu, R., Comparative
study of chemical and biochemical properties of different melon cultivars: Standard, hybrid, and grafted
melons. J. Agric. Food Chem., 58, 9764–9769, 2010.
26. Horax, R., Hettiarachchy, N., and Islam, S., Total phenolic contents and phenolic acid constituents in
4 varieties of bitter melons (Momordica charantia) and antioxidant activities of their extracts. J. Food
Sci., 70, C275–C280, 2005.
27. Kenny, O., Smyth, T.J., Hewage, C.M., and Brunton, N.P., Antioxidant properties and quantitative
UPLC-MS analysis of phenolic compounds from extracts of fenugreek (Trigonella foenum-graecum)
seeds and bitter melon (Momordica charantia) fruit. Food Chem., 141, 4295–4302, 2013.
28. Lucas, E.A., Dumancas, G.G., Smith, B.J., Clarke, S.L., and Arjmandi, B.H., Health benefits of bitter
melon (Momordica charantia), in Bioactive Foods in Promoting Health., Watson, R.R. and Preedy,
V.R., Eds., Academic Press, San Diego, CA, 2010, pp. 525–549.
29. Du, Q., Zhang, Q., and Ito, Y., Isolation and identification of phenolic compounds in the fruit of
Benincasa hispida by HSCCC. J. Liq. Chromatogr. Relat. Technol., 28, 137–144, 2005.
30. Palamthodi, S. and Lele, S.S., Nutraceutical applications of gourd family vegetables: Benincasa
hispida, Lagenaria siceraria, and Momordica charantia. Biomed. Prev. Nutr., 4, 15–21, 2014.
31. Zaini, N.A.M., Anwar, F., Hamid, A.A., and Saari, N., Kundur., Benincasa hispida (Thunb.) Cogn:
A potential source for valuable nutrients and functional foods. Food Res. Int., 44, 2368–2376, 2011.
32. Vouldoukis, I., Lacan, D., Kamate, C., Coste, P., Calenda, A., Mazier, D., Conti, M., and Dugas,
B., Antioxidant and anti-inflammatory properties of a Cucumis melo LC extract rich in superoxide
dismutase activity. J. Ethnopharmacol., 94, 67–75, 2004.
33. Carillon, J., Del Rio, D., Teissèdre, P.L., Cristol, J.P., Lacan, D., and Rouanet, J.M., Antioxidant c apacity
and angiotensin I converting enzyme inhibitory activity of a melon concentrate rich in superoxide
dismutase. Food Chem., 135, 1298–1302, 2012.
34. Pellegrini, N., Serafini, M., Colombi, B., Del Rio, D., Salvatore, S., Bianchi, M., and Brighenti, F., Total
antioxidant capacity of plant foods, beverages and oils consumed in Italy assessed by three different
in vitro assays. J. Nutr., 133, 2812–2819, 2003.
35. Shetty, B.V., Arjuman, A., Jorapur, A., Samanth, R., Yadav, S.K., Valliammai N., Tharian, A.D., Sudha K.,
and Rao, G.M., Effect of extract of Benincasa hispida on oxidative stress in rats with indomethacin
induced gastric ulcers. Indian J. Physiol. Pharmacol., 52, 178–182, 2008.
36. Naito, Y., Akagiri, S., Uchiyama, K., Kokura, S., Yoshida, N., Hasegawa, G., Nakamura, N. et al.,
Reduction of diabetes-induced renal oxidative stress by a cantaloupe melon extract/gliadin biopolymers,
oxykine, in mice. BioFactors, 23, 85–95, 2005.
37. Nerurkar, P.V., Lee, Y.K., Motosue, M., Adeli, K., and Nerurkar, V.R., Momordica charantia (bitter
melon) reduces plasma apolipoprotein B-100 and increases hepatic insulin receptor substrate and
phosphoinositide-3 kinase interactions. Br. J. Nutr., 100, 751–759, 2008.
38. Nerurkar, P.V., Lee, Y.K., and Nerurkar, V.R., Momordica charantia (bitter melon) inhibits primary
human adipocyte differentiation by modulating adipogenic genes. BMC Complement. Altern. Med., 10,
34–44, 2010.
39. Kaur, M., Deep, G., Jain, A.K., Raina, K., Agarwal, C., Wempe, M.F., and Agarwal, R., Bitter melon
juice activates cellular energy sensor AMP-activated protein kinase causing apoptotic death of human
pancreatic carcinoma cells. Carcinogenesis, 34, 1585–1592, 2013.
40. Chen, Q. and Li, E.T.S., Reduced adiposity in bitter melon (Momordica charantia) fed rats is associated
with lower tissue triglyceride and higher plasma catecholamines. Br. J. Nutr., 93, 747–754, 2005.
Melon Juice 397
41. Raza, H., Ahmed, I., Lakhani, M.S., Sharmu, A.K., Pallot, D., and Montague, W., Effect of bitter
melon (Momordica charantia) fruit juice on the hepatic cytochrome P450-dependent monooxygen-
ases and glutathione S-transferases in streptozotocin-induced diabetic rats. Biochem. Pharmacol., 52,
1639–1642, 1996.
42. Tan, M.J., Ye, J.M., Turner, N., Hohnen-Behrens, C., Ke, C.Q., Tang, C.P., Chen, T. et al., Antidiabetic
activities of triterpenoids isolated from bitter melon associated with activation of the AMPK pathway.
Chem. Biol., 15, 263–273, 2008.
43. Singh, J., Hundal, H.S., Wackerhage, H., Hope, M., Belle, M., Cummings, E., and Adeghate, E.,
Momordica charantia fruit juice stimulates glucose and amino acid uptakes in L6 myotubes. Mol. Cell.
Biochem., 261, 99–104, 2004.
44. Ahmed, I., Adeghate, E., Cummings, E., Sharma, A.K., and Singh, J., Beneficial effects and mechanism
of action of Momordica charantia juice in the treatment of streptozotocin-induced diabetes mellitus in
rat. Mol. Cell. Biochem., 261, 63–70, 2004.
45. Sitasawad, S.L., Shewade, Y., and Bhonde, R., Role of bittergourd fruit juice in STZ-induced diabetic
state in vivo and in vitro. J. Ethnopharmacol., 73, 71–79, 2000.
46. Ahmed, I., Lakhani, M.S., Gillett, M., John, A., and Raza H., Hypotriglyceridemic and hypocholes-
terolemic effects of anti-diabetic Momordica charantia (karela) fruit extract in streptozotocin-induced
diabetic rats. Diabetes Res. Clin. Pract., 51, 155–161, 2001.
47. Décordé, K., Agne, A., Lacan, D., Ramos, J., Fouret, G., Ventura, E., Feillet-Coudray, C., Cristol,
J.P., and Rouanet, J.M., Preventive effect of a melon extract rich in superoxide scavenging activity on
abdominal and liver fat and adipokine imbalance in high-fat-fed hamsters. J. Agric. Food Chem., 57,
6461–6467, 2009.
48. Chan, L.L.Y., Chen, Q., Go, A.G.G., Lam, E.K.Y., and Li, E.T.S., Reduced adiposity in bitter melon
(Momordica charantia)-fed rats is associated with increased lipid oxidative enzyme activities and
uncoupling protein expression. J. Nutr., 135, 2517–2523, 2005.
49. Nerurkar, P.V., Pearson, L., Efird, J.T., Adeli, K., Theriault, A.G., and Nerurkar, V.R., Microsomal
triglyceride transfer protein gene expression and ApoB secretion are inhibited by bitter melon in HepG2
cells. J. Nutr., 135, 702–706, 2005.
50. Ru, P., Steele, R., Nerurkar, P.V., Phillips, N., and Ray, R.B., Bitter melon extract impairs prostate cancer
cell-cycle progression and delays prostatic intraepithelial neoplasia in TRAMP model. Cancer Prev.
Res., 4, 2122–2130, 2011.
51. Ray, R.B., Raychoudhuri, A., Steele, R., and Nerurkar, P., Bitter melon (Momordica charantia) extract
inhibits breast cancer cell proliferation by modulating cell cycle regulatory genes and promotes apopto-
sis. Cancer Res., 70, 1925–1931, 2010.
52. Décordé, K., Ventura, E., Lacan, D., Ramos, J., Cristol, J.P., and Rouanet, J.M., An SOD rich melon
extract Extramel® prevents aortic lipid and liver steatosis in diet induced model of atherosclerosis. Nutr.
Metab. Cardiovasc. Dis., 20, 301–307, 2010.
53. Nakashima, M., Shigekuni, Y., Obi, T., Shiraishi, M., Miyamoto, A., Yamasaki, H., Etoh, T., and Iwai, S.,
Nitric oxide-dependent hypotensive effects of wax gourd juice. J. Ethnopharmacol., 138, 404–407,
2011.
54. Lin, J.Y. and Tang, C.Y., Strawberry, loquat, mulberry, and bitter melon juices exhibit prophylactic
effects on LPS-induced inflammation using murine peritoneal macrophages. Food Chem., 107, 1587–
1596, 2008.
55. Carillon, J., Romain, C., Bardy, G., Fouret, G., Feillet-Coudray, C., Gaillet, S., Lacan, D., Cristol, J.P.,
and Rouanet, J.M., Cafeteria diet induces obesity and insulin resistance associated with oxidative
stress but not with inflammation: Improvement by dietary supplementation with a melon superoxide
dismutase. Free Rad. Biol. Med., 65, 254–261, 2013.
56. Milesi, M.A., Lacan, D., Brosse, H., Desor, D., and Notin, C., Effect of an oral supplementation with a
proprietary melon juice concentrate Extramel® on stress and fatigue in healthy people: A pilot, double-
blind, placebo controlled clinical trial. Nutr. J., 8, 40–46, 2009.
57. Lallès, J.P., Lacan, D., and David, J.C., A melon pulp concentrate rich in superoxide dismutase reduces
stress proteins along the gastrointestinal tract of pigs. Nutrition, 27, 358–363, 2011.
58. Fonteles, T.V., Costa, M.G.M., de Jesus, A.L.T., de Miranda, M.R.A., Fernandes, F.A.N., and Rodriguez,
S., Power ultrasound processing of cantaloupe melon juice: Effects on quality parameters. Food Res.
Int., 48, 41–48, 2012.
59. Wolbang, C.M., Fitos, J.L., and Treeby, M.T., The effect of high pressure processing on nutritional value
and quality attributes of Cucumis melo L. Innov. Food Sci. Emerg. Technol., 9, 196–200, 2008.
60. Wang, Z., Ma, Y., Zhao, G., Liao, X., Chen, F., Wu, J., Chen, J., and Hu, X., Influence of gamma
irradiation on enzyme, microorganism, and flavor of cantaloupe (Cucumis melo L.) juice. J. Food Sci.,
71, M215–M220, 2006.
61. Fonteles, T., Costa, M.G.M., de Jesus, A.L.T., Lima Fontes, C.P.M., Fernandes, F.A.N., and Rodriguez, S.,
Optimization of the fermentation of cantaloupe juice by Lactobacillus casei NRRL B-442. Food
Bioprocess Technol., 5, 2819–2826, 2012.
62. Wegkamp, H.B.A., dos Santos, F.B., Smid, E.J., and Hugenholtz, J., Increased folate production levels
in Lactobacillus fermenting melon juice, U.S. Patent No. 8,524,297 B2, 2013.
63. Solval, K.M., Sundararajan, S., Alfaro, L., and Sathivel, S., Development of cantaloupe (Cucumis melo)
juice powders using spray drying technology. LWT—Food Sci. Technol., 46, 287–293, 2012.
33
Mulberry Juice
Meltem Türkyılmaz and Mehmet Özkan
CONTENTS
33.1 Introduction.................................................................................................................................... 399
33.2 Nutritional Characteristics............................................................................................................. 400
33.4 Health Effects................................................................................................................................. 403
33.5 Novel Products/Formulations and Future Trends.......................................................................... 405
33.6 Conclusion..................................................................................................................................... 406
References............................................................................................................................................... 406
33.1 Introduction
The genus Morus, commonly known as mulberry, is mostly grown in Asia, North Africa, Arabia, and
South Europe, and has more than 150 species [1,2]. Among them, Morus alba (white mulberry), M. rubra
(red mulberry), and M. nigra (black mulberry) are the best known species [1,3]. Harvest time and shelf
life of these popular species, similar to other mulberry species, are very short. Therefore, to benefit
from the fruits throughout the year, the fruits are processed to products such as juice, marmalade, and
wine, which have a longer shelf life. According to current consumer trends, mulberry juice is the most
preferred among the mulberry products. The juice yield of mulberry is no less than 50% for the common
varieties and more than 60% for the seedless mulberry [4]. Mulberry is one of the most suitable fruits for
juice production due to its high yield and appropriate sugar to acid ratio.
Red and black mulberries are often preferred species to white mulberry for the production of juice as
well as purée by fruit juice industry due to their attractive colors resulting from anthocyanins and their
higher juice yields. The yield of juice from red and black mulberries ranges from 3.3 to 11.0 L/tree, while
that from white mulberry 2.8 L/tree [4]. In recent years, there has been a steady increase in the produc-
tion and consumption of red and black mulberry juices. Especially in China, Japan, and Korea, mulberry
juice commercially produced as a health beverage has become very popular [4]. The high demand for
mulberry juice is the result of studies that have shown high contents of bioactive compounds in mulberry.
The primary bioactive compounds in mulberry juice are the colorless polyphenols, anthocyanins, and
ascorbic acid. Mulberry juice provides significant health benefits originating from the presence of the bio-
active compounds with high antioxidant activity [5]. In addition to their high antioxidant activity, mulberry
juice possesses antimicrobial [6], antiatherosclerotic, antiinflammatory [7], anti-HIV, antistress [8],
a nd antifatigue effects [5]. Moreover, although consumption of 100% fruit juice increases the risk
of obesity, a previous study indicated that mulberry juice could be used to help counteract obesity [7].
This chapter highlights the nutritional characteristics and phytochemical content of mulberry juice, its
antioxidant capacity, health benefits, as well as novel formulations that may improve the aforementioned
functional properties.
399

The nutritional characteristics of mulberry juices depend on mulberry species and environmental condi-
tions where mulberries are grown in China, India, Italy, and Turkey. However, despite the differences
between the nutritional characteristics, all species from growing areas include appreciable amount of
carbohydrate, minerals, and vitamins as well as low contents of lipid and protein [1,3,9,10] (Table 33.1).
Similar to mulberry fruits that contain a high amount of carbohydrate (8.3–12.2 g/100 g) [1], their
corresponding juices also contain a high percentage of carbohydrate (16–28 g/100 mL) [11,12]. Mulberry
juice obtained from black specie contained 28 g carbohydrate/100 mL [12], which provides about 9%
of the Recommended Dietary Allowances (RDA) of carbohydrate for adults (300 g, based on 2000 cal).
Invert sugar and sucrose contents of mulberry juices also depend on mulberry species. For example,
white mulberry has the highest contents of invert sugar (16.5 g/100 g) and sucrose (1.5 g/100 g), while
red mulberry has the lowest contents of invert sugar (11.9 g/100 g) and sucrose (0.9 g/100 g) [9].
Mulberry juice is also rich in potassium (159–1300 mg/100 mL), calcium (7.4–150 mg/100 mL), and
iron (1.14–40 mg/100 mL) [6,13]. On the contrary, low sodium (9.8 mg/100 mL) [14] content was reported
in mulberry juice. According to these values, 100 mL mulberry juice could supply 4%–37% of the RDA
for potassium for adults.
Ascorbic acid content of mulberry is also quite high (10.1–23.4 mg/100 mg) [3,6,15], 100 g of which
would supply 17%–39% of the RDA for adults and that of mulberry juice 4.3–23 mg/100 mL [6,13].
However, processing method used for mulberry juice production had a significant effect on its ascorbic
acid content. For example, 92% of ascorbic acid was retained in mulberry juice upon thermal pasteuriza-
tion (95°C, 1 min), while only 59% of it was retained after ultrahigh pressure homogenization (UHPH)
[16]. This study showed that ascorbic acid in mulberry juice was more sensitive to ultrahigh pressure
than thermal processing.
TABLE 33.1
Compositional and Nutritional Characteristics of Mulberry Juice (per 100 mL)
Unit RDA (mg, Based on 2000 cal) Content References
Water g na 93 [13]
Energy kcal na 66 [11]
Lipid g 65 0–2.15 [11,13]
Protein g 50 0–1.17 [11,13]
Carbohydrate g 300 16.0–28.0 [11]
Total sugar g 300 4.5–28.0 [11,13,23]
Total dietary fiber g 25 0–2.4 [11,12]
Minerals
Calcium mg 1000 7.4–150 [6,13]
Iron mg 15 1.14–40 [13,14]
Magnesium mg 400 130 [6]
Manganese mg 2.0 7 [14]
Potassium mg 3500 159–1300 [6,13]
Sodium mg 2400 9.8 [14]
Zinc mg 15 45 [14]
Vitamins
Ascorbic acid mg 60 4.3–23 [6,13]
Folic acid mg na 0.01 [13]
Pyridoxine mg 2.0 0.02 [13]
Riboflavin mg 1.6 0.08 [13]
Thiamine mg 1.4 0.04 [13]
Abbreviations: RDA, Recommended Dietary Allowances; na, not available.
Mulberry Juice 401
Mulberry juice had a low concentration of B complex vitamins such as nicotinic acid (1.2 mg/100 mL),
thiamine (0.04 mg/100 mL), and riboflavin (0.08 mg/100 mL).

Bioactive compounds are minor constituents of fruits and vegetables. The major bioactive compounds
in mulberry juices are colorless polyphenols, anthocyanins, and ascorbic acid. Although mulberries are
good sources of carotenoids and tocopherols, the bioactive compounds that are lipid soluble are low or
in some cases below the limits of detection in mulberry juices. The contents of bioactives in mulberries
and their juices are given in Table 33.2.
TABLE 33.2
Bioactives of Mulberries and Their Juices
Unit Mulberries Unit Mulberry Juices References
Total Polyphenol Contents mg GAE/100 g 181–2237 mg GAE/L 2398–2830 [18]
Total Flavonoid Contents mg QE/kg 290–2760 mg QE/L 86.6–1913.0 [3,18,19]
Quercetin 3-O-rutinoside mg QRE/kg 1270–3290 mg QRE/L 33.5–1330.4 [19]
Quercetin 3-O-glucoside mg QRE/kg 290–340 — — [21]
Quercetin 3-O-galactoside mg QRE/kg 0.9 — — [20]
Quercetin 3-O(6″-acetyl)glucoside mg QGE/kg 1.2 — — [20]
Quercetin 3-O(6″-acetyl)galactoside mg QGAE/kg 18.3 — — [20]
Kaempferol 3-O-rutinoside mg KRE/kg 150–270 — — [21]
Kaempferol 3-O-galactoside mg KGAE/kg 0.53 — — [21]
Kaempferol 3-O(6″-acetyl) mg KGE/kg 1.1 — — [22]
glucoside
Phenolic Acids
Gallic acid — — mg GAE/L 129 [16]
Protocatechuic acid — — mg PAE/L 202 [16]
Caffeic acid — — mg CAE/L 435 [16]
Coumaric acid — — mg COAE/L 82 [16]
Total Monomeric Anthocyanin mg CGE/L 22–3300 mg CGE/L 148–2745 [18,19,22]
Contents
Cyanidin 3-O-glucoside mg CGE/kg 1790 mg CGE/L 906–1233 [18,21]
Cyanidin 3-O-rutinoside mg CGE/kg 750 mg CRE/L 624–1056 [18,21]
Pelargonidin 3-O-glucoside mg CGE/kg 120 mg PGE/L 53–109 [18,21]
Pelargonidin 3-O-rutinoside mg CGE/kg 40 — — [18,21]
Carotenoids
Lutein mg/kg 13.2–28.9 — — [17]
β-Carotene mg/kg 4.7–7.7 — — [17]
Neoxanthin mg/kg 0.6–1.4 — — [17]
Tocopherols
δ-Tocopherol mg/kg 363–1140 — — [17]
γ-Tocopherol mg/kg 225–426 — — [17]
α-Tocopherol mg/kg 2.14–17.9 — — [17]
Abbreviations: CAE, caffeic acid equivalents; CGE, cyanidin-3-O-glucoside equivalents; COAE, coumaric acid equiva-
lents; CRE, cyanidin 3-O-rutinoside equivalents; GAE, gallic acid equivalents; KGAE, kaempferol
3-O-galactoside equivalents; KGE, kaempferol 3-O(6″-acetyl)glucoside equivalents; KRE, kaempferol
3-O-rutinoside equivalents; PAE, protocatechuic acid equivalents; PGE, pelargonidin 3-O-glucoside equiva-
lents; QE, quercetin equivalents; QGAE, quercetin 3-O(6″-acetyl)galactoside equivalents; QGE, quercetin
3-O(6″-acetyl)glucoside equivalents; QRE, quercetin 3-O-rutinoside equivalents.
Mulberry juice has a high amount of total polyphenol (2398–2830 mg gallic acid equivalents (GAE)/L)
[18] and total flavonoid (86.6–1913.0 mg quercetin equivalents (QE)/L) [19]. However, processing steps
affect the total polyphenol content of mulberry juice. While depectinization leads to slight increase (3%)
[18], clarification with bentonite, gelatin, and kieselsol may decrease the total polyphenol content of mul-
berry juice (8%) [18]. The reason for the reduction is that polyphenols are also removed from mulberry
juice as a result of the interaction of polyphenols with gelatin–kieselsol flocks. Therefore, to produce
mulberry juice with high polyphenol content, after depectinization, the dosages of clarification agents
should be carefully determined.
In the literature, there are a few studies investigating polyphenol profiles of mulberries [20,21]
and their juices [16], and there were significant variations between polyphenol profiles of mulber-
ries and their juices reported. Although only a hydroxycinnamic acid (5-O-caffeoylquinic acid)
and eight flavonoids (quercetin 3-O-rutinoside [1270–3290 mg quercetin 3-O-rutinoside equiv-
alents (QRE)/kg], quercetin 3-O-glucoside [290–340 mg QRE/kg], kaempferol 3-O-rutinoside
[150–270 mg QRE/kg], kaempferol 3-O-galactoside [0.53 mg kaempferol 3-O-galactoside equiva-
lents (KGAE)/kg], kaempferol 3-O(6″-acetyl)glucoside [1.1 mg kaempferol 3-O(6″-acetyl)glucoside
equivalents (KGE)/kg], quercetin 3-O-galactoside [0.9 mg QRE/kg], quercetin 3-O(6″-acetyl)gluco-
side [1.2 mg quercetin 3-O(6″-acetyl)glucoside equivalents (QGE)/kg], and quercetin 3-O(6″-acetyl)
galactoside [18.3 mg quercetin 3-O(6″-acetyl)galactoside equivalents (QGAE)/kg)]) were charac-
terized in mulberries, five different phenolic acids and a flavonoid were tentatively identified in
different mulberry juices [16]. Two phenolic acids in mulberry juice were hydroxybenzoic acids
(gallic acid [129 mg GAE/L] and protocatechuic acid [202 mg protocatechuic acid equivalents
(PAE)/L]), while the others were hydroxycinnamic acids (caffeic acid [435 mg caffeic acid equiva-
lents (CAE)/L], p-coumaric acid [82 mg p-coumaric acid equivalents (PCAE)/L], and unknown
phenolic acids with p-coumaric acid ultraviolet spectrum [161 mg PCAE/L]). The major phenolic
acid in mulberry juice was caffeic acid (435 mg CAE/L). Moreover, quercetin aglycone (81 mg/L)
was also determined in mulberry juice [16]. However, its content was lower than those of phenolic
acids in mulberry juice. Structures of common phenolic compounds found in mulberry juice are
shown in Figure 33.1.
There were significant effects of thermal pasteurization and UHPH on phenolic acid profiles of mul-
berry juice. A study investigating the effects of UHPH and thermal pasteurization on phenolic acids in
mulberry juice showed that the contents of gallic acid, protocatechuic acid, and caffeic acid increased
66%, 24%, and 40%, respectively, after UHPH [16], possibly due to the release of phenolic acids that
also exist as insoluble bound complexes, which are coupled to cell wall polymers through ester and
glycosidic linkages [16]. Contrary to UHPH, there were significant reductions (P < 0.05) in the contents
O
O
OH
OH OH
OH HO
Caffeic acid p-Coumaric acid
OH
OH
O
OH HO O OH
OH O
OH O OH
OH O
OH HO OH
OH OH O
OH
Gallic acid Protocatechuic acid Quercetin 3-O-glucoside
FIGURE 33.1 Structures of common phenolic compounds found in mulberry juice.

Mulberry Juice 403
of gallic acid (68%), protocatechuic acid (51%), caffeic acid (14%), and p-coumaric acid (23%) after
thermal pasteurization. Therefore, instead of thermal pasteurization, UHPH of mulberry juice was
suggested by Yu et al. [16].
The other important phenolic group in mulberry juice is anthocyanins with attractive colors. The con-
tent of total anthocyanin in mulberry juice varied depending on genotype, cultivar, and environmental
conditions. For example, total anthocyanin contents of different mulberry cultivars grown in China ranged
from 22 to 3300 mg cyanidin 3-O-glucoside equivalents (CGE)/L [19], while total anthocyanin contents
of raw black mulberry juices in Turkey and Iran were 1609 [18] and 164 mg CGE/L [22], respectively.
In addition to the aforesaid factors, juice processing also affects the total anthocyanin content in mulberry
juice since the stabilities of anthocyanins to enzymes (especially polyphenoloxidase), oxygen, heat, and light
are very low. In fact, depectinization (10%) [18], clarification (bentonite, gelatin, and kieselsol) (12%) [18],
thermal pasteurization (14%) [23], and storage (57% at 20°C for 138 days) [18] lead to decreases in total
monomeric anthocyanin contents of mulberry juices. Thus, to prevent the anthocyanins from degradation,
production and storage conditions of mulberry juices should also be carefully examined.
The major anthocyanin in black mulberry varieties and their juices is cyanidin 3-O-glucoside (1790 mg
CGE/kg fresh weight [FW] and 906–1233 mg CGE/L juice), followed by cyanidin 3-O-rutinoside (750 mg
CGE/kg FW and 624–1056 mg cyanidin 3-O-rutinoside equivalents [CRE]/L juice), pelargonidin
3-O-glucoside (120 mg CGE/kg FW and 53–109 mg pelargonidin 3-O-glucoside equivalents [PGE]/L
juice), and pelargonidin 3-O-rutinoside (40 mg CGE/kg FW and not determined in juice), respectively
[18,21]. Different from that of black mulberry varieties, anthocyanin profile of white mulberry varieties
consists of five anthocyanins, only two of which (cyanidin 3-O-glucoside and cyanidin 3-O-rutinoside)
are the same with those of black mulberry varieties. The others are cyanidin 3-O-rhamnosylgalactoside,
cyanidin 3-O-galactoside, and cyanidin 7-O-glucoside [24]. The contents of the individual anthocyanins
in white mulberry fruits have not yet been determined.
Mulberries and their juices have strong antioxidant activity (13.95–16.01 mM trolox equivalents
[TE]/mL) [18]. The high antioxidant activities of mulberries and their juices are attributed to their bioac-
tive compounds and phenolic compounds were the main antioxidant compounds in mulberry juices.
In fact, there are strong correlations between antioxidant activities with the contents of total polyphenols
(r = 0.9337) [24], total flavonoids (r = 0.9402) [5], and total anthocyanins ([r = 0.8450 for total anthocya-
nins] [24], [r = 0.8987 for cyanidin 3-O-rutinoside] [18], [r = 0.8795 for pelargonidin 3-O-glucoside] [16],
[r = 0.7451 for cyanidin 3-O-glucoside]) [18] of mulberry juices. The correlation constants indicated that
the strongest correlation (r = 0.9402) [5] existed between total flavonoids and antioxidant activity. Among
flavonoids of mulberry juices, quercetin had the highest TE antioxidant activity and its antioxidant activ-
ity was 3.5-fold higher than that of kaempferol [25]. Antioxidative effect of polyphenols in mulberries
and mulberry juices mainly results from glucosides of quercetin, especially quercetin 3-O-rutinoside.
Processing methods also affect the antioxidant activity of mulberry juice depending on differences in
the contents of bioactive compounds. During thermal pasteurization (11%) and UHPH (58%), the oxygen
radical absorbance capacity (ORAC) values of mulberry juices were significantly reduced [16]. The
reduction of ORAC value in mulberry juice processed by both methods was mainly attributed to the loss
of phenolic acids and anthocyanins due to oxidation and degradation. Similarly, depectinization (11%)
and clarification with bentonite, gelatin, and kieselsol (13%) also lead to reduction in antioxidant activ-
ity of mulberry juice because of anthocyanin degradation effects of β-glucosidase and β-galactosidase
activities in depectinization enzymes used in the process and loss of phenolic acids and anthocyanins [6].
33.4 Health Effects

Mulberries and mulberry juices are good sources of anthocyanins and other flavonoids that have
been suggested to be responsible for health benefits. White mulberry has been used exclusively
in Chinese medicine since 659 AD. Similarly, mulberry juice is also listed as an official drug in
the British Pharmacopoeia [1]. Mulberries and mulberry juices improve the health by enhanc-
ing immunity, balancing internal secretions, enriching the blood, calming nerves, and promoting
the metabolism of alcohol [1]. To date, antimicrobial [6], anti-HIV [8], antihyperglycemic [26],
antiatherogenic [7], antihyperlipidemic [7], antistress [8], hepatoprotective [19], nephroprotective [27],

and immunomodulatory [28] activities of mulberries and their juices have been revealed. In this
chapter, the effects of consumption of mulberry juice against the most common diseases among
people such as atherosclerosis, diabetes, and cancer are reviewed.
Anthocyanins and other flavonoids have significant effects on the prevention of coronary heart disease
(CHD). In fact, there is an inverse association between flavonoid intake and risk of CHD [29,30] and
cancer [30]. A high intake of flavonoids (approximately 30 mg/day) caused approximately 50% reduction
in CHD mortality rate compared with the individuals who had a low flavonoid intake (19 mg/day) [29].
Taking into consideration the high flavonoid intake, on average 20 mL/day of mulberry juice and 13 g/day
of mulberry fruits, may significantly reduce the mortality rate from CHD. Moreover, mulberry juice is
effective in stopping atherosclerosis, as it inhibits oxidative modification of low-density lipoprotein (LDL)
via antioxidant activity of their polyphenols such as anthocyanin and quercetin glycosides. Similarly,
dietary intake of mulberry juice also reduced the absorption of blood glucose [26]. The effects of black
and white mulberry juices on blood glucose, lipid profile, and oxidative stress of normal and diabetic
rats were investigated by Khalil [31]. In their study, diabetic rats were injected intraperitoneally with
100 mg/kg body weight of alloxan. Serum glucose in diabetic control groups was 3.8-fold higher than that
of normal control group. Mulberry juices from black, white, and their mixture led to 45%, 20%, and 37%
reductions, respectively, in serum glucose in diabetic rats.
The suppression of the increase in postprandial glucose is especially attributed to the combination
of polyphenols and 1-deoxynojirimycin (DNJ), which is a polyhydroxylated piperidine alkaloid present
in mulberry juice. DNJ is one of the most potent α-glycosidase inhibitors and helps to establish greater
glycemic control in type 2 diabetes [1]. Although white and black mulberries are very rich in DNJ (rang-
ing from 1389 to 3483 mg/kg), mulberry juice contains much less DNJ (34–346 mg/L) [19] than its fruit,
but it still showed 82% inhibition of α-glycosidase. The low concentration of DNJ in mulberry juice may
be attributable to the processing steps during juice production. In a study conducted by Yu et al. [16],
thermal pasteurization led to 14% reduction in α-glycosidase inhibitory activity of mulberry juice, while
no significant change (P > 0.05) in the α-glycosidase inhibitory activity was observed during UHPH
processing of mulberry juice. Thus, to produce mulberry juice with high DNJ content, UHPH processing
is preferred.
In recent years, the possibility of preventing the onset of obesity as well as diabetes using natural dietary
supplements containing DNJ has been investigated. For this purpose, there has been an intense interest
in mulberries as well as the mulberry juices. In a study investigating the effect of administration of mulberry
juice on the development of obesity in mice fed with a high fat diet, mulberry juice intake reduced body
weight by 9.8% [7]. Moreover, mulberry juice supplementation could significantly (P < 0.05) decrease
total cholesterol (26%), triacylglycerols (TAG) (36%), high-density lipoprotein (HDL) cholesterol (23%),
glucose (30%), aspartate aminotransferase (33%), and alanine aminotransferase (27%) in blood serum.
Thus, mulberry juice has considerable hypoglycemic and hypocholesterolemic effects and may also be
used to counteract obesity [7].
In addition to the preventive effect of mulberry juice on obesity development, mulberry juice treatment
also increases the antioxidative compounds. After mulberry juice treatment, glutathione peroxidase and
superoxide dismutase concentrations increased in diabetic rats [31]. Moreover, mulberry juice led to a
significant reduction in oxidative stress, which was manifested by the decrease in the level of malondial-
dehyde and nitric oxide. When mice were subjected to water immersion restraint stress at 25°C for 8 h,
the plasma lipid peroxide level was almost doubled. Mulberry juice treatment completely arrested lipid
peroxidation [32].
Sakagami et al. [8] separated mulberry juice into two fractions by centrifugation in order to examine
their antistress activities. The supernatant (containing approximately 22% of total fiber) fraction had a
similar preventive effect to mulberry juice, while the precipitate (containing approximately 78% of total
fiber) fraction was slightly less active. Thus, the results showed that the antistress activity was mainly
retained in the supernatant fraction. In the same study [8], the anti-HIV activity was recovered from the
precipitate fraction containing most of the fiber. Therefore, the precipitate was extracted by using dif-
ferent solvents (hot water, sodium hydroxide, ethanol, and acetic acid), and the anti-HIV effects of six
Mulberry Juice 405
different fractions were investigated. Lignin fractions among the six fractions of mulberry juice showed
the highest anti-HIV activity, although this activity was much lower than that of popular anti-HIV agents
(dextran sulfate, curdlan sulfate, 3′-azidothymidine, and 2′-3′dideoxycytidine) [8].
Another important functionality of mulberry juice is its antimicrobial effects [6]. Thus, black mul-
berry juice was especially shown to serve as a potential candidate for phytomedicine and may be an
effective antibiotic against certain bacteria [6].

Epidemiologic studies have indicated that many chronic diseases in humans are diet related. Therefore,
in recent years, consumers have demanded healthier food products containing bioactive compounds,
while food industry has researched novel products/formulations and processing technologies, which will
meet the consumer’s demand. The fruit juice industry has also shown much interest in novel products,
since the consumption of fruit or fruit-based products, including mulberries, is an important part of any
diet providing health benefits.
The contents of bioactive compounds in mulberry juice were significantly affected by juice processing.
Especially, anthocyanins (14%) [23] and ascorbic acid (8%) [16] in mulberry juice were negatively affected
by conventional thermal pasteurization, which is one of the most important steps in juice processing
because of its ability to inactivate microorganisms and spoilage enzymes (such as polyphenol oxidase
and pectin methylesterase). Contrary to conventional thermal pasteurization, it has been reported that
novel nonthermal processing methods, a good alternative to thermal pasteurization, provide high-quality
products with “fresh-like” characteristics [33]. Therefore, the effects of nonthermal pasteurization meth-
ods such as power ultrasound, irradiation, pulsed electric fields, ozone, and oscillating magnetic fields on
the bioactive compounds of mulberry juices should also be determined in future studies. The most suit-
able pasteurization method to minimize anthocyanin and ascorbic acid losses should also be revealed.
Today, the most important defect in commercially produced mulberry juice by conventional juice
processing methods is the precipitate formation in consumer packages. To prevent this formation, new
juice processing methods, especially new clarification agents, have been investigated. In the literature,
pectinase, chitosan, and polyvinylpolypyrrolidone for the clarification of mulberry juice were used [34].
Consequently, chitosan was recommended for the production of clear mulberry juice since the transmit-
tance of clarified mulberry juice was over 95%. However, the precipitate formation during storage could
not be totally prevented by the use of chitosan.
Another new method for investigating the prevention of precipitate formation is the deacidification of
mulberry juice by electrodialysis. Since the precipitate occurs at the pH of mulberry juice (pH 3.3–3.4),
Vera et al. [35] reported that the pH value of mulberry juice could be increased to 4.0–4.5 by using
electrodialysis, and this would significantly reduce the precipitate formation in the juice. With electrodi-
alysis, the pH of mulberry juice can be increased up to 7.9 [35]. At this alkaline pH, mulberry juice can
be added to high pH foods such as milk products where the precipitation of casein can otherwise occur
at acidic pHs. By deacidification of mulberry juice, a new functional beverage, such as milk containing
mulberry juice without casein precipitation, can be produced.
Consumer demand for the blend juices has significantly increased in recent years [36]. Up to now,
mulberry juice has not commonly been used in blend juice. However, due to consumer interest in health
benefits of fruit juices, it has been suggested that mulberry juice can be used in blended juice products.
Compared with the properties of individual juices, blend juice has not only different organoleptic prop-
erties but also different bioactives due to synergistic, additive, or antagonistic effects of different com-
pounds in each juice on total antioxidant activity. The synergistic effect of the combinations is important
for developing functional beverages with enhanced antioxidant activity [37]. However, the combination
of some foods may have antagonistic effect on the antioxidant activity. Therefore, to provide synergis-
tic effect, appropriate foods and their combinations should be strategically selected. For example, a
study showed that polyphenols could act synergistically with lutein, lycopene, and β-carotene [38]. Thus,
the blend of mulberry and purple carrot juices may be a very good source of antioxidants since both
polyphenol and carotenoid contents (176 µg lutein/100 g FW, 318 µg β-carotene/100 g FW, and 4 93 µg
total carotenoid/100 g FW) of the purple carrots were high [39].
Mulberry juice can also be used in making wine, fruit sauce, and cake and as food colorant in milk
products such as ice cream and yogurt [4]. In Europe, mulberry wine is very popular as a ladies drink.
In Azerbaijan, Georgia, and Armenia, a liqueur (Tut araghi) made from mulberry fruit juice is one of
the most preferred drinks. Moreover, in Greece, mulberry fruit juice is often used for the production
of the traditional aromatic mouro distillate [4].
Fruit juice powders are also another novel and remarkable products since they have many benefits and
economic potentials over their liquid counterparts such as reduced volume or weight, reduced packag-
ing, and easier handling and transportation [40]. Moreover, the spray-drying method used for fruit juice
powder production also provides much longer shelf life. One of the economically feasible methods for
the preservation of anthocyanins, whose stability is very low against many factors such as temperature,
light, oxygen, pH, and enzymes, is “microencapsulation” by using spray-drying.
The spray-drying conditions, for example, inlet air temperature, compressed airflow rate, and carrier
agent concentration, had a significant effect on mulberry juice powder drying yield, moisture content,
water activity, bulk density, and solubility. According to the results of the study conducted by Fazaeli
et al. [40], higher inlet air temperature caused an increase in mulberry juice powder yield and solubility
and a decrease in bulk density, moisture content, and water activity. Moreover, increasing the com-
pressed airflow rate had a positive effect on drying yield and bulk density and a negative effect on
solubility, moisture content, and water activity. Compared with maltodextrin 20DE and gum arabic,
maltodextrin 6DE was the best carrier agent for mulberry juice [40].
33.6 Conclusion
Mulberry juice can be considered as a functional beverage since it contains significant levels of bioactive
compounds that render functional and beneficial health effects. Mulberry juices are especially rich in colorless
polyphenols, anthocyanins, and alkaloids. These compounds have antimicrobial, anti-HIV, antihyperglyce-
mic, antiatherogenic, antihyperlipidemic, antistress, hepatoprotective, neophroprotective, neuroprotective, and
immunomodulatory properties. Due to the properties of the bioactive compounds, mulberry juices provide
protection for many diseases such as atherosclerosis, diabetes mellitus, cancer, neurodegenerative diseases,
and depression. Other than bioactive compounds, mulberry juices also contain high amounts of potassium
that is crucial for cardiovascular and nerve function. Thus, in order to increase acceptability of mulberry juice
products and to provide a wide spectrum of health benefits of this valuable fruit, new products such as blend
juice and standardized mulberry juice microcapsules should be produced.
REFERENCES
1. Kumar, V. and Chauhan, S., Mulberry: Life enhancer. J. Med. Plants Res., 2, 271–278, 2008.
2. Zafar, M.S., Muhammad, F., Javed, I., Akhtar, M., Khaliq, T., Aslam, B., Waheed, A., Yasmin, R.,
and Zafar, H., White mulberry (Morus alba): A brief phytochemical and pharmacological evaluations
account. Int. J. Agric. Biol., 15, 612–620, 2013.
3. Ercisli, S. and Orhan, E., Some physico-chemical characteristics of black mulberry (Morus nigra L.)
genotypes from Northeast Anatolia region of Turkey. Sci. Hortic., 116, 41–46, 2008.
4. Singhal, B.K., Khan, M.A., Dhar, A., Baqual, F.M., and Bindroo, B.B., Approaches to industrial exploi-
tation of mulberry (mulberry sp.) fruits. J. Fruit Ornam. Plant Res., 18, 83–99, 2010.
5. Jiang, D.Q., Guo, Y., Xu, D.H., Huang, Y.S., Yuan, K., and Lv, Z.Q., Antioxidant and anti-fatigue effects
of anthocyanins of mulberry juice purification (MJP) and mulberry marc purification (MMP) from dif-
ferent varieties mulberry fruit in China. Food Chem. Toxicol., 59, 1–7, 2013.
6. Khalid, N., Fawad, S.A., and Ahmed, I., Antimicrobial activity, phytochemical profile and trace miner-
als of black mulberry (Morus nigra L.) fresh juice. Pakistan J. Bot., 43, 91–96, 2011.
7. Wu, T., Tang, Q., Gao, Z., Yu, Z., Song, H., Zheng, X., and Chen, W., Blueberry and mulberry juice
prevent obesity development in C57BL/6 mice. PLoS One, 8, e77585, 2013.
Mulberry Juice 407
8. Sakagami, H., Asano, K., Satoh, K., Takahashi, K., Kobayashi, M., Koga, N., Takahashi, H. et al.,
Anti-stress, anti-HIV and vitamin C-synergized radical scavenging activity of mulberry juice fractions.
In Vivo, 21, 499–505, 2007.
9. Akbulut, M., Çetin, Ç., and Çoklar, H., Farklı dut çeşitlerinin bazı kimyasal özellikleri ve mineral
madde içeriklerinin belirlenmesi (Determination of selected chemical properties and mineral content
of mulberries from various varieties), Gerçekçioğlu, R., Ed., II. Ulusal Üzümsü Meyveler Sempozyumu
(National Symposium on Berries), Tokat, Turkey, 2006, pp. 176–180 (in Turkish).
10. Butt, M.S., Nazir, A., Sultan, M.T., and Schroën, K., Morus alba L. nature’s functional tonic. Trends
Food Sci. Technol., 19, 505–512, 2008.
11. Nutrition Rank, Fresh Mulberry Juice. Published online at: http://www.nutritionrank.com/calories-
fresh-mulberry-juice-423136 (accessed April 12, 2014).
12. Jiyao, D. and Longji, H., Processing technique of concentrated mulberry juice. J. Sci. Technol. Food
Ind., 4, 2, 1993.
13. Xueming, L., Gengsheng, X., Yujuan, X., Jiyun, W., and Weidong, C., Study on comprehensive develop-
ment and industrialization technology of mulberry, in Proceedings of International Workshop on Silk
Handcrafts Cottage Industries and Silk Enterprises Development in Africa, Europe, Central Asia, and
the Near East & Second Executive Meeting of Black, Caspian Seas, and Central Asia Silk Association
(BACSA), Kozabirlik Sericultural Cooperative, Bursa, Turkey, 2006, pp. 533–539.
14. Lim, T.K., Morus nigra, 2012. Published online at: http://springer.com/chapter/10.1007/978-94-007-2534-8_57/
fulltext (accessed November 13, 2013).
15. Gungor, N. and Sengul, M., Antioxidant activity, total phenolic content and selected physicochemical
properties of white mulberry (Morus alba L.) fruits. Int. J. Food Prop., 11, 44–52, 2008.
16. Yu, Y., Xu, Y., Wu, J., Xiao, G., Fu, M., and Zhang, Y., Effect of ultra-high pressure homogenisation
processing on phenolic compounds, antioxidant capacity and anti-glucosidase of mulberry juice. Food
Chem., 153, 114–120, 2014.
17. Isabelle, M., Lee, B.L., Ong, C.N., Liu, X., and Huang, D., Peroxyl radical scavenging capacity, poly-
phenolics, and lipophilic antioxidant profiles of mulberry fruits cultivated in southern China. J. Agric.
Food Chem., 56, 9410–9416, 2008.
18. Özkan, M., Küçüköner, E., Tağı, Ş., Erbay, B., and Türkyılmaz, M., Determination of the stabilities of
black mulberry anthocyanins against thermal processing and storage as well as their some chemical
properties and antimicrobial activities, (Project Number: 111R004), The Scientific and Technological
Research Council of Turkey (TÜBİTAK-TOVAG), Ankara, Turkey, 2012 (in Turkish).
19. Song, W., Wang, H.J., Bucheli, P., Zhang, P.F., Wei, D.Z., and Lu, Y.H., Phytochemical profiles of dif-
ferent mulberry (Morus sp.) species from China. J. Agric. Food Chem., 57, 9133–9140, 2009.
20. Mikulic-Petkovsek, M., Slatnar, A., Stampar, F., and Veberic, R., HPLC-MSn identification and quantifi-
cation of flavonol glycosides in 28 wild and cultivated berry species. Food Chem., 135, 2138–2146, 2012.
21. Pawlowska, A.M., Oleszek, W., and Braca, A., Quali-quantitative analyses of flavonoids of Morus nigra
L. and Morus alba L. (Moraceae) fruits. J. Agric. Food Chem., 56, 3377–3380, 2008.
22. Hojjatpanah, G., Fazaeli, M., and Emam-Djomeh, Z., Effects of heating method and conditions on the
quality attributes of black mulberry (Morus nigra) juice concentrate. Int. J. Food Sci. Technol., 46,
956–962, 2011.
23. Juan, C., Jianquan, K., Junni, T., Zijian, C., and Ji, L., The profile in polyphenols and volatile com-
pounds in alcoholic beverages from different cultivars of mulberry. J. Food Sci., 77, 430–436, 2012.
24. Du, Q., Zheng, J., and Xu, Y., Composition of anthocyanins in mulberry and their antioxidant activity.
J. Food Compos. Anal., 21, 390–395, 2008.
25. Rice-Evans, C.A., Miller, N.J., and Paganga, G., Structure-antioxidant activity relationships of flavo-
noids and phenolic acids. Free Radical Bio. Med., 20, 933–956, 1996.
26. Lee, J., Chae, K., Ha, J., Park, B.Y., Lee, H.S., Jeong, S., Kim, M.-Y., and Yoon, M., Regulation of obe-
sity and lipid disorders by herbal extracts from Morus alba, Melissa officinalis, and Artemisia capillaris
in high-fat diet-induced obese mice. J. Ethnopharmacol., 115, 263–270, 2008.
27. Shoaib-Zafar, M., Muhammad, F., Javed, I., Akhtar, M., Khaliq, T., Aslam, B., Waheed, A., Yasmin, R.,
and Zafar, H., White mulberry (Morus alba): A brief phytochemical and pharmacological evaluations
account. Int. J. Agric. Biol., 15, 612–660, 2013.
28. Venkatachalam, V.V., Kannan, K., and Ganesh, S., Preliminary immunomodulatory activities of aque-
ous extract of Morus alba Linn. Int. J. Chem. Sci., 7, 2233–2238, 2009.
29. Hertog, M.G.L., Feskens, E.J.M., Hollman, P.C.H., Katan, M.B., and Kromhout, D., Dietary antioxidant
flavonoids and risk of coronary heart disease: The Zutphen Elderly Study. Lancet, 342, 1007–1011,
1993.
30. Hertog, M.G.L., Kromhout, D., Aravanis, C., Blackburn, H., Buzina, R., Fidanza, F., Giampaoli, S.
et al., Flavonoid intake and long-term risk of coronary heart disease and cancer in the seven countries
study. Arch. Intern. Med., 155, 381–386, 1995.
31. Khalil, F.A., Impact of administration mulberry juice on blood glucose, lipid profile and oxidative stress
in normal and diabetic rats. Egypt. J. Biochem. Mol. Biol., 27, 45–62, 2009.
32. Sakagami, H., Asano, K., Satoh, K., Takahashi, K., Terakubo, S., Shoji, Y., Nakamura, H., and
Nakamura, W., Anti-stress activity of mulberry juice in mice. In Vivo, 20, 499–504, 2006.
33. Rawson, A., Patras, A., Tiwari, B.K., Noci, F., Koutchma, T., and Brunton, N., Effect of thermal and non
thermal processing technologies on the bioactive content of exotic fruits and their products: Review of
recent advances. Food Res. Int., 44, 1875–1887, 2011.
34. Hong-Mei, L.I.U., Comparative study on chitosan and pectinase clarifying mulberry fruit juice. J. Norm.
Univ. Sci. Technol., 1, 10, 2006.
35. Vera, E., Sandeaux, J., Persin, F., Pourcelly, G., Dornier, M., Piombo, G., and Ruales, J., Deacidification
of clarified tropical fruit juices by electrodialysis. Part II. Characteristics of the deacidified juices.
J. Food Eng., 78, 1439–1445, 2007.
36. Faye, S., Consumer Trends for Fruit and Vegetable Products. Strategic Information Services Unit,
Alberta Agriculture, Food and Rural Development, Edmonton, Alberta, Canada, 2004.
37. Wang, S., Meckling, K.A., Marcone, M.F., Kakuda, Y., and Tsao, R., Synergistic, additive, and antago-
nistic effects of food mixtures on total antioxidant capacities. J. Agric. Food Chem., 59, 960–968, 2011.
38. Zelkha, M., Levy, R., Paran, E., Sharoni, Y., and Levy, J., Synergistic combinations of carotenoids and
polyphenols. U.S. Patent Application 13/137,061, 2011. Published online at: http://www.google.com/
patents/WO2010082205A1?hl=tr&cl=en (accessed April 12, 2014).
39. Nicolle, C., Simon, G., Rock, E., Amouroux, P., and Rémésy, C., Genetic variability influences carot-
enoid, vitamin, phenolic, and mineral content in white, yellow, purple, orange, and dark-orange carrot
cultivars. J. Am. Soc. Hortic. Sci., 129, 523–529, 2004.
40. Fazaeli, M., Emam-Djomeh, Z., Kalbasi Ashtari, A., and Omid, M., Effect of spray drying conditions
and feed composition on the physical properties of black mulberry juice powder. Food Bioprod. Proces.,
90, 667–675, 2012.
34
Noni Fruit Juice
Johannes Westendorf
CONTENTS
34.1 Introduction................................................................................................................................... 409
34.4 Health Effects.................................................................................................................................414
34.6 Conclusion......................................................................................................................................417
References................................................................................................................................................418
34.1 Introduction
The noni plant (Morinda citrifolia L., family Rubiaceae) grows in almost all tropical areas of the world,
but its origin is most probably North Australia and New Guinea from where it was distributed natu-
rally and by human activity first over the Polynesian triangle and later worldwide [1,2]. The robust
fast-growing plant grows as shrub or tree on all kinds of soil from lime soil at beaches to dark volcanic
soil and even on solidified lava flows as long as the roots take hold. Noni plants bear fruits at all phases
of ripeness year around. The fruits start out green and very hard, but during maturation they become
slowly brighter and finally develop a yellowish-white color and become very soft. Young fruits are odor-
less, but at maturation, they develop a strong odor reminiscent of overripe cheese [3]. Although noni
fruits are edible, the unpleasant taste and odor reduces their use as food mainly to times of famine [4]. In
Polynesian folk medicine, fruits and leaves of the noni plant have traditionally been used as herbal medi-
cine [5]. Because of the importance of the noni plant for the ancient Polynesians, it was introduced into
every new habitat it discovered. This is the reason for the presence of noni plants on almost all islands
within the Polynesian triangle. After 1996, juice prepared from ripe noni fruit became very popular
worldwide as wellness drink. In 2003 and 2008, noni fruit juice was approved as a Novel Food by the
European Commission [6,7]. Extensive toxicological studies have been performed for the novel food
approval. No substantial adverse effects have been observed in these studies, including tissue culture,
animal experiments, and human clinical trials [8,9]. The beneficial effects of noni fruit juice known by
the ancient Polynesians and people drinking noni fruit juice worldwide after 1996 are currently a matter
of intensive research. Numerous experimental and clinical studies have been published during the last
15 years to confirm these benefits and to explain the molecular mechanisms behind it. This chapter high-
lights the nutritional characteristics, with special attention of biologically active compounds and health
benefits of noni fruit juice.

The first report of the use of noni fruit as food comes from Captain James Cook, who visited the island of
Tahiti in 1769 [10]. After arrival of the Europeans to Polynesian islands, the tradition of eating noni fruit
ceased, but young green noni fruit, cut into small pieces and mixed with other fruits, are still eaten as fruit
409
salad by indigenous Indonesian and Polynesian people [3]. Today, noni fruit is mainly appreciated because
of the health benefits of noni fruit juice. The main preparation procedure includes fermenting of ripe noni
fruits and separation of the liquid phase by filtering or decanting. Most noni fruit juice preparations
contain some amounts of fruit debris but clear juices are also available. It has to be pasteurized prior to
marketing in order to comply with food regulation. The nutritional characteristics of noni fruit juice are
shown in Table 34.1. With only 32.5 kcal/100 mL, noni fruit juice contains almost half of the energy of
orange juice [11]. The reason for this difference is the low content of digestible carbohydrate, which favors
its use for people suffering from diabetes. Noni fruit juice also contains a variety of polysaccharides,
which are nondigestible by the human gastrointestinal (GI) tract [12]. The water-soluble polysaccharide
fraction has a high content of galacturonic acid, which is responsible for the viscosity of noni fruit juice.
Noni fruit juice contains most of the proteogenic amino acids in concentrations between 20 and
40 mg/100 g, except glutamic acid and aspartic acid, which are present at 64 and 80 mg/100 g, respectively.
TABLE 34.1
Compositional and Nutritional Characteristics of Noni Fruit Juice (per 100 mL)
Nutrient Unit Noni Fruit Juice
Water g 91.6
Energy kcal 32.5
Protein g 0.55
Lipid (fat) g 0.10
Carbohydrate g 7.2
Total sugars g 2.5
Minerals
Calcium mg 48.2
Copper mg 0.064
Iron mg 0.7
Manganese mg 0.15
Magnesium mg 13
Molybdenum μg < 0.4
Potassium mg 214
Selenium μg 0.2
Sodium mg 17
Zinc mg 0.13
Vitamins
Biotin µg 2.0
Folic acid µg <6.0
Niacin mg 3.0
Riboflavin mg <2.0
Thiamine mg <2.0
Vitamin A (RAE) µg <30
Vitamin B5 mg <2.0
Vitamin B6 mg <2.0
Vitamin B12 µg <0.1
Vitamin C mg 110
Carotenoid
β-Carotene mg 1.9
Source: Adapted from EFSA, EFSA J., 998, 1, 2009.
Abbreviations: RAE, retinol activity equivalents; ATE, alpha-tocopherol equivalents.
Noni Fruit Juice 411
Noni fruit is not a rich source of vitamins, except for vitamin C (ascorbic acid) (Table 34.1). A 100 mL of
noni fruit juice contains 110 mg vitamin C, which contributes to more than 100% of the Recommended
Dietary Allowances (RDA). The other vitamins are present in noni fruit juice in low concentrations
and therefore cannot contribute much to human nutrition requirements [11]. Ripe noni fruit contains
about 0.35% saturated and unsaturated fatty acids with carbon chain lengths of 4–20 [13]. Most abundant
ones are hexanoic acid and octanoic acid, which contribute to 20% and 58% of the volatile fraction [14].
These acids are responsible for the odor of ripe noni fruit and the juice thereof, which resembles that of
old cheese. Unripe fruit is odorless and contains the acids in the form of glycosides. When fruit ripens,
glycosidases are liberated, which transform the glycosides to the free volatile acids. A rare unsaturated
fatty acid, [2E, 4Z, 7Z]-decatrienoic acid, was first identified in noni fruit by Basar and Westendorf [15].
The average concentration of it in noni fruit juice samples of fruits collected at 32 locations in French
Polynesia was 490 mg/L. This fatty acid is very rare in the plant kingdom and has not yet been observed
in any other fruit. It is, therefore, an ideal marker compound for the identity of noni. This is important,
because adulteration of noni juice is common. There are many products on the market that contain only
small amounts of noni fruit juice or noni fruit powder, although they are labeled as “pure noni juice.”
As most other fruit juices, noni fruit juice is rich in potassium. It contains approximately 2000 mg/L
[11] (Table 34.1). This should be taken into consideration by people with limited kidney function, who
have a reduced tolerance to potassium consumption. Although noni plants grow on volcanic soil, which
is usually rich in minerals and trace elements, the concentrations of these metals in noni fruit are com-
parable to those of most other fruits. However, the metal content of noni fruit from trees grown on lime
soil at beaches is generally an order of magnitude lower, compared to plants grown on lava soil. The
concentrations of most metals are considerably higher in noni fruit powder compared to noni fruit juice.
Nevertheless, the amounts present in 100 mL of noni fruit juice usually do not exceed more 10% of the
daily requirements of adult humans [16].

More than 200 chemical compounds have thus far been reported in noni plants [17–20]. Only com-
pounds occurring in noni fruit juice are discussed in this chapter (Table 34.2). A compound with a broad
spectrum of biological activities is scopoletin, a coumarin derivative that occurs in freshly prepared noni
fruit juice, mainly as glycoside. During the fermentation process, the glycoside is hydrolyzed, releasing
the free aglycone, which is the predominant form in noni fruit juice available on the market. Analysis
of the ratio of the free and glycosidic form of scopoletin is, therefore, a good indicator for a possible
fermentation of noni fruit juice. The average concentration of scopoletin in 32 noni fruit juice samples
prepared from freshly squeezed ripe fruits was 114 mg/L, whereas the concentrations in a variety of
16 commercial noni fruit beverages varied between 9 and 235 mg/L [16]. The reason why the content
in some commercial samples exceeds that of the standard is that they have been prepared by insufficient
rehydration of noni fruit juice concentrate. Although scopoletin belongs to the coumarin class, it does
not inhibit the blood clotting process and no interference with coumadin is expected. Scopoletin has
potent anti-inflammatory and pain-releasing activities, which has been demonstrated in vitro and in
animal models [22–25]. The potency of scopoletin to inhibit pain in mice was comparable to that of the
anti-inflammatory drug indometacin [23]. Mechanistically, the anti-inflammatory activity of scopoletin
depends on the inhibition of the formation of the proinflammatory molecules nitric oxide (NO), pros-
taglandine E2 (PGE2), and tumor necrosis factor-α (TNF-α), which was demonstrated in tissue culture
[24,25] as well as in laboratory animals [22,23]. Scopoletin is also a potent antioxidant, which was dem-
onstrated by the ability of scopoletin to scavenge superoxide radical anion [26,27]. A dose-dependent
restoration in the activity of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase
(GPx) was observed in the rat hind paw after injection of the inflammation inducer carrageenan, which
reduced the enzyme activities considerably compared to the control. Scopoletin has also spasmolytic
properties, which can be attributed to the mobilization of calcium ions in muscle cells [28]. It has been
shown that scopoletin inhibits xanthine oxidase. This enzyme converts the purine derivative xanthine
into uric acid, which is excreted via the kidneys, mostly by tubular secretion. This process is limited
TABLE 34.2
Concentrations of Bioactive Compounds in Noni Fruit Juice
Compound Concentration (mg/L) References
Flavonoids
Quercetin 2.9 [21]
Quercetin derivative 4.6 [21]
Rutin 46.3 [21]
Iridoids
Deacetylasperulosidic acid 1441 [49]
Deacetylasperulosidic acid 1591 [21]
Asperulosidic acid 218 [49]
Asperulosidic acid 716 [21]
Fatty Acids
Decatrienoic acid 492 [15]
Hexanoic acid 329 [13]
Decanoic acid 3058 [13]
Coumarins
Scopoletin 114 [15]
Esculetin 2.0 [21]
Phenolics
Vanillin 3.5 [21]
Vanillic acid 2.6 [21]
and high blood levels of uric acid may saturate this process. As a result, uric acid can form crystals in
joints and other parts of the body, which is known as gout. Inhibition of the enzyme xanthine oxidase
leads to a decrease of the blood level of uric acid and may be responsible for the positive effects of
noni fruit juice on gout [29]. Scopoletin also possesses antimicrobial activity by inhibiting the growth
of bacteria and fungi [30]. Finally, it is known that scopoletin induces a suicidal process (apoptosis)
in leukemia cells [31]. These examples of multiple pharmacological properties of scopoletin make it
very likely that it is involved in many of the known benefits of noni fruit juice. Noni fruit also contains
flavonoids such as quercetin and kaempferol as well as rutin, a glycosidic form of quercetin. These com-
pounds are present in many plants, which have been used by humans as food since the earliest time of
mankind. Because some of these plants contain flavonoids in a considerable concentration, it is feasible
that flavonoids have become a regulating part of the human physiology. In noni fruit juice, quercetin is
the dominating flavonoid [32]. As with scopoletin, fresh noni fruit juice prior to fermentation contains
both compounds mainly as glycosides that are hydrolyzed during fermentation. Quercetin has also been
proposed as a marker substance for the identity and quality of beverages prepared from noni fruit [18].
However, because of its widespread occurrence in plants, it is less specific compared to decatrienoic
acid, which was only found in noni fruit juice. In scopoletin, the pharmacological profiles of quercetin
and kaempferol are very broad. In plants, flavonoids function as antioxidants, and this property is also
responsible for many of the observed pharmacological properties [33,34]. Because of the inactivation
of reactive oxygen species (ROS), flavonoids protect unsaturated fatty acids located in cell membranes
against destruction by radicals and thus inhibit the formation of oxidized low-density lipoprotein (LDL),
a major promoter of arteriosclerosis [35–37]. Another protective effect on the cardiovascular system
is the inhibition of the aggregation of platelets, which inhibits the formation of clots in arteries and
veins. Additionally, a decrease of the muscle tonus in arteries reduces the blood pressure. Quercetin
also stabilizes the heart rate [38]. Epidemiological investigations are meanwhile available, suggesting
a relationship between the reduced risk of heart attack and the daily consumption of quercetin [39,40].
The anticancer activity of quercetin, demonstrated in animal experiments, also appears to be linked
to the antioxidative potential [41]. In contrast to the protective activity on the cardiovascular system,
the cancer-inhibiting activity could not yet be confirmed epidemiologically [42]. Another important
property of flavonoids is their modulating effect on the hormonal system. Different mechanisms seem to
be involved in this activity. One is the induction of the enzyme aromatase, which is responsible for the
conversion of testosterone into estrogen. As a result, the concentration of estrogen in the body increases
[43]. Flavonoids are also able to mimic the function of the natural hormone 17β-estradiol by binding to
intracellular estrogen receptors. Kaempferol has the highest affinity to estrogen receptors, which is only
two to three orders of magnitude lower than that of the natural hormone. Quercetin, on the other hand,
is able to function as an estrogen antagonist in the presence of an excess of this hormone. Together, the
two flavonoids appear to have a regulating effect on the hormonal homeostasis [44].
Ursolic acid and oleanolic acid occur in the leaves and fruits of the noni tree and are also present in
noni fruit juice. Both compounds are triterpenes and strong antioxidants [45]. Unfortunately, we do not
have information on their concentrations in noni fruit juice. In addition to their antioxidant capacity,
these compounds act as anticancer agents by inhibiting the vascularization of tumor tissue [46]. Anti-
inflammatory activity has also been demonstrated for both compounds. A possible mechanism is the
reduction of the transcription of cyclooxygenase-2 (COX-2) by inhibition of the signal pathways of PKC
and AP-1 [47].Iridoids are a group of structurally related monoterpenes containing hundreds of deriva-
tives that are distributed widely in the plant kingdom. About 16 different iridoids have been detected thus
far in noni fruit juice [48]. Deacetyl asperulosidic acid (DAA) and asperulosidic acid (AA) are the most
abundant iridoids present in noni fruit juice (Figure 34.1 and Table 34.2), making up more than 90% of
the total iridoid content [49].
R
OH
HO O
O O OH
OH
O OH O
Scopoletin R = H: Kaempferol
R = OH: Quercetin
H COOH
O H
HO
HO
(2E,4Z,7Z)-Decatrienoic acid Ursolic acid
O
O HO COOH
O O
H3COCOH2C ROH2C
O O
OH OH
O O
OH OH
OH OH OH OH
Asperulosid R = H: Deacetyl asperulosidic acid
R = CH3CO: Asperulosidic acid
FIGURE 34.1 Chemical structures of main bioactive compounds present in noni fruit juice.
Plants use iridoids as repellents against herbivorous animals. The compounds are bitter and many of
them are toxic to certain animals. The toxicity depends on the cleavage of the glycosidic bond by glycosi-
dases, which are present in the plant itself and released by chewing or in the GI tract of the animals. The
iridoid aglycones then form dialdehydes by opening of the lactone ring, which further reacts with free
amino groups in proteins causing denaturation of functioning macromolecules [50]. During evolution,
some animals including the ancestors of humans have developed strategies to overcome the toxicity of
some iridoids. If the iridoid aglycones possess certain stability, they are coupled with glucuronic acid or
sulfate and thus stabilized. In experiments with radio-labeled DAA performed on mice, we could show
that this iridoid, which forms a highly reactive aglycone, is absorbed without prior hydrolyzation and
excreted rapidly through the kidneys without further metabolism (own unpublished result).
While the toxicity of iridoids is not an issue for many higher developed animals and humans, the
compounds nevertheless may exert a variety of pharmacological effects, which can be used beneficially.
Thus far, there are only very few investigations available concerning the biological activities of DAA and
AA, the main iridoids in noni fruit juice; however, numerous studies have been performed with closely
related iridoids [51].
Antioxidant and anti-inflammatory properties have been observed for DAA and AA in vitro and in vivo.
In vitro inhibition of LDL oxidation was observed by DAA and AA at a concentration of 20 µg/mL [52].
After oral application of DAA to rats, an increase in the concentration of SOD was observed [53].
Induction of glutathione-S-transferase (GSH), an enzyme with an important function in the detoxifica-
tion of radicals, was observed after treatment of rat hepatocytes in culture with geniposide [54]. A variety
of iridoids, which are structurally related to DAA and AA, exhibited potent anti-inflammatory activity
in the tetraphorbol acetate (TPA)–induced mouse ear edema model after topical application and in the
carrageenan-induced rat hind paw edema model after oral application [55]. Comparison of pairs of iri-
doid acids and their corresponding methyl esters (loganin/loganic acid and geniposide/geniposidic acid)
showed that the esters exhibited stronger activity in the mouse ear edema assay after topical application,
whereas the free acids were more active in the rat hind paw edema assay after oral application. The reason
is certainly the higher lipophilicity of the esters, which enables a better permeation through the skin.
A variety of iridoids, which are closely related to DAA and AA, were investigated with respect to their
capacity to inhibit the formation of the inflammatory mediator thromboxane (TBX), PGE2, TNF-α, and
NO in cell lines derived from macrophages and erythroleukemia cells. The iridoids were active, however,
only after pretreatment with glucosidase. Geniposide was active after treatment with glucosidase, but the
aglycone genipin was inactive, showing that the activity depends on an intermediate during the enzy-
matic hydrolyzation of the iridoid glycoside and not on the aglycone itself [56]. Anti-inflammatory and
antinociceptive activity was also demonstrated for monotropeine, an iridoid glucoside, which is isomeric
to DAA. The compound inhibited the formation of hind paw edema in rats after injection of carrageenan
and reduced the pain perception of mice in the hot plate assay. The latency times were prolonged by 64%
and 96% after the daily oral application of 10 or 30 mg/kg body weight, respectively, for a total duration
of 1 week. The high-dose effect was comparable to that of the oral application of 10 mg/kg morphine [57].
Iridoids possess also anticarcinogenic and anticancer activities. Asperulosidic acid, isolated from noni
fruit, was demonstrated to inhibit AP-1 transactivation and cell transformation in mouse epidermal JB6
cells, treated with TPA or epidermal growth factor (EGF) [58]. A reduction of the mutagenic response of
Chinese hamster ovary cells in vitro and mice in vivo treated with the genotoxic anticancer compound
mitomycin C was observed after pretreatment with DAA, AA, and asperuloside, isolated from Tochu tea,
an aqueous extract of Eucommia ulmoides leaves [59]. Geniposide was shown to exhibit antiangiogenic
activity in the chick embryo chorioallantoic membrane assay and inhibited dose dependently the growth
of NIH-3T3 cells in culture at concentrations between 25 and 100 µM [60].
34.4 Health Effects

The juice prepared from fruits of the noni tree in fermented and nonfermented forms was used by indig-
enous inhabitants of the South Pacific since thousands of years. In fact, it was one of their most important
plant-derived medicines [1]. In 2005, an epidemiological survey was published about the traditional use
of noni in Fiji islands [61]. The treatment of all kinds of inflammatory diseases was the most frequent
reason for the use of noni, which is called “kura” by the people of Fiji.
In 2003, the year of the approval of noni fruit juice as novel food by the European Union, and 7 years
after the beginning of the worldwide marketing of noni, we performed an epidemiological survey among
consumers of noni fruit juice in Europe [3]. The reason for this investigation was to learn why people take
noni fruit juice and what the benefits of its use might be. A summary of the results published is shown in
Table 34.3 [3]. About 30% of the 1124 people participating in the investigation reported that noni fruit juice
increased their energy and 25% experienced an increase in overall well-being. The third most frequent
observation was a reduction of pain and discomfort in joints and the back. This was also the most frequent
benefit observed by Pande et al. [61] in their investigation of the traditional use of noni fruit juice on Fiji
islands. Other benefits reported in our study were fewer infections (12.4%), improved sleeping (8.6%), fewer
problems with stomach and digestion (8.2%), fewer allergies and asthma (7.0%), improved skin (5.9%),
fewer headaches (4.1%), and a number of other less frequently observed benefits. Together, the observed
benefits suggest that noni fruit juice acts as an adaptogen. The name “adoptogen’’ was first introduced by
the Russian scientist N.V. Lazarew in 1947 for a group of medicinal plants, which enhance the adaptation
capacity of the human body against stress without exerting toxicity, even at very high doses [62].
A variety of clinical studies and animal experiments have been performed with noni fruit juice in
order to scientifically confirm the benefits reported traditionally and by modern consumers.
Increase in energy is one of the most frequently reported benefits of noni fruit juice. In order to prove
this activity, two groups each of 20 young well-trained athletes of both sexes were running on a treadmill
until exhaustion. The time to fatigue was monitored and a blood sample was taken immediately after
termination of the exercise for the determination of the luminescence as an indicator for the concentra-
tion of free radicals. One group then drunk 100 mL of a commercial noni juice (Tahitian Noni™ Juice)
per day over a period of 2 weeks, and the other group received the same amount of blackberry juice for
the same period of time. The exercise on the treadmill was then repeated. No statistically significant
difference (P > 0.05) in the time to fatigue could be observed between the blackberry and the control
group, whereas a 20% increase was observed in the noni fruit juice group. Moreover, the athletes in
the noni fruit juice group had a 25% reduction in the concentration of free radicals in their blood after
TABLE 34.3
Benefits from Tahitian Noni Fruit Juice in the Order of Their Frequency
Number Gender Average Average Daily Average Duration
Benefit (%) % (M/F) Age (Year) Intake (mL) of Intake (Month)
All participants 1142 (100) 38/62 48.2 52.3 26.4
More energy 337 (29.5) 34/66 47.2 53.3 27.7
Better overall feeling 285 (25.0) 38/62 48.5 51.4 24.5
Less peripheral pain 184 (16.2) 31/69 51.6 55.9 29.6
Fewer infections 141 (12.4) 29/71 50.1 52.5 36.8
Better sleep 97 (8.6) 33/67 50.2 59.0 27.8
Less problems with digestion 94 (8.2) 27/73 50.0 55.0 26.6
Fewer allergies and asthma 79 (7.0) 37/63 45.6 58.8 32.4
Improved skin 67 (5.9) 27/73 45.9 51.9 23.5
Reduction of headache 46 (4.1) 17/83 46.9 48.9 25.7
Fewer gynecological problems 30 (4.4) 0/100 44.2 63.0 26.4
Improved growth of hair and nails 24 (2.1) 29/71 49.0 46.3 27.0
Reduction of BP and cholesterol 22 (1.9) 50/50 52.0 50.5 32.6
Beneficial effects on diabetes 16 (1.4) 62/38 49.1 54.4 32.2
Less gingival problems 9 (0.79) 22/78 48.1 18.9 50.0
Improved wound healing 5 (0.44) 40/60 47.0 48.0 19.8
Source: Adapted from Westendorf, J. and Mettlich, C., J. Med. Food Plants, 1, 64, 2009. With permission.
Note: Numbers significantly different (P < 0.05) from all participants are in bold.
Abbreviation: BP, blood pressure.
the exposure to the juice compared to the control group, whereas no such decrease was observable in
the blackberry juice group [63]. The concomitant increase of the endurance and decrease of the free
radical concentration in the blood suggests an enhancement of the antioxidant potential in body tissues.
An increase in endurance was also observed in an experiment with mice, running on a rotating rod until
exhaustion. Groups of 10 animals each received 0, 10, 20, or 40 mL of noni fruit juice in their drinking
water for 3 weeks. Noni fruit juice enhanced dose dependently the endurance of the mice on the rod [64].
A direct investigation of the antioxidant potential was performed in a study with 30 students of the
University of Ekatarinburg, Russia. A polarographic electrode, enabling the direct measurement of the
antioxidative potential in small volumes of blood, developed by scientists of the University of Ekatarinburg
was used [65]. The volunteers were divided into three groups of 10 persons each. At the beginning of the
experiment, blood samples were taken from each volunteer and the antioxidative potential was monitored.
The participants of the first group then received 200 mL of water, whereas those of the second and third
group received 200 mL orange juice or noni fruit juice (Tahitian NoniTM juice), respectively. After 90 min,
another blood sample was taken. The blood was separated into fractions of serum and red blood cells.
The antioxidant capacity was monitored in both subfractions, and the mean values and standard devia-
tions for each group were calculated (Figure 34.2). The mean values of the antioxidative potentials of the
volunteers drinking noni fruit juice were significantly increased compared to the water control in the red
blood cell fraction. Although orange juice resulted in an increase compared to the control group, it was
noticeably less compared to noni fruit juice. An increase of the antioxidant activity in the noni fruit juice
group compared to the control was also observed in the serum; however, it was not statistically significant
(P > 0.05). The antioxidant potentials of the red blood cells were more than 10-fold increased, compared
to those observed in the serum. It is not known from this experiment, whether compounds present in noni
juice migrate into the cells or whether they enhance the synthesis of endogenous antioxidants.
A double-blind, placebo-controlled clinical trial performed with 132 heavy smokers revealed that
drinking 30–180 mL noni juice (Tahitian Noni™ juice) improved the lipid serum profile and reduced
systemic inflammation caused by smoking. Noni juice significantly reduced the serum levels of total
and LDL cholesterols and increased the high-density lipoprotein (HDL) cholesterol. The serum concen-
tration of the inflammation marker C-reactive protein was decreased [66].
5 Erythrocyte 5
Serum
4 4
Delta AOA serum (mM eq/L ×10)
3 3
Delta AOA erythrocytes (mM eq/L)
2 2
1 1
0 0
–1 –1
–2 –2
–3 –3
Water Orange juice TNJ
FIGURE 34.2 Antioxidative potency of red blood cells and serum (1 h after intake of 200 mL of Tahitian Noni™ Juice
[TNJ] by 20 volunteers per group). The AOA values were derived by use of a potentiometric method. Values of the serum
in the right y-axis have been multiplied with 10 (unpublished data).
The anti-inflammatory and pain-releasing activity of noni fruit juice was investigated in the mouse hot
plate test. A group of 10 animals received 10% noni fruit juice (Tahitian NoniTM juice) in their drinking
water, for 3 days prior to the test. A control group remained untreated and a third group received the cen-
tral analgesic drug tramadol (30 mg/kg body weight) 1 h prior to the test by subcutaneous injection. Noni
fruit juice significantly prolonged the reaction time of the mice comparable to tramadol. Injection of the
central analgesic antagonist naloxone inhibited the effect of the noni fruit juice only partly, whereas the
effect of tramadol was completely abolished by naloxone. This indicates that central as well as peripheral
mechanisms are involved in the analgesic activity of noni fruit juice [67]. The anti-inflammatory activity
of noni fruit juice was also demonstrated in a clinical dentistry study with patients suffering from gin-
givitis. Two randomized groups of patients with different degrees of gingivitis were trained to perform
a standardized oral hygiene protocol. One group additionally performed a daily mouth wash with noni
fruit juice. After 2 weeks, the patients were monitored again and noni fruit juice reduced gum inflamma-
tion considerably compared to the control group [68].
Recently, clinical trials performed at the University Clinic Hamburg, Germany, demonstrated a posi-
tive effect of noni fruit juice on patients suffering from type II diabetes. A daily dose of 2 mL/kg body
weight reduced the morning blood sugar level and the HbA1c-value significantly. The level of the inflam-
matory cytokine interleukin-6 (IL-6), which is believed to be involved in the etiology of diabetes, was
also decreased, whereas the level of C-peptide, which is used as a marker of the insulin secretion, was
increased [69]. Recently, a significant decrease of the insulin requirement of patients suffering from type
I diabetes was achieved after consumption of noni fruit juice (own unpublished results).

The ancient Polynesians used fermented noni fruit juice because of its multifold beneficial effects on
health, and the tradition is still alive, not only in Polynesia but also in many other places in the trop-
ics. After the worldwide distribution of noni fruit juice in the 1990s, companies first tried to copy the
Polynesian tradition; however, the unpleasant taste of pure noni fruit juice was an inhibiting factor
for marketing. This problem could be solved by blending noni fruit juice with other fruit juices. Such
blended fermented noni fruit juice may become very famous and will soon be available throughout the
world. Because its great variety of beneficial effects [3], noni fruit juice will certainly be further used
by many people. After more and more knowledge became available about the chemical composition and
the biological activity of noni fruit juice, companies started to refine their products in order to increase
the potency. After 2010, iridoids have been recognized as possibly most important bioactive compounds
in noni fruit juice [70,71]. Consequently, products have been designed to increase the concentration of
iridoids. Furthermore, a literature survey showed that different iridoids can provide different health
benefits [51]. In order to increase the spectrum of benefits, noni fruit juice was mixed with other iridoid
containing plant products, such as Cornelian sherry juice (Cornus mas), blueberry juice (Vaccinium
corymbosum and Vaccinium myrtillus), and olive leaves extract (Olea europaea). Other combinations
may be introduced to the market in the next few years.
Until now, standard analytical methods to characterize the quality of noni fruit juice are not available.
The high market price of noni fruit juice makes it interesting for companies to adulterate noni fruit juice,
which may damage the image of this beverage because such products do not have the beneficial proper-
ties expected by the consumers. It will, therefore, be of great importance for noni fruit juice–producing
companies to develop proper quality standards and to enforce its compliance.
34.6 Conclusion
Noni fruit juice emerged from an old Polynesian natural remedy to a modern wellness drink with world-
wide distribution. Consumers prefer noni fruit juice because of its multiple benefits, which includes,
among others, increase of energy and overall well-being, reduction of pain and inflammation, and
enhancement of the immune defense system. This makes noni fruit juice a functional beverage with
benefits for almost everybody. The adaptogenic properties of noni fruit juice with the enhancement of the
body’s ability to adapt to different stress situations, occurring from normal daily life as well as a great
variety of diseases, make this beverage a useful tool for the maintenance of health and the treatment of
symptoms that accompany diseases, as an alternative or in combination with pharmaceuticals.
REFERENCES
1. Dixon, A.R., McMillan, H., and Atkin N.L., Ferment this: The transformation of noni, a traditional
Polynesian medicine (Morinda citrifolia, Rubiaceae). Econ. Bot., 53, 51–68, 1999.
2. Abbott, I. and Shimazu, C., The geographic origin of the plants most commonly used for medicine by
Hawaiians. J. Ethnopharmacol., 14, 213–222, 1985.
3. Westendorf, J. and Mettlich. C., The benefits of noni juice: An epidemiological evaluation in Europe.
J. Med. Food Plants, 1, 64–79, 2009.
4. Fahs, B., Noni needn’t taste nasty, in Proceedings of the 2002 Hawaii Noni Conference, Nelson, S.C.,
Ed., University of Hawaii at Manoa, College of Tropical Agriculture and Human Resources, Hilo, HI,
2003, pp. 17–19.
5. Dittmar, A., Morinda citrifolia L. use in indigenous Samonan medicine. J. Herbs Spices Med. Plants, 1,
77–92, 1993.
6. EU Comission, Decision of 5 June 2003 authorising the placing on the market of “noni juice” (juice of
the fruit of Morinda citrifolia L.) as a novel food ingredient under Regulation (EC No.258/97) of the
European Parliament and of the Council. Off. J. Eur. Union, L144, 2003.
7. EU Comission, Decision of 15 November 2008 authorising the placing on the market of leaves of
Morinda citrifolia L. as a novel food ingredient under Regulation (EC No. 258/97) of the European
Parliament and of the Council (document number: C(2008)8108) (2008/985/EC). Off. J. Eur. Union,
L352/46, 2008.
8. Westendorf, J., Effenberger, K., Iznaguen, H., and Basar, S., Toxicological and analytical investigations
of noni (Morinda citrifolia) fruit juice. J. Agric. Food Chem., 55, 529–537, 2007.
9. West, B.J., Jensen, C.J., Westendorf, J., and White, L.D., A safety review of noni fruit juice. J. Food.
Sci., 71, R100–R106, 2006.
10. Cheeseman, T.F., The flora of Rarotonga, the chief island of the Cook Group. Trans. Linnean Soc.
Lond., 6, 261–313, 1903.
11. EFSA., Scientific opinion of the panel on dietetic products nutrition and allergies on a request from
European Commission on the safety of “Morinda citrifolia (noni) fruit puree and concentrate” as a novel
food ingredient. EFSA J., 998, 1–9, 2009.
12. Bui, A.K.T., Bacic, A., and Pettolino, F., Polysaccharide composition of the fruit juice of Morinda citri-
folia (noni). Phytochemistry, 71, 1–5, 2006.
13. Pino, J.A., Marquez, E., and Castro, D., Volatile and non-volatile acids of noni (Morinda citrifolia L.)
fruit. J. Sci. Food Agric., 89, 1247–1249, 2009.
14. Farine, J.P., Legal, L., Moreteau, B., and Le Quere, J.L., Volatile components of ripe fruits of Morinda
citrifolia and their effects on Drosophila. Phytochemistry, 41, 433–438, 1996.
15. Basar, S. and Westendorf, J., Identification of (2E,4Z,7Z)-decatrienoic acid in noni fruit and its use in
quality screening of commercial noni products. Food Anal. Method., 4, 57–65, 2010.
16. Basar, S. and Westendorf, J., Mineral and trace element concentrations in Morinda citrifolia L. (noni)
leaf, fruit and fruit juice. Food Nutr. Sci., 3, 1176–1188, 2012.
17. Su, B.N., Pawlus, A.D., Jung, H.A., Keller, W.J., McLaughlin, J.L., and Kinghorn, A.D., Chemical
constituents of the fruits of Morinda citrifolia (noni) and their antioxidant activity. J. Nat. Prod., 68,
592–595, 2005.
18. Potterat, O. and Hamburger, M., Morinda citrifolia (noni) fruit, phytochemistry, pharmacology and
safety. Planta Med., 73, 191–199, 2007.
19. Pawlus, A.D. and Kinghorn, A.D., Review of the ethno botany, chemistry, biological activity and safety
of the botanical dietary supplement Morinda citrifolia (noni). J. Pharm. Pharmacol., 59, 1587–1609,
2007.
20. Deng, S., West, B.J., and Jensen, J.C., Simultaneous characterisation and quantitation of flavonol glyco-
sides and aglycones in noni leaves using a validated HPLC-UV/MS method. Food Chem., 111, 526–529,
2008.
21. Dussossoy, E., Brat, P., Bony, E., Boudard, F., Poucheret P., Mertza, C., Giaimis, J., and Michel, A.,
Characterization, anti-oxidative and anti-inflammatory effects of Costa Rican noni juice (Morinda citri-
folia L.). J. Ethnopharmacol., 133 108–115, 2011.
22. Ding, Z., Dai, Y., Hao, H., Pan, R., Yao, X., and Wang, Z., Anti-inflammatory effects of scopoletin and
underlying m., echanisms. Pharmaceut. Biol., 46, 854–860, 2008.
23. Chang, T.N., Deng, J.S., Chang, Y.C., Lee, C.Y., Jung-Chun, L., Lee, M.M., Peng, W.H., Huang, S.S.,
and Huang, G.J., Ameliorative effects of scopoletin from Crossostephium chinensis against inflam-
mation pain and its mechanisms in mice. Evid. Based Complement. Alternat. Med., 2012 (article ID
595603, Epub).
24. Kim, H.J., Jang, S.I., Kim, Y.J., Chung, H.T., Yun, Y.G., Kang, T.H., Jeong, O.S., and Kim, Y.C.,
Scopoletin suppresses pro-inflammatory cytokines and PGE2 from LPS-stimulated cell line, RAW
264.7 cells. Fitoterapia, 75, 261–266, 2004.
25. Moon, P.D., Lee, B.H., Jeong, H.J., An, H.J., Park, S.J., Kim, H.R., Ko, S.G., Um, J.Y., Hong, S.H., and
Kim, H.M., Use of scopoletin to inhibit the production of inflammatory cytokines through inhibition of
the IκB/NF-κB signal cascade in the human mast cell line HMC-1. Eur. J. Pharmacol., 555, 218–225,
2007.
26. Shaw, C.Y., Chen, C.H., Hsu, C.C., Chen, C.C., and Tsai, Y.C., Antioxidant properties of scopoletin
isolated from Sinomonium acutum. Phytother. Res., 17, 823–825, 2003.
27. Lee, S.H., Ding, Y., Yan X.T., Kim Y.H., and Jang H.D., Scopoletin and scopolin isolated from Artemisia
iwayomogi suppress differentiation of osteoclastic macrophage RAW 264.7 cells by scavenging reactive
oxygen species. J. Nat. Prod., 76, 615–620, 2013.
28. Oliveira, E.J., Romero, M.A., Silva, M.S., Silva, B.A., and Medeiros, I., Intracellular calcium mobiliza-
tion as a target for the spasmolytic action of scopoletin. Planta Med., 67, 605–608, 2001.
29. Ding, Z., Dai, Y., and Wang, Z., Hypouricemic action of scopoletin arising from xanthine oxidase inhi-
bition and uricosuric activity. Planta Med., 71, 183–185, 2005.
30. Carpiella, M.C., Ferrayoli, C.G., and Palacios, S.M., Antifungal synergistic effect of scopoletin a
hydroxycoumarin isolated from Melia azedarach L. fruits. J. Agric. Food Chem., 53, 2922–2927, 2005.
31. Kim, E.K., Kwon, K.B., Shin, B.C., Seo, E.A., Lee, Y.R., Kim, J.S., Park, J.W., Park, B.H., and Ryu,
D.G., Scopoletin induces apoptosis in human promyeloleukemic cells, accompanied by activations of
nuclear factor κ-B and caspase-3. Life Sci., 77, 824–836, 2005.
32. Sang, S., Cheng, X., Zhu, N., Stark, R.E., Badmaev, V., Ghai, G., Rosen, R.T., and Ho, C.-T., Flavonol
glycosides and novel iridoid glycoside from the leaves of Morinda citrifolia. J. Agric. Food Chem., 49,
4478–4481, 2001.
33. De Groot, H. and Rauen, U., Tissue injury by reactive oxygen species and the protective effects of flavo-
noids. Fundam. Clin. Pharmacol., 12, 249–255, 1998.
34. Hanasaki, Y., Ogawa, S., and Fukui, S., The correlation between active oxygen scavenging and antioxi-
dative effects of flavonoids. Free Rad. Biol. Med., 16, 845–850, 1994.
35. Hollmann, P.C.H. and Katan, M.B., Absorption, metabolism and health effects of dietary flavonoids in
man. Biomed. Pharmacother., 51, 305–310, 1997.
36. Peng, I.W. and Kuo, S.M., Flavonoid structure affects the inhibition of lipid peroxidation in Caco-2
intestinal cells at physiological concentrations. J. Nutr., 133, 2184–2187, 2003.
37. Sakanashi, Y., Oyama, K., Matsui, H., Oyama, T.B., Oyama, T.M., Nishimura, Y., Sakai, H., and
Oyama, Y., Possible use of quercetin an antioxidant for protection of cells suffering from overload of
intracellular Ca2+: A model experiment. Life Sci., 83, 164–169, 2008.
38. Formica, J.V. and Regelson, W., Review of the biology of quercetin and related bioflavonoids. Food
Chem. Toxicol., 33, 1061–1080, 1995.
39. Knekt, P., Jarvinen, R., Reunanen, A., and Maatela, J., Flavonoid intake and coronary mortality in
Finland: A cohort study. Br. Med. J., 312, 478–481, 1996.
40. Hertog, G.L., Antioxidative flavonoids in food: Risk of heart attack and carcinoma (in German). Forsch.
Komplementärmed., 2, 283–288, 1995.
41. Wang, H.K., Xia, Y., Yang, Z.Y., Natschke, S.L., and Lee, K.H., Recent advances in the discovery and
development of flavonoids and their analogues as antitumor and anti-HIV agents. Adv. Exp. Med. Biol.,
439, 191–225, 1998.
42. Hertog, M.G., Kromhout, D., and Aravanis, C., Flavonoid intake and long-term risk of coronary heart
disease and cancer in the seven countries study. Arch. Intern. Med., 155, 381–386, 1995.
43. Senderson, J.T., Hordijk, J., Denison, M.S., Springsteel, M.F., Nantz, M.H., and van den Berg, M.,
Induction and inhibition of aromatase (CYP19) activity by natural and synthetic flavonoid compounds
in H295R human adrenocortical carcinoma cells. Toxicol. Sci., 82, 70–79, 2004.
44. Zeglin, A., Investigation of the estrogenic potential of Morinda citrifolia L. with a combination of a
receptor binding assay and alkaline phosphatase induction assay in Ishikawa cells (in German), PhD
thesis, Medical University, Hamburg, Germany, 2009.
45. Ahmad, V.U. and Bano, S., Isolation of β-sitosterol und ursolic acid from Morinda citrifolia L. J. Chem.
Soc. Pak., 2, 71, 1980.
46. Shafaei, A., Muslim, N.S., Nassar, Z.D., Aisha, A.F.A., Abdul M.A., and Ismail, Z., Antiangiogenic
effect of Ficus deltoidea Jack standardised leaf extracts. Trop. J. Pharm. Res., 13, 761–768, 2014.
47. Dzubak, P., Haiduch, M., Vydra, D., Hustova, A., Kvasnica, M., Biedermann, D., Markova, L., Urban,
M., and Sarek, J., Pharmacological activities of natural triterpenoids and their therapeutical implica-
tions. Nat. Prod. Rep., 23, 394–411, 2006.
48. Kamiya, K., Tanaka, Y., Endang, H., Umar, M., and Satake, T., New anthraquinone and iridoid from the
fruits of Morinda citrifolia. Chem. Pharm. Bull., 53, 1597–1599, 2005.
49. Deng, S., West, B.J., Palu, A.K., and Jensen, C.J., Determination and comparative analysis of major iri-
doids in different parts and cultivation sources of Morinda citrifolia. Phytochem. Anal., 22, 26–30, 2011.
50. Dobler, S., Petschenk, G., and Pankoke, H., Coping with toxic plant compounds—The insect perspective
on iridoid glycosides and cardenolides. Phytochemistry, 72, 1593–1604, 2011.
51. Tundis, R., Loizzo, M.R., Menichini, F., Statti, G.A., and Menichini, F., Biological and pharmacological
activities of iridoids: Recent developments. Mini-Rev. Med. Chem., 8, 399–420, 2008.
52. Kim, D.H., Lee, H.J., Oh, Y.J., Kim, M.J., Kim, S.H., Jeong, T.S., and Baek, N.I., Iridoid glycosides
isolated from Oldenia diffusa inhibit LDL-oxidation. Arch. Pharm. Res., 28, 1156–1160, 2005.
53. De-Lu Ma, D., Chen, M., Su, C., and West, B., In vivo antioxidant activity of deacetylasperulosidic acid
in noni. J. Anal. Meth. Chem., 2013 (article ID 804504, Epub).
54. Kuo, W.S., Wang, C.J., Young, S.C., Sun, Y.C., Chen, Y.J., and Chou, F.P., Differential induction of the
expression of GST subunits by geniposide in rat hepatocytes. Pharmacology, 70, 15–22, 2004.
55. Recio, M.C., Giner, R.M., Máñez, S., and Rios, L., Structural considerations on the iridoids as anti-
inflammatory agents. Planta Med., 60, 232–234, 1994.
56. Park, K.S., Kim, B.H., and Chang, I.M., Inhibitory potencies of several iridoids on cyclooxygenase-1,
cyclooxygnase-2 enzymes activities, tumor necrosis factor-α and nitric oxide production in vitro. Evid.
Based Complement. Alternat. Med., 7, 41–45, 2010.
57. Choi, J., Lee, K.T., Choi, M.Y., Nam, J.H., Jung, H.J., Park, S.K., and Park, H.J., Antinociceptive anti-
inflammatory effect of monotropein isolated from the root of Morinda officinalis. Biol. Pharm. Bull.,
28, 1915–1918, 2005.
58. Liu, G., Bode, A., Ma, W.Y., Sang, S., Ho, C.-T., and Dong, Z., Two novel glycosides from the fruits of
Morinda citrifolia (noni) inhibit AP-1 transactivation and cell transformation in the mouse epidermal
JB6 cell line. Cancer Res., 61, 5749–5756, 2001.
59. Nakamura, T., Nakazawa, Y., Onizuka, S., Satoh, S., Chiba, A., Sekihashi, K., Miura, A., Yasugahira,
N., and Sasaki, F., Antimutagenicity of tochu tea (an aqueous extract of Eucomia ulmoides leaves: 1.
The clastogen-suppressing effects of Tochu tea in CHO cells and mice. Mut. Res., 388, 7–20, 1997.
60. Koo, H.J., Lee, S., Shin, K.H., Kim, B.C., Lim, C.J.M., and Park, E.H., Geniposide, an antiangiogenic
compound from the fruits of Gardenia jasmoides. Planta Med., 70, 467–469, 2004.
61. Pande, M., Naiker, N., Mills, G., Singh, N., and Voro, T., The Kura files: Qualitatively social survey.
Pac. Health Surv. Resp., 12, 85–93, 2005.
62. Davydov, M. and Krikorian, A.D., Eleutherococcus senticosus (Rupr. & Maxim.) Maxim. (Araliaceae)
as an adaptogen: A closer look. J. Ethnopharmacol., 72, 345–393, 2000.
63. Palu, A., Seifulla, R., and West, B., Morinda citrifolia L. (noni) improves athlete endurance: Its mecha-
nisms of action. J. Med. Plant Res., 2, 154–158, 2008.
64. Ma, D.I., West, B.J., Su, C.X., Gao, J.H., Liu, T.Z., and Liu, Y.W., Evaluation of the ergogenic potential
of noni juice. Phytother. Res., 21, 1100–1101, 2007.
65. Brainina, K.Z., Gerasimova, E.L., Varzakova, D.P., Balezin, S.L., Portnov, I.G., Makutina, V.A., and
Tyrchaninova, E.V., Potentiometric method for evaluating the oxidant/antioxidant activity of semi-
nal and follicular fluids and clinical significance of this parameter for human reproductive function.
Open Chem. Biomed. Meth. J., 5, 1–7, 2012.
66. Wang, M.Y., Peng, L., Weidenbacher-Hoper, V., Deng, S., Anderson, G., and West, B.J., Noni juice
improves serum lipid profiles and other risk markers in cigarette smokers. Sci. World J., 2012
(Doi:10.1100/2012/594657).
67. Basar, S., Uhlenhut, K., Högger, P., Schöne, F., and Westendorf, J., Analgesic and antiinflammatory
activity of Morinda citrifolia L. (noni) fruit. Phytother. Res., 24, 38–42, 2010.
68. Glang, J., Falk, W., and Westendorf, J., Effect of Morinda citrifolia L. fruit juice on gingivitis/
periodontitis. Mod. Res. Inflamm., 2, 21–27, 2013.
69. Bartnicka, A., Seidel, A., John, M., Basar-Maurer, S., Westendorf, J., and Algenstaed, P., Noni juice
decreases significantly blood HbA1c of patients with diabetes type 2. Diabetologie und Stoffwechsel, 8,
P121, 2013 (in German).
70. Deng, S., West, B.J., Palu, A.K., and Jensen, C.J., Determination and comparative analysis of major
iridoids in different parts of Morinda citrifolia. Phytochem. Anal., 22, 26–30, 2013.
71. West, B.J., Palmer, S.K., Deng, S., and Palu, A.K., Antimicrobial activity of an iridoid rich extract from
Morinda citrifolia fruit. Curr. Res. J. Biol. Sci., 4, 52–54, 2012.
35
Orange Juice
Rita Maria Velázquez-Estrada, José Afid Chávez-Ocegueda,

Ma. Manuela Hernández-Herrero, and Artur Xavier Roig-Sagués
CONTENTS
35.1 Introduction................................................................................................................................... 423
35.3.1 l-Ascorbic Acid................................................................................................................ 425
35.3.2 Carotenoids....................................................................................................................... 425
35.3.3 Polyphenols....................................................................................................................... 427
35.3.4 Antioxidant Efficacy......................................................................................................... 429
35.4 Health Effects................................................................................................................................ 430
35.6 Conclusion..................................................................................................................................... 434
References............................................................................................................................................... 434
35.1 Introduction
Citrus fruits are one of the most consumed fruit products in the world due to their appealing organoleptic
characteristics and nourishing value. Among them, orange juice is the most widely consumed. Actually,
fresh orange consumption is declining in the developed countries, and is being replaced by orange juice
consumption since juice is a product that offers many advantages in the lifestyles of people who are
health conscious, demand convenience, and place a premium on food safety [1].
Fresh orange juice is generally perceived to be more wholesome and to have a better flavor than pro-
cessed juice, although the consumption of frozen and concentrated juice has increased steadily over the
years. The consumption of natural juices is increasing as a consequence of the search for a healthier life.
Thus, it is important to know the positive or negative effects of oran juice products on health.
Orange juice is a rich source of vitamin C and carotenoids as well as the major source of flavanones
(mainly hesperidin and narirutin) in the diet of those in the developed countries and is therefore a natu-
ral source of several antioxidants [2–4]. There is also a biomedical interet in citrus fruits because their
consumption appears to be associated with a lower risk of cardiovascular disease (CVD) as well as of
colorectal, esophageal, and gastric cancers due to their chemical composition [5,6]. This chapter high-
lights the nutritional characteristics of orange juice and its benefits on health, reviewing especially the
contribution of bioactive substances, the effect of processing, and the future trends, with remarks on the
use of new technologies to obtain more stable fresh-like juices.

Orange juice is the liquid extract from mature oranges of the species Citrus sinensis and hybrids thereof.
It is made by squeezing the fresh orange, drying, and later rehydrating the juice, or concentration of the
juice and later adding water to the concentrate. Almost, all oranges in the world are consumed as fresh
423
TABLE 35.1
Compositional and Nutritional Characteristics of Orange Juices
Nutrient Unit Orange Juice (per 100 g) Orange Juice (per Cup, 248 g) Percentage of RDAa
Water g 88.30 218.98 0
Protein g 0.70 1.74 3
Fat (lipid) g 0.20 0.50 1
Carbohydrate g 10.40 25.79 9
Total dietary fiber g 0.20 0.50 2
Total sugars g 8.40 20.8 0
Minerals
Calcium mg 11 27 3
Sodium mg 1 2 0
Vitamins
Folate (DFE) μg 30 74 19
Niacin mg 0.400 0.992 5
Pantothenic acid mg 0.190 0.471 5
Riboflavin mg 0.030 0.074 4
Thiamin mg 0.090 0.223 15
Vitamin B6 mg 0.040 0.099 5
Vitamin C mg 50 124 207
Reference Release 27, 2014, Published online at: http://ndb.nal.usda.gov/ndb/ (accessed January 9, 2015).
a Values are based on 2000 cal reference diet for adults and children aged >4 years.
Abbreviations: DFE, dietary folate equivalents; RDA, Recommended Dietary Allowances.
fruit, fresh squeezed juice, pasteurized juice, and juice reconstituted from concentrate, with at least 85%
of the oranges produced by most producers being processed into juice [7].
It is well known that orange is one of the most abundant sources of vitamin C; however, it also provides
a variety of vitamins and minerals. A cup serving of fresh orange juice (248 g) provides more than 100%
of the Recommended Dietary Allowances (RDA) of vitamin C, 14% of potassium, 15% of thiamin, and
19% of folate (Table 35.1). Because of its citric acid content, orange juice is acidic, with a typical pH of
around 3.5.
The samples of orange juices marketed as straight processed and as single-strength, γ-aminobutyric
acid, arginine, asparagine, aspartic acid, glutamic acid, proline, and serine were present in high amounts,
accounting for some 89% of total free amino acids [9]. In general, single-strength juices from con-
centrates showed lower mean content of free amino acids (2740 mg/L) than straight processed juices
(3835 mg/L).

Orange juice is a rich source of phytochemicals with antioxidant activities. Phytochemicals can be
defined as substances found in plant sources, including fruits and vegetables that, when ingested on a
daily basis, may modulate human metabolism in a manner favorable for the prevention of chronic and
degenerative diseases [6]. According to some epidemiological studies, high consumption of orange juice
is associated with a reduced risk of free radical–related oxidative damage and suffering from diseases
Orange Juice 425
such as certain types of cancer or cardiovascular and neurological ailments [10]. The most important
antioxidants naturally present in orange juice are l-ascorbic acid, carotenoids, flavonoids, and other
polyphenolic compounds [11].
35.3.1 l-Ascorbic Acid

Orange juice is appreciated and consumed, partly, because of its content of vitamin C, which is an
essential nutrient for humans, and because of its high antioxidant power it provides protection against
the presence of free radicals participating in the prevention of many diseases, but orange juice has other
prominent properties. Vitamin C is highly bioavailable and consequently is one of the most important
water-soluble antioxidants in cells, efficiently scavenging reactive oxygen species (ROS) such as O2•−,
HO•, peroxyl radicals, and singlet oxygen [12]. The term vitamin C is used as the generic descriptor for
all compounds exhibiting qualitatively the biological activity of ascorbic acid. The principal natural
compound with vitamin C activity is l-ascorbic acid, which is easily and reversibly oxidized to dehy-
droascorbic acid (DHAA), forming ascorbyl radical anion (also known as semidehydroascorbate) as an
intermediate. DHAA possesses full vitamin C activity and in vivo it is readily taken up by erythrocytes
and other cells and reduced to ascorbic acid. However, DHAA is not readily absorbed across the intes-
tinal mucosa, and supplemental DHAA has little antiscorbutic activity. Thus, oxidized ascorbic acid
content of orange juice can be used as an indicator of oxidative stress but cannot be considered a dietary
source of vitamin C [13].
35.3.2 Carotenoids
Orange juice contains other compounds, such as carotenoids, that also contribute significantly to its anti-
oxidant capacity. The carotenoids profile and their total content in orange juices depends on the variety/
cultivar, stage of maturity of the fruit, climatic factors, industrial processing methods (pasteurization,
freezing, and concentration), and storage conditions, among others (Table 35.2) [14].
Orange juice is a complex source of carotenoids that include those with pro–vitamin A activ-
ity (β-carotene, α-carotene, and β-cryptoxanthin), as well as antioxidant carotenoids (zeaxanthin and
lutein). Structures of some common carotenoids found in orange juices are shown in Figure 35.1. The
characteristic color of both the peel and pulp of oranges are mainly due to carotenoid pigments. However,
the reddish color of juices from certain varieties, such as blood oranges, is due to anthocyanins [15] and
in the exceptional case of the navel orange Cara Cara color is due to lycopene [16].
Gama and Sylos [17] determined 13 carotenoid pigments in Brazilian Valencia orange juice, among
which violaxanthin, lutein, zeaxanthin, β-cryptoxanthin, ζ-carotene, α-carotene, and β-carotene were
the most abundant. The total carotenoid content reported was 12 mg/L and the major carotenoids were
lutein (23%), β-cryptoxanthin (21%), and zeaxanthin (20%).
The industrial processing influences the profile and content of carotenoids in orange juice. Some
studies have demonstrated that thermal pasteurization of orange juice leads to decreased level of
some carotenoids such as violaxanthin, antheraxanthin [18], and lutein [19] as well as pro–vitamin A
carotenoids (β-carotene, α-carotene, and β-cryptoxanthin) [20]. Nevertheless, other losses of total
TABLE 35.2
Total Carotenoids in Some Kinds of Orange Juices
Orange Juice Reported Major Carotenoids Total Carotenoids (mg/L) References
Freshly squeezed juice β-cryptoxanthin and lutein 4.93 [22]
Fresh juice from Brazilian Lutein, β-cryptoxanthin, and 12 [16]
valencia orange zeaxanthin
Made-from-concentrate juice Auroxanthin and mutatoxanthin 1.37–5.89 [20]
Ultra frozen juice Violaxanthin and 17.2–29.4 [21]
antheraxanthin
Commercial pasteurized juice β-Cryptoxanthin 1.49–2.87 [22]
α-Carotene
β-Carotene
HO
β-Cryptoxanthin
OH
HO
Lutein OH
O
O
HO
Violaxanthin
OH
HO
Zeaxanthin
FIGURE 35.1 Chemical structures of some typical carotenoids found in orange juice. (Adapted from Meléndez-Martínez,
A.J. et al., J. Food Comp. Anal., 20, 638, 2007. With permission.)
carotenoids, pro–vitamin A, and the amount of zeaxanthin may not be significant after thermal pasteuri-
zation and concentration of orange juice [19]. Around 30 carotenoids have been detected in orange
juices from concentrated, which indicate that such foodstuff is one of their most intricate sources of
those pigments. In this kind of orange juices, the absence or occurrence at low levels of certain isomers
of the 5,6-epoxycarotenoids antheraxanthin and violaxanthin, which are major carotenoids in fresh or
slightly processed juices, suggests that the intrinsic acidity of juice and the long shelf life contribute to
the isomerization of 5,6-epoxycarotenoids into their corresponding 5,8-epoxycarotenoids auroxanthin
and mutatoxanthin. The total contents of carotenoids for orange juices from concentrates are in the
range of 1.37–5.89 mg/L [21], being clearly lower than those reported for ultra-frozen orange juices
(17.2–29.4 mg/L) [22].
In a comparison based on the concentration of carotenoids between factory-produced concentrates
with those obtained from the Brazilian retail markets and with authentic hand-squeezed juices, it was
observed that the total carotenoids present in samples of frozen concentrated orange juice from factories
contained 0.26 mg/L, while retail samples contained a slightly greater amount (0.46 mg/L). Frozen con-
centrated orange pulp wash presented much lower concentrations, ranging from 0.04 to 0.08 mg/L, and
hand-squeezed juices ranged from 0.11 to 1.21 mg/L. Moreover, in hand-squeezed juices, zeaxanthin
was present at the highest concentration and β-carotene was present in all orange juice samples analyzed,
while in frozen concentrated orange juice and frozen concentrated orange pulp wash neither α-carotene
nor β-carotene was detected. Finally, the profile of the carotenoids in retail freshly squeezed orange juice
matched with authentic hand-squeezed juices in which zeaxanthin was found in the highest concentration
and β-carotene was present in similar amounts [14].
Orange Juice 427
Another study comparing commercial orange juices consumed in Spain with their freshly squeezed
counterparts showed that the main antioxidant carotenoids of the commercial orange juices were
β-cryptoxanthin, α-cryptoxanthin, zeaxanthin, lutein, β-carotene, and α-carotene with β-cryptoxanthin
being present at the highest concentration. Similar carotenoids’ profile was also found in freshly squeezed
orange juice. Moreover, the orange juices studied in this work showed a high content of pro–vitamin A
carotenoids (α-carotene, β-carotene, and β-cryptoxanthin). The concentration of total antioxidant
carotenoid compounds in the commercial orange juice studied varied from 4.01 to 9.17 mg/L, where the
orange juice processed by freezing without previous pasteurization showed the highest total carotenoid
content, as well as the highest content of all carotenoids, with the exception of β-cryptoxanthin [23].
35.3.3 Polyphenols
Flavonoids constitute the largest group of phenolics in the plant kingdom. To date, more than 6000
flavonoids have been identified, although a smaller number of them are important from a dietary point
of view. Structurally, flavonoids are usually characterized by a C6 –C3–C6 carbon skeleton. Flavonoids
usually occur as aglycones (without sugar moieties) and as glycosides (with sugar moieties). The six
selected classes of flavonoids described (flavones, flavanones, flavonols, isoflavones, anthocyanidins, and
catechins) vary in their structural characteristics around the heterocyclic oxygen ring [24]. Flavonoids
identified in citrus fruits cover over 60 types, particularly; the presence of anthocyanins is typical of
blood orange varieties (Moro, Tarocco, and Sanguinello varieties) [15].
In citrus fruits, flavanones are present in either the glycoside or aglycone forms (Figure 35.2). Among the
aglycone forms, naringenin and hesperetin are the most important flavanones. Of the glycoside forms, two
types are classified: neohesperidosides and rutinosides. Neohesperidosides, flavanones, naringin, neohes-
peridin, and neoeriocitrin are compounds consisting of a flavanone with a neohesperidose (rhamnosyl-α-1,2
glucose), and they have a bitter taste, while rutinosides (flavanones, hesperidin, narirutin, and didymin)
have a flavanone and a disaccharide residue (e.g., rutinose [ramnosyl-α-1,6 glucose]) and are tasteless.
Flavanones are usually present in diglucoside form, conferring the typical taste to citrus fruits [25].
Flavonoids are powerful antioxidants and exhibit their antioxidant activity in several ways [26]. The
chemical nature of phenolics depends on their structural class, degree of hydroxylation, or other sub-
stitutions and conjugations, and degree of polymerization [27]. The surveys about flavonoid contents in
orange juice are vast, encompassing different orange types and also including both fresh and concen-
trated orange juices from several commercial brands or processing technologies (Table 35.3).
Hand-squeezed navel orange juice contains 839 mg/L phenolics, including flavanones, flavones, and
hydroxycinnamic acid derivatives [4]. The flavanones are the main phenolics in the soluble fraction
(649 mg/L) and are also present in the cloud fraction (105 mg/L). Commercial orange juices contain only
81–200 mg/L of soluble flavanones (15%–33%), but the content in the cloud is higher (206–644 mg/L)
(62%–85%), showing that during industrial processing and storage the soluble flavanones precipitate and
are included in the cloud. An in vitro simulation of orange juice digestion shows that a serving of fresh
orange juice (240 mL) provides 9.7 mg of soluble hesperidin (4′-methoxy-3′,5,7-trihydroxyflavanone-
7-rutinoside) and 4.7 mg of the C-glycosyl flavone vicenin 2 (apigenin, 6,8-di-C-glucoside), whereas
pasteurized commercial juices provide 3.7 mg of soluble hesperidin and a higher amount of vicenin
2 (6.3 mg) [4]. Red oranges are an important dietary source of anthocyanins, including cyanidin
3-glucoside and an acylated derivative, cyanidin 3-(6″-malonyl) glucoside [15].
Kawaii et al. [29] determined 24 flavonoids from 66 citrus species and near-citrus relatives grown in the
same field and year; hesperidin (548 μg/100 mg of dry weight) and narirutin (186 μg/100 mg of dry weight)
were the major flavonoids in the case of the oranges. In sour and sweet orange juices, the highest content
of 48 mg/100 g of total flavanones was observed in sour orange (C. aurantium), while in sweet orange
(C. sinensis) it was 17 mg/100 g. Sour orange has a flavanone profile dominated by naringin and neohesperi-
din, while sweet orange had a flavanone glycoside pattern consisting predominantly of hesperidin and nariru-
tin. No differences were observed (P > 0.05) between whole fruit (pulp and vesicle) and the juice (hand and
commercial) for narirutin. There is a slightly significant difference for hesperidin, but only between the juice
squeezed by hand and the whole fruit (pulp and vesicle). No differences (P > 0.05) existed between com-
mercial juices and juices squeezed by hand, or between commercial juices and whole fruit for hesperidin [5].
R3
Flavanone Aglycone Forms R4
Naringenin R1 —
— OH; R2 —
— OH; R3 —
—H; R4 —
—OH R1 O
Hesperetin R1—
—OH; R2 —
— OH; R3 —
— OH; R4 —
— OCH3
Isosakuranetin —OH; R2 —
R1 — —OH; R3 —
—H; R4 —
— OCH3
Heridictyol R1 —
—OH; R2 —
—OH; R3 —
—OH; R4 —
— OH R2 O
Flavone and Flavonol Aglycone Forms R3

Apigenin R1 —
—OH; R2 —
—OH; R3 — — OH; R5 —
—H; R4 — —H R4
Luteolin R1 — —OH; R3 —
—OH; R2 — — OH; R4 —
—OH; R5 —
—H R1 O
Diosmetin R1—
—OH; R2 —
—OH; R3 —
— OH; R4 —
—OCH3; R5—
—H
Quercetin R1—
—OH; R2 —
— OH; R3 —
— OH; R4 —
—OH; R5 —
—OH R5
Kaempferol R1 —
— OH; R2 — —H; R4 —
—OH; R3 — —OH; R5 —
— OH R2 O
R3
Flavanone Neohesperidoside Forms
CH2OH R4
Naringin R2 —
—OH; R3 —
— H; R4 —
—OH H O O
R2 — OH
Neohesperidin —OH; R3 —
—OH; R4 —
—OCH3
H
Poncirin R2 —
—OH; R3 — — OCH3
—H; R4 — OH OH O
CH3 R2 O
H
OHOH
Flavanone Rutinside Forms
R3
Narirutin R3 —
—H; R4 —
—OH R2
OH OO—CH2
Hesperidin R3 —
—OH; R4 —
—OCH3
CH3 O O
Didymin — H; R4—
R3 — —OCH3 OH
H
Eriocitrin — OH; R4 —
R3 — —OH OH
OH
Diosmin — OH; R4 —
R3 — —OCH3
R2 O
FIGURE 35.2 Structural characteristics of orange juice flavonoids in the aglycone and glycoside form. (Adapted from
Tripoli, E. et al., Food Chem., 104, 466, 2007. With permission.)
In juices obtained from citrus varieties (sweet oranges) from China, hesperidin and narirutin were the
major flavanone glycosides observed, whereas naringin and neohesperidin were not detectable. In these
juices, total phenolic acids ranged from 14 to 72.61 mg/L [30]. In the case of orange juices available in
the U.S. market, Vanamala et al. [31] evaluated 12 products made from concentrated and 14 made from
nonconcentrated juices, where hesperidin was found to be the major flavonoid followed by narirutin and
didymin. Total flavonoid content in samples made from concentrated orange juice (53 mg/100 mL) was
significantly (P < 0.05) higher than those made from nonconcentrated orange juice (36.5 mg/100 mL),
being mainly influenced by the hesperidin content. It has been reported that the concentration process may
duplicate the total flavanone content in the cloud portion of the juice (pulp) due to precipitation from the
soluble fraction [32]. In commercial orange juices obtained from C. sinensis L. Valencia late, naringenin
and hesperitin were the main flavanones identified, being in traditional pasteurized juices at higher content.
The concentration of total flavanone content varied from 36.6 to 137.03 mg/L, averaging 99.82 mg/L [23].
In addition to blond oranges, other important varieties of sweet oranges (C. sinensis L. Osbeck) are
blood (pigmented or red) oranges (Moro, Tarocco, and Sanguinello) whose juice is a typical Italian
product. These cultivars are distinguished because of the presence of anthocyanins in the flesh, as well
as a higher content of vitamin C, flavanones (being hesperidin and narirutin the most abundant), and
Orange Juice 429
TABLE 35.3
Reported Major Polyphenols in Some Kinds of Orange Juices
Orange Juice Polyphenol Content (mg/L) References
Freshly squeezed orange juice Hesperidin 76.61–78.4 [22,57]
Narirutin 20.4
Naringenin 18.8–30.12
Fresh juice from orange varieties of China Hesperidin 305–534 [29]
Narirutin 24.42–288
Made from concentrated juice Hesperidin 329–548 [30]
Narirutin 44–80
Didymin 11.7–25.7
Made from nonconcentrated juice Hesperidin 180–428 [30]
Narirutin 29.5–54.1
Didymin 11.6–31.4
Commercial pasteurized juice Hesperidin 100–101 [22,57]
Naringenin 17–36.54
Ultra high pressure homogenized juice (300 MPa) Hesperidin 188 [57]
Narirutin 19.1
Blood orange juice Hesperidin 113–143 [69]
Narirutin 28.8–32.6
TABLE 35.4
Anthocyanins in Blood Orange Juices
Blood Orange Juice Reported Major Anthocyanins Total Anthocyanins (mg/L) References
Freshly Squeezed Juices
Moro Cyanidin 3-glucoside 72.3–291.3 [34,68,69]
Tarocco Cyanidin 3-(6″-malonyl glucoside) 1.2–103
Sanguinello 6.4–52.6
Processed Juices
NFC 48.3–97 [34]
RFC 49–104 [34]
Commercial Juices
NFC 65.7 [34]
Source: Rapisarda, P. et al., J. Agric. Food Chem., 47, 4718, 1999.
Abbreviations: NFC, not from concentrated; RFC, reconstituted from concentrated.
hydroxycinnamic acids, compared to those of blond orange varieties [33]. The content of anthocyanin
pigments in the juice depends on the varieties, season, or processing applied. The main anthocyanins
present in blood orange juice are cyanidin 3-glucoside and cyanidin 3-(6″-malonyl glucoside) (Table 35.4),
varying in their level in the order of Moro > Sanguinello > Tarocco [35].

Regarding the antioxidant activity of citrus juices (Table 35.5), it has been demonstrated that the total
antioxidant activity is not due to a single phytochemical, though there are some divergences as which
compound is the major contributor [2,11,37]. Ascorbic acid, a nonphenolic compound, is the principal
contributor to the total antioxidant activity of citrus juices [10,30,37]. However, Wang et al. [2] found that
the contribution of vitamin C to the total antioxidant activity was less than 15%. Miller and Rice-Evans
[38] have emphasized the significant contributory role of polyphenols (particularly hesperidin and nariru-
tin) in the total antioxidant activity of long-life orange juice, even when vitamin C was the most abundant
antioxidant. In the case of blood orange juices, the higher antioxidant activity appears to be correlated
TABLE 35.5
Antioxidant Activities in Some Kinds of Orange Juices
Orange Juices Unit Content References
Freshly squeezed juice mmol TE/L 9.70 [57]
mmol TE/L 6.04 [57]
Fresh juice from orange varieties of China mg AAE/L 12.61–899.31 [29]
% 47–60.24 [29]
Commercial ready-to-drink juice mmol TE/L 0.58–2.97 [67]
mg AAE/L 74.8–402.9 [67]
Commercial pasteurized juice mmol FeSO4/L 7.1 [51,57]
% 47.5 [51,57]
mmol TE/L 8.71 [51,57]
mmol TE/L 5.58 [51,57]
Ultra high pressure homogenized juice (300 MPa) mmol TE/L 8.11 [57]
mmol TE/L 6.12 [57]
Freshly squeezed blood orange juice mmol TE/L 1.03–7.05 [34,68]
Commercial blood orange juice mmol TE/L 4.18 [34]
Abbreviations: TE, trolox equivalents; AAE, ascorbic acid equivalents.
with their anthocyanin content. Contrary to this, Arena et al. [37] observed that in blood orange juices,
contribution of anthocyanins to total antioxidant activity does not exceed 10%. Concerning carotenoids,
its contribution to the antioxidant potential of orange juice seems to be negligible.
Sánchez-Moreno et al. [23] found that vitamin C was the major contributor to the antioxidant potential,
followed by flavonoid and carotenoid compounds in commercial traditional pasteurized orange juices,
and freshly squeezed orange juices. Ascorbic acid, total vitamin C, and β-cryptoxanthin content cor-
related positively with the free radical scavenging parameters. Gil-Izquierdo et al. [32], evaluating the
effect of processing techniques from an industrial scale (squeezing, mild pasteurization, standard pas-
teurization, concentration, and freezing) on orange juice antioxidant and bioactive compounds (phenolic
compounds and vitamin C), observed that the content of ascorbic acid provided 77%–96% of the total
antioxidant activity of orange juice. Processing did not affect the total antioxidant activity of the whole
juice, but was reduced by 47% in the pulp.
There are evidences that many antioxidants can act synergistically to provide an effective barrier
against oxidation. In this sense, vitamins E, C, and β-carotene exhibited cooperative synergistic effects
scavenging reactive nitrogen species, while vitamin C regenerates vitamin E in lipid systems. Antioxidant
activity of carotenoid mixtures was assayed in multilamellar liposomes, measuring the inhibition of the
formation of thiobarbituric acid-reactive substances (TBARS). Mixtures are more effective than single
compounds, and the synergistic effect is most pronounced when lycopene or lutein coexist [39].
35.4 Health Effects

Epidemiological studies have suggested associations between the consumption of foods or beverages
rich in polyphenols and a decreased risk of CVD and certain forms of cancer [40,41]. Silvio et al. [42]
studied the effects of red orange juice intake on endothelial function and inflammatory markers in adult
subjects with increased cardiovascular risk and discovered that endothelial function, which was mea-
sured as flow-mediated dilation, significantly improved after 1 week of consumption. Red orange juice
had no significant effect on nitric oxide plasma concentrations and concluded that 7 days consumption
of red orange ameliorates endothelial function and reduces inflammation in nondiabetic subjects with
increased cardiovascular risk. Napoleone et al. [43] investigated if the intake of red or blond orange
juice, enriched or free of anthocyanins, could affect the whole blood procoagulant activity in healthy
volunteers. Orange juice intake decreased the procoagulant activity, possibly through mechanisms inde-
pendent of its anthocyanin content.
Orange Juice 431
Milenkovic et al. [44] studied the effects of orange juice on blood leukocytes in human volunteers
consuming 500 mL of orange juice, 500 mL control drink plus hesperidin or 500 mL control drink on
a daily basis, and placebo throughout a 4-week period. Both orange juice and hesperidin consumption
significantly affected leukocyte gene expression. Orange juice consumption induced changes in expres-
sion of 3422 genes, while hesperidin intake modulated the expression of 1819 genes. Between the orange
juice and hesperidin consumption groups, 1582 regulated genes were in common. Many of these genes
are implicated in chemotaxis, adhesion, infiltration, and lipid transport, which is suggestive of lower
recruitment and infiltration of circulating cells to vascular wall and lower lipid accumulation, and con-
cluded that regular consumption of orange juice for 4 weeks alters leukocyte gene expression to an anti-
inflammatory and antiatherogenic profile, and hesperidin displays a relevant role in the genomic effect
of this beverage. Perche et al. [45] investigated on healthy volunteers the effect of a daily consumption
of hesperidin as a purified compound and orange juice on a panel of immune cell functions, including
pro- and anti-inflammatory cytokines secretion (IL-2 and IL-4, respectively) by leukocytes, lytic activity
of natural killer (NK) cells, and polymorphonuclear neutrophil cells (PMN), cell-induced ROS burst and
found that whatever the intake was, no variations were recorded on leukocyte subset distributions (PMN,
B and T lymphocytes, NK cells, and monocytes). ROS production by stimulated PMNs, lytic activity of
NK cells, or cytokine production capacity of leukocytes in well-nourished healthy volunteers demonste
that consumption within the usual daily intake range of orange juice and its major polyphenol hesperidin
does not induce immunomodulation of cell immune function in healthy well-nourished humans.
Christine et al. [46] discovered that diastolic blood pressure (DBP) was significantly lower after a
4-week consumption of orange juice or control drink plus hesperidin (CDH), than after consumption of
control drink plus placebo (CDP), whereas microvascular endothelium-related reactivity was not sig-
nificantly affected when measured after an overnight fast. However, both orange juice and CDH inges-
tion significantly improved the postprandial microvascular endothelial reactivity compared with CDP
(P < 0.05) when measured at the peak of plasma hesperetin concentration and concluded that in healthy,
middle-aged, moderately overweight men, orange juice decreased DBP when regularly consumed and
postprandially increased endothelium-dependent microvascular reactivity.
On the other hand, in recent years, there has been a dramatic increase in the incidence of overweight
and obesity in children and adolescents, due these health problems. In this sense, O’Neil et al. [47] exam-
ined the association of 100% orange juice consumption on the intake of select nutrients, diet quality,
body weight status, and other physiologic parameters and associated risk factors in children and adoles-
cents. Consumers had higher energy intake than nonconsumers, but there were no differences in weight
or body mass index (BMI), and there was no significant difference in the risk of being overweight or
obese between consumers and nonconsumers. Consumers had a higher percentage of population meeting
the estimated average requirement for vitamin A, vitamin C, folate, and magnesium.
Aptekmann and Cesar [48] investigated how the association of orange juice consumption with aerobic
training affected serum lipids and physical characteristics of overweight and middle-aged women. The
study consisted of 13 women who consumed 500 mL/day of orange juice and did 1 h aerobic training
three times a week for 3 months. The control group consisted of another 13 women who did the same
aerobic training program but did not consume orange juice. It was observed that consumption of orange
juice by the experimental group was associated with increased dietary intake of vitamin C and folate by
126% and 61%, respectively. Serum low-density lipoprotein (LDL) cholesterol decreased by 15% (P <
0.05) and high-density lipoprotein (HDL) cholesterol increased by 18% (P < 0.05) in the experimental
group, but no significant change was observed in the control group, concluding that consumption of
500 mL/day of orange juice associated with aerobic training in overweight women decreased the CVD
risk by reducing LDL cholesterol levels and increasing HDL cholesterol levels. This association also
decreased blood lactate concentration and increased anaerobic threshold, showing some improvement
in the physical performance. The same authors [33] investigated the hypothesis that a daily long-term
consumption of orange juice is associated with an improvement of lipid profile, helping to lower the risk
factors of CVD in normolipidemic and moderately hypercholesterolemic adults, and found that orange
juice consumers with both normal serum lipid levels and moderate hypercholesterolemia had signifi-
cantly lower total cholesterol, LDL cholesterol, apolipoprotein B (Apo B), and LDL/HDL ratio in com-
parison with nonconsumer counterparts. A similar study was made by Titta et al. [49] who observed that
dietary supplementation with Moro orange juice significantly reduced body weight gain and fat accumu-
lation regardless of the increased energy intake because of sugar content. Furthermore, mice drinking
Moro juice were resistant to high fat diet–induced obesity with no alterations in food intake. Only the
anthocyanin extract, but not the purified cyaniding 3-glucoside, slightly affected fat accumulation. Gene
expression analysis of fat tissues confirmed that Moro juice could entirely rescue the high fat‒induced
transcriptional reprogramming.

Orange juice is produced in numerous forms such as frozen concentrate, orange juice from concentrate,
and pasteurized juice. Although these kinds of presentations of orange juice must conform to strict
guidelines that prevent unnatural change in the juice, concern for diet and nutrition has led consumers to
seek products with not only a long shelf life but also high quality as freshly prepared flavor, texture, and
color, with minimal processing and no chemical preservatives added.
The inactivation of spoilage and pathogenic microorganisms as well as the inactivation of endog-
enous pectin methylesterase (PME) are prerequisites for the extension of the shelf life of the juice [50].
Today, refrigerated juices that are not obtained from concentrates and have been subjected to mild pas-
teurization partly satisfy the requirements of higher quality demanded by consumers. The shelf life of
these juices ranges between 28 and 45 days in refrigeration and their quality approaches that of freshly
squeezed juices [51]. However, during storage, the orange juice suffers an important number of deteriora-
tion reactions (ascorbic acid degradation, cloud loss, microbial spoilage, development of off flavors, and
changes in color, among others) that cause an important quality loss [10,52].
In case of vitamin C, its content in orange juice is highly variable and depends on, for example, the
variety and maturity of the oranges, fruit freshness, and processing factors including packaging [18,32].
In this connection, Gil-Izquierdo et al. [32] determined the effects of individual orange juice processing
techniques on an industrial scale (pasteurization, concentration, and freezing) and two juice squeezing
methods, observing that orange juice produced by commercial squeezing presented 25% more vitamin C
than domestic squeezing, because of the additional extraction of vitamin C from the solid parts of the
orange caused by the commercial squeezing system. Mild and standard pasteurization treatments also
slightly increased the l-ascorbic acid and DHAA contents, possibly due to a better extraction from the
solid parts as a consequence of temperature. These authors suggested that the concentration process
of juice could be considered as a positive method to maintain the antioxidant activity since this tech-
nique favored the conversion of DHAA to l-ascorbic acid. Another processing step in the orange juice
production is the reduction of bitterness (debittering). Bitter or astringent tastes tend to be rejected by
consumers, and for this reason, early-season orange juice or orange juice obtained from immature fruits
must be subjected to an appropriate debittering step, to remove bitter components in the orange by physi-
cal adsorption in a resin, a process that causes a 26% reduction in the ascorbic acid content of orange
juice [53].
Routine handling and storage of orange juice postproduction likely reduces the vitamin C. Lee and
Coates [18] observed a 19.2% loss of vitamin C after 24 months of storage at −23°C in frozen, fresh
squeezed, unpasteurized, and polyethylene-bottled orange juice. Nevertheless, the % retention of vitamin
C after 24 months of storage at −23°C was still sufficient to supply 100% RDA from 240 mL of orange
juice. Orange juices from frozen concentrates contained 86 mg reduced vitamin C per fluid cup at initial
preparation and 39–46 mg/100 mL after 4 weeks of storage at 4°C, while ready-to-drink juices contained
27–65 mg/100 mL at opening and 0–25 mg/100 mL at expiration 4 weeks later. Ready-to-drink orange
juices had two- to threefold higher concentrations of oxidized vitamin C than the reconstituted from
frozen orange juices. The decomposition rate of reduced vitamin C was similar for all juices, about 2%
per day, once opened. Thus, ready-to-drink orange juices should be purchased 3–4 weeks before expi-
ration date and consumed within a week of opening to assure the necessary daily supply of vitamin C
[13]. Esteve et al. [51] observed that l-ascorbic acid in commercial orange juice decreased faster at 10°C
than at 4°C. A period of at least 42 days at 4°C and 35 days at 10°C was established as the shelf life of
the juices considering the minimum values recommended for processed orange juice (40 mg/100 mL).
Orange Juice 433
In the same line, Klimczak et al. [54] observed that after 6 months of storage at 18°C, 28°C, and 38°C,
the content of vitamin C decreased by 21%, 31%, and 81%, respectively. Kabasakalis et al. [3] and Arena
et al. [37] also reported a decrease in l-ascorbic acid content in commercial orange juices after 6 months
of storage at room temperature by 29% and by about 25% in reconstituted from concentrate blood orange
juice stored at 25°C during 60 days.
At present, there is a renewed interest within the citrus industry in the development of innovative
practices to meet the demand of orange juices of the highest quality, as a consequence of which a
noticeable rise in the consumption of direct orange juices not subjected to thermal treatments has taken
place in the last years [22]. New technologies, such as high hydrostatic pressure (HHP), pulsed electric
fields (PEF), and ultra-high pressure homogenization (UHPH) are being introduced by the food indus-
try as an alternative or complementary process to traditional thermal treatments. HHP and PEF have
demonstrated potential to improve the quality and freshness character of processed food while assuring
its microbiological safety [10,55–57]. Meanwhile, UHPH allows to process foodstuffs in continuous
and its great potential to inactivate pathogenic and spoilage microorganisms [58], inhibit the activity
of enzymes [59], and minimize the adverse effects of heat on constituents in orange juice has been
demonstrated [60].
Sánchez-Moreno et al. [61] showed that application of HHP orange juice conditions (400
MPa/40°C/1 min) with respect to a freshly squeezed juice had higher content of β-cryptoxanthin,
α-cryptoxanthin (as β-cryptoxanthin), zeaxanthin, lutein, β-carotene, α-carotene (as β-carotene), total
carotenoids, and vitamin A.
Yeom et al. [62] showed that orange juice treated with PFE at 35 kV/cm for 59 µs retained significantly
higher content of ascorbic acid than heat-pasteurized orange juice during storage at 4°C. The concentra-
tion of ascorbic acid in PEF-treated orange juice reached 25 mg/100 mL (the concentration of ascorbic
acid that orange juice should at least have at the time of expiration date to supply 100% of RDA of vita-
min C) at 4°C after 47 days, which is significantly longer than the 31 days of heat-pasteurized orange juice.
Regarding UHPH, Welti-Chanes et al. [63] observed that vitamin C content of orange juice was not
affected by this technology when treated between 50 and 250 MPa at 22°C, although these treatments
were insufficient to guaranty a permanent inactivation of the pectin methylesterase after a single pass.
Velázquez-Estrada et al. [60] evaluated the effect of UHPH (100, 200, and 300 MPa at 10°C and 20°C)
on bioactive compounds and antioxidant activity of orange juice. They observed that the remaining
content of ascorbic acid in the UHPH treated samples at any pressure was significantly higher than that
in the thermal pasteurized one. Moreover, flavanone content increased with the UHPH treatments, more
specifically the amounts of hesperidin, achieving its highest concentration on the samples treated at 200
and 300 MPa. The total polyphenol content and antioxidant capacity values were significantly lower in
the thermal pasteurized one.
Concerning others technologies, Vikram et al. [64] studied the application of ohmic heating system,
infrared heating system, microwave heating system, and conventional processing observing that the
degradation of vitamin C was higher during microwave heating due to uncontrolled temperature gener-
ated during processing. Ohmic heating gave the best result facilitating better vitamin retention. Zerdin
et al. [65] studied the storage of orange juice packed in oxygen scavenging and oxygen barrier film to
determine the extent of ascorbic acid loss due to oxygen as a function of time and temperature. The initial
concentration of ascorbic acid in the orange juice was 374 mg/L and decreased by 74 and 104 mg/L after
3 days of storage at 25°C in the oxygen scavenging and oxygen barrier film, respectively.
The development of innovative bioactive functional food products is also getting concentrated focus
from fruit juices industry. Several technologies and innovations have been devised to conserve and uti-
lize the orange juice in food formulations. Smoothies have rapidly increased in popularity (e.g., product
growth rising 2.39 times from 2002 to 2007 according to food merchandisers) due to their healthy and
convenient features [66]. These beverages are prepared by blending fruit, fruit juice, ice, and yogurt or
milk. Orange juice tends to be a common ingredient in the formulation, and consequently contain a high
quantity of phytochemicals, in particular, polyphenolic compounds [67].
Dehydrated orange juice is a product which shows many benefits and economic potentials over their
liquid counterparts such as reduced volume or weight, reduced packaging, easier handling and trans-
portation, and much longer shelf life. In addition their physical state provides a stable, natural, and
easily dosable ingredient, which generally finds usage in many foods and pharmaceutical products such
as flavoring and coloring agents. There are studies which include the blend of diverse ingredients with
concentrated orange juice in order to improve the physical, chemical, reconstitution, and sensory char-
acteristics of the dehydrated orange juice (e.g., the addition of maltodextrin or skim milk). The potential
target industries for this orange juice product would be ice cream, yogurt, and nonalcoholic beverage
manufacturing [68].
35.6 Conclusion
Orange juice is the most widely consumed fruit juice in the world due to its appealing organoleptic
characteristics and its nourishing value, being a rich source of a great variety of phytochemicals with
antioxidant activities, such as ascorbic acid, carotenoids, flavonoids, and other polyphenolic compounds,
including flavanones, flavones, and hydroxycinnamic acid derivatives. Consumption of orange juice
decreases the risk of CVD, due to an improvement of the lipid profile in blood, and certain forms of
cancer due to its chemical composition. Thermal processing of juice is necessary in order to ensure sta-
bility during shelf life and the elimination of pathogen microorganism, but can reduce the nutritive and
sensorial properties. New technologies, such as HHP, PEF, and especially UHPH, are being tested by the
food industry as an alternative to traditional thermal treatments since they can better preserve or even
increase the antioxidant activity during the shelf life of juices.
REFERENCES
1. Spreen, T.H., Projections of World Production and Consumption of Citrus to 2010, China/FAO Citrus
Symposium, 2003. Published online at: http://www.fao.org/docrep/003/x6732e/x6732e02.htm (accessed
December 1, 2014).
2. Wang, H., Cao, G., and Prior, R.L., Total antioxidant capacity of fruits. J. Agric. Food Chem., 44,
701–705, 1996.
3. Kabasakalis, V., Siopidou, D., and Moshatou, E., Ascorbic acid content of commercial fruit juices and
its rate of loss upon storage. Food Chem., 70, 325–328, 2000.
4. Gil-Izquierdo, A., Gil, M.I., Ferreres, F., and Tomás-Barberán, F.A., In vitro availability of flavonoids
and other phenolics in orange juice. J. Agric. Food Chem., 49, 1035–1041, 2001.
5. Peterson, J.J., Dwyer, J.T., Beecher, G.R., Bhagwat, S.A., Gebhardt, S.E., Haytowitz, D.B., and Holden,
J.M., Flavanones in oranges, tangerines (mandarins), tangors, and tangelos: A compilation and review
of the data from the analytical literature. J. Food Comp. Anal., 19, S66–S73, 2006.
6. Tripoli, E., Guardia, M.L., Giammanco, S., Majo, D.D., and Giammanco, M., Citrus flavonoids:
Molecular structure, biological activity, and nutritional properties: A review. Food Chem., 104, 466–479,
2007.
7. Grigelmo-Miguel, N. and Martin-Belloso, O., Characterization of dietary fiber from orange juice extrac-
tion. Food Res. Int., 31, 355–361, 1999.
8. U.S. Department of Agriculture (USDA), National Nutrient Database for Standard Reference Release
27, 2014. Published online at: http://ndb.nal.usda.gov/ndb/ (accessed January 9, 2015).
9. Del Castillo, M.D., Santa-María, G., Pueyo, E., Corzo, N., and Olano, A., Differences in amino acid
composition in commercial orange juices. J. Sci. Food Agric., 46, 2329–2331, 1998.
10. Polydera, A., Stoforos, N., and Taoukis, P., Effect of high hydrostatic pressure treatment on post process-
ing antioxidant activity of fresh Navel orange juice. Food Chem., 91, 495–503, 2005.
11. Gardner, P.T., White, T.A., McPhail, D.B., and Duthie, G.G., The relative contributions of vitamin C,
carotenoids and phenolics to the antioxidant potential of fruit juices. Food Chem., 68, 471–474,
2000.
12. Halliwell, B., Antioxidants in human health and disease. Ann. Rev. Nutr., 16, 33–50, 1996.
13. Johnston, C.S. and Bowling D.L., Stability of ascorbic acid in commercially available orange juices.
J. Am. Dietetic Assoc., 102, 525–529, 2003.
14. Pupin, A., Dennis, M., and Toledo, M., HPLC analysis of carotenoids in orange juice. Food Chem., 64,
269–275, 1999.
Orange Juice 435
15. Maccarone, E., Campisi, S., Fallico, B., Rapisarda, P., and Sgarlata, R., Flavor components of Italian
orange juices. J. Agric. Food Chem., 46, 2293–2298, 1998.
16. Lee, H.S., Characterization of carotenoids in juice of red navel orange (Cara Cara). J. Agric. Food
Chem., 49, 2563–2568, 2001.
17. Gama, J.J.T. and de Sylos, C.M., Major carotenoid composition of Brazilian Valencia orange juice:
Identification and quantification by HPLC. Food Res. Int., 38, 899–903, 2005.
18. Lee, H.S. and Coates, G.A., Effect of thermal pasteurization on Valencia orange juice color and
pigments. LWT—Food Sci. Technol., 36, 153–156, 2003.
19. Gama, J.J.T. and de Sylos, C.M., Effect of thermal pasteurization and concentration on carotenoid
composition of Brazilian Valencia orange juice. Food Chem., 100, 1686–1690, 2007.
20. Lessin, W.J., Catignani, G.L., and Schwartz, S.J., Quantification of cistrans isomers of provitamin A
carotenoids in fresh and processed fruits and vegetables. J. Agric. Food Chem., 45, 3728–3732, 1997.
21. Meléndez-Martínez, A.J., Britton, G., Vicario, I.M., and Heredia, F.J., The complex carotenoid pattern
of orange juices from concentrate. Food Chem., 109, 546–553, 2008.
22. Meléndez-Martínez, A.J., Vicario, I.M., and Heredia, F.J., Review: Analysis of carotenoids in orange
juice. J. Food Comp. Anal., 20, 638–649, 2007.
23. Sánchez-Moreno, C., Plaza, L., De Ancos, B., and Cano, M.P., Quantitative bioactive compounds assess-
ment and their relative contribution to the antioxidant capacity of commercial orange juices. J. Sci. Food
Agric., 83, 430–439, 2003.
24. Peterson, J. and Dwyer, J., Flavonoids: Dietary occurrence and biochemical activity. Nutr. Res., 12,
1995–2018, 1998.
25. Macheix, J.J., Fleuriet, A., and Billot, J., Fruit Phenolics, CRC Press/Taylor & Francis Group, Boca
Raton, FL, 1990.
26. Bombardelli, E. and Morazzoni, P., The flavonoids: New perspectives in biological activities and thera-
peutics. Chimica Oggi, 11, 25–28, 1993.
27. Rice Evans, C., Miller, N.J., and Paganga, G., Structure—Antioxidant activity relationships of flavo-
noids and phenolic acids. Free Radic. Biol. Med., 20, 933–956, 1996.
28. Kelebek, H., Canbas, A., and Selli, S. Determination of phenolic composition and antioxidant capacity
of blood orange juices obtained from CVS. Moro and Sanguinello [Citrus sinensis (L.) Osbeck] grown
in Turkey. Food Chem., 107, 1710–1716, 2008.
29. Kawaii, S., Tomono, Y., Katase, E., Ogawa, K., and Yano, M., Quantitation of flavonoid constituents in
citrus fruits. J. Agric. Food Chem., 47, 3565–3571, 1999.
31. Vanamala, J., Reddivari, L., Yoo, K.S., Pike, L.M., and Patil, B.S., Variation in the content of bioac-
tive flavonoids in different brands of orange and grapefruit juices. J. Food Comp. Anal., 19, 157–166,
2006.
32. Gil-Izquierdo, A., Gil, M.I., and Ferreres, F., Effect of processing techniques at industrial scale on
orange juice antioxidant and beneficial health compounds. J. Agric. Food Chem., 50, 5107–5114, 2002.
33. Aptekmann, N.P. and Cesar, T.B., Long-term orange juice consumption is associated with low LDL-
cholesterol and apolipoprotein B in normal and moderately hypercholesterolemic subjects. Lipids
Health Dis., 12, 119, 2013.
34. Rapisarda, P., Tomaino, A., Lo Cascio, R., Bonina, F., De Pasquale, A., and Saija A., Antioxidant effec-
tiveness as influenced by phenolic content of fresh orange juices. J. Agric. Food Chem., 47, 4718–4723,
1999.
35. Rapisarda, P., Fanella, F., and Maccarone, E., Reliability of analytical methods for determining antho-
cyanins in blood orange juices. J. Agric. Food. Chem., 48, 2249–2252, 2000.
36 Stella, S.P., Ferrarezi, A.C., dos Santos, K.O., and Monteiro, M., Antioxidant activity of commercial
ready-to-drink orange juice and nectar. J. Food Sci., 76, C392–C397, 2011.
37. Arena, E., Fallico, B., and Maccarone, E., Evaluation of antioxidant capacity of blood orange juices as
influenced by constituents, concentration process and storage. Food Chem., 74, 423–427, 2001.
38. Miller, N.J. and Rice-Evans, C.A., The relative contributions of ascorbic acid and phenolic antioxidants
to the total antioxidant activity of orange and apple fruit juices and blackcurrant drink. Food Chem., 60,
331–337, 1997.
39. Stahl, W. and Sies, H., Antioxidant activity of carotenoids. Mol. Aspects Med., 24, 345–351, 2003.
40. Benavente-García, O., Castillo, J., Marin, F.R., Ortuño, A., and Del Río, J.A., Uses and properties of
citrus flavonoids. J. Agric. Food Chem., 45, 4505–4515, 1997.
41. Cesar, T.B., Aptekmann, N.P., Araujo, M.P., Vinagre, C.C., and Maranhão, R.C., Orange juice decreases
low-density lipoprotein cholesterol in hypercholesterolemic subjects and improves lipid transfer to high-
density lipoprotein in normal and hypercholesterolemic subjects. Nutr. Res., 30, 689–694, 2010.
42. Silvio, B., Giuseppe, R., Gioacchina, A., Alessandro, M., Baldassare, C., Maria, M., Salvatore, V., and
Giovanbattista, R., Effects of red orange juice intake on endothelial function and inflammatory markers
in adult subjects with increased cardiovascular risk. Am. J. Clin. Nutr., 95, 1089–1095, 2012.
43. Napoleone, E., Cutrone, A., Zurlo, F., Di Castelnuovo, A., D’Imperio, M., Giordano, L., De Curtis, A.
et al., Both red and blond orange juice intake decreases the procoagulant activity of whole blood in
healthy volunteers. Thrombosis Res., 132, 288–292, 2013.
44. Milenkovic, D., Deval, C., Dubray, C., Mazur, A., and Morand, C., Hesperidin displays relevant role in
the nutrigenomic effect of orange juice on blood leukocytes in human volunteers: A randomized con-
trolled cross-over study. PLoS One, 6, e26669, 2011.
45. Perche, O., Vergnaud-Gauduchon, J., Morand, C., Dubray, C., Mazur, A., and Vasson, M.P., Orange
juice and its major polyphenol hesperidin consumption do not induce immunomodulation in healthy
well-nourished humans. Clin. Nutr., 33, 130–135, 2014.
46. Christine, M., Claude, D., Dragan, M., Delphine, L., Jean François, M., Augustin, S., and Andrzej, M.,
Hesperidin contributes to the vascular protective effects of orange juice: A randomized crossover study
in healthy volunteers. Am. J. Clin. Nutr., 93, 73–80, 2011.
47. O’Neil, C.E., Nicklas, T.A., Rampersaud, G.C., and Fulgoni Iii, V.L., One hundred percent orange juice
consumption is associated with better diet quality, improved nutrient adequacy, and no increased risk
for overweight/obesity in children. Nutr. Res., 31, 673–682, 2011.
48. Aptekmann, N.P. and Cesar, T.B., Orange juice improved lipid profile and blood lactate of overweight
middle-aged women subjected to aerobic training. Maturitas, 67, 343–347, 2010.
49. Titta, L., Trinei, M., Stendardo, M., Berniakovich, I., Petroni, K., Tonelli, C., Riso, P., Porrini, M.,
Minucci, S., and Pelicci, P., Blood orange juice inhibits fat accumulation in mice. Int. J. Obes., 34,
578–588, 2009.
50. Katsaros, G.I., Tsevdou, M., Panagiotou, T., and Taoukis, P.S., Kinetic study of high pressure microbial
and enzyme inactivation and selection of pasteurisation conditions for Valencia orange juice. Int. J.
Food Sci. Technol., 45, 1119–1129, 2010.
51. Esteve, M., Frigola, A., Rodrigo, C., and Rodrigo, D., Effect of storage period under variable conditions
on the chemical and physical composition and colour of Spanish refrigerated orange juices. Food Chem.
Toxicol., 43, 1413–1422, 2005.
52. Bezman, Y., Rouseff, R.L., and Naim, M., 2-Methyl-3-furanthiol and methional are possible off-flavors
in stored orange juice: Aroma-similarity, NIF/SNIF GC-O, and GC analyses. J. Agric. Food. Chem., 49,
5425–5432, 2001.
53. Stinco, C.M., Fernández-Vázquez, R., Hernanz, D., Heredia, F.J., Meléndez-Martínez, A.J., and Vicario
I.M., Industrial orange juice debittering: Impact on bioactive compounds and nutritional value. J. Food
Eng., 116, 155–161, 2013.
54. Klimczak, I., Malecka, M., Szlachta, M., and Gliszczynska-Swiglo, A., Effect of storage on the con-
tent of polyphenols, vitamin C and the antioxidant activity of orange juices. J. Food Comp. Anal., 20,
313–322, 2007.
55. De Ancos, B., Sgroppo, S., Plaza, L., and Cano, M.P., Possible nutritional and health-related value pro-
motion in orange juice preserved by high-pressure treatment. J. Sci. Food Agric., 82, 790–796, 2002.
56. Elez-Martínez, P., Soliva-Fortuny, R.C., and Martín-Belloso, O., Inactivation of Lactobacillus brevis in
orange juice by high-intensity pulsed electric fields. Food Microbiol., 22, 311–319, 2005.
57. Odriozola-Serrano, I., Soliva-Fortuny, R., Hernández-Jover, T., and Martín-Belloso, O., Carotenoid
and phenolic profile of tomato juices processed by high intensity pulsed electric fields compared with
conventional thermal treatments. Food Chem., 112, 258–266, 2009.
58. Velázquez-Estrada, R., Hernández-Herrero, M., López-Pedemonte, T., Briñez-Zambrano, W., Guamis-
López, B., and Roig-Sagués, A., Inactivation of Listeria monocytogenes and Salmonella enterica
serovar Senftenberg 775W inoculated into fruit juice by means of ultra-high pressure homogenisation.
Food Control, 22, 313–317, 2011.
Orange Juice 437
59. Velázquez-Estrada, R.M., Hernandez-Herrero, M.M., Guamis-López, B., and Roig-Sagués, A.X.,
Impact of ultra-high pressure homogenization on pectin methylesterase activity and microbial charac-
teristics of orange juice: A comparative study against conventional heat pasteurization. Innov. Food Sci.
Emerg. Technol., 13, 100–106, 2012.
60. Velázquez-Estrada, R.M., Hernandez-Herrero, M.M., Rüffer, C.F., Guamis-López, B., and Roig-Sagués,
A.X., Influence of ultra-high pressure homogenization processing on bioactive compounds and antioxi-
dant activity of orange juice. Innov. Food Sci. Emerg. Technol., 18, 89–94, 2013.
61. Sánchez-Moreno, C., Plaza, L., De Ancos, B., and Cano, M.P., Vitamin C, provitamin A carotenoids,
and other carotenoids in high-pressurized orange juice during refrigerated storage. J. Agric. Food
Chem., 51, 647–653, 2003.
62. Yeom, H.W., Streaker, C.B., Zhang, Q.H., and Min, D.B., Effects of pulsed electric fields on the quality
of orange juice and comparison with heat pasteurization. J. Agric. Food Chem., 48, 4597–4605, 2000.
63. Welti-Chanes, J., Ochoa-Velasco, C., and Guerrero-Beltrán, J., High-pressure homogenization of orange
juice to inactivate pectinmethylesterase. Innov. Food Sci. Emerg. Technol., 10, 457–462, 2009.
64. Vikram, V., Ramesh, M., and Prapulla, S., Thermal degradation kinetics of nutrients in orange juice
heated by electromagnetic and conventional methods. J. Food Eng., 69, 31–40, 2005.
65. Zerdin, K., Rooney, M.L., and Vermuë, J., The vitamin C content of orange juice packed in an oxygen
scavenger material. Food Chem., 82, 387–395, 2003.
66. Lal, G.G., Getting specific with functional beverages. Food Technol., 61, 24–28, 2007.
67. Rastogi, N.K., Raghavarao, K.S.M.S., Balasubramaniam, V.M., Niranjan, K., and Knorr, D.,
Opportunities and challenges in high pressure processing of foods. Crit. Rev. Food Sci. Nutr., 47,
69–112, 2007.
68. Shrestha, A.K., Ua-Arak, T., Adhikari, B.P., Howes, T., and Bhandari, B.R., Glass transition behavior
of spray dried orange juice powder measured by differential scanning calorimetry (DSC) and thermal
mechanical compression test (TMCT). Int. J. Food Propert., 10, 661–673, 2007.
36
Papaya Juice
Sui Kiat Chang and Cesarettin Alasalvar
CONTENTS
36.1 Introduction................................................................................................................................... 439
36.3 Bioctives and Antioxidant Efficacy.............................................................................................. 442
36.3.2 Polyphenols....................................................................................................................... 442
36.4 Health Effects................................................................................................................................ 445
36.4.1 Bioavailability of Bioactives from Papaya Juice in Relation to Its Intake....................... 445
36.4.2 Results from In Vitro, Animal, and Human Intervention Studies................................... 445
36.6 Conclusion..................................................................................................................................... 450
References................................................................................................................................................451
36.1 Introduction
Carica papaya belongs to the family Caricaceae, having only four genera in the world. The genus Carica
Linn. is represented by four species [1]. Papaya is one of the major fruit crops cultivated in tropical and
subtropical zones. The 2010 world production of papaya was over 11.2 million tons, and these were pro-
duced in an area of 438,588 hectares in 60 countries [2]. Approximately, 47% of the papaya production
was in Central and South America (mainly in Brazil), 30% in Asia, and 20% in Africa. Other papaya
growing nations include Venezuela, China, Peru, Congo, and Ethiopia, all of which contribute less than
3% to the papaya production. There are also other genotypes of papaya with importance in the world [3].
Papaya is a climacteric fruit, which grows year round, and is an elongated berry of various sizes with
a smooth thin skin and a greenish yellow color. Its flesh is thick with a color ranging from yellow to red
and offers a pleasant, sweet, and mellow flavor, containing numerous black peppery seeds [4,5]. It is
marketed mainly as a fresh fruit, but interest in its processed products, such as canned nectar and puree,
has increased [4]. Today, there are many types of fruit juices available commercially on the market. The
consumption of various fruit juices is increasing rapidly as they are convenient, nutritious, and ready to
drink for cooling, refreshing, and detoxifying purposes [6,7]. They may be particularly useful in places
where there is malnutrition, which could lead to diseases related to nutritional deficiency [6–8].
The quality of papaya juice, such as its bioactive compounds and health benefits, have scarcely been
reviewed in the past years. Hence, this chapter highlights the nutritional characteristics, phenolic con-
tents, antioxidant activity, health effects, novel formulations, and future trends of papaya juice.

Table 36.1 summarizes the proximate composition and nutritional characteristics of papaya (C. papaya L.)
products found in the U.S. market [9]. Papaya nectar is an important beverage prepared on a commer-
cial scale. The beverage contains 12%–20% pulp. However, the nutritional composition of its freshly
439
TABLE 36.1
Compositional and Nutritional Characteristics of Papaya Products (per 100 g)
Papaya Nectar (Canned,
Nutrient Unit Papaya Nectar (Canned) Heavy Syrup, Drained) Raw Papaya (Pulp)
Water g 85.02 43.33 88.06
Protein g 0.17 0.14 0.47
Lipid (fat) g 0.15 0.55 0.26
Total sugars g 13.91 52.20 7.82
Minerals
Calcium mg 10 21 20
Iron mg 0.34 0.29 0.25
Magnesium mg 3 6 21
Phosphorus mg nd 6 10
Sodium mg 5 9 8
Zinc mg 0.15 0.05 0.08
Vitamins
Folate (DFE) μg 2 nd 37
Niacin mg 0.15 0.06 0.669
Riboflavin mg 0.004 0.015 0.027
Thiamin mg 0.006 0.015 0.023
Vitamin A IU 361 6 950
Vitamin B12 μg nd 0.00 0.00
Vitamin B6 mg 0.009 0.015 0.038
Vitamin C mg 3.0 3.5 60.9
Vitamin E (ATE) mg 0.24 nd 0.30
Vitamin K μg 0.8 0.3 2.6
Abbreviations: DFE, dietary folate equivalents; IU, international unit; ATE, alpha-tocopherol equivalents; nd, not
detected.
made juice is rarely found in the literature. Although fresh papaya pulp has 1.7 g dietary fiber/100 g, a
small amount of it is present in papaya nectar (0.6 g/100 g) and papaya nectar heavy syrup (1.5 g/100 g).
Papaya nectar contains a low protein content (0.17 g/100 g) compared to its fresh pulp (0.47 g/100 g).
Carbohydrate and total sugar contents of papaya nectar are 14.51 and 13.91 g/100 g, respectively. The
energy value of the papaya nectar (57 kcal/100 g) is higher than that of the papaya pulp (43 kcal/100 g) [9].
Papaya nectar contains a low amount of potassium (31 mg/100 g) even though its fresh pulp has a
much higher amount of 182 mg/100 g. In addition, it also has a low amount of sodium (5 mg/100 g).
Both papaya nectar and papaya nectar heavy syrup contain a very low amount of vitamin C (3.0 and
3.5 mg/100 g, respectively) compared to fresh papaya pulp (60.9 mg/100 g). Papaya nectar is a good
source of vitamin A (361 IU), but at a significantly lower amount than that of its fresh pulp (950 IU).
Significant amounts of vitamins A and C are lost during processing of papaya nectar from its pulp [9].
Meanwhile, trace amounts of folate, niacin, riboflavin, thiamin, vitamins B6, E, and K are present in
papaya nectar (Table 36.1).
Falguera et al. [10] reported that total sugar content of fresh papaya juice is 67.1 g glucose equiva-
lents (GE)/L, with reducing and nonreducing sugar contents of 60.0 and 7.1 g GE/L, respectively.
Papaya Juice 441
The sugar content of fresh papaya juice is considered very low compared to fresh banana passion
fruit and mango juices (90 and 110 g GE/L, respectively). Hence, fresh papaya juice can be considered
as a healthy fruit juice for people choosing a low-sugar drink. El-Mansy et al. [11] reported the total
sugar, fat, protein, ash, and crude fiber contents of a papaya puree (53.4% pulp content) at 7.12%,
0.5%, 0.76%, 0.74%, and 1.19%, respectively. They reported that reducing and nonreducing sugars
of papaya puree were at 2.88% and 4.23%, respectively. Papaya puree contains 93.09 mg/100 g of
vitamin C and 22.6 mg/L of carotenoids [11]. Papaya juice is usually sold as 100% puree or nectar,
or blended with other fruit juices, such as orange, peach, and pineapple.
The pulp of papaya has been shown to contain tocopherol, lycopene, benzyl isothiocyanate [12], pro-
teolytic enzymes such as papain and chymopapain, cystatin, ascorbic acid, cyanogenic glucosides, glu-
cosinolates [13], alkaloids such as carpain and carpasemine, triterpenes, organic acids, and flavonoids,
among others [1,14]. However, the presence of enzymes such as papain, chymopapain, alkaloids, and
glucosinolates has not been determined in papaya juice. Future research should be carried out to fill
this research gap. There is only one study reporting the presence of carotenoids, such as α-carotene,
β-carotene, lycopene, β-cryptoxanthin, and zeaxanthin (Figure 36.1) in freshly made papaya juice at
368, 3427, 5081, 1885, and 83 µg/g (w/w) of juice made from 523 g of the papaya pulp [15] (Table 36.2).
Thus, papaya nectar provides a sufficient amount of vitamin A, some other vitamins as well as min-
erals. Furthermore, it also has a lower energy value compared to other fruit juices. This makes papaya
nectar a potential functional beverage. There is, however, some knowledge gap in the nutritional compo-
sitions (proximate, vitamins, and minerals) of fresh papaya juice. Moreover, the nutritional composition
of the papaya juice produced from different papaya cultivars needs to be investigated.
HO
β-Cryptoxanthin
Lycopene
β-Carotene
α-Carotene
FIGURE 36.1 Structures of major carotenoids detected in papaya juice.
TABLE 36.2
Carotenoids in Fresh Papaya Juice (per µg/g)
Juice α-Carotene β-Carotene Lycopene β-Cryptoxanthin Zeaxanthin
Fresh papaya juice 368 3427 5081 1885 83
Source: Adapted from Gouado, I. et al., Eur. J. Clin. Nutr., 61, 1180, 2007. With permission.
36.3 Bioctives and Antioxidant Efficacy

A recent study reported the production of papaya juice from ripe flesh of C. papaya cultivar Batu Arang
from Malaysia with a yield of 47.5%. This is a low value compared to watermelon juice (71.67%) and star
fruit (64.17%) juice [16]. The total phenolic content of fresh papaya juice is 377 µg gallic acid equiva-
lents (GAE)/g sample while its vitamin C content is 268 µg ascorbic acid equivalents (AAE)/g sample.
The antioxidant activities of the freshly made papaya juice using 2,2-diphenyl-1-picrylhydrazyl (DPPH)
radical scavenging, ferric-reducing antioxidant power (FRAP), and metal chelating assays were 203 µg
AAE/g sample, 469 µg trolox equivalents (TE)/g sample, and 7.89%, respectively. The antioxidant activ-
ity of the freshly made papaya juice correlated well with its total phenolic and vitamin C contents [16].
Another study reported that DPPH radical scavenging activity of the papaya juice extract was 26.5% [17].
Falguera et al. [10] reported that the activity of polyphenol oxidase (PPO) in freshly made papaya juice was
low (0.0125 unit/mL of PPO activity), compared to fresh mangosteen juice (0.1435 unit/mL of PPO activity).
This is not surprising since papaya juice has a low total phenolic content (244 mg GAE/L). A low PPO activity
of papaya juice indicates that the phenolic compounds of fresh papaya juice do not easily oxidize by external
factors, such as oxygen and high temperature; hence, this will not easily affect its antioxidant capacity [10].
A recent study described a new blend of lemon (Citrus limon [L.] Burm. f.) juice beverage enriched with
papaya (C. papaya L.) juice (LP). The antioxidant activities were evaluated by DPPH radical scavenging
activity, superoxide radical (O2•−), hydroxyl radical (•OH), and hypochlorous acid (HOCl) assays (Table 36.3)
[18]. Papaya juice showed a lower DPPH radical scavenging activity (high IC50) compared to LP juice blend.
In terms of superoxide radical scavenging activity, the addition of LP juice blend resulted in the highest
increase when compared to the control papaya juice. Papaya juice exhibited a good hydroxyl radical scav-
enging activity, which did not differ significantly from that of the LP juice blend. A significant negative
correlation existed between total xanthones glycosides and •OH scavenging activity [18]. In terms of HOCl
scavenging activity, the addition of lemon juice to the LP juice blend resulted in the highest increase when
compared to the control papaya juice alone. In short, the antioxidant activity of LP juice blend increased sig-
nificantly after adding lemon into papaya juice. This increased antiradical activity of the blend with respect
to the control is probably due to the effect of the higher content of phytochemicals in the new bever-
age [18]. These results suggest that phenolic compounds are not the only phytochemicals indicative of the
antioxidant potential, as other components which possibly exist in papaya and lemon, but the interactions
between them in the food matrix may also play an essential role in the scavenging of reactive species [18].
36.3.2 Polyphenols
The polyphenol contents of many foods have appeared in nutrient databanks such as the Phenol- Explorer
[19,20] and USDA Flavonoid and Proanthocyanidin Databases [21]. However, there are no data on the
polyphenol content of papaya juice in both databases, except for papaya fruit pulp. Data on the content
and composition of polyphenols from Phenol-Explorer [19,20] and USDA flavonoids database [21] are
also inadequate to cover the polyphenol contents of papaya from different varieties or cultivars.
TABLE 36.3
Antioxidant Activities (IC50 in mg/mL) of Fresh Papaya and Lemon + Papaya Juices
Juice Blends DPPH O2•− • OH HOCl
Fresh papaya juice 29.38a 6.72a 3.96a 40.85a
Lemon + papaya juice 7.73b 2.39b 3.32a 19.58b
blend
Source: Adapted from Gironés-Vilaplana, A. et al., Food Chem., 170, 16, 2015. With permission.
Abbreviations: DPPH, 2,2-diphenyl-2-picrylhydrazyl; O2•−, superoxide radical; •OH, hydroxyl radical; HOCl,
hypochlorous acid.
Means (n = 3) in the same columns followed by different letters are significantly different at P < 0.05.
Papaya Juice 443
The major polyphenol classes in papaya pulp are flavones and flavonols. The flavones in papaya pulp are
apigenin and luteolin (0.01 and 0.02 mg/100 g, respectively), while the flavonols present are kaempferol
and myricetin (0.01 and 0.02 mg/100 g, respectively) [21–23].
The phenolic compositions of papaya juice extract (5%), reported only in one study [18], where man-
giferin (a xanthone glucoside) and the galloylated forms of mangiferin and isomangiferin (Table 36.4 and
Figure 36.2) were higher in LP juice blend compared to the fresh. The presence of xanthone glycosides in
TABLE 36.4
Reported Phenolic Compounds in Papaya Juice Extract and Lemon + Papaya Juice Blend (mg/100 mL)
Compounds Papaya Juice Extract (5%) Lemon + Papaya Juice Blend (5%)
Xanthones Glycosides
Mangiferin 0.28 0.28
Mangiferin gallate 0.08 0.11
Isomangiferin gallate 0.25 0.34
Total 0.61 0.73
Flavonols Glycosides
Quercetin 3-O-rutinoside-7-O-hexoside nd 0.19
Quercetin 3-O-rutinoside-7-O-pentoside nd nd
Quercetin 3-O-rutinoside 0.18 0.60
Quercetin 3-O-galactoside 0.06 0.08
Quercetin 3-O-glucoside 0.07 0.15
Kaempferol 3-O-rutinoside nd nd
Total 0.31 1.03
Flavones Glycosides
Apigenin 6,8-di-C-glucoside nd 0.65
Diosmetin 6,8-di-C-glucoside nd 4.94
Diosmetin 7-O-rutinoside nd 2.10
Total nd 7.69
Flavanones Glycosides
Eriodictyol 7-O-rutinoside nd 2.76
Hesperidin 7-O-rutinoside nd 2.92
Total nd 5.68
Source: Adapted from Gironés-Vilaplana, A. et al., Food Chem., 170, 16, 2015. With permission.
R1
OH
R2 O
— H, R2—
R1— — OH: Kaempferol
— OH, R2—
R1— — OH: Quercetin
OR3 — Glycoside moiety
R3—
OH O
General structure and substitution pattern of flavonol glycosides
HO O OH
OH
O
OH
OH O
HO OH
OH
Mangiferin
FIGURE 36.2 Structures of major xanthones and flavonols glycosides detected in papaya juice. (Continued)
OH
HO OH
HO O OH
O O
O
OH
OH O
HO OH
OH
Mangiferin gallate
OH
HO OH
O
HO
HO O OH
HO
OH O
Isomangiferin
OH
HO
O OH
O
HO
O O OH
HO
OH OH
Quercetin 3-O-galactoside
OH
OH
HO
HO
O
O OH
HO O
O O
OH
O OH
HO OH
OH
Quercetin 3-O-rutinoside
FIGURE 36.2 (Continued) Structures of major xanthones and flavonols glycosides detected in papaya juice.
papaya juice extract is interesting since they have been reported as potent hypolipidemic and antidiabetic
agents [24]. Mangiferin is the major flavonoid present in papaya juice (0.28 mg/100 mL). Higher concen-
tration of quercetin 3-O-rutinoside is found in LP juice blend (0.60 mg/100 mL) due to the contribution
of lemon juice, since this compound is also present in lemon [18] (Figure 36.2). Both the number and the
concentration of flavonols and xanthone glycosides of the LP juice blend are notably higher compared
Papaya Juice 445
to the fresh papaya juice extract alone [18]. However, flavone glycosides and flavanone glycosides are
not detected in papaya juice extract compared to the LP juice blend due to the contribution from lemon
juice (Table 36.4). Future research should identify and quantify the phenolic compounds of papaya juice
produced from pulp of different cultivars to fill the existing knowledge gap in the Phenol-Explorer and
USDA database.
36.4 Health Effects

36.4.1 Bioavailability of Bioactives from Papaya Juice in Relation to Its Intake
Papaya and its products are popular due to their high nutraceutical and pharmaceutical values [4,5].
However, little information is available on the absorption, distribution, metabolism, and excretion of
polyphenols in humans, especially the bioaccessibility and bioavailability of papaya juice. A study
determined the systemic levels of various carotenoids and their bioavailability in papaya consumed as
juice and fresh pulp [15]. Lycopene and β-carotene were present in the highest amount in both papaya
pulp and juice. The quantities of the carotenoids of papaya juice absorbed systemically are at an opti-
mum where its bioavailability lasts until 8 h of the experiment. A significant positive correlation existed
between major carotenoids (lutein, α-, and β-carotenes) found in chylomicrons at different periods (0, 4,
and 8 h), supporting the long-lasting bioavailability of these carotenoids in vivo [15]. Papaya juice
contributes greatly to the levels of lutein, α-, and β-carotenes in the chylomicrons of the subjects. There
was also a slight increase in serum retinol after 8 h of drinking papaya juice. However, the contribution
of β-cryptoxanthin and zeaxanthin was lower compared to other carotenoids, such as lutein, α-, and
β-carotenes [15]. It has been reported that fruit carotenoids are in the chromoplast of plant cells, hence
limiting their liberation during digestion. Thus, transformation of fruits to juice before consumption
may be beneficial. The particle sizes of the juice are reduced, hence facilitating digestion as compared
to fresh papaya pulp that needs to be broken down by the digestive system [15]. This could be the rea-
son why saturation of the cells still occurs after 4 h of ingestion of papaya juice. Lycopene, which is
abundant in papaya, administered to subjects is detected at very low levels in the chylomicrons. This
happens because lycopene could have been chemically degraded by the acid in the stomach or modified
enzymatically at the level of mucosal cells [15].
In another single-blind crossover study, the bioavailability of β-cryptoxanthin esters extracted from
papaya puree in human subjects was determined [25]. Results showed that the plasma β-cryptoxanthin
ester and nonesterified β-cryptoxanthin concentrations increased significantly (P < 0.05) and peaked even
after 6–12 h. The maximum plasma β-cryptoxanthin concentration obtained was 205 nmol (113 µg/L) [25].
Based on the assumption that about 4% of the human body weight is plasma (2–4 L plasma/60 kg), this
concentration corresponds to an absolute amount of 271 µg β-cryptoxanthin, amounting to approximately
one-quarter of the applied dose of the papaya puree [25].
36.4.2 Results from In Vitro, Animal, and Human Intervention Studies

Papaya juice and fermented papaya juice extract have been reported to render various health ben-
efits, such as antiobesity, antioxidant, immune-stimulating property, prevention of cytotoxicity, and
cardiotoxicity. In 2002, Rahmat et al. [26] screened the antiproliferative activity of the juice, pure, and
extracted lycopene from papaya on human breast and liver cancer cell lines. It was shown that papaya
juice and pure lycopene caused cell death in the liver cancer (HepG2) cell line with the half maximal
inhibitory concentration (IC50) of 20 mg/mL and 22.8 µg/mL, respectively. However, both papaya juice
and pure lycopene did not show any effect on the cell viability of breast cancer cell MDA-MB-231 [26].
On the other hand, the extracted lycopene from papaya juice did not show any effect on the prolifera-
tion of either cell lines. In short, fresh fruit juice seems to be better than extracted lycopene samples in
inhibiting cancer cell growth [26].
Another study reported that among 14 plant foods commonly consumed in Mexico (avocado, black
sapote, guava, mango, prickly pear cactus [nopal], pineapple, tomato, pear, grape, tomato, and papaya),
only papaya exhibited a significant inhibitory effect on breast cancer cell growth [27]. The juice extracts
from papaya flesh at all five tested concentrations (0.01%, 0.5%, 1%, 2%, and 4%) inhibited prolifera-
tion of MCF-7 cells after treatment for 72 h, in which the extract at 2% and 4% exerted 30% and 53%
inhibition of cell proliferation, respectively. Interestingly, the antiproliferative effect in cancer cells did
not correlate with total phenolic content or with antioxidant activity of the papaya juice extracts [27].
An ethanolic extract from papaya flesh inhibited cancer cell growth and scavenged nitric oxide (NO)
(about 35% of NO has been scavenged by the extract at the concentration of 640 µg/mL) [28].
The potential benefits of papaya juice in comparison to LP juice blend on cognitive function as ace-
tylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibitors have also been reported [18].
The AChE and BuChE inhibitory activities (IC50) of papaya juice were 20.47 and 19.8 mg/mL,
respectively, while the AChE and BuChE inhibitory activities (IC50) of LP juice blend were 7.64 and
7.54 mg/mL, respectively. These inhibitory activities increased significantly after the addition of lemon
juice to papaya juice compared to papaya juice alone [18].
Morimoto et al. [29] patented the extremely high effectiveness of a water extract of papaya pulp for the
prevention, treatment, or improvement of several cancer types, such as those of stomach, lung, pancre-
atic, colon, liver, ovarian, neuroblastoma, lymphoma, and leukemia. Although papaya is a major tropical
fruit, only a few pharmacological studies have been conducted on it compared to other fruits [30]. To the
best of our knowledge, no human clinical trials have been carried out, and no in vivo cancer studies have
been conducted with extracts from any part of papaya. Only several case studies have been reported in a
patent with very limited data [30,31].
Table 36.5 summarizes selected animal or human studies about the health benefits of papaya or fer-
mented papaya juice. A study reported the toxicological and antioxidant potential of C. papaya juice
in vitro and in vivo [32]. The oral lactate dehydrogenase 50 (LD50) activity of the juice of C. papaya was
determined, and the antioxidant activities were monitored by DPPH and FRAP tests. In vivo examina-
tion was performed after oral administration of papaya juice to rats for 2 weeks at doses of 100, 200, and
400 mg/kg. Blood thiobarbituric acid reactive substances (TBARS) and FRAP assays have been used
to determine the potential of the juice to act against oxidative stress in vivo [32]. The acute toxicity test
(LD50) demonstrated that papaya juice was not cytotoxic up to a dose of 1500 mg/kg after oral admin-
istration, and thus it is considered nontoxic. In treated groups, no sign of toxicity were observed [32].
In vitro evaluation of the antioxidant activities of papaya juice demonstrated that the highest antioxidant
activity (80%) was achieved at a concentration of 17.6 mg/mL. Blood lipid peroxidation levels decreased
significantly after administration of various doses of papaya juice (100, 200, and 400 mg/kg/day) to
35.5%, 39.5%, and 40.86% of the control, respectively, compared with a value of 28.8% for vitamin E
[32]. The blood total antioxidant power was increased significantly by all doses of papaya juice (100, 200,
and 400 mg/kg/day) to 11.11%, 23.58%, and 23.14% of the control, respectively. The value for vitamin E
was 18.44%. This study indicates the safety and antioxidant activity of the juice of C. papaya, which was
comparable to the standard antioxidant compound α-tocopherol [32]. In this relation, Rajkapoor et al.
[33] demonstrated that the LD50 value of the papaya juice extract was 2516 mg/kg, indicating that papaya
juice extract is nontoxic even at high concentrations.
Hassan et al. [17] evaluated the protective role of papaya juice extracts (27%, w/w) against aflatoxins
(AFs)-induced liver damage and oxidative stress in vivo. Their results showed that the papaya juice extract
alone did not induce any significant change in alanine aminotransferase (ALT) or aspartate aminotrans-
ferase (AST). However, it decreased the alkaline phosphatase (ALP) activity significantly compared to
the control group [17]. Papaya juice extract has also induced a significant decrease in total cholesterol,
triacylglycerols (TAG), and low-density lipoprotein (LDL)-cholesterol and increased high-density lipo-
protein (HDL) cholesterol of the rats insignificantly. These results demonstrated that the papaya juice
extract could normalize total antioxidant activity as well as improving malondialdehyde (MDA) and
sodium/potassium adenosine triphosphatase (N+/K+-ATPase) [17]. Furthermore, histological examina-
tion of the liver in the group fed (AFs)-contaminated diet and treated with papaya juice extract showed a
marked improvement in liver cells around the central veins [17]. The authors proposed a mechanism for
the protective role of papaya extracts against AFs-induced liver damage that may be through enhanced
production of Th1 type cytokines such as interleukin-12 (IL-12), interferon-gamma (IFN-γ), and tumor
necrosis factor-alpha (TNF-α) or through inducing a shift from Th2 to Th1 type immune response [17].
Papaya Juice 447
TABLE 36.5
Selected Studies on the Possible Health Effects of Papaya or Fermented Papaya Juice
Possible Health Effects Experimental Model Mode of Action References
Prevention of AFB1-induced rats Restoration of endogenous antioxidant enzymes, [14]
cardiotoxicity induced reducing MDA, and NO formation.
by AFB1 Reduction of Ca2+, ATPase, and normalized TAC.
Hepatoprotective, Aflatoxin-induced Improvement in liver function indices, lipid profile, [17]
antioxidative, and obese rats antioxidant status, lipid peroxidation, histological,
lipid-lowering effects and histochemical picture of the liver of the rats.
Improvement of the structural integrity of liver cell
membrane.
Hepatoprotective effects Obese rats Improvement in liver function indices, lipid profile, [34]
antioxidant status, lipid peroxidation, histological,
and histochemical picture of the liver of the rats.
Anti-obesity effects and High-fat cafeteria Reduction of body weight and relative weights of [37]
related disorders diet—obese rats fat in liver, kidney, and spleen.
Improvement in antioxidant status, liver function
indices, lipid profile, and lipid peroxidation.
Antioxidant and immune Acrylamide toxicity- Enhanced liver function indices and immune status [38]
stimulation induced rats in stomach, liver, and kidney.
Reduced lipid peroxidation in vivo.
Antioxidant defense Elderly patients with Enhancement of individual’s antioxidant defense [39]
chronic atrophic system.
gastritis Modulation of atrophic and metaplastic changes of
gastric mucosa.
Prevention of brain SHRs Upregulation of the redox defense activity in the [40]
antioxidative damage brain of SHRs.
Protection on SHRs Reduction of DNA damage in vivo. [41]
neurodegenerative Inhibition of the decay rate of the MC-PROXYL
diseases ESR signal in the brain.
Prevention of contact FITC (Th1 type) and Increased plasma immune function status. [45]
hypersensitive immune oxazolone (Th2 type) Increased IgA, dendritic, and inflammatory cells in
response induced ear and colon colon.
oedema rats model Reduced events induced by FITC or oxazolone.
Abbreviations: AFB1, aflatoxin B1; ATPase, adenosine triphosphatase; FITC, fluorescein isothiocyanate; IgA, immunoglob-
ulin A; MC-PROXYL ESR, 3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine-1-oxyl electron spin reso-
nance; MDA, malondialdehyde; NO, nitric oxide; SHRs, spontaneously hypertensive rats; TAC, total
antioxidant capacity.
Similarly, Rajkapoor et al. [33] also proposed that papaya juice extract protected liver against hepatotox-
icity by maintaining the normal hepatic physiology. Papaya juice extract also decreased the levels of liver
enzymes, improving the structural integrity on hepatocyte cell membrane, or regeneration of damaged
liver cells. In short, papaya juice extract demonstrated significant improvement in liver function indices,
lipid profile, antioxidant status, lipid peroxidation, and histological and histochemical profile of the liver
of tested rats [33].
Fresh papaya juice, however, did not show any inhibitory activity on α-glucosidase in vitro compared
to fruit juices produced from other indigenous fruits in Malaysia [16]. The effect of papaya juice extract
on the activities of the essential hepatic P450 enzymes, including CYP1A1, CYP1A2, CYP2E1 and
CYP3A11, was determined in vitro in mouse microsomes using their responsive enzymatic reactions,
namely ethoxyresorufin O-deethylation (EROD), methoxyresorufin O-demethylation (MROD), aniline
hydroxylation (ANH), erythromycin N-demethylation (ENDM), respectively [34]. Results indicated that
papaya juice extract decreased EROD and ANH activities, with no effect on the activity of ENDM.
However, pineapple and mangosteen juices demonstrated strong inhibitory activities on EROD, MROD,
ANH, and ENDM compared to papaya juice [34]. Wang et al. [35] showed that the addition of papaya
juice to human liver microsomes reduced the level of CYP2C9 diclofenac residual metabolism activity to
less than 20%, and the residual activity of tolbutamide metabolism to less than 40%, although not being
the strongest inhibitor of CYP2C9 drug metabolism in vitro.
In addition to these limited data, by indirect means, several studies have claimed health benefits of
C. papaya for protection against cancer due to the antioxidant properties of papaya extract [31,35].
However, there is continuous argument about whether a high antioxidant activity is a good indicator
of high anticancer activity, and no conclusive proof has been drawn until now [35]. Therefore, further
investigation is required to assess the underlying mechanism of action rather than attributing the putative
anticancer effects to antioxidant properties of bioactive compounds in papaya [31,35].
Athesh et al. [36] evaluated the antiobesity potential of aqueous fruit extract of C. papaya L.
(AFECP) on high-fat cafeteria diet–induced obese rats. Results demonstrated that the body weights
of the rats treated with high fat diet (HFD)+AFECP extract decreased in a dose-dependent man-
ner as compared to the HFD group. In addition, the relative weights of fat in the liver, kidney,
and spleen were significantly lower in the AFECP-treated groups as compared to the HFD group
[36]. Specifically, serum glucose, TAG, total-, LDL-, and very-low density lipoprotein (VLDL)-
cholesterols decreased significantly, while HDL-cholesterol increased in the treated groups in a
dose-dependent manner as compared to the HFD group. Hepatic TAG, total cholesterol, free fatty
acids (FFA), and phospholipid levels were also reduced significantly in the treated groups [36].
Furthermore, administration of AFECP produced a significant inhibition of MDA production and
a significant increase in reduced glutathione (GSH) and activity of superoxide dismutase (SOD). In
short, these results suggest that AFECP could be useful in the treatment of obesity and related dis-
orders [36]. The authors suggest that future studies should be carried out to determine the possible
mechanism of actions of AFECP.
Mannaa et al. [14] reported the ability of papaya juice extract in preventing cardiotoxicity induced
by aflatoxin B1 (AFB1) in rats. Papaya juice extract (27%, w/w) protected against the oxidative
stress caused by AFB1 intoxication by restoring the endogenous antioxidant enzymes, and decreased
MDA and NO formation that were induced by AFB1 intoxication, suggesting the antioxidant poten-
tials of papaya juice extract [14]. Papaya juice extract also reduced Ca2+ levels and adenosine
triphosphatase (ATPase) and normalized total antioxidant capacity in animals treated with AFB1.
The authors suggested the possible role of papaya juice extract as a chain breaking antioxidant
against lipid peroxidation in vivo.
Another study [37] determined the antioxidant and immune-stimulant activities of papaya juice
extract against acrylamide toxicity in rats. Papaya juice extract resulted in a significant increase in the
content of reduced GSH, SOD, catalase (CAT), immunoglobulin G (IgG), and immunoglobulin M
(IgM) in stomach, liver, and kidney, while this decreased the levels of MDA and lipid peroxidation
in vivo [37].
In a small double-blind, placebo-controlled study, a fermented papaya juice was administered to
elderly patients without major diseases, where the fermented papaya juice extract‒supplemented group
demonstrated a significant enhancement of the individual’s antioxidant defense system by modulat-
ing the atrophic and metaplastic changes of gastric mucosa in chronic atrophic gastritis patients [38].
This shows that fermented papaya juice extract could be a promising functional beverage for improving
antioxidant defense in elderly patients even without any overt antioxidant deficiency state [38]. Similarly,
another study [39] reported the supplementation with fermented papaya juice extract (50 mg/rat/day)
(5–7 months) significantly increased the decay of the electron spin resonance (ESR) images of the
3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine-1-oxyl (MC-PROXYL) in spontaneously hyperten-
sive rats (SHR), suggesting that fermented papaya juice may have upregulated the redox defense activity
in the brain of SHR [39].
The fermented papaya juice (FPJ) derived from C. papaya L. has also been investigated for its abil-
ity to modulate oxidative DNA damage due to hydrogen peroxide (H2O2) intoxication in rat pheochro-
mocytoma (PC12) cells and protection of brain oxidative damage in hypertensive rats [40]. The ability
of FPJ to modulate oxidative stress in the brain of SHR has been determined using the MC-PROXYL
(a blood brain barrier permeable nitroxyl spin probe)-L-band ESR technique [40]. A significant reduc-
tion (P < 0.05) of DNA damage was observed at concentrations ≥10 μg/mL in FPJ. Supplementation
Papaya Juice 449
with FPJ was shown to significantly inhibit the increased decay rate constant of the MC-PROXYL ESR
signal in the spontaneously hypertensive rat brain, suggesting modulation of oxidative stress in vivo.
These results indicate that FPJ can modulate oxidative injury supporting the view that prophylactic
potentials in neurodegenerative diseases could be facilitated by FPJ [40].
Previous studies have indicated that FPJ has antioxidant functions in vitro and in vivo by its abil-
ity to inhibit lipid peroxidation and protect supercoiled plasmid DNA against ferric nitrilotriacetate
(Fe-NTA) in combination with H2O2-induced single- and double-strand breaks as well as protecting
human T-lymphocytes challenged with Fe-NTA/H2O2 [41]. In another scenario, FPJ has been shown to
demonstrate both immunomodulatory and antioxidant activity in the macrophage cell line RAW 264.7
[42]. It is interesting to note that low- and high-molecular-weight fractions (LMF and HMF, respectively)
of FPJ demonstrated different activity patterns. In the presence of IFN-γ, both FPJ fractions stimu-
lated TNF-α (a central regulatory cytokine in macrophage antimicrobial activity) secretion. However, in
nonstimulated macrophages, TNF-α secretion was enhanced only by HMF. This shows that FPJ could
act as a macrophage activator as a result of its augmentation of NO synthesis and secretion of TNF-α
[42]. Furthermore, FPJ was shown to attenuate β-amyloid-mediated copper neurotoxicity in β-amyloid
precursor protein and β-amyloid precursor protein Swedish mutation overexpressing SH-SY5Y cells
[43]. FPJ posttreatment increased cell viability and decreased the intracellular calcium ion, reactive oxy-
gen species (ROS) generation such as hydroxyl radical, superoxide radical anion, and NO accumulation
in SH-SY5Y cells. These results also show that FPJ prevents cell apoptosis through bax/bcl-2 sensitive
pathway [43].
Hiramoto et al. [44] determined whether the oral administration of FPJ to mice restrained two
types of contact hypersensitivity models, fluorescein isothiocyanate (FITC) (Th2 type) and oxazolone
(Th1 type)-induced ear and colon oedema [44]. The sensitization of FITC or oxazolone increased the
plasma levels of IL-10, IFN-γ, and TNF-α. Meanwhile, histological examinations demonstrated an
obvious increase of immunoglobulin A (IgA), dendritic cells, and inflammatory cells in the colon [44].
When the animals were given FPJ before sensitization by FITC or oxazolone, all the events induced
by either FITC or oxazolone were decreased significantly. In short, these results suggest that the oral
administration of the FPJ may have a therapeutic potential for the prevention of contact hypersensitive
immunoresponse [44].
The antioxidant beverage effective microorganism-X (EM-X) formulated from the fermentation of
unpolished rice, papaya, and seaweeds was examined in another study [45]. Results demonstrated that
EM-X inhibited the release of IL-8 at the transcriptional level in A549 cells. EM-X also decreased the
iron-/ascorbate-dependent peroxidation of ox-brain phospholipids in a concentration-dependent manner.
Results showed that the anti-inflammatory and antioxidant properties of EM-X were due to the flavonoid
contents of the beverage EM-X [45].

Papaya juice is a good source of carotenoids, vitamins, and minerals. However, it is not commonly pro-
duced as a functional juice as compared to other fruit juices such as kiwi, orange, grape, pomegranate,
guava, and various berries [7,8]. Only a few published studies are available using papaya juice for the for-
mulation of functional fruit juices. A recent study described the formulation of a spiced aloe gel–papaya
blend (SAGPB) functional beverage [46]. The physicochemical characteristics, antioxidant potential,
microbial quality, and sensory acceptability of the newly developed SAGPB in comparison to spiced
papaya juice (control) were evaluated. The SAGPB had a vitamin C, total polyphenol, and total flavonoid
contents of 5.31, 240, and 115 mg/100 mL, respectively. These values do not differ significantly with
those of the spiced papaya juice. Thus, aloe gel does not contribute much to the phenolic and flavonoid
contents of SAGPB juice blend [46].
Both SAGPB and spiced papaya juice have been found to be free from microbial proliferation until
the end of their storage period in terms of standard plate count and yeast and mold count, thus indicat-
ing that they are fit for consumption even after 5 months of storage [46]. This could be attributed to the
addition of citric acid which acts as a preservative, and also due to the addition of spices, which possess
good antimicrobial and antioxidant activities. SAGPB also received better sensory acceptability in terms
of its taste and flavor compared to spiced papaya juice (control) [46]. The spiced papaya juice scored
lower in terms of its taste and aroma mainly due to the sour/acidic taste. This shows that SAGPB could
be promoted as a nutraceutical with multiple benefits to the consumer [46].
A recent study described a new blend of lemon juice beverage enriched either with noni juice or
papaya juice [18]. It was demonstrated that LP juice blend is the most interesting blend in terms of
antioxidant activity, and also in terms of inhibition of cholinesterases (AChE and BuChE), mainly due
to the combination with lemon juice [18]. This new beverage presented phytochemical profiles rich in
bioactive phenolics (flavonols, xanthone, flavones, flavanones, and anthraquinone derivatives). New data
on scavenging abilities of a beverage made of lemon enriched with papaya suggest that phenolic com-
pounds are not the only compounds indicative of the antioxidant potential in papaya juice and LP juice
blend, as other components which possibly exist in papaya and other fruits. The interactions between
them in the food matrix may also play important roles in the scavenging of reactive species [18]. Hence,
using papaya juice enriched with lemon, as well as other fruits, for dietary interventions and nutrition
programs for aging adults is of much interest. The authors recommended that further research in formu-
lation, bioaccessibility, bioavailability, and bioefficacy of papaya juice, papaya juice blend, and papaya
is needed [18]. The development of functional foods with cholinesterases inhibitors, with potential
application in preserving cognitive function, and managing age-related conditions (e.g., Alzheimer’s
and Parkinson’s diseases, among others) is possible with healthy fruit-based beverages to facilitate their
appropriate daily intake [18].
Papaya juice produced from its fruit pulp has been proposed as a potential renewable energy source
for industrial alcohol production because of its low cost and easy availability [47]. Utilization of papaya
juice for winemaking offers an alternative means of adding value where papaya wine receives good
rating and acceptability among tested subjects, comparable to the popular fruit wines, such as grape
wine [47,48]. A total of 118 volatiles have been detected, and 97 of them have been positively identified
in papaya wine. These included 53 esters, 22 alcohols, nine acids, seven phenols and derivatives, seven
sulfur-containing compounds, five lactones, five terpenes, three ketones, two aldehydes, and five miscel-
laneous compounds [48]. The major compounds identified were 3-methylbutan-1-ol, 2-methylbutan-1-ol,
and 2-phenylethanol. Six odorants were considered as odor-active volatiles, namely, ethyl octanoate,
(E)-β-damascenone, 3-methylbutyl acetate, benzyl isothiocyanate, ethyl hexanoate, and ethyl butanoate
[48]. The authors suggested that sensory studies need to be carried out to determine the actual contribu-
tion of these volatile compounds to the acceptability of papaya wine [48].
Balasubramanian et al. [49] recently patented a new fruit and vegetable-based beverage made by
coproduction of juice from various fruits and vegetables, including papaya juice, for use in beverage and
food products in the United States. This new functional beverage may enhance the metabolic and gut
health benefits, including an enhanced feeling of satiety, a reduction of postprandial glucose and insulin
response, and an increased fermentability by colonic microflora and short-chain fatty acids production
in the colon for the consumer upon consumption [49]. Future research should be carried out to determine
which preservation technique is the best to retain fresh-like quality of the beverage since some preserva-
tion techniques have been shown to affect the phytochemical or nutrient contents of the juice.
36.6 Conclusion
Papaya juice is rich with carotenoids and antioxidant phenolic compounds (flavonols, flavones, and
xanthones). Furthermore, papaya juice products such as its puree, nectar, or fermented juice extract
have good aromatic and sensory qualities. Further research should be carried out so that more novel
papaya juice functional beverages can be developed using new preservation technologies. Papaya
juice has been shown to render various health benefits. Hence, more clinical trials and human inter-
vention studies should also be carried out to ascertain the beneficial effects of papaya juice in health
promotion in relation to its stability, bioaccessibility, and bioavailability after metabolism in the
human body.
Papaya Juice 451
REFERENCES
1. Vij, T. and Prashar, Y., A review on medicinal properties of Carica papaya Linn. Asian Pac. J. Trop.
Dis., 5, 1–6, 2015.
2. Food and Agriculture Organization of the United Nation Statistics Division (FAOSTAT), Food and
Agricultural Commodities Production, 2013. Published online at http://faostat.fao.org/site (accessed
February 7, 2015).
3. Chan, Y.K. and Paull, R.E., Papaya (Carica papaya L.), in Encyclopedia of Fruit and Nuts, Janick, J.
and Paull, R.E., Eds., CABI, Wallingford, UK, 2008, pp. 237–247.
4. Krishna, K.L., Paridhavi, M., and Patel, J.A., Review on nutritional, medicinal and pharmacological
properties of papapa (Carica papaya Linn.). Nat. Prod. Radiance, 7, 364–373, 2008.
5. De Oliveira, J.G. and Vitoria, A.P., Papaya: Nutritional and pharmacological characterization and qual-
ity loss due to physiological disorders. An overview. Food Res. Int., 44, 1306–1313, 2011.
6. Zulueta, A., Esteve, M.J., Frasquet, I., and Frígola, A., Vitamin C, vitamin A, phenolic compounds
and total antioxidant capacity of new fruit juice and skim milk mixture beverages marketed in Spain.
Food Chem., 103, 1365–1374, 2007.
7. Sun-Waterhouse, D., The development of fruit-based functional foods targeting the health and wellness
market: A review. Int. J. Food Sci. Technol., 46, 899–920, 2011.
8. Corbo, M.R., Bevilacqua, A., Petruzzi, L., Casanova, F.P., and Sinigaglia, M., Functional beverages:
The emerging side of functional foods. Compr. Rev. Food Sci. F., 13, 1192–1206, 2014.
10. Falguera, V., Sánchez-Riaño, A.M., Quintero-Cerón, J.P., Rivera-Barrero, C.A., Méndez-Arteaga, J.J.,
and Ibarz, A., Characterization of polyphenol oxidase activity in juices from 12 underutilized tropical
fruits with high agroindustrial potential. Food Bioprocess Technol., 5, 2921–2927, 2012.
11. El-Mansy, H.A., Sharoba, A.M., Bahlol, H.E.L.M., and El-Desouky, A.I., Rheological properties of
mango and papaya nectar blends. Ann. Agric. Sci., 43, 665–686, 2005.
12. Basu, A. and Haldar, S., Dietary isothiocyanate mediated apoptosis of human cancer cells is associated
with Bcl-xL phosphorylation. Int. J. Oncol., 33, 657–663, 2008.
13. Seigler, D.S., Pauli, G.F., Nahrstedt, A., and Leen, R., Cyanogenic allosides and glucosides from
Passiflora edulis and Carica papaya. Phytochemistry, 60, 873–882, 2002.
14. Mannaa, F.A., Abdel-Wahhab, K.G., and Abdel-Wahhab, M.A., Prevention of cardiotoxicity of aflatoxin
B1 via dietary supplementation of papaya fruit extracts in rats. Cytotechnology, 66, 327–334, 2014.
15. Gouado, I., Schweigert, F.J., Ejeh, R.A., Tchouanguep, M.F., and Camp, J.V., Systemic levels of carot-
enoids from mangoes and papaya consumed in three forms (juice, fresh, and dry slice). Eur. J. Clin.
Nutr., 61, 1180–1188, 2007.
16. Sulaiman, S.F. and Ooi, K.L., Antioxidant and α-glucosidase inhibitory activities of 40 tropical juices
from Malaysia and identification of phenolics from the bioactive fruit juices of Barringtonia racemosa
and Phyllanthus acidus. J. Agric. Food Chem., 62, 9576–9585, 2014.
17. Hassan, N.S., Abdel, K.G.A.W.G., Khadrawy, Y.A.K.A., Aziza, A., Mannaa, F.A.M.A., and Abdel-
Wahhab, M.A., Evaluation of radical scavenging properties and the protective role of papaya fruits
extracts against oxidative stress in rats fed aflatoxin-contaminated diet. Comunicata Sci., 4, 43–57, 2013.
18. Gironés-Vilaplana, A., Valentão, P., Andrade, P.B., Ferreres, F., Moreno, D.A., and García-Viguera, C.,
Beverages of lemon juice and exotic noni and papaya with potential for anticholinergic effects. Food
Chem., 170, 16–21, 2015.
19. Pérez-Jiménez, J., Neveu, V., Vos, F., and Scalbert, A., Systematic analysis of the content of 502 poly-
phenols in 452 foods and beverages: An application of the Phenol-Explorer Database. J. Agric. Food
Chem., 58, 4959–4969, 2010.
metabolism and pharmacokinetics in humans and experimental animals. Database, doi: 10.1093/data-
base/bas031, 2012.
Foods, Release 3.1, 2014. Published online at: http://www.ars.usda.gov/nutrientdata/flav (accessed
November 2, 2014).
22. Franke, A.A., Custer, L.J., Arakaki, C., and Murphy, S.P., Vitamin C and flavonoid levels of fruits and
vegetables consumed in Hawaii. J. Food Comp. Anal., 17, 1–35, 2004.
23. Lako, J., Trenerry, V.C., Wahlqvist, M., Wattanapenpaiboon, N., Sotheeswaran, S., and Premier, R.,
Phytochemical flavonols, carotenoids and the antioxidant properties of a wide selection of Fijian fruit,
vegetables and other readily available foods. Food Chem., 101, 1727–1741, 2007.
24. Sellamuthu, P.S., Muniappan, B.P., Perumal, S.M., and Kandasamy, M., Antihyperglycemic effect of
mangiferin in streptozotocin induced diabetic rats. J. Health Sci., 55, 206–214, 2009.
25. Breithaupt, D.E., Weller, P., Wolters, M., and Hahn, A., Plasma response to a single dose of dietary
β-cryptoxanthin esters from papaya (Carica papaya L.) or non-esterified β-cryptoxanthin in adult
human subjects: A comparative study. Br. J. Nutr., 90, 795–801, 2003.
26. Rahmat, A., Rosli, R., Wan Mohd Zain, W.N.I., Endrini, S., and Abdullah Sani, H., Antiproliferative activ-
ity of pure lycopene compared to both extracted lycopene and juices from watermelon (Citrullus vulgaris)
and papaya (Carica papaya) on human breast and liver cancer cell lines. J. Med. Sci., 2, 55–58, 2002.
27. Garcia-Solis, P., Yahia, E.M., Morales-Tlalpan, V., and Diaz-Munoz, M., Screening of antiproliferative
effect of aqueous extracts of plant foods consumed in Mexico on the breast cancer cell line MCF-7.
Int. J. Food Sci. Nutr., 60, 32–46, 2009.
28. Jayakumar, R. and Kanthimathi, M.S., Inhibitory effects of fruit extracts on nitric oxide-induced prolif-
eration in MCF-7 cells. Food Chem., 126, 956–960, 2011.
29. Morimoto, C., Dang, N.H., Dang, N., YS Therapeutic Co Ltd (YSTH-Non-standard), Toudai Tlo Ltd
(TOUD Non-standard), Morimoto C (MORI-Individual), Dang NH (DANG-Individual), Cancer pre-
vention and treating composition for preventing, ameliorating, or treating solid cancers, e.g., lung or
blood cancers and lymphoma, comprises components extracted from brewing papaya. Patent number-
WO2006004226-A1; EP1778262- A1; JP2008505887-W; US2008069907-A1, 2008.
30. Pierson, J.T., Dietzgen, R.G., Shaw, P.N., Roberts-Thomson, S.J., Monteith, G.R., and Gidley, M.J.,
Major Australian tropical fruits biodiversity: Bioactive compounds and their bioactivities. Mol. Nutr.
Food Res., 56, 357–387, 2012.
31. Nguyen, T.T., Shaw, P.N., Parat, M.O., and Hewavitharana, A.K., Anticancer activity of Carica papaya:
A review. Mol. Nutr. Food Res., 57, 153–164, 2013.
32. Mehdipour, S., Yasa, N., Dehghan, G., Khorasani, R., Mohammadirad, A., Rahimi, R., and Abdollahi,
M., Antioxidant potentials of Iranian Carica papaya juice in vitro and in vivo are comparable to
α-tocopherol. Phytother. Res., 20, 591–594, 2006.
33. Rajkapoor, B., Jayakar, B., Kavimani, S., and Murugesh, N., Effect of dried fruits of Carica papaya linn
on hepatotoxicity. Biol. Pharm. Bull., 25, 1645–1646, 2002.
34. Chatuphonprasert, W. and Jarukamjorn, K., Impact of six fruits—Banana, guava, mangosteen, pine-
apple, ripe mango and ripe papaya—On murine hepatic cytochrome P450 activities. J. Appl. Toxicol.,
32, 994–1001, 2012.
35. Wang, S., Meckling, K.A., Marcone, M.F., Kakuda, Y., and Tsao, R., Can phytochemical antioxidant
rich foods act as anti-cancer agents? Food Res. Int., 44, 2545–2554, 2011.
36. Athesh, K., Karthiga, D., and Brindha, P., Anti-obesity effect of aqueous fruit extract of Carica papaya L.
in rats fed on high fat cafeteria diet. Int. J. Pharm. Pharm. Sci., 4, 327–330, 2012.
37. Mohamed Sadek, K., Antioxidant and immunostimulant effect of Carica papaya Linn. aqueous extract
in acrylamide intoxicated rats. Acta Inform. Med., 20, 180–185, 2012.
38. Marotta, F., Weksler, M., Naito, Y., Yoshida, C., Yoshioka, M., and Marandola, P., Nutraceutical supple-
mentation: Effect of a fermented papaya preparation on redox status and DNA damage in healthy elderly
individuals and relationship with GSTM1 genotype. Ann. N. Y. Acad. Sci., 1067, 400–407, 2006.
39. Yoshino, F., Kobayashi, K., Hayashi, Y., and Aruoma, O.I., Assessment of the effect of fermented papaya
preparation on oxidative damage in spontaneously hypertensive rat brain using electron spin resonance
(ESR) imaging and L-band ESR spectroscopy. J. Funct. Foods, 1, 375–380, 2009.
40. Aruoma, O.I., Colognato, R., Fontana, I., Gartlon, J., Migliore, L., Koike, K., Coecke, S. et al., Molecular
effects of fermented papaya preparation on oxidative damage, MAP kinase activation and modulation of
the benzo[a]pyrene mediated genotoxicity. Biofactors, 26, 147–159, 2006.
41. Virgili, F. and Packer, L., Ferric nitrilotriacetate induced DNA and protein damage: Inhibitory effect of
a fermented papaya preparation. Anticancer Res., 20, 2907–2914, 2000.
Papaya Juice 453
42. Rimbach, G., Park, Y.C., Guo, Q., Moini, H., Qureshi, N., Saliou, C., Takayama, K., Virgilli, F., and
Packer, L., Nitric oxide synthesis and TNF-α secretion in RAW 264.7 macrophages: Mode of action of
a fermented papaya preparation. Life Sci., 67, 679–694, 2000.
43. Zhang, J., Mori, A., Chen, Q., and Zhao, B., Fermented papaya preparation attenuates β-amyloid precur-
sor protein: β-amyloid–mediated copper neurotoxicity in β-amyloid precursor protein and β-amyloid
precursor protein Swedish mutation overexpressing SH-SY5Y cells. Neuroscience, 143, 63–72, 2006.
44. Hiramoto, K., Imao, M., Sato, E.F., Inoue, M., and Mori, A., Effect of fermented papaya preparation on
dermal and intestinal mucosal immunity and allergic inflammations. J. Sci. Food Agric., 88, 1151–1157,
2008.
45. Deiana, M., Dessi, M.A., Ke, B., Liang, Y.F., Higa, T., Gilmour, P.S., Jen, L.-S., Rahman, I., and
Aruoma, O.I., The antioxidant cocktail effective microorganism X (EM-X) inhibits oxidant-induced
interleukin-8 release and the peroxidation of phospholipids in vitro. Biochem. Biophys. Res. Commun.,
296, 1148–1151, 2002.
46. Ramachandran, P. and Nagarajan, S., Quality characteristics, nutraceutical profile, and storage stability
of aloe gel-papaya functional beverage blend. Int. J. Food Sci., Article ID 847013, 2014.
47. Lee, P.R., Ong, Y.L., Yu, B., Curran, P., and Liu, S.Q., Evolution of volatile compounds in papaya wine
fermented with three Williopsis saturnus yeasts. Int. J. Food Sci. Technol., 45, 2032–2041, 2010.
48. Pino, J.A. and Queris, O., Characterisation of odour-active compounds in papaya (Carica papaya L.)
wine. Int. J. Food Sci. Technol., 47, 262–268, 2012.
49. Balasubramanian, S., Bordenave, N., Harkness, L., Hitchcock, B.W., Hsieh, M., Jordan, R.L., Mathews,
J.D. et al., Preparation and incorporation of co-products into beverages to achieve metabolic and gut
health benefits. U.S. Patent Application 14/262,213, 2014.
37
Passion Fruit Juice
Chin-Kun Wang
CONTENTS
37.1 Introduction.....................................................................................................................................455
37.3.1 Flavonoids......................................................................................................................... 456
37.3.2 Carotenoids....................................................................................................................... 458
37.3.3 Alkaloids........................................................................................................................... 458
37.4 Health Effects................................................................................................................................. 459
37.6 Conclusion..................................................................................................................................... 460
References............................................................................................................................................... 460
37.1 Introduction
Passion fruit (Passiflora edulis), a vine species, originated from the edges of South American rain forests
in the Amazon region of Brazil and possibly in Paraguay and North Argentina. It is round, dark purple
or yellow at maturity, with a juicy pulp and numerous seeds [1]. Passion fruit juice is often added to other
fruit juices to enhance the aroma [2]. It is cultivated in warm, frost-free areas and becomes naturalized
in most of the tropical and subtropical world including South America, South Africa, Hawaii, California,
Kenya, Sri Lanka, India, Taiwan, and some Asian countries. The most important genus Passiflora has
about 400 species, which are mostly native to tropical America and about 40 species in Asia, Australia,
and the South Pacific and one in Madagascar [3].
In most countries, passion fruit production is based on cultivars of the yellow passion fruit (P. edulis
forma flavicarpa). The major exceptions are South Africa, Kenya, and New Zealand where production is
dependent on lines of the purple passion fruit (P. edulis Sims) and in Australia and Taiwan where hybrids
between the two forms are exploited. Passion fruit juice is enjoyed worldwide due to its pleasant unique
aroma and flavor. This chapter highlights the nutritional characteristics and bioactive substances as well
as health benefits of passion fruit juice.

Compositional and nutritional characteristics of passion fruit (purple) and two major commercial spe-
cies of passion fruit juices (purple and yellow) are presented in Table 37.1 [4]. Passion fruit contains
high levels of carbohydrate (23.38 g/100 g) and dietary fiber (10.4 g/100 g). During juice processing, the
peel, fiber, and seeds are removed. Therefore, carbohydrate and dietary fiber levels are greatly reduced
(Table 37.1). Both passion fruit and juices are good sources of minerals (e.g., potassium, phosphorus, and
magnesium) and vitamins (e.g., vitamin A, vitamin C, and folate) [4]. Total sugars range from 11.20 in
passion fruit to 13.4–14.25 g/100 g in passion fruit juices.
455
TABLE 37.1
Compositional and Nutritional Characteristics of Passion Fruit and Passion Fruit Juices (per 100 g Fruit
or Juice)
Passion Fruit Juice
Nutrient Unit Passion Fruit (Purple) Passion Fruit Juice (Purple) (Yellow)
Water g 72.93 85.62 84.21
Protein g 2.20 0.39 0.67
Lipid (fat) g 0.7 0.05 0.18
Total sugars g 11.20 13.4 14.25
Minerals
Calcium mg 12 4 4
Iron mg 1.6 0.24 0.36
Sodium mg 28 6 6
Zinc mg 0.10 0.05 0.06
Vitamins
Folate (DFE) µg 14 7 8
Niacin mg 1.5 1.46 2.24
Riboflavin mg 0.13 0.131 0.101
Thiamin mg 0.00 0.00 0.00
Vitamin A (RAE) µg 64 36 47
Vitamin B6 mg 0.1 0.05 0.06
Vitamin C mg 30 29.8 18.2
Vitamin E (ATE) mg 0.02 0.01 0.01
Vitamin K µg 0.7 0.4 0.4
Reference, Release 26, 2013, Published online at: http://www.nal.usda.gov/fnic/foodcomp/search/ (accessed
May 28, 2014).
In general, the main components in passion fruit juice are sugars (e.g., fructose, glucose, and sucrose)
and organic acids (e.g., ascorbic, citric, tartaric, and malic) [5–7]. Passion fruit is acidic (pH ~ 3.2) due
to its predominance of citric acid (~93%–96% of total) and malic acid (3%–6% of total). The acids and
sugars add to the unique taste and serve as a preservative nature for this tropical fruit.

Carotenoids make the juice very attractive with a golden color (Figure 37.1). There are three pri-
mary groups of phytochemicals present in passion fruit juice: flavonoids, carotenoids, and alkaloids
(Table 37.2).
37.3.1 Flavonoids
Flavonoids, which are a major group of phenolic compounds, have been found to possess strong antioxi-
dant activity [8–10] as well as anticancer properties [11]. Phenolic compounds in P. edulis and P. alata
Passion Fruit Juice 457
FIGURE 37.1 Ripen passion fruit and its golden juice.
TABLE 37.2
Phytochemicals Present in Passion Fruit Juices
Nutrient Unit Passion Fruit Juice References
Flavonoids mg/L 435 [8]
Isoorientin mg/L 158 [16]
Carotenoids mg/L 9.32 [8]
Total Alkaloids % 0.012–0.7 [21]
OH
OH
HO O
O OH
HO
HO
OH
OH O
FIGURE 37.2 Chemical structure of isoorientin found in passion fruit juice.
mainly belong to the flavones C-glucoside class [12]. Isoorientin (Figure 37.2), a C-glucoside flavone
found in P. edulis [12], was also found to be the major flavonoid in pulp of this species. In fact, the total
flavonoid content in P. edulis juice is reported to be quite significant in comparison with other beverages
that are good sources of flavonoids, such as orange juice and sugarcane juice [13].
Passion fruit is rich in phenolics and acts as antioxidants [14]. Ichimura et al. [15] showed the potential of
passion fruit and its rind for several purposes, such as the antihypertensive effect of passion fruit rind
attributed partially to the vasodilatory effect of polyphenols, especially the flavonoid especially luteolin.
Recently, the antioxidant activity from P. edulis and P. alata pulps, and P. edulis rinds is investigated in
phorbol 12-myristate 13-acetate-stimulated neutrophils and purified myeloperoxidase. P. edulis rinds
α-Carotene
β-Carotene
ζ-Carotene
HO
β-Cryptoxanthin
Lycopene
FIGURE 37.3 Chemical structures of carotenoids found in passion fruit juice.
exhibited higher antioxidant activity, possibly correlated with the isoorientin content in P. edulis, as
determined by high-performance liquid chromatography–diode array detector (HPLC-DAD) [16].
37.3.2 Carotenoids
In passion fruit, 13 different carotenoids have been identified, including ζ-, β-, and α-carotenes [17]. The
chemical structures of most common carotenoids found in passion fruit juice are shown in Figure 37.3.
Three individual carotenoids, α-carotene (35 μg/100 g of edible portion), β-carotene (525 μg/100 g
of edible portion), and β-cryptoxanthin (46 μg/100 g edible portion), have been quantified in passion
fruit [18]. Passion fruit is richer in α- and β-carotenes than other popular tropical fruits such as papaya,
mango, and banana. However, when compared to other rich sources of carotenoids such as tomato and
carrot, passion fruit contains higher α-carotene than tomato and carrot but lower α- and β-carotenes [18].
Carotenoids are widely regarded as effective quenchers of singlet oxygen, triplet oxygen, and peroxyl
radicals [19,20]. The total carotenoid concentration of passion fruit juice is 9.32 mg/L (Table 37.2) [8].
37.3.3 Alkaloids
Harmala alkaloids are a group of β-carboline compounds. These are present in passion fruit juice, which
include harman, harmine, harmol, harmalol, and harmaline (Figure 37.4). The content of total alkaloids
in passion fruit juice ranges from 0.012% to 0.7% (Table 37.2) [21].
N
N
H
Harman
O N
N
H
Harmine
OH
N
N
H 3C H
Harmol
HO N
N
H
Harmalol
O N
N
H
Harmaline
FIGURE 37.4 Harmala alkaloids found in passion fruit juice.
37.4 Health Effects

The health effects of passion fruit juice are mainly from their phytochemicals and micronutrients. The
health effects of passion fruit juice are briefly discussed here.
Passion fruit juice is rich in potassium and contains alkaloids, which can help regulate heart rate and
blood pressure. The juice, but mainly the leaves of passion fruit, contains alkaloids, including harman,
which has blood pressure lowering, sedative, and antispasmodic action [5,22]. The antihypertensive effect
of passion fruit juice has been found from the inhibitory activity on angiotensin-converting enzyme [23].
The use of passion fruit juice improves the lipid and glycemic profile of offspring from diabetic
and nondiabetic mothers of Wistar rats. This result suggests that passion fruit juice has potential
as an adjuvant in the prevention and treatment of dyslipidemia and hyperglycemia [24]. The phyto-
chemicals found in passion fruit juice responsible for this anticancer effect are carotenoids and certain
polyphenols [25].
Knee osteoarthritis is a common degenerative joint disorder and a major cause of pain and disability.
The passion fruit juice, a flavonoid-rich dietary supplement, substantially alleviates osteoarthritis symp-
toms. This beneficial effect may be due to its antioxidant and anti-inflammatory properties [26].
When Spanish explored South America, they discovered that passion fruit was used in native folk
medicine as a sedative. Many plants in the genus Passiflora have long been used in traditional folk medi-
cines as a remedy for many neurogenic diseases in many countries. Studies showed anxiety relieving at
low dose, but sedative effect at high dose. Flavonoids are important active constituents [27,28].
Passion fruit juice is a rich source of vitamin C, carotenoids, and flavonoids [5,21]. These are power-
ful antioxidants and help the body develop resistance against flu-like infectious agents and scavenge
harmful, proinflammatory free radicals and anti-inflammation [8–10]. Inflammation is an indicator of
various diseases, including cancer. Anti-inflammatory activity has been found in the C-glucosylflavones
of passion fruit [29–31].
Animal study convinces that the treatment of passion fruit juice clearly lowers the total cholesterol,
low-density lipoprotein (LDL) cholesterol, and increases high-density lipoprotein (HDL) cholesterol
[24]. Pilot clinical studies show that the consumption of passion fruit juice could reduce the serum
cholesterol [32]. This effect is probably from the pectin and high levels of fiber from the seeds of the
pulp [32,33]. In addition, the blood coagulation could be suppressed by passion fruit juice to prevent the
cardiovascular disease (CVD) [34].
Passion fruit juice contains alkaloids, an endogenous β-carboline, which influence learning and
memory [35].
Passion fruit juice is rich in phytochemicals, in addition to the aforementioned health benefits. Ripa
et al. [36] also found that it exerted antibacterial activity on 12 microorganisms.
Sleep problem in modern times due to social changes and development is experienced, and Passiflora
provides subjective sleep quality [37].

Although passion fruits are mainly collected by local people in forests, they are now grown in vineyards
in several countries. They are concentrated, frozen, and shipped worldwide. Fresh fruits are cut in half
and the succulent pulp is scooped out with a spoon for mixing with other fruits in making a salad. The
extracted juice is a tasty drink. Cake icing, candy, ice cream, jelly, mousses, sauces, sherbets, syrups, and
pies are some of the value-added products made from the fruit.
Passion fruit juice contains alkaloids and a number of other phytochemicals [34,35]. A cyanogenic gly-
coside is found in the pulp of all passion fruits, but it is only present in trace amounts in the ripened fruit
juice [13,19,21]. The future development for alkaloids and flavonoids in whole passion fruit is worthwhile
for the health benefits and for increasing the economic value of products.
37.6 Conclusion
Passion fruit juice is rich in phytochemicals and provides several health benefits. It is also known as a
natural medicinal food in tropical and subtropical countries. There is a need for further development and
research in order to promote the use of passion fruit and its juice as healthful products.
REFERENCES
1. Boning, C.R., Florida’s Best Fruiting Plants: Native and Exotic Trees, Shrubs, and Vines, Pineapple
Press Inc., Sarasota, FL, 2006.
2. U.S. Department of Agriculture (USDA), Passiflora edulis Sims, Germplasm Resources Information
Network, 2007. Published online at: http://www.ars-grin.gov/cgi-bin/npgs/html/taxon.pl?26962
(accessed May 28, 2014).
3. Department of Agriculture (Government of Sri Lanka), Passion Fruit, 2006. Published online at: http://
www.agridept.gov.lk/index.php/en/crop-recommendations/1099 (accessed May 27, 2014).
Release 26, 2013. Published online at: http://www.nal.usda.gov/fnic/foodcomp/search/ (accessed May
28, 2014).
5. Chen, C.C., Kuo, M.C., Hwang, L.S., Wu, J.S.B., and Wu, C.M., Headspace components of passion fruit
juice. J. Agric. Food Chem., 30, 1211–1215, 1982.
6. Casimir, D.J., Kefford, J.F., and Whitfield F.B., Technology and flavor chemistry of passion fruit juices
and concentrates, in Advances in Food Research, vol. 27, Chichester, C., Ed., Academic Press Inc.,
New York, 1981, pp. 243–290.
6. Strauss, D., The microscopy of foreign fruits. VII. Passion fruit Passiflora edulis Sims. Deut. Lebensm.
Rundsc., 70, 144–146, 1974.
7. da Fonseca, J.L.F., Concentrated passion fruit juice. Bel. Tec. CEPED., 3, 31–66, 1976.
8. Talcott, S.T., Percival, S.S., Pittet-Moore, J., and Celoria, C., Phytochemical composition and antioxi-
dant stability of fortified yellow passion fruit (Passiflora edulis). J. Agric. Food Chem., 51, 935–941,
2003.
9. Rice-Evans, C. and Miller, N.J., Antioxidant activities of flavonoids as bioactive components of food.
Biochem. Soc. Trans., 24, 790–795, 1996.
10. Salah, N., Miller, N.J., Paganga, G., Tijburg, L., Bolwell, G.P., and Rice-Evans, C., Polyphenolic fla-
vanols as scavengers of aqueous phase radicals and as chain-breaking antioxidants. Arch. Biochem.
Biophys., 322, 339–346, 1995.
11. Yoshida, M., Yamamoto, M., and Nikaido, T., Quercetin arrest human leukemic cells in late G1 phase of
the cell cycle. Cancer Res., 52, 6679–6681, 1992.
12. Dhawan, K., Dhawan, S., and Sharma, A., Passiflora: A review update. J. Ethnopharmacol., 94, 1–23,
2004.
13. Zeraik, M.L. and Yariwake, J.H., Quantification of isoorientin and total flavonoids in Passiflora edulis
fruit pulp. Microchem. J., 96, 86–91, 2010.
14. Vasco, C., Ruales, J., and Kamal-Eldin, A., Total phenolic compounds and antioxidant capacities of
major fruits from Ecuador. Food Chem., 111, 816–823, 2008.
15. Ichimura, T., Yamanaka, A., Ichiba, T., Toyokawa, T., Kamada, Y., and Tamamura, T., Antihypertensive
effect of an extract of Passiflora edulis rind in spontaneously hypertensive rats. Biosci. Biotechnol.
Biochem., 70, 718–721, 2006.
16. Zeraik, M.L., Serteyn, D., Deby-Dupont, G., Wauters, J.N., Tits, M., Yariwake, J.H., Angenot, L., and
Franck, T., Evaluation of the antioxidant activity of passion fruit (Passiflora edulis and Passiflora alata)
extract on stimulated nutraphils and myeloperoxidase activity assays. Food Chem., 128, 259–265, 2011.
17. Mercadante, A., Britton, G., and Rodriguez-Amaya, D., Carotenoids from yellow passion fruit (Passiflora
edulis). J. Agric. Food. Chem., 46, 4106–4106, 1998.
18. U.S. Department of Agriculture (USDA), Nutrition Coordinating Center Carotenoid Database for U.S.
Foods, Department of Agriculture/Agriculture Research Service, 1998. Published online at: http://
naldc.nal.usda.gov/download/22707/PDF (accessed on May 28, 2014).
19. Beutner, S., Bloedorn, B., Frixel, S., Blanco, I.H., Hoffmann, T., Martin, H.D., Mayer, B. et al.,
Quantitative assessment of antioxidant properties of natural colorants and phytochemicals: Carotenoids,
flavonoids, phenols and indigoids. The role of α−carotene in antioxidant functions. J. Sci. Food Agric.,
81, 559–568, 2001.
20. Lowe G.M. and Young, A.J., Antioxidant and prooxidant properties of carotenoids. Arch. Biochem.
Biosyn., 385, 20–27, 2001.
21. Copeland, L. and Slaytor, M., The excretion of the β-carboline alkaloid harman in passion fruit. Physiol.
Plantarum, 31, 327–329, 1974.
22. Konta, E.M., Almeida, M.R., do Amaral, C.L., Darin, J.D., de Rosso, V.V., Mercadante, A.Z., Antunes,
L.M., and Bianchi, M.L., Evaluation of the antihypertensive properties of yellow passion fruit pulp
(Passiflora edulis Sims f. flavicarpa Deg.) in spontaneously hypertensive rats. Phytother. Res., 28,
28–32, 2014.
23. Restrepo, R.A., Loango, N., Moncada, M.V., and Landazuri, P., Angiotensin-converting enzyme inhibi-
tory activity of Passiflora edulis f. flavicarpa and Petroselinum crispum (Mill) fuss. Br. J. Pharm. Res.,
3, 776–785, 2013.
24. Barbalho, S.M., Damasceno, D.C., Spada, A.P., Lima, I.E., Araujo, A.C., and Guiguer, E.L., Effects of
Passiflora edulis on the metabolic profile of diabetic Wistar rat offspring. J. Med. Food, 14, 1490–1495,
2011.
25. Cindy, M., The effects of yellow passion fruit, Passiflora edulis Flavicarpa, phytochemicals on cell
cycle arrest and apoptosis of leukemia lymphoma molt-4 cell line, MSc thesis, University of Florida,
Gainesville, FL, 2003.
26. Farid, R., Rezaieyazdi, Z., Mirfeizi, Z., Hatef, M.R., Mirheidari, M., Mansouri, H., Esmaelli, H. et al.,
Oral intake of purple passion fruit peel extract reduces pain and stiffness and improves physical func-
tion in adult patients with knee osteoarthritis. Nutr. Res., 30, 601–606, 2010.
27. Deng, J., Zhou, Y., Bai, M., Li, H., and Li, L., Anxiolytic and sedative activities of Passiflora edulis
f. flavicarpa. J. Ethnopharmacol., 128, 148–153, 2010.
28. Li, H., Zhou, P., Yang, Q., Shen, Y., Deng, J., and Li, L., Comparative studies on anxiolytic activi-
ties and flavonoid compositions of Passiflora edulis ‘edulis’ and Passiflora edulis ‘flavicarpa’.
J. Ethnopharmacol., 133, 1085–1090, 2011.
29. Silva, D.C., Freitas, A.L., Pessoa, C.D., Paula, R.C., Mesquita, J.X., and Leal, L.K., Pectin from
Passiflora edulis shows anti-inflammatory action as well as hypoglycemic and hypotriglyceridemic
properties in diabetic rats. J. Med. Food, 14, 1118–1126, 2011.
30. Zucolotto, S.M., Goulart, S., Montanher, A.B., Reginatto, F.H., Schenkel, E.P., and Frode, T.S.,
Bioassay-guided isolation of anti-inflammatory C-glucosylflavones from Passiflora edulis. Planta Med.,
75, 1221–1226, 2009.
31. Vargas, A.J., Geremias, D.S., Provensi, G., Fornari, P.E., Reginatto, F.H., and Gosmann, G., Passiflora
alata and Passiflora edulis spray-dried aqueous extracts inhibit inflammation in mouse model of pleu-
risy. Fitoterapia, 78, 112–119, 2007.
32. Ramos, A.T., Cunha, M.A.L., Sabaasrur, A.U.O., Pires, V.C.F., and Cardoso, A.A., Use of Passiflora
edulis f. flavicarpa on cholesterol reduction. Braz. J. Pharmacog., 17, 592–560, 2007.
33. Chau, C.F. and Huang, Y.L., Characterization of passion fruit seed fibres—A potential fibre source.
Food Chem., 85, 189–194, 2004.
34. Sato, A.C., Andrade, S.A., Brito, M.V., Miranda, A., Sampaio, M.U., and de Abreu Maffei, F.H., Effects
of compounds from Passiflora edulis Sims f. flavicarpa juice on blood coagulation and on proteolytic
enzymes. Protein Pept. Lett., 19, 501–508, 2012.
35. Celikyurt, I.K., Utkan, T., Gocmez, S.S., Hudson, A., and Aricioglu, F., Effect of harmane, an endog-
enous β-carboline, on learning and memory in rats. Pharmacol. Biochem. Behav., 103, 666–671, 2013.
36. Ripa, F.A., Haque, M., Nahar, L., and Islam, M.M., Antibacterial, cytotoxic and antioxidant activity of
Passiflora edulis Sims. Eur. J. Sci. Res., 31, 592–598, 2009.
37. Ngan, A. and Conduit, R., A double-blind, placebo-controlled investigation of the effects of Passiflora
incarnata (passionflower) herbal tea on subjective sleep quality. Phytother. Res., 25, 1153–1159, 2011.
38
Peach Juice
Emilio Alvarez-Parrilla, Laura A. de la Rosa, Joaquín Rodrigo-García,

Gustavo A. González-Aguilar, and Jesús F. Ayala-Zavala
CONTENTS
38.1 Introduction................................................................................................................................... 463
38.3.1 Phenolic Compounds........................................................................................................ 465
38.3.2 Carotenoids....................................................................................................................... 467
38.3.3 Dietary Fiber.................................................................................................................... 468
38.4 Health Effects................................................................................................................................ 469
38.6 Conclusion..................................................................................................................................... 472
References............................................................................................................................................... 472
38.1 Introduction
Fruit juice has become one of the main sources of dietary phytochemicals, and according to different
organizations, the consumption of a glass (~200 mL) of juice can be considered as one portion of fruit or
vegetable [1]. In 2009, the global world juice consumption was estimated at 40,580 million liters, with
Europe (27.8%) and the United States (23.5%) as the largest consumers [1]. Worldwide, orange juice is
the main fruit juice consumed, representing 54% of the juice consumed in 2010 in the United States [2].
Consumption of peach juice or nectar is highly dependent on cultural and ethnic traditions. For
instance, China represents 50% of world’s peach production, and peach juice is the second most con-
sumed juice just after orange juice [3]. Similar trends are observed in southern and central European
countries such as Italy, Spain, Bulgaria, Greece, Hungry, and Malta [1]. In contrast, in the United States
and northern European countries, peach juice consumption is scarce [1,2].
Due to technological problems, derived from the large stone and thick peel, peach is initially extracted
as peach pulp or puree, and then the puree is diluted to prepare peach juice or nectar [4]. Peach juice
production involves washing, peeling (steam or lye treatment), cutting, squeezing, pectinase treatment,
filtration and clarification (optional), concentration (optional), and pasteurization [5]. It is known that
some processing operations induce a decrease in the phytochemical content of fruit juices. Therefore, in
this chapter, the effect of processing on peach juice phytochemicals and antioxidant capacity is described
and its health benefits are discussed.

As previously stated, peach pulp or puree are normally used to produce peach juice (pulp with added
water) or nectar (pulp with added water and sugar). Complete compositional data on nutritional proper-
ties of peach juice and nectar are published by the USDA [6]. Table 38.1 summarizes the compositional
and nutritional characteristics of raw peach and some of the main juices and nectars commercialized in
463
TABLE 38.1
Compositional and Nutritional Characteristics of Fresh Peach and Peach Juice Products (per 100 g Fruit
or Juice)
Nutrient Unit Fresh Fruit Baby Juice Nectar Nectara
Water g 88.87 89.00 85.64 85.64
Protein g 0.91 0.20 0.27 0.27
Lipid (fat) g 0.25 0.10 0.02 0.02
Carbohydrate g 9.54 10.50 13.92 13.92
Total sugars g 8.39 9.59 13.32 13.32
Minerals
Calcium mg 6 3 5 5
Iron mg 0.25 0.56 0.19 0.19
Phosphorus mg 20 4 6 6
Potassium mg 190 97 40 40
Sodium mg 0 0 7 7
Zinc mg 0.17 0.03 0.08 0.08
Vitamins
Folate (DFE) µg 4 1 1 1
Niacin mg 0.806 0.213 0.288 0.288
Riboflavin mg 0.031 0.011 0.014 0.014
Thiamin mg 0.024 0.008 0.003 0.003
Vitamin A (RAE) µg 16 3 13 13
Vitamin B6 mg 0.025 0.022 0.007 0.007
Vitamin C mg 6.6 58.5 5.3 26.8
Vitamin E (ATE) mg 0.73 0.15 0.29 nr
Vitamin K µg 2.6 0.5 1.0 nr
Source: Adapted from the U.S. Department of Agriculture (USDA), USDA National Nutrient Database for Standard Reference,
Release 26, 2013, Published online at: http://www.nal.usda.gov/fnic/foodcomp/search/ (accessed April 28, 2014).
a Fortified with vitamin C.
Abbreviations: DFE, dietary folate equivalents; RAE, retinol activity equivalents; ATE, alpha-tocopherol equivalents; nr,
not reported.
the United States [6]. The water content remains practically unchanged for fresh peach and baby juice,
while peach nectar has lower water content. Furthermore, a decrease in total fiber and an increase in total
sugars are noted in baby juice, with a subsequent increase in energy value. These variations are conse-
quences of pectinase treatment of the peach pulp in order to obtain a clear peach juice [4]. Peach nectar
showed a lower fiber content than fresh peach, as well as a 45% increase in total sugar content due to the
pectinase treatment and the addition of high-fructose syrup during nectar formulation. Both peach juice
and nectar showed an increase on their energy value, as compared to fresh fruit, due to the increase in
total sugars. Considering that peach fiber is of high quality due to its soluble to insoluble fiber ratio [7],
several attempts have been carried out to supplement peach juice/nectar with peach by product fiber [8],
as will be further described in other sections of this chapter.
Protein content of peach juice decreased during processing and storage, compared to fresh peach
(Table 38.1). A threefold decrease in protein content was observed in both peach juice and nectar. Buedo
et al. [9,10] reported that the decrease in amino acid content was a result of nonenzymatic browning
reactions, mainly Maillard reactions, and that degradation degree was temperature dependent, both dur-
ing processing and storage. Aspartic acid showed the biggest decrease, about sixfold for both nectar
and juice, compared to fresh peach. Garza et al. [11] reported that nonenzymatic reactions also affected
Peach Juice 465
sucrose, glucose, and fructose concentration during storage at different temperatures. These authors
reported that while sucrose hydrolysis followed a pseudo-first-order reaction, increase of glucose and
fructose could not be adjusted to a pseudo-first-order appearance reaction because these two molecules
are consumed during Maillard reactions to produce brown pigment products, evidenced by a decrease
in Hunter b* parameter (blue color) and an increase in a* value (red color). They also concluded that
the effect of temperature on the nonenzymatic browning reactions could be described by the Arrhenius
model. Enzymatic and nonenzymatic browning reactions have been described as detrimental for peach
juice not only due to a loss in nutritional quality but also loss of sensory quality. Several attempts have
been made to reduce both enzymatic [12–14] and nonenzymatic [15] browning in peach juice.
Potassium, phosphorus, manganese, calcium, iron, and zinc are the main minerals found in fresh
peach, peach juice, and nectars, according to USDA (Table 38.1) [6]. Similar zinc and iron values were
reported for Turkish peach juice and nectars [16]. In general terms, a decrease in all mineral concentra-
tion, except sodium, was observed in peach juice and nectar, compared to fresh peach.
Fresh peach contains a variety of lipid- and water-soluble vitamins, including vitamins A, C, E, K, and
B-complex and folate (Table 38.1). Most of these vitamins are labile, and consequently, their concentra-
tion decreases during juice/nectar processing, due to high temperatures and the presence of some metals.
Vitamin C concentration of peach nectar showed a 20% decrease, compared to fresh peach. Vitamin C is
known to prevent oxidation during processing; however, no protective effect of it on the rest of the vita-
mins was observed in fortified nectar or baby juice, even though 5–10 times more vitamin C was added
to the juice. Vitamin A content showed a moderate decrease (21%) for both nectar and fortified nectar;
however, baby juice, which has been treated with pectinase enzyme and filtrated to clarify it, showed low
vitamin A content (81% decrease).

Peach is a rich source of health-promoting phytochemicals, including phenolic compounds, carotenoids,
and dietary fiber. These are the main bioactive compounds found in peach and its products. As previously
described for nutrients, there is usually a loss in phytochemical content during peach juice processing.
In this section, phenolic compounds, carotenoids, and dietary fiber content in peach juice are described.
38.3.1 Phenolic Compounds

Phenolic compounds are secondary plant metabolites of both flavonoid and nonflavonoid classes
(Figure 38.1). The main flavonoids found in peach are catechin, epicatechin, procyanidin B1, and
isoquercetin (quercetin-3-glucoside), while chlorogenic acid is the main nonflavonoid compound present.
Total phenolic content in fresh peach ranges from 38 to 300 mg gallic acid equivalents (GAE)/100 g fresh
fruit [17–22]. These wide variations can be explained due to cultivar variety, ripening stage, geographic
area, agronomic, and postharvest practices, among others [23].
Peach juice processing drastically modifies phenolic content, due to heating and blending. Lavelli
et al. [24] stated that the main losses were observed during peach puree preparation, while no further
degradation occurred during the processing of puree into nectar. Rababah et al. [19] reported a large
increment in total phenolics and anthocyanins content in peach puree compared to fresh fruit, prob-
ably due to lower moisture content. In another study, a 15% reduction in total phenolic content due to
pasteurization process was reported [18]. Talcott et al. [25] demonstrated that peach periderm removal
reduced the content of phenolics in the purees without improving its sensory quality. In the same study,
in agreement with Rababah et al. [19], the authors observed an increase of some phenolic compounds
during processing (steam blanching, blending, and pasteurization). It has been reported that losses in
phenolic compounds, compared to fresh fruit during processing, varied from 4% (chlorogenic acid, from
72 mg/kg in whole peach to 69 mg/kg in freshly prepared nectar) to 82% (cyanidin-3-O-glucoside, from
17.3 mg/kg in whole peach to 3.1 mg/kg in freshly prepared nectar), depending on the specific pheno-
lic compound, peach variety, and processing conditions [26]. Other polyphenolic compounds found in
peach nectar in this study were neochlorogenic acid (38–49 mg/kg), catechin (21–45 mg/kg), quercetin
OH OH
OH OH
HO O HO O
OH OH
OH OH
Catechin Epicatechin
OH
OH
HO O
OH OH
HO O OH OH
OH
O O HO O
OH OH
OH O
HO OH OH
OH
OH
Isoquercetin Procyanidin B1
HO OH
HO O
OH
HO O
HO O
Chlorogenic acid
FIGURE 38.1 Chemical structures of main phenolic compounds found in peach juice.
glycosides (7–23 mg/kg), and procyanidins (161–253 mg/kg) [26]. Within the same study, the authors
observed a pseudo-first-order phenolic degradation kinetic in accelerated degradation studies (37ºC).
In these conditions, cyanidin-3-O-glucoside was the least stable compound with a half-life of 13 days,
followed by procyanidins, catechin, isoquercetin, ascorbic acid, and chlorogenic acids, which was the
most stable compound with a half-life of 58 months. In addition, Lavelli et al. [26] observed that extrac-
tion of phenolic compounds during puree and juice production was temperature dependent. Authors
attributed this lower phenolic content at low temperature that could be due to a lower extraction rate and
higher polyphenol oxidase (PPO) and peroxidase activity, since at room temperature these enzymes are
not inactivated.
Various authors have determined phenolic compounds in commercial juices and nectars [27] and in
nectars from different peach varieties [22], showing a wide variation in the concentrations of individual
phenolic compounds (Table 38.2). In general, flavanols (catechin and procyanidins), chlorogenic acids
(chlorogenic and neochlorogenic), and flavonol (quercetin) glycosides are the major compounds found in
peach juice. Fernández de Simón [27] observed a higher content of flavonols in peach juice, compared
to apple, grape, pear, pineapple, and orange juices. In the same study, they observed an increase in fla-
vonols, in mixed fruit juice that included peach (peach–apple and peach–grape) compared to single fruit
juice. Considering that phenolic degradation may occur due to oxidation of phenolic compounds by PPO,
several attempts to reduce its activity using ascorbic acid [26] or cyclodextrins [12,13] have been carried
out, observing a drastic reduction on the degradation of phenolic compounds in model systems.
Antioxidant capacity of vitamin C‒fortified peach juice was evaluated with different methods [28],
such as trolox equivalent antioxidant capacity (TEAC), total peroxyl radical-trapping antioxidant
parameter (TRAP), and ferric-reducing antioxidant power (FRAP). They found a slight increase in anti-
oxidant capacity in peach juice with respect to fresh peach; addition of ascorbic acid during processing
Peach Juice 467
TABLE 38.2
Content of Polyphenolic Compounds in Peach Juice or Nectar
Polyphenolic Compounds Peach Juice or Nectar (mg/kg) References
Chlorogenic acid 0.22–7.12 [27]
12–19 [22]
47–96 [26]
20–51 [24]
Neochlorogenic acid 38–49 [26]
20–25 [24]
Caffeic acid 0.02–0.31 [27]
0.5–1.8 [22]
Catechin 0.12–4.64 [27]
20–34 [22]
21–45 [26]
13–24 [24]
Epicatechin 0.41–3.60 [27]
Procyanidins 161–253 [26]
Quercetin (+ glycosides) 0.12–0.54 [27]
5.2–7.1 [22]
7–23 [26]
3.9–10.6 [24]
Other flavonols (myricetin and kaempferol) 0.95–1.03 [27]
Cyanidin glycosides 1.2–17.0 [26]
nd-9.4 [24]
was contemplated. Lavelli et al. [26] observed a reduction in both phenolic content and antioxidant
capacity of peach puree and juice during storage and determined a pseudo-first order rate constants
for the loss of individual phenolic compounds and antioxidant capacity. Antioxidant capacity showed a
half-life of 6 months at 37ºC, while individual phenolic compounds have a half-life ranging from 13 to
1740 days, as previously mentioned.
38.3.2 Carotenoids
Other important functional constituents of peach are carotenoids, which can be divided in two groups. One
of them is formed by precursors of vitamin A, which include β-carotene, α-carotene, and β-cryptoxanthin.
The other is mainly integrated by compounds with antioxidant activity but no pro–vitamin A activity and
includes lutein, zeaxanthin, and lycopene. Peach is rich in β-carotene (43.0 μg/100 g fresh weight [FW]),
β-cryptoxanthin (7.0 μg/100 g FW), and lutein (20 μg/100 g) (Figure 38.2) [29]. Higher contents of these
carotenoids have been reported for yellow peach as compared to white cultivar [30]. This fact shows
the importance of the cultivar chosen for peach juice and for potential health benefits. Another impor-
tant decision is to keep whole fruit in juice, which will provide both components from peel and flesh.
Carotenoids are mainly detected in peel, so inclusion of peel in juice increases carotenoid content [26].
As in the case of phenolic compounds, carotenoid content of peach juice and intermediate products has
been reported (Table 38.3). A 66% reduction in total carotenoids of pasteurized peach has been reported
[18]. Lavelli et al. [26] found a 48%–65% reduction in carotenoid concentration of peach puree, observ-
ing that this reduction was higher in varieties, which were more susceptible to enzymatic browning. They
also observed higher carotenoid content in heat-processed purees and juices, compared to products at
room temperature, due to higher extraction rates at high temperatures. In contrast, Guiffrida et al. [31]
observed that carotenoid content remained practically unchanged during peach juice processing, when
both free and esterified carotenoids were taken into account. They observed that β-carotene was the
main free carotenoid found in peach juice, while among the monoesters, β-cryptoxanthin myristate was
H3C CH3 H3C

CH3 CH3
CH3 CH3 H3C CH3

CH3
β-Carotene
H3C OH
H3C CH3 CH3 CH3
CH3 CH3 H3C CH3

CH3
β-Cryptoxanthin
H3C CH3 H 3C OH
CH3 CH3
CH3 CH3 H3C CH3

HO CH3
Lutein
FIGURE 38.2 Chemical structures of main carotenoids found in peach juice.
TABLE 38.3
Carotenoids Content in Peach Fruit and Juice
Carotenoid Fresh Peach (μg/100 g) Peach Juice (μg/100 g)
Zeaxanthin 27 19
β-Carotene 198 161
β-Cryptoxanthin 12 26
β-Cryptoxanthin laureate 9 26
β-Cryptoxanthin myristate 44 52
β-Cryptoxanthin palmitate 7 20
Source: Adapted from Guiffrida, D. et al., Fruits, 68, 39, 2013. With permission.
Note: Data expressed as means (n = 3) on a fresh weight basis.
most abundant. Interestingly, these authors observed higher monoester concentrations in the juice com-
pared to fresh peach, which could partly explain the fact that previous studies had found lower carotenoid
content in peach juices than in fresh fruits.
38.3.3 Dietary Fiber

Dietary fiber is another functional ingredient in peach, although its content is usually drastically reduced
in peach juice/nectar. According to the USDA National Nutrient Database for Standard Reference [6],
total dietary fiber decreases from 1.5% in fresh peach fruit to 0.1% and 0.6% in baby peach juice and
peach nectar, respectively (Table 38.1). These large reductions in fiber content are the result of pro-
cessing during peach juice/nectar production, which include pectinase treatment and/or filtration and
clarification (optional), concentration (optional), and pasteurization [5]. Grigelmo-Miguel et al. [7] inves-
tigated the nutrient composition of “Sudanell” peach cultivar and found that total dietary fiber consti-
tuted 30.7%–36.1% dry weight (DW), from which 66% was insoluble fiber (cellulose, hemicellulose, and
lignin) and 34% soluble fiber (pectin, β-glucans, and arabinoxylans). Interestingly, peach fiber showed
higher insoluble/soluble ratio (near to 2) compared to other matrices, such as cereals, fruits, and vegeta-
bles, which made them a good source of fiber with beneficial health effects. Other interesting properties
Peach Juice 469
of peach fiber are its high water retention (9–12 g of water/g of fiber), low energy (less than 4 kcal/g
of fiber), and the insoluble to soluble fiber ratio (66:34), which made peach fiber an interesting functional
constituent [7]. Considering these functional properties of peach fiber, recently, Augusto et al. [8] studied
the rheological properties of peach fiber‒fortified juices (0%–12% fiber addition), observing that peach
juice passed from a Newtonian fluid without the addition of fiber, to viscoelastic behavior with 12.5%
fiber addition. It was concluded that the addition of fiber resulted in a more consistent product, and that
this modification on the rheological behavior of peach juice could be explained in terms of interactions
between peach fiber polysaccharides with natural sugars and acids present in the juice, as well as the
degree of methoxylation of the polysaccharides present (in the case of pectin) [8].
38.4 Health Effects

Health benefits of peach juice are attributed to the presence of different classes of bioactive compounds,
which include phenolic compounds, carotenoids, and indigestible polysaccharides. The healthy prop-
erties of phenolic compounds are widely attributed to their antioxidant capacity, which is dependent
on their chemical structure, mainly on the number and position of hydroxyl substituents. Free radical
theory of aging states that oxidative stress is one of the most important causes of aging and age diseases.
It is known that oxidized low-density lipoprotein (LDL) cholesterol is one of the initial mechanisms to
promote atheromatous plaque formation, directly related to the development of cardiovascular disease
(CVD) [32]. Therefore, consumption of peach juice can raise the antioxidant capacity of a diet, poten-
tially lowering the formation of oxidized LDL and decreasing CVD risk.
Ko et al. [33] studied the in vivo effect of peach juice consumption in human plasma antioxidant capac-
ity, observing that drinking of 120 mL of peach juice induced a peak in the plasma antioxidant capacity
at 30 min, which was lost 2 h after consumption. In another study, peach juice was used in a 12-week
intervention with Japanese immigrants in Brazil. Participants (37 people) were divided in two groups,
and both groups received 200 mL peach juice daily. One of them drank juice enriched with 7.5 g of soy
protein and 10 mg of isoflavones. Placebo group just got peach juice. After intervention, the enriched-
juice group showed a significant decrease in body weight, body mass index, systolic blood pressure,
and total and LDL cholesterol. High-density lipoprotein (HDL) cholesterol also showed a tendency to
increase in this group. Interestingly, values for glycosylated hemoglobin (HbA1c) significantly decreased
in both groups, which revealed an effect of peach juice by itself [34]. This study also showed that peach
juice could be a perfect vehicle for other functional ingredients that can be potentiated by the bioactive
ingredients naturally found in peach juice. However, the processing operations involved in juice and nec-
tar production induce loss of some nutrients and bioactive compounds; juices also show increased sugar
and reduced fiber content compared to whole fruits. As a consequence, juices have recently been viewed
as less healthy options for fruit intake. Novel juice formulations, with added fiber, combination with
other fruit juices or herbal extracts, and/or use of innovative technologies to control microbial spoilage
are advisable to increase health benefits and market share of peach-based beverages.
Formerly, interest in carotenoids was only related to their pro–vitamin A activity, but recently, other
activities such as antioxidant activity, immune system enhancement, modulation of gene transcription,
and visual function have become more important. There are some contradictory results related to the
potential protective effect of carotenoids in some kinds of cancer such as lung cancer, especially in smok-
ers. This fact has raised concerns about the indiscriminate consumption of β-carotene supplements, and
the Institute of Medicine of the United States does not recommend their use for general population and
has only approved it for vitamin A–deficit cases [35]. However, consumption of food such as peach juice
that is rich in β-carotene is still recommended.
Even though there is controversy of their potential benefits against chronic-degenerative diseases,
many authors have pointed to evidences that showed potential health benefits of carotenoids. These
potential benefits are due to their antioxidant capacity and/or their ability to be precursors of vitamin A.
In addition, β-cryptoxanthin in combination with lycopene suppressed levels of tumor necrosis factor α,
which leads to lung inflammation prevention [36]. This potential synergic effect would be desirable in
smokers, therefore inclusion of peach and tomato juices in their regular diet could be another tool for
lung cancer prevention and/or treatment. A study conducted in Singapore was set to evaluate if plasma
levels of carotenoids could be useful as a predictive tool of acute myocardial infarction (AMI). A group
of 280 people diagnosed and registered with AMI was compared with a control (560 people) of similar
ages. Authors were able to find statistically significant correlation between prediagnostic plasma levels
of β-cryptoxanthin and lutein and risk of AMI. They also found that the inverse relation was independent
of different coronary heart disease (CHD) risk factors, such as smoking, diabetes, obesity, hypertension,
and hyperlipidemia [32].
Pectin is one of the most studied fiber components in peach products. It can be considered as soluble
fiber and, therefore, is capable of forming high viscosity solutions that can help to lower plasma choles-
terol and postprandial glucose. This fact is in agreement with Moriguchi et al. [34], who found that daily
consumption of 200 mL of peach juice decreased glycosylated hemoglobin. Moreover, fiber from peach
juice can be fermented by intestinal microorganisms that could generate short-chain fatty acids (SCFA),
with several biological activities, and also allow the growth of selected beneficial microorganisms, which
could prevent pathogens to colonize the intestine. Among SCFA, the most important are acetate, pro-
pionate, and butyrate. It has been demonstrated that increasing fiber levels in diet could have potential
benefits in obesity control and prevention. This beneficial effect is mainly due to the ability of fiber to
prevent full nutrient absorption by making the nutrients unavailable to digestive enzymes, resulting in a
lower energy intake that favors a fixed body weight [37]. Taking into account all the benefits of fiber, it is
very important to include it in peach juice, as much as the sensory attributes allow.

These days, the market is turning toward the commercialization of food and ingredients that reduce the
risk of diseases such as CVD, cancer, and atherosclerosis. Moreover, the juice market is saturated with
traditional products, and consumers are eager for products with improved and novel sensory characteris-
tics. In this context, the development of new products is as important for the fresh peach fruit sector as it
is for manufactured beverages. The current trends regarding innovation in the juice industry are mainly
influenced by consumer demands, and responding to these preferences, producers and governments are
taking action. In this respect, consumers are demanding more flavor and nutrient-rich juice, and gov-
ernmental agencies around the world are aiming at reducing chronic diseases (obesity, diabetes, etc.) by
controlling the consumption of sugar-rich juice products. Consequently, flavorful and nutritious peach
juice products can be contemplated to be formulated as innovative products to compete in the beverage
market. The mixture of peach juice with other beverages and foods could be contemplated. For instance,
due to consumer demand on health products, there are fruit–milk beverage mixtures on the market.
Zulueta et al. [38] analyzed 17 fruit–milk beverage mixtures commercialized in Spain, which contained
in their formulation different amounts of orange, pineapple, apple, mango, strawberry, banana, apricot,
lemon, passion fruit, kiwi fruit, or peach juices. In this study, authors reported a high variability in the
beverage carotenoid content, observing that those mixtures containing peach juice contained the highest
total carotenoid. β-Cryptoxanthin, β-carotene (and its isomers), and lutein were the main carotenoids
found in these mixtures. The addition of fiber to peach juice has also been contemplated as a promising
alternative for novel product development [8].
The development and marketing of new products is a complex process. In the food industry, sensory
and consumer data are frequently produced and applied as the basis for decision making worldwide.
The trends in the fruit juice industry mark the potential success of formulations based on peach juice,
such as smoothies, tea mixtures, and exotic fruit flavor mixtures. For this reason, it is thought that
the characteristic sweet aroma and taste of peach juice could be used to complement other compatible
aromas and flavors, so their sensory evaluation becomes essential. Robust knowledge on whether or
not consumers would like a novel flavor/taste is important in reaching a go/no-go decision to proceed
with the development. New acetic fruit juices (pineapple juice with 10 g/kg of wine vinegar and peach,
orange, and apple juices with 50 g/kg of wine vinegar), previously optimized by a panel of experts, were
subsequently tasted by a group of consumers, who evaluated the olfactory and gustatory impression, pre-
ferring beverages made from peach and pineapple juices [39]. The gustatory impression was revealed as
Peach Juice 471
the most influential descriptor of the new product, according to a correlation study of “purchase intent”
with “olfactory impression” and “gustatory impression.” Considering the flavor acceptance popularity
and health benefits of peach juice on the market, this can be an excellent option for accomplishing the
innovation goals in this area.
The U.S. government promoted the “National School Lunch Program and School Breakfast Program:
Nutrition Standards for All Foods Sold in School as Required by the Healthy, Hunger-Free Kids Act of
2010” [40]. This act affected the juice industry, by limiting the caloric content offered to young con-
sumers; therefore, producers need to develop new products that fit this bill. In this context, producers
will start to reduce the available portions, eliminate added sugar, or contemplate sugar substitution.
Parpinello et al. [41] studied the suitability of stevioside as a sweetener in peach juice, observing that
160 mg/L of stevioside could replace 34 g/L sucrose with a 25% reduction in calories, without affecting
sensory characteristics of the juice. However, consumers consider more than just the sugar content of
the juice products they purchase [42]. Among the factors that are most often assumed to be important
to the consumer are the taste of the product and the specific nature of the health benefit to be achieved.
The taste of the product has been viewed as a particularly important variable, since many nutraceutical
compounds have natural bitter, astringent, or other off flavors that can lead to an unacceptable flavor in
these products [43].
The microbiological quality and addition of additives to peach juice is another factor that should be
considered for new formulations. The refrigerated juice market is getting popular among consumers, for
convenience, flavor, and higher content of bioactive compounds from the fruit; however, safety problems
are being reported in this kind of products, especially for low-pH juices (<4.6). Considering the market
trends of preservation of foods, the use of natural antimicrobial compounds can be an option to reduce
microbial problems in new peach juice formulations, with the purpose of providing natural products
acceptable to consumers.
Manufacturing peach juice/nectar by conventional preservation methods (heat treatments) maintains
its safety but reduces its sensory and nutritional qualities. Different novel techniques have been employed
to improve microbiological quality without impairing nutritional quality. Pulsed electric field (PEF)
treatment has successfully been used to inactivate microorganisms in peach juice, without losing large
amounts of heat-sensitive bioactives; in fact, only a 10% decrease in ascorbic acid was observed after
PEF treatment [14]. Altuntaş et al. [44] observed that a 30 kV/cm electric field strength PEF treatment
significantly inactivated Escherichia coli O157:H7, Staphylococcus aureus, Listeria monocytogenes,
Erwinia carotovora, Pseudomonas syringae, Botrytis cinerea, and Penicillium in peach nectar with-
out affecting peach quality parameters (pH, ºBrix, titration acidity, color, ascorbic acid, and β-carotene
content). Supercritical CO2 and N2O pasteurization (15 min of CO2 ⁄ N2O treatment, 10 MPa, and 35ºC)
of peach juice showed total inactivation of Saccharomyces cerevisiae, without affecting nutritional or
sensory quality [45].
Due to the growing concern about food allergy, and considering that peach juice consumption is asso-
ciated with oral allergy syndrome (oral itching, mucosal edema, labial itching, and papulas), due to the
presence of the lipid transfer protein (Pru p1), mainly in the epicarp, Brenna et al. [46] studied the effect
of peeling on the allergenic effect of peach juice. For this, authors lyed peeled peaches (10% NaOH,
60ºC, 90 s) before puree and nectar preparation, and observed a drastic reduction on peach allergenicity,
which was related to Pru p1 denaturalization at high-alkaline conditions, without any loss in peach juice
quality. Similar results were obtained by ultrafiltration using 10 kDa cutoff membranes.
Finally, studies on the use of peach for fruit wine manufacture have been carried out, since peach
wine is highly consumed in some countries such as the United States. Recently, Davidović et al. [47] pro-
duced peach wine from Redhaven peach, which is a yellow-fleshed, freestone cultivar, and compared its
total phenolic and flavonoid contents, antioxidant activity assays (1,1-diphenyl-2-picryl-hydrazyl [DPPH]
and FRAP), and sensory acceptability against those of white grape wines (Riesling and Chardonnay).
Results showed that peach wine presented higher total phenolics, flavonoids, and antioxidant capacity,
as well as lower individual and total sugar and alcohol content, and good acceptability among regular
wine consumers. Chlorogenic acid, caffeic acid, and catechin were the main phenolic compounds found
in peach wine, all at concentrations higher than those in grape wines. These results suggest that peach
wine could be an interesting functional product among white wine consumers.
38.6 Conclusion
Peach juice and nectar are popular beverages in some countries where they represent an important
source of nutrients and bioactive compounds. The major phytochemicals in peach juice are polyphenols
such as chlorogenic and neochlorogenic acids, catechin, quercetin glycosides, and carotenoids such as
β-carotene and β-cryptoxanthin. All of these compounds have been linked, mainly by in vitro, animal,
and epidemiological studies, to the prevention of CVD and tumor growth prevention or suppression,
among others.
REFERENCES
1. European Fruit Juice Association (AIJN), 2010 Market Report, Brussels, Belgium, 2010.
2. U.S. Department of Agriculture (USDA), Item clusters, percent of consumption, and representative foods
for all USDA food patterns, food groups or subgroups, August 2010. Published online at: http://www.
cnpp.usda.gov/Publications/USDAFoodPatterns/ItemClustersAndRepFoods.pdf (accessed August 10,
2013).
3. U.S. Department of Agriculture (USDA), China-People Republic of- Stone Fruit Annual Report,
July 2013. Published online at: http://gain.fas.usda.gov/Recent%20GAIN%20Publications/Stone%20
Fruit%20Annual_Beijing_China%20-%20Peoples%20Republic%20of_7–1–2013.pdf (accessed August
9, 2013).
4. Food and Agriculture Organization (FAO), Principles and Practices of Small- and Medium-Scale Fruit
Fuice Processing, Agricultural Services Bulletin 146, FAO, Rome, Italy, 2001.
5. Siddiq, M., Peach and nectarine, in Handbook of Fruits and Fruit Processing, Hui, Y.H., Barta, J., Cano,
J.M., Gusek, T.K., Sidhu, J.S., and Sinha, N.K., Eds., Wiley-Blackwell, Oxford, UK, 2006, pp. 519–531.
Release 26, 2013. Published online at: http://www.nal.usda.gov/fnic/foodcomp/search/ (accessed April
28, 2014).
7. Grigelmo-Miguel, N., Gorinstein, S., and Martin-Belloso, O., Characterisation of peach dietary fibre
concentrate as a food ingredient. Food Chem., 65, 175–181, 1999.
8. Augusto, P.E.D., Fulguera, V., Cristianini, M., and Ibarz, A., Influence of fibre addition on the rheologi-
cal properties of peach juice. Int. J. Food Sci. Technol., 46, 1086–1092, 2011.
9. Buedo, A.P., Elustondo, M.P., and Urbicain, M.J., Amino acid loss in peach juice concentrate during
storage. Innov. Food Sci. Emerg., 1, 281–288, 2001.
10. Buedo, A.P., Elustondo, M.P., and Urbicain, M.J., Non-enzymatic browning of peach juice concentrate
during storage. Innov. Food Sci. Emerg., 1, 255–260, 2001.
11. Garza, S., Ibarz, A., Pagán, A., and Giner, J., Non-enzymatic browning in peach puree during heating.
Food Res. Int., 32, 335–343, 1999.
12. de la Rosa, L.A., Mercado-Mercado, G., Rodrigo-García, J., González-Aguilar, G.A., and Alvarez-
Parrilla, E., Peach polyphenol oxidase inhibition by β-cyclodextrin and 4-hexylresorcinol is substrate
dependent. CYTA. J. Food, 8, 87–93, 2010.
13. López-Nicolás, J.M., Pérez-López, A.J., Carbonell-Barrachina, A., and García-Carmona, F., Use of
natural and modified cyclodextrins as inhibiting agents of peach juice enzymatic browning. J. Agric.
Food Chem., 55, 5312–5319, 2007.
14. Meneses, N., Jaeger, H., and Knorr, D., Minimization of thermal impact by application of electrode
cooling in a co-linear PEF treatment chamber. J. Food Sci., 76, E536–E543, 2011.
15. Arslanoğlu, F.N., Kar, F., and Arslan, N., Adsorption of dark coloured compounds from peach pulp by
using powdered-activated carbon. J. Food Eng., 71, 156–163, 2005.
16. Acar, O., Determination of lead, copper, iron, and zinc levels in fruit jams, nectars, juices and beverages
by electrothermal and flame atomic absorption spectrometry. Eurasian J. Anal. Chem., 6, 114–128,
2011.
17. Di Vaio, C., Graziani, G., Marra, L., Cascone, A., and Ritieni, A., Antioxidant capacities, carotenoids
and polyphenols evaluation of fresh and refrigerated peach and nectarine cultivars from Italy. Eur. Food
Res. Technol., 227, 1225–1231, 2008.
Peach Juice 473
18. Oliveira, A., Pintado, M., and Almeida, D.P.F., Phytochemical composition and antioxidant activity of
peach as affected by pasteurization and storage duration. LWT—Food Sci. Technol., 49, 202–207, 2012.
19. Rababah, T.M., Ereivej, K.I., and Howard, L.R., Effect of ascorbic acid and dehydration on concentra-
tions of total phenolics, antioxidant capacity, anthocyanins, and color in fruits. J. Agric. Food Chem.,
53, 4444–4447, 2005.
20. Rodrigo-Garcia, J., de la Rosa, L.A., Herrera-Duenez, B., Gonzalez-Barrios, A.G., Lopez-Diaz, J.A.,
González-Aguilar, G.A., Ruiz-Cruz, S., and Alvarez-Parrilla, E., Cuantificacion de polifenoles y capa-
cidad antioxidante en duraznos comercializados en Ciudad Juarez, Mexico. Tecnociencia Chihuahua,
5, 67–75, 2011.
21. U.S. Department of Agriculture (USDA), Database for the flavonoid content of selected foods, Release 3,
September 2011. Published online at: http://www.ars.usda.gov/Services/docs.htm?docid=6231 (accessed
October 28, 2013).
22. Versari, A., Castellari, M., Parpinello, G.P., Riponi, C., and Galassi, S., Characterisation of peach juices
obtained from cultivars Redhaven, Suncrest and Maria Marta grown in Italy. Food Chem., 76, 181–185,
2002.
23. Alvarez-Parrilla, E., de la Rosa, L.A., González-Aguilar, G.A., and Ayala-Zavala, J.F., Phytochemical
composition and health aspects of peach products, in Dried Fruits: Phytochemicals and Health Effects,
Alasalvar, C. and Shahidi, F., Eds., Wiley-Blackwell, Oxford, UK, 2013, pp. 309–324.
24. Lavelli, V., Pompei, C., and Casedei, M.A., Quality of nectarine and peach nectars as affected by lye-
peeling and storage. Food Chem., 115, 1291–1298, 2009.
25. Talcott, S.T., Howard, L.R., and Brenes, C.H., Contribution of periderm material and blanching time to
the quality of pasteurized peach puree. J. Agric. Food Chem., 48, 4590–4596, 2000.
26. Lavelli, V., Pompei, C., and Casedei, M.A., Optimization of color and antioxidant activity of peach and
nectarine puree: Scale-up study from pilot to industrial plant. J. Agric. Food Chem., 56, 7091–7099, 2008.
27. Fernández de Simón, B., Pérez-Ilzarbe, J., Hernández, M.T., Gómez-Cordovés, C., and Estrella, I.,
Importance of phenolic compounds for the characterization of fruit juices. J. Agric. Food Chem., 40,
1531–1535, 1992.
28. Pellegrini, N., Serafini, M., Colombi, B., del Rio, D., Salvatore, S., Bianchi, M., and Brighenti, F., Total
antioxidant capacity of plant foods, beverages and oils consumed in Italy assessed by three different
in vitro assays. J. Nutr., 133, 2812–2819, 2003.
29. O´ Shea, N., Arendt, E., and Gallagher, E., Dietary fibre and phytochemical characteristics of fruit and
vegetable by-products and their recent applications as novel ingredients in food products. Innov. Food
Sci. Emerg., 16, 1–10, 2012.
30. Gil, M.I., Tomás-Barberán, F.A., Hess-Pierce, B., and Kader, A.A., Antioxidant capacities, pheno-
lic compounds, carotenoids, and vitamin C contents of nectarine, peach, and plum cultivars from
California. J. Agric. Food Chem., 50, 4976–4982, 2002.
31. Guiffrida, D., Torre, G., Dugo, P., and Dugo, G., Determination of the carotenoid profile in peach fruits,
juice and jam. Fruits, 68, 39–44, 2013.
32. Koh, W.P., Yuan, J.M., Wang, R., Lee, Y.P., Yu, M.C., and Ong, C.N., Plasma carotenoids and risk of
acute myocardial infarction in the Singapore Chinese Health Study. Nutr. Metab. Cardiovasc. Dis., 21,
685–690, 2011.
33. Ko, S.H., Choi, S.W., Ye, S.K., Kim, H.S., and Chung, M.H., Comparison of the antioxidant activities of
nine different fruits in human plasma. J. Med. Food, 8, 41–46, 2005.
34. Moriguchi, E.H., Yamori, Y., Mori, M., Sagara, M., Mori, H., Sakuma, T., Ishikawa, P.M., and
Moriguchi, Y., New beverage for cardiovascular health, proposal based on oriental and occidental food
culture from world-wide epidemiological study. Geriatr. Gerontol. Int., 8, S3–S7, 2008.
35. Olmedilla-Alonso, B. and Granado-Lorencio, F., Componentes bioactivos, in Alimentos Funcionales:
Aproximación a una Nueva Alimentación, Barberá Mateos, J.M. and Marcos, A. Eds., Dirección
General de Salud Pública y Alimentación, Madrid, Spain, 2008, pp. 170–192.
36. Tang, F.Y., The silver bullet for cancer prevention: Chemopreventive effects of carotenoids. Biomedical,
2, 117–121, 2012.
37. Zarzuelo-Zurita, A. and Gálvez-Peralta, J., Fibra, in Alimentos Funcionales: Aproximación a una
Nueva Alimentación, Barberá Mateos, J.M. and Marcos, A. Eds., Dirección General de Salud Pública
y Alimentación, Madrid, Spain, 2008, pp. 104–127.
38. Zulueta, A., Esteve, M.J., and Frígola, A., Carotenoids and color of fruit juice and milk beverage
m ixtures. J. Food Sci., 72, C457–C463, 2007.
39. Cejudo-Bastante, M.J., Dodero, M.C.R., Guerrero, E.D., Mejias, R.C., Marin, R.N., and Barroso, C.G.,
Development and optimisation by means of sensory analysis of new beverages based on different fruit
juices and sherry wine vinegar. J. Sci. Food Agric., 93, 741–748, 2013.
40. Crawford, P.B., Gosliner, W., and Kayman, H., Peer reviewed: The ethical basis for promoting nutri-
tional health in public schools in the United States. Prev. Chronic Dis., 8, 1–6, 2011.
41. Parpinello, G.P., Versari, A., Castellari, M., and Galassi, S., Stevioside as a replacement of sucrose in
peach juice: Sensory evaluation. J. Sens. Stud., 16, 471–484, 2001.
42. Institute of Food Technologists (IFT), State of the industry: Juice & juice drinks, juices reformulate to
meet consumer preferences, July 2013. Published online at: http://www.bevindustry.com/articles/86556
(accessed August 20, 2013).
43. Cardello, A.V. and Schutz, H.G., The importance of taste and other product factors to consumer interest
in nutraceutical products: Civilian and military comparisons. J. Food Sci., 68, 1519–1524, 2003.
44. Altuntaş, J., Evrendilek, G.A., Sangun, M.K., and Zhang, H.Q., Processing of peach nectar by pulsed
electric fields with respect to physical and chemical properties and microbial inactivation. J. Food
Process Eng., 34, 1506–1522, 2011.
45. Spilimbergo, S. and Ciola, L., Supercritical CO2 and N2O pasteurisation of peach and kiwi juice. Int. J.
Food Sci. Tech., 45, 1619–1625, 2010.
46. Brenna, O., Pompei, C., Ortolani, C., Pravettoni, V., Farioli, L., and Pastorello, E.A., Technological pro-
cesses to decrease the allergenicity of peach juice and nectar. J. Agric. Food Chem., 48, 493–497, 2000.
47. Davidović, S.M., Veljović, M.S., Pantelić, M.M., Baošić, R.M., Natić, M.M., Dabić, D.Ĉ., Pecić, S.P.,
and Vukosavljević, P.V., Physicochemical, antioxidant and sensory properties of peach wine made from
Redhaven cultivar. J. Agric. Food Chem., 61, 1357–1363, 2013.
39
Pear Juice
Dulcineia Ferreira Wessel, Elisabete Coelho, and Manuel A. Coimbra
CONTENTS
39.1 Introduction....................................................................................................................................475
39.4 Health Effects................................................................................................................................ 481
39.6 Conclusion..................................................................................................................................... 485
References............................................................................................................................................... 485
39.1 Introduction
The pear tree (Pyrus) is a member of the subfamily Maloideae, family Rosaceae. In Europe, the main
cultivated species is Pyrus communis L., the European pear. This pear is the pomaceous fruit of the trees
within the genus Pyrus. There are about 2000 P. communis cultivars, but only a few are commercial-
ized on a large scale [1]. In East Asia, the main cultivated species are P. pyrifolia, P. ussuriensis, and
P. bretschneideri Rehd [2]. The Korean pear from the species P. pyrifolia, also known as the Nashi pear,
is consumed for its medicinal properties.
After China, the European Union (EU) is the world’s largest producer of pears, followed by the United
States and Argentina [3]. The volume of pears used for processing, which includes pear juice production,
within the European Union was around 260,000 tons for the marketing year 2011–2012, and was esti-
mated at 778,000 tons for the United States. The production of pear juice is low when compared to other
fruits [4] but eventually can be exploited in order to create new products and new markets.
The designation “pear juice” is used several times ambiguously, since there are some differences in the
technological process of preparing the juice: clarified or turbid juices. The product names authorized in
the EU are fruit juice, fruit juice from concentrate, concentrated fruit juice, water extracted fruit juice,
dehydrated/powdered fruit juice, and fruit nectar [5]. Pear juice is the fermentable but unfermented prod-
uct obtained from the edible part of the fruit, which is sound and ripe, fresh or preserved by chilling or
freezing, of one or more kinds mixed together, having the characteristic color and flavor typical of the
juice of the fruit from which it originates. Flavor, pulp, and cells obtained by suitable physical means
from the same species of fruit may be restored to the juice. The mixing of fruit juice with fruit purée
is also authorized in the production of the fruit juice. Pear nectar is obtained by adding water, with or
without the addition of sugar and/or honey, to products such as fruit juice, fruit juice from a concentrate,
concentrated fruit juice, and fruit purée, fulfilling the requirements of a minimum juice and/or purée
content of 50% by volume of the finished product [5]. The data on production and consumption of pear
juice are limited. To have an idea of pear juice consumption in Europe, the situation in Germany can
be used as an example. This is a country that heads up the EU juice and nectars market ranking with
a 26% share of consumption in the EU, 10.7 billion liters in 2011 [6]. According to the USDA Foreign
Agriculture Service 2011 report [3], the annual average per capita consumption of pear juice in Germany
in the years 2007–2010 was 0.23 L far below the 8.49 and 9.33 L of apple and orange juices, respectively.
475
Taking into account the phytochemical components that constitute the pear, the biological activity
they may have as constituents of pear juice, and the beneficial effects to health, one may predict that the
pear juice and nectar will be a segment of the beverage market to be explored in the future. This chapter
highlights the nutritional characteristics, bioactive compounds, as well as antioxidant efficacy, health
effects, and novel formulations of pear juice.

The nutritional quality of pear juice may be influenced by various factors such as cultivars, soil, and climate
conditions during fruit development [7]. For each cultivar, the nutritional content of pear and pear juice
depends on the harvest time and storage conditions of the fruits and processing technology [7,8].
Pear juice has a low content of protein and fat and high content of total sugars (Table 39.1). It is a natu-
ral source of vitamin C (1.1 mg/100 g), an essential antioxidant for metabolism and tissue repair, and
involved in the prevention of free radical damage. With respect to minerals, potassium is the highest
(13 mg/100 g) in pear juice [9]. The other minerals (e.g., calcium, sodium, magnesium, and phosphorus)
are present in low concentrations (Table 39.1).
TABLE 39.1
Compositional and Nutritional Characteristics of Pear Nectar (per 100 g)
Nutrient Unit Pear Nectar
Water g 84.01
Energy kcal 60
Protein g 0.11
Lipid (fat) g 0.01
Minerals
Calcium mg 5
Copper mg 0.067
Iron mg 0.26
Magnesium mg 3
Manganese mg 0.030
Phosphorus mg 3
Potassium mg 13
Sodium mg 4
Zinc mg 0.07
Vitamins
Choline mg 2.0
Niacin mg 0.128
Riboflavin mg 0.013
Thiamin mg 0.002
Vitamin C mg 1.1
Vitamin B6 mg 0.014
Reference, Release 26, 2013, Published online at: http://ndb.nal.usda.gov/ndb/search/list (September 9, 2014).
Abbreviation: ATE, alpha-tocopherol equivalents.
Pear Juice 477
Pear juice contains a high amount of fructose (6.3–6.4 g/100 mL) and low concentrations of glucose
(2.3 g/100 mL) and sorbitol (2.0 g/100 mL), and only a minor amount of sucrose (0.90–0.92 g/100 mL) [10].
The concentration of these compounds may vary depending on the juice preparation and the particular
cultivar of fruit employed. Not all fruit juices contain sorbitol, as for example, the white grape juice. Pear
juice is known as a highly fermentable oligo-, di-, and monosaccharides and polyols (FODMAP) food
source [11]. They comprise fructose, lactose, fructo- and galactooligosaccharides (fructans and galactans),
and polyols (e.g., sorbitol, mannitol, xylitol, and maltitol), all of which claimed to have the following
common functional properties [11]: (1) poorly absorbed in the small intestine, (2) osmotically active, and
(3) rapidly fermented by bacteria.
Some pear cultivars may also contain fructooligosaccharides (FOS). In Bosc and d’Anjou cultivars,
small amounts of 1-kestose (0.8 and 0.3 mg/g on dry weight [DW], respectively) and 1F-β-fructo
furanosylnystose (0 and 1.1 mg/g on DW, respectively) have been identified [12]. The dietary fiber in
pear juices with pulp is similar to that present in the fruit. The juice from one medium size pear (166 g)
contains 5.1 g of total dietary fiber [13], providing about 25% of the daily need of dietary fiber [14].
Fresh pear pulp contains soluble (1.5 g) and insoluble (3.6 g) dietary fiber [13].
The free amino acids of d’Anjou pear juice showed that the most abundant free amino acid is aspartic
acid (128 mg/L), followed by cysteine (38 mg/L) and serine (25 mg/L) [15]. Pear juice is also character-
ized by a predominance of malic and quinic acids, 371 and 220 mg/100 mL of juice, respectively.

Bioactive compounds are considered as nonnutritional but vital ingredients for the maintenance of human
health [16]. In recent years, several bioactive compounds have been investigated with respect to their
potential antioxidant, anticancer, and anti-inflammatory activities using different in vitro and in vivo mod-
els as well as clinical trials. Among these bioactive compounds, pear juice can be a good source of phe-
nolic compounds. Plant phenolic compounds, a large group of natural antioxidants, are serious candidates
that may explain the protective effects of fruits against cancer and cardiovascular disease (CVD) [17,18].
To evaluate the potential beneficial health effects of pear juice, the bioactive compounds of pear and
its juice need to be examined. Thus, elucidation of the structures of bioactive compounds of pear juice
and their quantification is essential. The study of antioxidant capacity in vitro is usually the first step to
understand the potential bioactivity of these compounds. Bioavailability is also a key step to ensure the
bioefficacy of the bioactive compounds [19]. The literature on bioactive compounds in pear and pear
juice is scarce. However, it is possible to find a slight increase in the interest of the researchers on study-
ing pears of various cultivars and pear juice. The research work that is reported in the scientific literature
about characterization and quantification of bioactive compounds from pear juice and its antioxidant
properties is presented and discussed in this section. Reports on strategies to improve antioxidant effi-
cacy of bioactive compounds of pear juice are also discussed.
The pear naturally undergoes changes in its chemical composition during juice preparation. The pro-
cessing may cause modifications in the pear phenolic compounds resulting from oxidation and reaction
with other components [20]. In order to improve juice yield, industrial production of pear juice uses mash
maceration with pectinolytic enzymes to facilitate the extraction of the juice by degradation and solubiliza-
tion of the cell wall polysaccharides. Pectinolytic enzymes can also enhance the extraction of antioxidant
phenols [21]. These enzymes are also used in order to avoid turbidity, especially during the concentration
of clarified juice. Using the comparison of the wines, where the addition of pectinolytic enzyme prepara-
tions at the vinification step resulted in higher levels of phenolic compounds, especially low-molecular-
weight phenols and flavan-3-ol derivatives [22], it is possible that the same could occur in pear juice.
Although pear is classified in the group of fruits with low antioxidant capacity by the oxygen radi-
cal absorbance capacity (ORAC) assay using a peroxyl radical generator [23], it was suggested that
the pear peel is an excellent source of antioxidants [24]. Thus, pear juice industrially prepared from
whole fruit can be a good source of phenolic compounds. Phenolic compounds such as arbutin and the
hydroxycinnamic acids such as 5-caffeoylquinic (chlorogenic), caffeic, p-coumaroylquinic, p-coumaric,
and p-coumaroylmalic acids (Figure 39.1), along with a number of proanthocyanidins and glycosides of
OH
R R—
—H p-Coumaric acid
O R—
—OH Caffeic acid
HO
OH
O —H
R— 5-p-Coumaroylquinic acid
R
—OH
R— 5-Caffeoylquinic acid
O OH
5 7
1 COOH
HO
HO
FIGURE 39.1 Chemical structures of major hydroxycinnamic acids found in pear and pear juice.
TABLE 39.2
Concentration of the Major Pear Juice Phenolic Compounds (mg/L)
Phenolic Compound Pear Juice References
Hydroxycinnamic Acids
5-Caffeoylquinic acid 1.1–131 [26,27]
p-Coumaroylquinic acid 0.2–2.1 [26,27]
p-Coumaric acid 0.1–2.0 [26,27]
Caffeic acid 0.1–8.8 [26,27]
Arbutin 7.9–16.1 [26]
Flavonols
Quercetin and isorhamnetin glycosides 1.8–10.4 [26]
Proanthocyanidins
Catechin 1.5–3.1 [26]
Epicatechin 4.3–19.0 [26]
Total procyanidinsa 11.3–74.2 [26]
Procyanidin DP 25b Qualitative [29]
Monogalloylated procyanidins DP 25b Qualitative [29]
a Total procyanidins contains dimers (B1, B2, B3, and B4), trimers, tetramers, and other unknown procyanidins.
b Degree of polymerization up to 25.
Abbreviation: DP, degree of polymerization.
quercetin, kaempferol, and isorhamnetin, have been reported in pear [20,25,26]. Considering the pear
juice, 5-caffeoylquinic, caffeic, p-coumaroylquinic, and p-coumaric acids were reported as well as arbu-
tin and glycosides of quercetin and isorhamnetin (Table 39.2) [27,28]. The compound 5-caffeoylquinic
acid (Figure 39.1) was the most abundant hydroxycinnamic acid found in pear and pear juice [20,27,29].
In pome fruits such as pear [20,29] and pear juice [30], the proanthocyanidins are essentially pro-
cyanidins, referred also as condensed tannins, composed mainly of (–)-epicatechin units. In pear, the
procyanidins are the predominant class of phenolic compounds known as flavan-3-ols [31]. Pear procy-
anidins are oligomers or polymers, and their average size differs among varieties with an average degree
of polymerization of 6–24 [20].
Several studies dealing with the biological activities of proanthocyanidins have suggested that the anti-
oxidant, antifungal, and antitumor properties may be correlated to their degree of polymerization [32].
The structural elucidation of polymeric proanthocyanidins is difficult because of their heterogeneous
character. Proanthocyanidins are formed from flavan-3-ol monomers, which are linked through C4–C8
or C4–C6 linkages (Figure 39.2).
Pear Juice 479
OH
3΄
4΄
OH
2΄
HO 8
O 1΄ B
7 8a 2 5΄
A C H 6΄
6 R1
4a 3
5 4 R2
OH
R1 —H and R2— OH (+)-(2R,3S)-catechin

R1 —OH and R2 —H (–)-(2R,3R)-epicatechin
C OH
OH
B
HO O
A C
OH
4 OH
OH OH
HO 8 B
O
HO OH
6 A C
B OH
4 OH
HO OH
C OH
O OH B
A HO O
A C
C OH OH
OH
FIGURE 39.2 Chemical structures of flavan-3-ol monomers and substructure of procyanidin type B found in pear (show-
ing the phenolic rings A and B and the heterocyclic ring C).
In pear juice, two series of condensed tannin oligomers have been identified by matrix-assisted laser
desorption–ionization time-of-flight mass spectrometry analysis [30]. Supporting the observations from
nuclear magnetic resonance spectroscopy, the first series consist of tannins identified as procyanidin
polymeric units with chain lengths of up to 25. A second series of monogalloyl flavan-3-ols polymers
with a degree of polymerization of up to 25 were also detected. The polymeric fraction exhibited a potent
antioxidant power compared to that of (+)-catechin and B3 dimeric procyanidin, using 2,2-diphenyl-
1-picrylhydrazyl (DPPH) radical, suggesting that the increasing degree of polymerization enhances the
effectiveness of proanthocyanidins against radicals.
Very few studies about the antioxidant activity of pear juice are available. One way of evaluating the
antioxidant power of a system is to study its singlet oxygen quenching activity. Singlet oxygen is com-
monly formed by the excitation of triplet oxygen in the presence of a sensitizer and light [33]. The singlet
oxygen quenching activity has been determined in pear juice, and data were compared with those of other
juices prepared from vegetables and fruits commonly consumed in Korea [34]. Pear juice exhibited the
highest antioxidant activity among the juices tested in singlet oxygen-induced oxidation of a photosen-
sitizer compound, rubrene (5,6,11,12-tetraphenyltetracene), showing 59% inhibition. However, pear juice
showed low radical scavenging activities when compared with most of the juices tested by 2,2′-azino-
bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) method. There was poor correlation between the
singlet oxygen quenching activity and the ABTS radical scavenging activity of the juices (r 2 = 0.11).
Moreover, there was no relationship between the content of total phenolics in juices and singlet oxygen
quenching activities (r 2 = 0.02), suggesting that the phenolic compounds in pear juice are not the major
components governing the whole singlet oxygen quenching activities. No correlation existed between
total phenolics and DPPH-linked antioxidant activity for pear juice as demonstrated in another study [35].
The pear juice that showed the highest singlet oxygen quenching activities did not contain a large quantity
of total phenolics (187.1 μg gallic acid equivalents [GAE]/mL). There was also no relationship between
total phenolics in red cabbage and spinach juices and singlet oxygen quenching activities. Red cabbage
and spinach juices also showed strong singlet oxygen quenching activities (approximately 40% inhibi-
tion); however, the amount of total phenolic compounds was relatively high, 432 and 1805 μg /mL, respec-
tively. The phenolic content ranged from 165 to 1804 μg GAE/mL for all juices, the highest value being
for spinach. It seems that other antioxidant components of the pear, red cabbage, and spinach juices that
are not phenolic compounds may play a role in the singlet oxygen quenching activities. It has also been
demonstrated that the singlet oxygen quenching abilities of pear juice may explain the protective activities
of pear juice against both human erythrocyte lysis and protein oxidation induced by photodynamic means
[34]. It is expected that the protective effects of pear juice on singlet oxygen–induced biological damages
may contribute to their beneficial effects against singlet oxygen–involved pathogenesis.
To improve the contents of bioactive compounds and antioxidant activity of pear juice its fermentation
may be considered. Although the definition described earlier for juice leaves out the fermented juice, it
makes sense to discuss bioactive compounds and the antioxidant activity of fermented pear juice, since
new products based on pear juice can be featured, including fermented pear juice (discussed later in this
chapter). Pear juice fermented with Lactobacillus acidophilus has been investigated with respect to its
total phenolic compounds, antioxidant activity, key enzymes related to carbohydrate metabolism, and
control of blood pressure using in vitro assay models [35]. Before fermentation, for all pear juices of the
various cultivars (Anjou, Red Anjou, Bosc, and Comice), the DPPH-linked radical scavenging activity
was high (73%–80%) for all tested samples at 0 h, indicating that the whole pear juice has a good poten-
tial for free radical inhibition [35]. These results are in contrast to what was suggested for pear that was
classified in the group of low-antioxidant fruits [23]. For Red Anjou pear juice, despite an initial decrease,
there was a significant increase in total phenolics at 74 h of fermentation. The total soluble phenolics
present in a fermented pear substrate is due to the balance between the formation/degradation of large
polymeric phenolics and the formation/degradation of low-molecular-weight phenolic compounds [36].
The degradation of simple soluble monomeric phenolics could be a possible detoxification mechanism
for bacteria and yeast [36]. With fermentation, the DPPH-linked antioxidant activity increased until 48 h,
and then it decreased at 72 h [35]. In case of Anjou, a high inverse correlation (r = –0.81) existed between
total phenolics and DPPH-linked antioxidant activity. An inverse relationship has also been reported
during soy milk fermentation with lactic acid bacteria [36].
In order to improve the antioxidant bioefficacy of pear juice, microemulsions rich in bioactive com-
pounds have been used. The microemulsions were obtained by mixing different ratios of oil, surfactant,
and cosurfactant. Pear juice enriched with microemulsions containing plant extracts of oregano and wild
thyme showed high levels of antioxidant activity [37]. These results give evidence of the possibility of
using natural antioxidants instead of synthetic ones. The juice with oregano improved the antioxidant
capacity approximately 1.45 times compared to the juice with synthetic antioxidants such as butylated
hydroxyanisole and approximately 1.67 times compared to the juice with butylated hydroxytoluene.
This study demonstrates the potential use of microemulsions in enriching pear juice with antioxidant
compounds. The use of microemulsions can be a strategy to enhance the antioxidant capacity of pear
juice, considering, for example, the incorporation of antioxidant compounds derived from by-products
resulting from industrial transformation of pear.
One approach to enhance the quality and antioxidant efficacy of pear juice is to use cyclodextrins to
avoid juice browning. This browning is closely connected to the phenolic composition and monophe-
nol dihydroxyphenylalanine: oxygen oxidoreductases, EC 1.14.18.1 polyphenol oxidase (PPO) activity.
Cyclodextrins have been used in order to preserve the natural antioxidant capacity of pear juice [38], as
the enzymatic browning by PPO is the main process involved in pear juice browning at room tempera-
ture. Cyclodextrins were employed as substrate sequestering agents, and it has been demonstrated that
cyclodextrins are able to complex the PPO substrates, thereby preventing their oxidation to quinones and
subsequent polymerization to brown pigments. In this way, cyclodextrins can offer protection against
oxidation of ascorbic acid (AA) by o-quinones (o-Qs). In the absence of cyclodextrins, the total concen-
tration of PPO substrate is available to be oxidized by PPO in the presence of O2 to o-Qs. However, in the
Pear Juice 481
presence of cyclodextrins, the AA is protected due to the complexation of PPO substrates in the hydro-
phobic cavity of cyclodextrins. Cyclodextrins slow down the production of o-Qs and, hence, the oxida-
tion of AA, because only free substrates, in equilibrium with cyclodextrin-bound phenol, are oxidized
by PPO in the presence of O2 to o-Qs [39]. In this way, the reaction is slowed down and the antioxidant
efficacy of pear juice improved.
Another aspect influencing the bioefficacy of phenolic antioxidant compounds in pear juice relates to
possible interactions between these compounds and the cell wall polysaccharides. These interactions can
impact on dietary quality of pear juice. If phenolic compounds bound to the cell wall polysaccharides are
not available for absorption in the gastric or small intestinal environments, their interactions may be ben-
eficial in protecting and transporting polyphenol compounds to the large intestine where fermentation
of fibrous material occurs by gut bacteria releasing the phenolic compounds where they are important
[40,41]. Understanding the mechanisms behind these interactions could be useful in choosing the ideal
technological processing of producing pear juice in order to obtain a juice with potential benefits to the
human health.
In addition, evidence of the effect of food matrix and noncovalent interactions between procyanidin
and cell wall on the metabolism of procyanidins by human microflora has been found [42]. The conver-
sion rate of procyanidins in microbial metabolites was much lower with isolated procyanidins than with
whole fruit [42]. Through the studies reported in the literature for other fruits, it becomes obvious that
the study of the interactions between phenolic compounds of pear juice and cell wall polysaccharides is
of great importance for understanding the antioxidant efficacy of the juice.
39.4 Health Effects

The most important health effects related to the ingestion of pear juice are due to phenolic compounds,
triterpenes, and sugars. The health effects reported relate to the alleviation of severe hangover [43,44],
the decrease of total cholesterol and low-density lipoprotein (LDL) cholesterol in smokers [45], and the
increase of iron bioavailability from FeCl3 [46].
Reactive oxygen species (ROS), generated by alcohol consumption [47], may be considered as a
major mechanism of alcohol toxicity, developing the hangover symptoms. Korean pear (P. pyrifolia) has
been used as a traditional prophylactic agent for alcohol hangover. Comparing the pear of the species
P. pyrifolia with the one of P. communis with respect to its composition, the greater difference is the
higher content of total dietary fiber and potassium of the former, 3.1 versus 2.8 g and 171 versus 119 mg,
respectively, per 100 g of whole pear [47].
In order to understand the mechanism involved in alcohol toxicity, healthy young men were used to
examine the effects of Korean pear juice on alcohol hangover. Korean pear juice may alleviate not only
some hangover symptoms but also total hangover severity. The hangover severity was alleviated by
Korean pear juice at 15 h after the alcohol consumption, with 20% reduction in the total hangover sever-
ity. Particularly, “trouble concentrating” was significantly improved by the pear juice intake. Moreover,
levels of blood acetaldehyde were decreased by the pear juice ingestion [43]. However, the totality of the
antioxidant compounds present in pear juice was not responsible for the reduction in the ROS g enerated
by alcohol consumption. This health effect seems to be related only with some of the pear antioxidant
compounds that stimulate alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). The
water-soluble compounds from peel highly stimulate these enzymes when compared with other parts
of the pear. The water-soluble compounds such as arbutin and catechins are expected to be the main
functional compounds for alcohol detoxification. The ADH activity increased with the pear juice con-
centration [44]. The concentration of arbutin and catechins needed for increased activity of the ADH
should be 10 and 8 mg, respectively, considering the concentration in fresh pulp of 30 and 26 mg of
arbutin and catechins, respectively [20]. Thus, the active compounds of the Korean pear may alleviate
alcohol hangover severity via stimulation of alcohol metabolism rather than antioxidative mechanisms.
However, there are differences of alcohol metabolism based upon genetic variations of acetaldehyde
dehydrogenase-2 (ALDH2) [44]. The alcohol hangover alleviation symptoms, impaired memory and
sensitivity to light and sound, by pear juice drinking are genotype dependent. Deficient ALDH2
subjects were not alleviated significantly from alcohol hangover by Korean pear juice treatment, but
normal subjects were [43].
Pear juice containing pulp and skin can be compared with all the edible parts of the pear in order to
see if a supplemented diet with pear juice has comparable effects. Pear supplemented diet exhibited a
hypocholesterolemic effect in rats fed with cholesterol and positively influenced the plasma antioxidant
potential in rats fed with and without added cholesterol [48]. A supplemented diet with one pear, one
apple, and orange juice also induced a low level of plasma lipid in humans. In smokers, a decrease in
total and LDL cholesterols was observed, but without any increase in total plasma antioxidant capacity.
However, in nonsmokers an increase in total and LDL cholesterols as well as total plasma antioxidant
capacity was noted [45].
Another health effect of pear juice is related to the improvement of iron bioavailability from FeCl3 in
caco-2 cells. Results showed that caco-2 cells ferritin formation increased 10-fold after in vitro digestion
of pear juice. Among the fruit juices tested (apple, orange, grapefruit, and white grape), the maximum
increase in ferritin was due to pear juice. Moreover, pear juice also promoted iron uptake from infant
cereals, since iron bioavailability from the cereal is relatively low due to the presence of phytic acid and
phenolic compounds that act as inhibitors of the uptake [46].
Considering pear juice prepared from the whole fruit, the health effect of the pulp and peel compounds
may be taken into account after ingestion of the juice. Constituents of pear from the P. pyrifolia showed
inhibitory activity on the production of nitric oxide (NO), a proinflammatory mediator, and on the expres-
sion of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) in macrophages and microglia.
Microglia are a type of glia cells that act as the first and main form of active immune defense in the
central nervous system [49]. The triterpenes lupeol, betulin, betulinic acid, β-sitosterol, and daucosterol
(Figure 39.3) showed inhibition of NO production in macrophages and microglia cell lines [49]. However,
these compounds are mainly located in the peel, as the extracts of the pulp and core did not show a sig-
nificant anti-inflammatory effect on the cell lines. We can assume that if pear juice is prepared from whole
unpeeled fruit, it can show inhibitory activity on NO production and on the expression of iNOS and COX-2
in macrophages and microglia. Similar triterpenes were found in P. pyrifolia pear peel, namely, betu-
linic aldehyde, lupeol, betulinic acid, 3-O-cis-caffeoylbetulinic acid, 3-O-trans-caffeoylbetulinic acid, and
3-O-trans-caffeoyloleanolic acid, with antioxidant activity and the ability to inhibit cholesteryl ester hydro-
peroxide formation in copper ion‒induced lipid peroxidation in rat blood plasma [50]. These results allow
inferring that for a potential beneficial effect on health, the pear juice must somehow contain pulp and skin.
Certain types of sugars present in pear juice may also provide a beneficial effect on human health
due to the presence in pear juice of the FODMAP and in some pear cultivars to the presence of FOS.
The beneficial effects of FOS have been attributed to their malabsorption in the small intestine and
delivery of carbohydrates to the large bowel, where they undergo rapid fermentation by bacteria with
the subsequent expansion of bacterial populations, especially of probiotic bifidobacteria and lacto-
bacilli [51]. These bacteria seem to mediate a wide range of responses including suppressing the growth
HO R
O
R—
— CH3 Lupeol R—
—H β-Sitosterol
— CH2OH
R— Betulin —Glc
R— Daucosterol
R—
— CO2H Betulinic acid
— CHO
R— Betulinic aldehyde
FIGURE 39.3 Chemical structures of major triterpenoids found in pear.

Pear Juice 483
of putrefactive pathogens in the colon [51], alleviating diarrhea [52], increasing the absorption of
calcium [53], and alleviating allergic inflammation [54].
Despite the health benefits of FODMAP for adults, some disadvantages may exist for young children.
An incomplete carbohydrate absorption or carbohydrate malabsorption in infants after pear juice inges-
tion has been reported by several authors [10,55–57]. Most infants consume fruit juices by the age of
6 months. However, fruit juices containing sorbitol may be associated with carbohydrate malabsorption
without clinical symptoms. The consequences of incomplete absorption of carbohydrates were associ-
ated with an increase on physical activity and in metabolic rate [55]. The efficiency of carbohydrate
absorption in childhood increases with age, and decreased carbohydrate absorption occurs more fre-
quently with juices containing more fructose than glucose and/or sorbitol, such as apple or pear juice,
than with juices that contain equal amounts of fructose and glucose and are sorbitol free (grape juice),
in children from 6 months to 5 years of age [57]. Sorbitol not only is incompletely absorbed in the small
bowel but also interferes with fructose absorption [10]. Fructose absorption is facilitated by equimolar
amounts of glucose [58]. The fructose that escapes absorption in the small intestine is fermented in the
large intestine, explaining why pear juices are used to treat constipation in children [13]. The pear juice
administered in clinical studies was always in the form of commercial product preparations without any
information on the pear variety, but all juices had similar sugar composition [10,55]. The carbohydrate
malabsorption after ingestion of pear juice in infants could be minimized or even avoided if an amount
of juice lower than the amount usually tested was ingested (15–20 mL/kg). Pediatricians have recognized
that excessive consumption of fruit juice is associated with diarrhea, gassiness, stomachache, and other
abdominal complaints [56]. It was observed that when carbohydrate is well absorbed, it produces no
adverse gastrointestinal symptoms and has no effect on stool water in healthy infants when the pear juice
is administrated in a lower quantity such as 10 mL/kg/day [10].
The fresh pear pulp has a high content of total sugars and dietary fiber ranging between 12% and
15% DW. Therefore, whole pear juice may serve as a potential source of dietary fiber. Dietary fiber
is the part of plant material in the diet that is resistant to enzymatic digestion. It comprises cellu-
lose, noncellulosic polysaccharides, usually referred to as hemicelluloses and pectic polysaccharides.
Gums, mucilages, and lignin, a noncarbohydrate material, are also components of dietary fiber. Diets
rich in fiber have a positive effect on health since their consumption decreases the incidence of several
diseases [59]. A generous intake of dietary fiber may reduce the risk for developing coronary heart dis-
ease (CHD), stroke, hypertension, diabetes, obesity, and certain gastrointestinal disorders. Increased
consumption of dietary fiber improves serum lipid concentrations, lowers blood pressure, improves
blood glucose control in diabetes, promotes regularity, aids in weight loss, and appears to improve
immune function [59].
The contribution of beverages to the intake of soluble dietary fiber, considering the Spanish diet, is
25%, where fruit juices are responsible for 6% [60]. These dietary fibers contain pectic polysaccharides
rich in uronic acids and xylose-rich hemicellulosic polysaccharides (glucuronoxylans and xyloglucans).
Pear pulp is mainly composed of pectic polysaccharides (36%) and cellulose (31%), followed by hemicel-
lulose, which includes glucuronoxylans (27%), xyloglucans (4%), and mannans (2%) [61]. Pectic poly-
saccharides are a type of soluble fiber that binds to fatty substances in the digestive tract and promotes
their elimination, which seems to help in lowering blood cholesterol levels. Pectic polysaccharides from
other sources tested indicated that consumption of 12–24 g/day in divided amounts is associated with
a 13% reduction in LDL cholesterol levels [59]. Diets high in fiber may reduce the risk of CHD and
certain types of cancer [59,62]. Concerning breast carcinoma cells, the tumor growth, angiogenesis, and
spontaneous metastasis were reduced in mice fed with pectic polysaccharides that specifically inhibit
the carbohydrate-binding protein galectin-3 [62]. Moreover, meals containing pectic polysaccharides
resulted in delayed gastric emptying and enhanced satiety. Fibers are necessary components for a healthy
diet as they help to sustain blood sugar levels and control the intestinal transit by their action on fecal
volume [59]. Increased intake of dietary fiber is commonly used for the prevention and management of
constipation or hemorrhoids. The pectic polysaccharides increase fecal weight in 1.2 g, whereas cellulose
increases in 3.5 g of administered fiber [59]. Concerning the whole pear juice, its intake represents an
increase of the fecal weight in threefold the weight ingested in fiber.

Pear juice has been studied in order to incorporate probiotics in its formulation [35,63]. Fruit juices could
serve as an ideal medium for functional health ingredients because they inherently contain essential
nutrients such as vitamins, contain antioxidants and polyphenols, have good flavor profiles, and attract
all age groups as they are perceived healthy and refreshing. Pear juice with probiotics could be one
good alternative to probiotics in dairy products, overcoming drawbacks related to lactose intolerance,
cholesterol content, and allergenic milk proteins. L. acidophilus in fermented pear juice produced a new
product with antihyperglycemia and antihypertensive activities. Sensory analysis of the fermented pear
juice had a sour taste with a typically fermented flavor but with a pleasant smell. The pH of natural pear
juice and the final fermented juice is not different; however, the lactic acid produced during fermentation
changes the flavor profile. It had a slight tangy but identifiable pear-like taste, maintaining the same con-
sistency. The juice also became cloudy because of the growth of the probiotic bacterium. Although the
taste of the fermented juice was not the same as the unfermented, it was an acceptable new product for
consumption considering the beneficial health properties enhanced by the presence of viable probiotic
bacterium. Probiotic dairy-free drinks are available on the market with several flavor blends, including
pear juice concentrate. This type of products extends the possibilities of a novel acceptable source of
lactic acid bacteria for lactose-intolerant individuals [35]. There are several factors such as pH, storage
temperature, oxygen levels, and water activity that create a great challenge of incorporating probiotics in
fruit juices. Bacteria may need protection against acidic medium to maintain high viability. The use of
encapsulated L. rhamnosus GG and L. acidophilus NCFM was tested in some fruit food products [63].
However, encapsulation of these two bacteria did not significantly enhance survivability but decreased
the pH of the fruit food products from 3.77 to 3.12. For pear fruit‒based foods, the encapsulation of
L. rhamnosus GG reduced acidification at 25°C; however, at 4°C, the difference in pH between the free
and encapsulated cells was not significant [63].
New formulations of pear juice have been proposed to reduce enzymatic browning and to improve
antioxidant capacity during storage [38,64]. The use of cyclodextrins as secondary antioxidants was
proposed to improve the color of fresh pear juice, because cyclodextrins can act as browning inhibi-
tors. Maltosyl-β-cyclodextrins enhance the ability of ascorbic acid to prevent enzymatic browning
because of the protection offered against ascorbic acid oxidation. Cyclodextrins are widely used as
browning inhibitors in different fruit juices. However, the effect of addition of α- and β-cyclodextrins
on the sensory quality, volatile compounds, and color parameters of fresh pear juice must be mini-
mal. The addition of α-cyclodextrin at 15 mM led to a pear juice with an acceptable color and at
same time with a high intensity of fruity and pear-like odors, making it the best appreciated juice
by sensory analysis [64]. Comparing the effect on volatile compounds of pear juice by adding α-, β-,
and γ-cyclodextrins at a concentration of 15 mM can be seen that the content of volatile components
decreases in all cases, although being less pronounced for α-cyclodextrin [65]. These three cyclodex-
trins are able to form inclusion complexes with the main volatiles present in pear juice. However, an
increase in the inner diameter of cyclodextrin produced a higher retention of the volatile compounds
present in the pear juice [65]. Concerning global analysis of pear juices, the best results were obtained
for α-cyclodextrin, followed by β-cyclodextrin and γ-cyclodextrin juices. The sensory analysis of
samples treated with 15 mM of α-cyclodextrin had the highest scores of global quality (6.6), followed
by β-cyclodextrin and control samples (5.9), and finally γ-cyclodextrin samples (4.3). The addition of
α-cyclodextrin to pear juice may significantly increase the global quality and reduce browning without
producing significant decreases in aroma [65]. Nevertheless, it can economically increase the price by
0.2–0.6 €/L of pear juice.
Novel trends in pear juice industry are related to the enhancement of clarification and stability of the
juice [66,67]. Microfiltration (MF) and ultrafiltration (UF) have been widely used for clarification of fruit
juices such as pear and apple [68]. However, MF and UF juices are susceptible to postbottling haze due to
incomplete elimination of all potential haze precursors, products of polyphenol, and protein interactions.
This suggests that MF and UF, due to large pore size, are unable to retain all constituents and colored
compounds potentially responsible for imparting haze and brown color formation. The use of smaller
Pear Juice 485
pore size or nanofiltration (NF) membranes may overcome this problem [67]. In order to differentiate
the membranes manufactured by casting in a single layer, having higher tolerance to chlorine, the term
“loose nanofiltration” has been used in order to reflect a separate class NF membranes. Thus, loose NF
was preferably used to enhance the stability and shelf life of pear juices compared with UF-treated
juices. Loose NF juices compared to the control (UF juices) showed only a minor increase in the absor-
bance values at 425 nm when tested for stability at 80°C. The loose NF-treated juice showed a significant
increase in lightness (L*), improvement from brownish tinge to clear light greenish tinge (a*), and an
improvement in overall color difference (ΔEab) values. Additionally, a significant reduction occurred
in the concentration of polyphenolic compounds of loose NF compared to the control (UF). Moreover,
loose NF was able to retain most of the polyphenolic and colored organic compounds, allowing produc-
tion of a very-light-colored pear juice with improved quality [67]. Quality enhancement of UF-clarified
pear juice was tested using adsorbent and weak base resins. Treated juice showed 85% reduction in both
color and titratable acidity and also showed a significant reduction in the content of polyphenolic com-
pounds and organic acids [66].
39.6 Conclusion
Nutritionally, pear juice contains a high amount of total sugars combined with a low content of protein
and fat. It is also a good source of vitamin C and potassium. Pear juice contains compounds with antioxi-
dant properties that have been investigated by studying the DPPH and ABTS radical scavenging activi-
ties and singlet oxygen quenching activity. Phenolic compounds may explain, in part, the antioxidant
activity of pear juice. Innovative products originating from pear juice, such as fermented pear juice, can
provide a good approach to enhance the antioxidant potential. Other strategies have been employed to
improve the antioxidant efficacy of pear juice such as the use of cyclodextrins incorporated in the juice
and application of microemulsions.
Beneficial health effects can potentially be derived from the consumption of pear juice, and some stud-
ies provide evidence for the alleviation of sever hangover, the decrease of total and LDL-cholesterols in
smokers, and the increase of iron bioavailability. Other potential health effect of pear juice ingestion is
related with the presence of FODMAP that may be responsible for stimulating the growth of bifidobac-
teria in the human colon.
Novel products have been studied such as fermented pear juice with L. acidophilus, and experimental
data demonstrated that the resulting juice had antihyperglycemia and antihypertensive activities. With
respect to the pear juice industry, the current trends show interest in the improvement of processes of
clarification and filtration. The use of nanofiltration has been proposed with positive effect on the stabil-
ity of pear juice.
REFERENCES
1. Deckers, T. and Schoofs, H., Status of the pear production in Europe, in Proceedings of the 10th
International Pear Symposium, Vols. 1 and 2, Webster, A.D. and Oliveira, C.M., Eds., ISHS Acta
Horticulturae 800, Acta Horticulture, Leuven, Belgium, 2008, pp. 95–105.
2. U.S. Department of Agriculture, Pyrus Crop Germplasm Committee: Report and Genetic Vulnerability
Statement, September 2004, Germ Resources Information Network (GRIN), USDA, Washington, DC, 2004.
3. U.S. Department of Agriculture (USDA), Good Prospects for EU-27 Apple and Pear Production,
Foreign Agricultural Service, USDA, Washington, DC, 2011.
4. U.S. Department of Agriculture (USDA), Fresh Deciduous Fruit (Apples, Grapes, & Pears): World
Markets and Trade, Foreign Agricultural Service, Office of Global Analysis, Washington, DC, 2013.
5. European Parliament and Council, Amending Council Directive 2001/112/EC Relating to Fruit Juices
and Certain Similar Products Intended for Human Consumption, Directive 2012/12/EU, Brussels,
Belgium, 2012.
6. AIJN, Association of the Industry of Juices and Nectars from Fruits and Vegetables of the European
Union, 2012 Market Report, European Fruit Juice Association, Brussels, Belgium, 2012.
7. Bates, R.P., Morris, J.R., and Crandall, P.G., FAO Agricultural Services, Principles and Practices of
Small and Medium Scale Fruit Juice Processing, Bulletin 146, Rome, Italy, 2001.
8. Kevers, C., Pincemail, J., Tabart, J., Defraigne, J.-O., and Dommes, J., Influence of cultivar, harvest
time, storage conditions, and peeling on the antioxidant capacity and phenolic and ascorbic acid con-
tents of apples and pears. J. Agric. Food Chem., 59, 6165–6171, 2011.
26, 2013. Published online at: http://ndb.nal.usda.gov/ndb/search/list (September 9, 2014).
10. Lifschitz, C.H., Carbohydrate absorption from fruit juices in infants. Pediatrics, 105, e4, 2000.
11. Gibson, P.R. and Shepherd, S.J., Evidence-based dietary management of functional gastrointestinal
symptoms: The FODMAP approach. J. Gastroenterol. Hepatol., 25, 252–258, 2010.
12. Campbell, J.M., Bauer, L.L., Fahey, G.C., Hogarth, A., Wolf, B.W., and Hunter, D.E., Selected fructooli-
gosaccharide (1-kestose, nystose, and 1(F)-β-fructofuranosylnystose) composition of foods and feeds.
J. Agric. Food Chem., 45, 3076–3082, 1997.
13. Slavin, J.L. and Lloyd, B., Health benefits of fruits and vegetables. Adv. Nutr., 3, 506–516, 2012.
14. Barda, N., Nutritional and compositional characterization of Argentinean ‘Williams Bon Chretien’
pears. Acta Hortic., 909, 587–594, 2011.
15. Van Gorsel, H., Li, C., Kerbel, E.L., Smits, M., and Kader, A.A., Compositional characterization of
prune juice. J. Agric. Food Chem., 40, 784–789, 1992.
16. Biesalski, H.-K., Dragsted, L.O., Elmadfa, I., Grossklaus, R., Müller, M., Schrenk, D., Walter, P.,
and Weber, P., Bioactive compounds: Definition and assessment of activity. Nutrition, 25, 1202–1205,
2009.
17. Clere, N., Faure, S., Martínez, M.C., and Andriantsitohaina, R., Anticancer properties of flavonoids:
Roles in various stages of carcinogenesis. Cardiovasc. Hematol. Agents Med. Chem., 9, 62–77, 2011.
18. Visioli, F. and Davalos, A., Polyphenols and cardiovascular disease: A critical summary of the evidence.
Mini-Rev. Med. Chem., 11, 1186–1190, 2011.
19. Rein, M.J., Renouf, M., Cruz-Hernandez, C., Actis-Goretta, L., Thakkar, S.K., and Pinto, M.D.S.,
Bioavailability of bioactive food compounds: A challenging journey to bioefficacy. Br. J. Clin.
Pharmacol., 75, 588–602, 2013.
20. Ferreira, D., Guyot, S., Marnet, N., Delgadillo, I., Renard, C., and Coimbra, M.A., Composition of
phenolic compounds in a Portuguese pear (Pyrus communis L. var. S. Bartolomeu) and changes after
sun-drying. J. Agric. Food Chem., 50, 4537–4544, 2002.
21. Landbo, A.-K. and Meyer, A.S., Effects of different enzymatic maceration treatments on enhance-
ment of anthocyanins and other phenolics in black currant juice. Innov. Food Sci. Emerg. Technol., 5,
503–513, 2004.
22. Revilla, I. and González-San José, M.L., Compositional changes during the storage of red, wines treated
with pectolytic enzymes: Low molecular-weight phenols and flavan-3-ol derivative levels. Food Chem.,
80, 205–214, 2003.
23. Prior, R.L. and Cao, G.H., Antioxidant phytochemicals in fruits and vegetables: Diet and health implica-
tions. HortScience, 35, 588–592, 2000.
24. Sánchez, A.C.G., Gil-Izquierdo, A., and Gil, M.I., Comparative study of six pear cultivars in terms of
their phenolic and vitamin C contents and antioxidant capacity. J. Sci. Food Agric., 83, 995–1003, 2003.
25. Lin, L.-Z. and Harnly, J.N., Phenolic compounds and chromatographic profiles of pear skins (Pyrus
spp.). J. Agric. Food Chem., 56, 9094–9101, 2008.
26. Schieber, A., Keller, P., and Carle, R., Determination of phenolic acids and flavonoids of apple and pear
by high-performance liquid chromatography. J. Chromatogr. A, 910, 265–273, 2001.
27. Spanos, G.A. and Wrolstad, R.E., Influence of variety, maturity, processing, and storage on the phenolic
composition of pear juice. J. Agric. Food Chem., 38, 817–824, 1990.
28. De Simón, B.F., Pérez-Ilzarbe, J., Hernández, T., Gómez-Cordovés, C., and Estrella, I., Importance of
phenolic compounds for the characterization of fruit juices. J. Agric. Food Chem., 40, 1531–1535, 1992.
29. Spanos, G.A. and Wrolstad, R.E., Phenolics of apple, pear, and white grape juices and their changes
with processing and storage—A review. J. Agric. Food Chem., 40, 1478–1487, 1992.
30. Es-Safi, N.-E., Guyot, S., and Ducrot, P.-H., NMR, ESI/MS, and MALDI-TOF/MS analysis of pear juice
polymeric proanthocyanidins with potent free radical scavenging activity. J. Agric. Food Chem., 54,
6969–6977, 2006.
Pear Juice 487
31. Le Bourvellec, C., Gouble, B., Bureau, S., Loonis, M., Plé, Y., and Renard, C.M.G.C., Pink discoloration
of canned pears: Role of procyanidin chemical depolymerization and procyanidin/cell wall interactions.
J. Agric. Food Chem., 61, 6679–6692, 2013.
32. Santos-Buelga, C. and Scalbert, A., Proanthocyanidins and tannin-like compounds—Nature, occur-
rence, dietary intake and effects on nutrition and health. J. Sci. Food Agric., 80, 1094–1117, 2000.
33. Choe, E. and Min, D.B., Chemistry and reactions of reactive oxygen species in foods. Crit. Rev. Food
Sci. Nutr., 46, 1–22, 2006.
34. Oh, Y.S., Jang, E.S., Bock, J.Y., Yoon, S.H., and Jung, M.Y., Singlet oxygen quenching activities of vari-
ous fruit and vegetable juices and protective effects of apple and pear juices against hematolysis and
protein oxidation induced by methylene blue photosensitization. J. Food Sci., 71, C26–C268, 2006.
35. Ankolekar, C., Pinto, M., Greene, D., and Shetty, K., In vitro bioassay based screening of antihypergly-
cemia and antihypertensive activities of Lactobacillus acidophilus fermented pear juice. Innov. Food
Sci. Emerg. Technol., 13, 221–230, 2012.
36. McCue, P.P. and Shetty, K., Phenolic antioxidant mobilization during yogurt production from soymilk
using Kefir cultures. Process Biochem., 40, 1791–1797, 2005.
37. Miron, T.L. and Dima, S., Enriched antioxidant activity of pear juice by supplementation with oregano
and wild thyme extracts. Ann. Univ. Dunarea de Jos of Galati, Fascicle VI: Food Technol., 36, 81–91,
2012.
38. López-Nicolás, J.M. and García-Carmona, F., Use of cyclodextrins as secondary antioxidants to improve
the color of fresh pear juice. J. Agric. Food Chem., 55, 6330–6338, 2007.
39. López-Nicolás, J.M., Núñez-Delicado, E., Sánchez-Ferrer, A., and García-Carmona, F., Kinetic model
of apple juice enzymatic browning in the presence of cyclodextrins: The use of maltosyl-β-cyclodextrin
as secondary antioxidant. Food Chem., 101, 1164–1171, 2007.
40. Bindon, K.A., Smith, P.A., and Kennedy, J.A., Interaction between grape-derived proanthocyanidins
and cell wall material. 1. Effect on proanthocyanidin composition and molecular mass. J. Agric. Food
Chem., 58, 2520–2528, 2010.
41. Padayachee, A., Netzel, G., Netzel, M., Day, L., Zabaras, D., Mikkelsen, D., and Gidley, M.J., Binding
of polyphenols to plant cell wall analogues—Part 2: Phenolic acids. Food Chem., 135, 2287–2292, 2012.
42. Bazzocco, S., Mattila, I., Guyot, S., Renard, C.G.C., and Aura, A.-M., Factors affecting the conversion
of apple polyphenols to phenolic acids and fruit matrix to short-chain fatty acids by human faecal micro-
biota in vitro. Eur. J. Nutr., 47, 442–452, 2008.
43. Lee, H.-S., Isse, T., Kawamoto, T., Baik, H.W., Park, J.Y., and Yang, M., Effect of Korean pear
(Pyruspyrifolia cv. Shingo) juice on hangover severity following alcohol consumption. Food Chem.
Toxicol., 58, 101–106, 2013.
44. Lee, H.-S., Isse, T., Kawamoto, T., Woo, H.-S., Kim, A.K., Park, J.Y., and Yang, M., Effects and action
mechanisms of Korean pear (Pyrus pyrifolia cv. Shingo) on alcohol detoxification. Phytother. Res., 26,
1753–1758, 2012.
45. Alvarez-Parrilla, E., De La Rosa, L.A., Legarreta, P., Saenz, L., Rodrigo-García, J., and González-
Aguilar, G.A., Daily consumption of apple, pear and orange juice differently affects plasma lipids and
antioxidant capacity of smoking and non-smoking adults. Int. J. Food Sci. Nutr., 61, 369–380, 2010.
46. Boato, F., Wortley, G.M., Liu, R.H., and Glahn, R.P., Red grape juice inhibits iron availability: applica-
tion of an in vitro digestion/Caco-2 cell model. J. Agric. Food Chem., 50, 6935–6938, 2002.
47. Del Rio, D., Stewart, A.J., and Pellegrini, N., A review of recent studies on malondialdehyde as toxic
molecule and biological marker of oxidative stress. Nutr. Metab. Cardiovasc. Dis., 15, 316–328, 2005.
48. Leontowicz, H., Gorinstein, S., Lojek, A., Leontowicz, M., Čı ž́ , M., Soliva-Fortuny, R., Park, Y.-S.,
Jung, S.-T., Trakhtenberg, S., and Martin-Belloso, O., Comparative content of some bioactive com-
pounds in apples, peaches and pears and their influence on lipids and antioxidant capacity in rats.
J. Nutr. Biochem., 13, 603–610, 2002.
49. Yoo, J.H. and Yang, K.S., Constituents of Pyrus pyrifolia with inhibitory activity on the NO production
and the expression of iNOS and COX-2 in macrophages and microglia. Nat. Prod. Sci., 18, 183–189,
2012.
50. Cho, J.-Y., Kim, C.M., Lee, H.J., Lee, S.-H., Cho, J.-A., Kim, W.-S., Park, K.-H., and Moon, J.-H.,
Caffeoyl triterpenes from pear (Pyrus pyrifolia Nakai) fruit peels and their antioxidative activities
against oxidation of rat blood plasma. J. Agric. Food Chem., 61, 4563–4569, 2013.
51. Gibson, G.R., McCartney, A.L., and Rastall, R.A., Prebiotics and resistance to gastrointestinal infec-
tions. Br. J. Nutr., 93(Suppl.), S31–S34, 2005.
52. Szajewska, H., Ruszczyński, M., and Radzikowski, A., Probiotics in the prevention of antibiotic-
associated diarrhea in children: Ameta-analysis of randomized controlled trials. J. Pediatr., 149,
367–372.e1, 2006.
53. Abrams, S.A., Griffin, I.J., Hawthorne, K.M., Liang, L., Gunn, S.K., Darlington, G., and Ellis, K.J.,
A combination of prebiotic short- and long-chain inulin-type fructans enhances calcium absorption and
bone mineralization in young adolescents. Am. J. Clin. Nutr., 82, 471–476, 2005.
54. Kirjavainen, P.V., Arvola, T., Salminen, S.J., and Isolauri, E., Aberrant composition of gut microbiota of
allergic infants: A target of bifidobacterial therapy at weaning? Gut, 51, 51–55, 2002.
55. Cole, C.R., Rising, R., and Lifshitz, F., Consequences of incomplete carbohydrate absorption from fruit
juice consumption in infants. Arch. Pediatr. Adolesc. Med., 153, 1098–1102, 1999.
56. Klish, W.J., Use of fruit juice in the diets of young children. Pediatrics, 95, 433–433, 1995.
57. Nobigrot, T., Chasalow, F.I., and Lifshitz, F., Carbohydrate absorption from one serving of fruit juice in
young children: Age and carbohydrate composition effects. J. Am. Coll. Nutr., 16, 152–158, 1997.
58. Truswell, A.S., Seach, J.M., and Thorburn, A.W., Incomplete absorption of pure fructose in healthy-
subjects and the facilitating effect of glucose. Am. J. Clin. Nutr., 48, 1424–1430, 1988.
59. Anderson, J.W., Baird, P., Davis Jr, R.H., Ferreri, S., Knudtson, M., Koraym, A., Waters, V., and
Williams, C.L., Health benefits of dietary fiber. Nutrition Rev., 67, 188–205, 2009.
60. Diaz-Rubio, M.E. and Saura-Calixto, F., Beverages have an appreciable contribution to the intake of
soluble dietary fibre: A study in the Spanish diet. Int. J. Food Sci. Nutr., 62, 715–718, 2011.
61. Ferreira, D.S.M., Estudo das Transformações Bioquímicas e Químicas da Pêra de S. Bartolomeu
Durante o Processo de Secagem—Recurso Endógeno da Região de Viseu, PhD thesis, University of
Aveiro, Aveiro, Portugal, 2003.
62. Nangia-Makker, P., Hogan, V., Honjo, Y., Baccarini, S., Tait, L., Bresalier, R., and Raz, A., Inhibition of
human cancer cell growth and metastasis in nude mice by oral intake of modified citrus pectin. J. Natl.
Cancer Inst., 94, 1854–1862, 2002.
63. Sohail, A., Turner, M.S., Prabawati, E.K., Coombes, A.G.A., and Bhandari, B., Evaluation of
Lactobacillus rhamnosus GG and Lactobacillus acidophilus NCFM encapsulated using a novel imping-
ing aerosol method in fruit food products. Int. J. Food Microbiol., 157, 162–166, 2012.
64. Andreu-Sevilla, A.J., Carbonell-Barrachina, A.A., López-Nicolás, J.M., and García-Carmona, F.,
Sensory quality, volatile compounds and color of pear juice treated with β-cyclodextrin. J. Incl. Phenom.
Macrocycl. Chem., 70, 453–460, 2011.
65. Andreu-Sevilla, A.J., López-Nicolás, J.M., Carbonell-Barrachina, A.A., and García-Carmona, F.,
Comparative effect of the addition of α-, β-, or γ-cyclodextrin on main sensory and physico-chemical
parameters. J. Food Sci., 76, S347–S353, 2011.
66. Vivekanand, Ajlouni, S. and Iyer, M., Quality enhancement of UF-clarified pear juice using adsorbent
and weak-base resins at different temperatures. J. Food Sci., 68, 333–338, 2003.
67. Vivekanand, V., Iyer, M., and Ajlouni, S., Clarification and stability enhancement of pear juice using
loose nanofiltration. J. Food Process. Technol., 3, 162–168, 2012.
68. Girard, B. and Fukumoto, L.R., Membrane processing of fruit juices and beverages: A review. Crit. Rev.
Biotechnol., 20, 109–175, 2000.
40
Pineapple Juice
Nauman Khalid, Hafiz Ansar Rasul Suleria, and Iftikhar Ahmed
CONTENTS
40.1 Introduction................................................................................................................................... 489
40.4 Health Effects................................................................................................................................ 493
40.6 Conclusion..................................................................................................................................... 497
References............................................................................................................................................... 498
40.1 Introduction
The pineapple tree belongs to the Bromeliaceae family. Its fruit can be processed into several prod-
ucts such as canned pineapple slices, pineapple pulp, dried pineapple, pasteurized pineapple juice,
and concentrate. Fresh pineapple juice is a popular product due to its pleasant aroma, flavor, and
numerous functional properties [1]. Pineapple juice satisfies the “5 A Day” dietary requirement of
fruits and vegetables set by many health agencies [2]. Pineapple juice contains a variety of minerals,
especially manganese, as well as amino acids, various sugars, vitamins, and polyphenols [3]. It is
considered as a functional drink due to its health-promoting properties and has anti-inflammatory,
antiatherosclerotic, antiaging, and many other healing properties, which are briefly described in this
chapter.
The pineapple juice market has increased fourfold worldwide since 1984 from 1.3 to 5.6 million tons
(fresh fruit equivalent) [4]. The United States and the European Union account for 90% of the global
market for pineapple juice and concentrate, while Russia, Japan, and Middle Eastern countries account
for 6% of the market share [5].
There are different types of pineapple juice available on the market. Some single-strength juice
is obtained from pineapple parts that are squeezed with the help of mills and screw presses. Other
types include juice from concentrate, blended juice with other fruits, clear juice, and many others.
Approximately, 10%–25% of pineapple juice is obtained from canning industry, which is not suit-
able for the production of single-strength or concentrate juice, due to its high acidity [6]. The acidity
is neutralized by adding sweetening agents or by employing a variety of processing techniques [6].
Figure 40.1 shows the simplified process flowchart for pineapple juice production. The pineapple juice
is processed by many advanced processing techniques to reduce bacterial contamination with improved
shelf life and preservation of antioxidant compounds, vitamins, and minerals. Pasteurization, ultrafil-
tration, high-pressure homogenization, ultraviolet irradiation, reverse osmosis, freeze drying, and many
other techniques are used to improve the quality of pineapple juice [1,7–10]. This chapter provides
insight into pineapple juice composition, phytochemical profile, potential health benefits, and future
perspectives of this industry.
489
Reception of pineapple
Sorting
Removing the crown and leaves
Washing
Washing with sterilized water
Peeling
Juice extraction Cutting
Juice pasteurization
Juice filtration Fresh juice is boiled at 80°C for
30 s
Diluted juice: Fresh boiled juice is added Juice syrups: Fresh boiled juice is added
Boiled mixture of sugar and water up to the cooling Boiled mixture of sugar and water up to the cooling
peak of 30°C (1 L of boiled fresh juice + 1.05 kg peak of 30°C (1 L of boiled fresh juice + 1.05 kg
sugar + 7 L of water + 26 g citric acid + 2.5 g sugar + 7 L of water + 26 g citric acid + 2.5 g
sodium benzoate) sodium benzoate)
Cooling
Juice is cooled
up to 30°C
FIGURE 40.1 Flowchart of pineapple juice preparation at small-scale level.

The composition of pineapple juice varies with geography, culture, harvest season, and processing time.
Only a few studies have reported the general composition of pineapple juice such as minerals, sugars, organic
acids, and amino acids, as well as physical properties such as pH, Brix, ash, and titratable acidity [3,11–13].
The typical composition (Table 40.1) of canned pineapple juice (100 mL) includes 0.36 g protein, 0.12 g
lipid, 12.87 g carbohydrate, and 0.1 g dietary fiber [14,15]. The major minerals in fresh pineapple juice are
Pineapple Juice 491
TABLE 40.1
Compositional and Nutritional Characteristics of Raw Pineapple and Pineapple Juices (per 100 g)
Pineapple Juice Pineapple Juice
Pineapple Pineapple (Undiluted) Frozen (3× Dilution) Frozen
Unit Raw Canned Juice Concentrated Concentrated
Water g 86.0 86.37 53.1 86.6
Energy kcal 50.0 53.0 179.0 51.0
Protein g 0.54 0.36 1.3 0.4
Lipid (fat) g 0.12 0.12 0.1 0.03
Carbohydrate g 13.12 12.87 44.3 12.67
Total sugars g 9.85 9.98 43.60 12.47
Ash g 0.29 0.3 1.2 0.3
Minerals
Calcium mg 13.0 13.0 39.0 13
Copper mg 0.11 0.09 0.313 0.09
Iron mg 0.29 0.31 0.9 0.16
Magnesium mg 12.0 12.0 35.0 14.0
Manganese mg 1.649 0.99 3.439 0.99
Phosphorus mg 8.0 8.0 28.0 9.0
Potassium mg 109.0 130.0 472.0 132.0
Sodium mg 1.0 2.0 3.0 1.0
Zinc mg 0.12 0.11 0.4 0.08
Vitamins
Ascorbic acid mg 47.8 10.0 42.0 12.0
Folate (DFE) μg 18.0 18.0 37.0 11.0
Niacin mg 0.50 0.199 0.90 0.20
Vitamin B6 mg 0.112 0.100 0.255 0.074
Riboflavin mg 0.032 0.021 0.06 0.02
Thiamin mg 0.079 0.055 0.23 0.07
Vitamin A IU 58.0 5.0 50.0 10.0
Vitamin E (ATE) mg 0.02 0.0 0.03 0.01
Thiamin mg 0.087 0.058 0.255 0.074
Vitamin K μg 0.7 0.3 1.0 0.3
(phylloquinone)
Sources: Adapted from the U.S. Department of Agriculture (USDA), National Nutrient Database for Standard
Reference, Release 26, 2013, Published online at: http://ndb.nal.usda.gov/ (assessed April 07, 2014),
except ash, copper, and manganese, which were adapted from Gebhardt, S.E. et al., Composition of
foods—Fruits and fruit juices, in Agriculture Handbook No 8-9, Consumer Nutrition Center—Human
Nutrition Information Service, USDA, Washington, DC, 1982, pp. 283.
Abbreviations: DFE, dietary folate equivalents; ATE, alpha tocopherol equivalents; IU, international unit.
potassium (124–130 mg/100 mL), magnesium (12–15.4 mg/100 mL), phosphorus (3.1–8.0 mg/100 mL),
iron (0.2–0.31 mg/100 mL), and manganese (0.3–0.99 mg/100 mL) [3,15,16]. Pineapple juice is rich in
vitamin C (ascorbic acid), an effective antioxidant [17]. The vitamin C content of fresh pineapple juice
is reported to range from 9.2 to 93.8 mg/100 mL [18,19]. Cárnara et al. [19] found the vitamin C content
of fresh pineapple juice to be 84.2 mg/100 mL, while that for commercial pineapple juice made from
concentrate varied between 12 and 42 mg/100 mL [14,15]. A 100 g of pineapple juice and concentrate
contains 5 and 50 IU of vitamin A, respectively [14,15].
Major free amino acids identified in the pineapple juice include asparagine, proline, aspartic acid, ser-
ine, glutamic acid, tyrosine, valine, and isoleucine [20]. The juice contains ample amounts of asparagine,
serine, threonine, and glycine [3]. The composition of free amino acids is significantly higher in pine-
apple juice in comparison with grape juice and apple juice [21].
Pineapple juice also contains sucrose (4.1 g/100 mL), followed by fructose (2.5 g/100 mL), and glu-
cose (2.3 g/100 mL) [22]. The amino acids and sugars have an important impact on Maillard reaction
that in turn produces hydroxymethylfurfural at pH below 7 and in the range of 0.1–22.0 mg/100 mL of
fresh pineapple juice [23]. The Millard reaction is considered to be an important quality parameter in
pineapple juice processing.

Pineapple juice is a major commercial product, but its phenolic profile has not been well charac-
terized, and limited research data are available on this subject. Only nonflavonoid phenolics were
reported for pineapple fruit [24], juice [25], and shell fibers [26], with the exception of myricetin in
fiber phenolics.
The pineapple juice contains phenolic acids such as p-coumaric acid, caffeic acid, ferulic acid, sinapic
acid, p-coumaroylquinic acid, feruloyl glucose, p-hydroxybenzoic acid, p-hydroxybenzaldehyde, and
syringic acid. The phenolic composition of pineapple juice analyzed by high-performance liquid
chromatography (HPLC) showed nine major peaks, representing tyrosine, serotonin, dimethylhy-
droxylfuranone, dimethylhydroxylfuranone-β-glucoside, tryptophan, S-sinapyl-l-cysteine, N-γ-l-
glutamyl-S-sinapyl-l-cysteine, S-sinapyl glutathione, and p-coumaric acid (Figure 40.2) [27]. The
detail compositions of pineapple phenolics are presented in Table 40.2. The other minor nonflavonoid
compounds include 4-O-caffeoylquinic acid [28], p-hydroxybenzoic acid, p-hydroxybenzaldehyde,
ferulic acid, caffeic acid, and p-coumaroylquinic acid [25]; no flavonoid-type phenolic compounds
have been reported in pineapple juice [25]. In the aqueous system, pineapple juice sinapyl derivatives
(Figure 40.2) have antioxidant activity of around 1.22–1.56 trolox equivalents (TE)/100 mL [27]. The
antioxidant activity is possibly caused by its S-allyl-l-cysteine, which is the major antioxidant in
numerous fruits and vegetables [29].
The major carotenoid mixture of pineapple juice constitutes violaxanthin (50%), leuteoxanthin (8%),
β-carotene (9%), and neoxanthin (8%) [30]. Less abundant carotenoids include ζ-carotene, hydroxyl
α-carotene, cryptoxanthins, lutein, auroxanthin, and neochrome [30]. The total carotenoid concentra-
tion is proportional to the degree of yellow color in pineapple flesh. Carotenoid content is higher in
the flesh than in the juice of Del Monte Hawaii Gold variety (1.36 in flesh and 0.25 µg/g in juice) and
Smooth Cayenne (0.45 in flesh and 0.07 µg/g in juice) [31]. Pineapple also contains alkaloid 6-hydroxy-
1-methyl-1,2,3,4-tetrahydro-α-carboline (0.62 in fruit and 1.69 mg/g in juice), which acts as an anti-
oxidant [32].
Gardner et al. [33] investigated the relationship between composition and antioxidant activity of sev-
eral juices such as orange, apple, pineapple, grapefruit, and vegetables. They found that the antioxidant
activity of citrus juices was mainly (>66%) attributed to ascorbic acid, whereas ascorbic acid contrib-
uted less than 5% to the other products and 0.8% to pineapple juice. Most of the antioxidant activity is
believed to be due to phenolic compounds in pineapple juice. In addition to the phenolic compounds,
pineapple juice contains both l-ascorbic acid and dehydroascorbic acid.
Laorko et al. [10] studied the effect of membrane filtration on total phenolic contents and
antioxidant capacity of pineapple juice. The highest total phenolic content (69.34 mg gallic acid
equivalents [GAE]/100 mL) and free radical scavenging capacity (25.76 mg ascorbic acid equivalents
(AAE)/100 mL) were obtained with 0.2 µm membrane, and the antioxidant activity decreased with the
pore size. Pineapple juice exhibited intermediate antioxidant activity in comparison with other fruit
juices (pineapple juice > orange juice > cherry juice) [34]. The 2,2-diphenyl-1-picrylhydrazyl (DPPH)
radical scavenging capacity of pineapple juice decreased with incubation time from 78.1% to 71.9%
after 120 min of incubation [34]. Mahdavi et al. [35] determined and compared the total polyphenols
in natural fresh and commercial packaged fruit juice. They found that the total polyphenol content in
natural fresh pineapple juice was 36.2 against 35.7 mg GAE/100 mL in commercial pineapple juice,
indicating no significant difference (P > 0.05).
Pineapple Juice 493
40.4 Health Effects

The pineapple has been extensively used as a folk remedy for several health ailments including diges-
tive problems. Recent research has shown that pineapple fruit, peel, and juice exhibit robust effects of
antioxidant capacity, phenolic content, and polysaccharide [36,37]. Pineapple juice inhibits cytochrome
P450 2C9 (these enzymes play role in oxidation and metabolize many therapeutic drugs) activity. Hidaka
et al. [38] reported that the major component extracted from pineapple could reduce CD25 expression
OH
HO
p-Coumaric acid
NH2
O
HO
OH
N
NH2 H
HO
Tyrosine Serotonin
H3C O CH3
O OH
OH
HO
OH
O
HO
O
O O
CH3
O
H3C
Dimethylhydroxylfuranone Dimethylhydroxylfuranone-β-glucoside
O
HO C CH NH2
CH2
OH O O
H3C
NH NH2 OH
CH3
Tryptophan S-Sinapyl-L-cysteine
FIGURE 40.2 Phenolic and sinapyl composition of authentic pineapple juice. (Continued)
OH O O NH2
O=C
CH2 NH C CH NH C CH2 CH2 CH
O
C=
CH2 HO
O O
H3C
OH
CH3
S-Sinapyl glutathione
OH O
O=C
CH2 NH C CH NH2
CH2
O O
H3C
OH
CH3
N-γ-L-Glutamyl-S-sinapyl-L-cysteine
FIGURE 40.2 (Continued) Phenolic and sinapyl composition of authentic pineapple juice.
(trans-membrane protein present on activated T cells) and inhibit cyclooxygenase-2 (COX-2) expression
via anti-inflammation and antitumor activities [39,40]. Pineapple juice has been associated with a lower
incidence of degenerative diseases [41]. It can be taken to alleviate sore throat and sea sickness [42].
Sonicated pineapple juice can serve as substrate for producing probiotic beverage by Lactobacillus
casei NRRL B442 [43]. Fresh pineapple juice containing bromelain enzyme (Figure 40.3) in a clinical
study has a healing pathway for HIV/AIDS. In a recent study by Pandjaitan et al. [44], HIV-positive
human serums were incubated with bromelain at different concentrations (4 h, 37°C). These yielded
negative results at bromelain concentrations of >10 mg/mL. Following this, seven HIV patients were
given two glasses/day of fresh pineapple juice. The results showed that within 4 months, all seven
patients achieved substantial improvement in their CD4 + counts. Three of them already reached normal
CD4 + counts. Moreover, two of them, showed that the viral counts in their system were below detection
limit (<400 copies/mL).
The bromelain in pineapple juice was able to correct menstrual disorders and providing relief from
painful periods [45]. Some of the other pineapple juice benefits for females included a reduction of exces-
sive water buildup in the body especially during pregnancy or menstruation [46]. Bromelain in pineapple
juice has been found to help suppress cough and loosen mucus. Recently, protein molecules from bro-
melain, such as piperazine chlorcyclizine (CCZ) (factor responsible for immunostimulant activity) and
calpain–calpastatin system (CCS) (factor responsible for immunomodulatory activity), have been identi-
fied as powerful anticancer agents and could lead to a new class of cancer-fighting drugs.
Pineapple Juice 495
TABLE 40.2
Nonflavonoids Phenolic Compounds in Pineapple Juice
Food Description Bioactive Compounds mg/100 mL References
Fresh pineapple juice Tyrosine 3.6 [27]
Serotonin 1.8 [27]
Dimethylhydroxylfuranone 1.4 [27]
Dimethylhydroxylfuranone-β-glucoside 6.2 [27]
Tryptophan 2.2 [27]
S-sinapyl-l-cysteine 1.1 [27]
N-γ-Glutamyl-S-sinapyl-l-cysteine 2.3 [27]
S-sinapyl glutathione 5.4 [27]
p-Coumaric acid 0.5 [27]
Commercial clear pineapple 4-O-Caffeoylquinic acid 4.1 [28]
Hydroxymethylfurfural 0.2 [25]
p-Hydroxybenzoic acid 0.07 [25]
p-Hydroxybenzoic aldehyde 0.01 [25]
Syringic aldehyde 0.04 [25]
Ferulic acid 0.18 [25]
Sinapic acid 0.06 [25]
p-Coumaroylquinic acid 0.04 [25]
Feruloyl glucose 0.01 [25]
OH
HO OH
OH
O O
OH
HO HO
O
O
HO O OH
OH O
OH
O OH
O O
HO O NH CH3
H3C NH O
OH
O O
OH
OH
Bromelain
FIGURE 40.3 Chemical structure of bromelain found in pineapple juice.
Bromelain enzymes present in fresh pineapple juice have anti-inflammatory activities. Hale et al.
[47] reported that long-term dietary supplementation with fresh or unpasteurized frozen pineapple juice
with proteolytically active bromelain enzymes was safe and decreased inflammation severity and the
incidence and multiplicity of inflammation-associated colonic neoplasia in murine model of inflam-
matory bowel disease. Fresh pineapple juice has healing power against acute tendon injuries. Recently,
Aiyegbusi et al. [48] compared the effects of commercial bromelain and fresh pineapple juice on teno-
cyte proliferation and the malondialdehyde (MDA) level in the early stage of healing in a crush injury
to the Achilles tendon of Sprague Dawley rats. They concluded that pineapple juice significantly low-
ered the MDA level compared with both the control and bromelain-treated groups. Pineapple bromelain
appears to be a promising adjuvant to antiarthritic drugs. Majeed and Borole [49] evaluated the anti-
inflammatory effect of pineapple juice in rheumatoid and osteoarthritic models in rats and concluded
weak anti-inflammatory activity, which can be used as dietary adjuvant to anti-inflammatory drugs.
Berger and Asenjo [50] reported anthelmintic activity of pineapple juice. They believed that the brome-
lain of pineapple juice might exhibit anthelmintic activity similar to the ficin of the latexes.
Vitamin C found in pineapple juice also helps as a great remedy for oral health and can reduce the risk
of gingivitis and periodontal disease. It also helps the body to fight against the bacteria and the toxins
that invade human gum tissues [51] and help in repairing damaged tissues and in keeping the lymphatic
system working healthy.
Pineapple is a good source of manganese, which is an essential cofactor in a number of enzymes
important in energy production and antioxidant defense. This high level of manganese in pineapple
benefits the skin, collagen, cartilage, and bone material [52]. Studies have also indicated that pineapple
juice is good for the health of pharynx and also the larynx. A combination of glucosamine, chondroitin
sulfate, and manganese may significantly improve the symptoms of mild to moderate osteoarthritis of
the knee [53]. Pineapple enzymes have been used with success to treat rheumatoid arthritis and to speed
up tissue repair as a result of injuries, diabetic ulcers, and general surgery.
In addition to helping to break down the proteins in food, the enzyme bromelain found in pineapple
juice also aids in destroying harmful bacteria in the stomach and intestine because it can improve poor
appetite resulting from insufficient gastric juice. Fresh pineapple juice is often prescribed as atonic and
body-building drink for convalescents and for cancer patients undergoing treatment. It promotes the
digestive processes by destroying the proteins in the stomach and intestine; consequently, broken protein
molecules can be absorbed by the intestine and enter the bloodstream more rapidly. Pineapple juice bro-
melain also cures urinary tract infections [54].
Pineapple juice has the capacity to relieve suffering from heart conditions. This is mainly because
pineapple juice can help in reducing blood clots in the bloodstream. Daher et al. [55] observed that pine-
apple juice significantly decreased plasma triacylglycerols (TAG) and chylomicron in normolipidemic
rats. Since pineapple juice helps in the metabolism of fats and cholesterol, it is one of the main drinks
that are recommended by dietitians for those trying to lose weight. Pineapple juice is also useful in quick
absorption of iron into the human body. One vital point is for those who are following the raw juice
therapy, they should ensure that they are using the ripe fruits. Ripe pineapple has a sweet aroma and is
heavier to hold. Another important use of pineapple juice is its ability to dissolve mucus and thus help
one in a quick recovery from diseases such as tuberculosis [56].

Pineapple juice production has increased significantly in recent years. These days, pineapple juice is
largely consumed around the world as canning industry by-product in the form of single-strength or
concentrated juice. To improve consumer preference, it must be reconstituted in blended composition to
obtain new flavors in beverages and other products [57]. The novel formulations of pineapple juice include
aseptic pineapple juice concentrates, natural pineapple pulp formulations, frozen pineapple concentrates,
sulfated pineapple pulps and purees, and ready-to-serve pineapple drinks. All of these formulations have
numerous applications in dairy and food industries. Recently, Jan and Masih [58] formulated pineapple
juice blended with carrot and orange juices. The aforementioned formulation increased the nutritional
profile and shelf life of pineapple juice because of the addition of extra carotenoids from carrots. In addi-
tion to formulation techniques, processing method and extraction yield also play an important role in
increasing the viability of product. Introducing new varieties, alternate processing methods that preserve
the nutritional profile over long period are highly desirable in modern industry. These new trends are
briefly discussed in this section.
In juice processing, extraction yield is a critical technological parameter [59]. Recently, Sreenath et al.
[60] increased the juice recovery from pineapple pulp/residue using cellulases and pectinases. Cellulase,
Pineapple Juice 497
pectinase, or their mixture, at an enzyme concentration of 0.02% at 27°C–30°C for 30 min, increased
juice recovery to 81%–86% compared to 72% in the conventional methods. Similarly, application of
xylanases from Aspergillus niger DFR-5 could be of great importance to the pineapple juice clarification
industries. Recent trends in pineapple juice industry also include ultrasonic treatments [61] for efficient
juice extraction with considerably enhanced yield and short processing time.
Tran and Le [62] studied the impact of ultrasound on the catalytic activity of pectinase preparation.
This process increased the extraction yield by 5.6% in comparison to no ultrasonic treatment. They used
Pectinex Ultra SP-L solution with an enzyme concentration of 63.3 polygalacturonase units/mL with
ultrasonic treatment for 60 s. The aforementioned treatment had a positive effect on the catalytic activity
of pectinase. The synergistic effect of this technique increased the levels of sugars, polyphenols, organic
acids, and l-ascorbic acid in pineapple juice. Costa et al. [7] studied the influence of ultrasound process-
ing on the physicochemical characteristics of pineapple juice. The polyphenol oxidase activity in the
pineapple juice was reduced by 20% at 376 W/cm2 ultrasonic treatment for 10 min. This process had no
significant effect (P > 0.05) on phenolic compounds compared to the fresh pineapple juice. Ultrasound
processing also enhanced the juice color and its stabilization for 42 days of storage, in comparison with
nonsonicated pineapple juice.
Another advancement in retaining quality of pineapple juice involves the use of ultra-high-pressure
processing [63]. High hydrostatic pressure is a novel technology for minimal processing of pineapple
products. In this technique, a pressure of about 300 MPa for 5 min was applied to pineapple at room
temperature, and the pineapple purée was diluted with water based on the optimum dilution rate. This
technology benefits in terms of reducing bacterial load. Total yeast and fungi counts decreased with
increasing processing pressure in fresh-cut pineapple chunks packed in heat-sealed polyethylene pouches
and treated under various ultra-high pressure, temperature, and time combinations [64]. Water and solute
of pressure-pretreated pineapple have been reported to render a significantly higher (P < 0.05) diffusion
rate during osmotic dehydration [65].
Clarification and concentration of pineapple juice is required in numerous dairy and beverage indus-
tries. Recent techniques in this process include the use of ultra- and microfiltration process. Jaeger de
Carvalho et al. [66] studied the clarification of pineapple juice by ultrafiltration and microfiltration with
0.22 µm ceramic membrane and 50 kDa polysulfone membrane. This ceramic membrane performs
better with respect to soluble solids, sugars, and acidity, but the recovery was less in ceramic membrane
compared to 50 kDa polysulfone membrane. Laorko et al. [10] studied the processing parameters and
quality profile of pineapple juice treated with microfiltration (pore size 0.1 and 0.2 µm) and ultrafiltra-
tion (membrane molecular weight cutoff of 30 and 100 kDa). The microfiltration process has no effect
on pH, reducing sugar, and acidity of clarified juice, whereas the suspended solids and microorganisms
were completely removed. The 0.2 µm membrane gave the highest permeate flux, vitamin C content,
total phenolic content, and antioxidant capacity as well as the highest value of irreversible fouling.
They concluded that membrane with a pore size of 0.2 µm was most suitable for clarification of pine-
apple juice [10].
Electrodialysis is another technique used to reduce the acid content of pineapple juice. In this tech-
nique, electrically charged membranes with electrical potential difference are used to separate ionic spe-
cies in the pineapple juice. This process showed considerable increase in sweetness and reduction in the
tartness, but the mineral content was reduced in comparison with fresh juice with no electrodialysis [6].
40.6 Conclusion
Pineapple juice and its blended formulations are used for their unique taste and aroma attributes; how-
ever, these are also a good source of dietary antioxidants and contribute significantly to daily dietary
requirements. Pineapple juice has many variations in health-promoting activities. It improves the blood
profile and also reduces constipation problems. The best advantage of drinking pineapple juice is that it
has low calories and thus is considered the best drink for diabetes. Although much research is available
for pineapple cultivation and other management techniques, its formulation (juices and other concen-
trates) advantages need extensive research.
REFERENCES
1. Rattanathanalerk, M., Chiewchan, N., and Srichumpoung, W., Effect of thermal processing on the qual-
ity loss of pineapple juice. J. Food Eng., 66, 259–265, 2005.
2. Food and Agriculture Organization (FAO), Principles and Practices of Small-and Medium-Scale Fruit
Juice Processing, Agricultural Services Bulletin 146, FAO, Rome, Italy, 2001.
3. Elkins, E.R., Lyon, R., Huang, C.J., and Matthys, A., Characterization of commercially produced pine-
apple juice concentrate. J. Food Comp. Anal., 10, 285–298, 1997.
4. De Vasconcelos Facundo, H.V., De Souza Neto, M.A., Maia, G.A., Narain, N., and Dos Santos Garruti, D.,
Changes in flavor quality of pineapple juice during processing. J. Food Process Preser., 34, 508–519,
2010.
5. Center for the Promotion of Imports from Developing Countries (CBI), CBI Market Survey: Fruit Juice
and Concentrates, Ministry of Foreign Affairs (CBI), The Hague, the Netherlands, 2009.
6. Sairi, M., Yih, L.J., and Sarmidi, M.R., Chemical composition and sensory analysis of fresh pineapple
and deacidified pineapple juice using electrodialysis, presented at Symposium on Membrane Science
and Technology, Johor Bahru, Malaysia, April 21–25, 2004, ID code 6174.
7. Costa, M., Fonteles, T., Jesus, A., Almeida, F., Miranda, M., Fernandes, F., and Rodrigues, S., High-
intensity ultrasound processing of pineapple juice. Food Bioprocess Technol., 69, 97–1006, 2013.
8. Zheng, H. and Lu, H., Use of kinetic, Weibull and PLSR models to predict the retention of ascorbic acid,
total phenols and antioxidant activity during storage of pasteurized pineapple juice. LWT—Food Sci.
Technol., 44, 1273–1281, 2011.
9. Shamsudin, R., Ling, C.S., Adzahan, N.M., and Daud, W.R.W., Rheological properties of ultraviolet-
irradiated and thermally pasteurized Yankee pineapple juice. J. Food Eng., 116, 548–553, 2013.
10. Laorko, A., Li, Z., Tongchitpakdee, S., Chantachum, S., and Youravong, W., Effect of membrane prop-
erty and operating conditions on phytochemical properties and permeate flux during clarification of
pineapple juice. J. Food Eng., 100, 514–521, 2010.
11. Hodgson, A.S. and Hodgson, L.R., Pineapple juice, in Fruit Juice Processing Technology, Nagy, S.,
Chen, C.S., and Shaw, P.E., Eds., Agscience, Inc., Auburndale, FL, 1993, pp. 334–371.
12. Pilando, L.S. and Wrolstad, R.E., The effectiveness of pattern recognition, sugar, nonvolatile acid, and
13C/12C analyses for detecting adulteration in apple juice. J. Food Comp. Anal., 5, 10–24, 1992.
13. Wallrauch, S. and Faethe, W., Amino acids: Criteria for the evaluation of fruit juices, in Adulteration
of Fruit Juices, Nagy, S., Attaway, J.A., and Rhodes, M.E., Eds., Marcel Dekker, Inc., New York, 1988,
pp. 21–48.
14. Gebhardt, S.E., Cutrufelli, R., and Matthews, R.H., Composition of foods—Fruits and fruit juices, in
Agriculture Handbook No 8–9, Consumer Nutrition Center—Human Nutrition Information Service,
USDA, Washington, DC, 1982, p. 283.
26, 2013. Published online at: http://ndb.nal.usda.gov/ (assessed April 07, 2014).
16. Krueger, D.A., Krueger, R., and Maciel, J., Composition of pineapple juice. J. AOAC Int., 75, 280–282,
1992.
17. Luximon-Ramma, A., Bahorun, T., and Crozier, A., Antioxidant actions and phenolic and vitamin C
contents of common Mauritian exotic fruits. J. Sci. Food Agric., 83, 496–502, 2003.
18. Kabasakalis, V., Siopidou, D., and Moshatou, E., Ascorbic acid content of commercial fruit juices and
its rate of loss upon storage. Food Chem., 70, 325–328, 2000.
19. Cárnara, M., Díez, C., and Torija, E., Chemical characterization of pineapple juices and nectars.
Principal components analysis. Food Chem., 54, 93–100, 1995.
20. Gawler, J.H., Constituents of canned Malayan pineapple juice I. Amino-acids, non-volatile acids, sug-
ars, volatile carbonyl compounds and volatile acids. J. Sci. Food Agric., 13, 57–61, 1962.
21. Dizy, M., Martín-Alvarez, P.J., Cabezudo, M.D., and Carmen Polo, M., Grape, apple and pineapple juice
characterisation and detection of mixtures. J. Sci. Food Agric., 60, 47–53, 1992.
22. Cámara, M.M., Díez, C., and Torija, M.E., Free sugars determination by HPLC in pineapple products.
Eur. J Food Sci. Technol., 20, 2233–2237, 1996.
23. Ooghe, W. and Dresselaerts, D., Quality parameters for pineapple juice. Fruit Process., 5, 11–17,
1995.
24. Macheix, J., Fleuriet, A., and Billot, J., Fruit Phenolics, CRC Press, Boca Raton, FL, 1990.
Pineapple Juice 499
25. Fernandez de Simon, B., Perez-Ilzarbe, J., Hernandez, T., Gomez-Cordoves, C., and Estrella, I., Importance
of phenolic compounds for the characterization of fruit juices. J. Agric. Food Chem., 40, 1531–1535, 1992.
26. Larrauri, J.A., Rupérez, P., and Calixto, F.S., Pineapple shell as a source of dietary fiber with associated
polyphenols. J. Agric. Food Chem., 45, 4028–4031, 1997.
27. Wen, L. and Wrolstad, R.E., Phenolic composition of authentic pineapple juice. J. Food Sci., 67, 155–161,
2002.
28. Mullen, W., Marks, S.C., and Crozier, A., Evaluation of phenolic compounds in commercial fruit juices
and fruit drinks. J. Agric. Food Chem., 55, 3148–3157, 2007.
29. Wen, L., Wrolstad, R.E., and Hsu, V.L., Characterization of sinapyl derivatives in pineapple (Ananas
comosus [L.] Merill) juice. J. Agric. Food Chem., 47, 850–853, 1999.
30. Morgan, R.C., Chemical studies on concentrated pineapple juice 1. Carotenoid composition of fresh
pineapples. J. Food Sci., 31, 213–217, 1966.
31. Ramsaroop, R.E.S. and Saulo, A.A., Comparative consumer and physicochemical analysis of Del Monte
Hawaii gold and smooth cayenne pineapple cultivars. J. Food Quality, 30, 135–159, 2007.
32. Herraiz, T. and Galisteo, J., Tetrahydro-beta-carboline alkaloids occur in fruits and fruit juices. Activity
as antioxidants and radical scavengers. J. Agric. Food Chem., 51, 7156–7161, 2003.
33. Gardner, P.T., White, T.A.C., McPhail, D.B., and Duthie, G.G., The relative contributions of vitamin C,
carotenoids and phenolics to the antioxidant potential of fruit juices. Food Chem., 68, 471–474, 2000.
34. Ramadan-Hassanien, M.F., Total antioxidant potential of juices, beverages and hot drinks consumed in
Egypt screened by DPPH in vitro assay. Grasas Y Aceites, 59, 254–259, 2008.
35. Mahdavi, R., Nikniaz, Z., Rafraf, M., and Jouyban, A., Determination and comparison of total polyphe-
nol and vitamin C contents of natural fresh and commercial fruit juices. Pak. J. Nutr., 99, 68–972, 2010.
36. de Oliveira, A.C., Valentim, I.B., Silva, C.A., Bechara, E.J.H., Barros, M.P.D., Mano, C.M., and Goulart,
M.O.F., Total phenolic content and free radical scavenging activities of methanolic extract powders of
tropical fruit residues. Food Chem., 115, 469–475, 2009.
37. Guo, Q.L. and Zhang, L.H., Study on polysaccharides extraction and ability to scavenge hydroxyl radi-
cals from pineapple. Chinese Agric. Sci. Bull., 25, 122–125, 2009.
38. Hidaka, M., Nagata, M., Kawano, Y., Sekiya, H., Kai, H., Yamasaki, K., Okumura, M., and Arimori, K.,
Inhibitory effects of fruit juices on cytochrome P450 2C9 activity in vitro. Biosci. Biotechnol. Biochem.,
72, 406–411, 2008.
39. Bhui, K., Prasad, S., George, J., and Shukla, Y., Bromelain inhibits COX-2 expression by blocking the
activation of MAPK regulated NF-kappa B against skin tumor-initiation triggering mitochondrial death
pathway. Cancer Lett., 282, 167–176, 2009.
40. Secor, E.R., Jr., Singh, A., Guernsey, L.A., McNamara, J.T., Zhan, L., Maulik, N., and Thrall,
R.S., Bromelain treatment reduces CD25 expression on activated CD4 + T cells in vitro. Int.
Immunopharmacol., 9, 340–346, 2009.
41. Bajpai, V.K., Yoon, J.I., and Chul Kang, S., Antioxidant and antidermatophytic activities of essential oil
and extracts of Metasequoia glyptostroboides Miki ex Hu. Food Chem. Toxicol., 47, 1355–1361, 2009.
42. Mendiola, J.A., Marin, F.R., Senorans, F.J., Reglero, G., Martin, P.J., Cifuentes, A., and Ibanez, E.,
Profiling of different bioactive compounds in functional drinks by high-performance liquid chromatog-
raphy. J. Chromatogr. A., 1188, 234–241, 2008.
43. Costa, M.G., Fonteles, T.V., de Jesus, A.L., and Rodrigues, S., Sonicated pineapple juice as substrate
for L. casei cultivation for probiotic beverage development: Process optimisation and product stability.
Food Chem., 139, 261–266, 2013.
44. Pandjaitan, M., Nugraha, T., and Pamudja, K.H., Bromelain enzyme in fresh pineapple juice as a healing
pathway for HIV/AIDS. Adv. Sci. Eng. Med., 6, 119–123, 2014.
45. Zhang, C.Z., Wang, S.X., Zhang, Y., Chen, J.P., and Liang, X.M., In vitro estrogenic activities of Chinese
medicinal plants traditionally used for the management of menopausal symptoms. J. Ethnopharmacol.,
98, 295–300, 2005.
46. Zava, D.T., Dollbaum, C.M., and Blen, M., Estrogen and progestin bioactivity of foods, herbs, and
spices. Proc. Soc. Exp. Biol. Med., 217, 369–378, 1998.
47. Hale, L.P., Chichlowski, M., Trinh, C.T., and Greer, P.K., Dietary supplementation with fresh pineapple
juice decreases inflammation and colonic neoplasia in IL-10-deficient mice with colitis. Inflamm. Bowel
Dis., 16, 2012–2021, 2010.
48. Aiyegbusi, A.I., Olabiyi, O.O., Duru, F.I., Noronha, C.C., and Okanlawon, A.O., A comparative study of
the effects of bromelain and fresh pineapple juice on the early phase of healing in acute crush achilles
tendon injury. J. Med Food, 14, 348–352, 2011.
49. Majeed, M. and Borole, K., Evaluation of anti-inflammatory effect of pineapple juice in rheumatoid and
osteo arthritis models in rats. Ind. J. Pharmacol., 45, S23, 2013.
50. Berger, J. and Asenjo, C.F., Anthelmintic activity of fresh pineapple juice. Science, 90, 299–300,1939.
51. Chapple, I.L.C., Milward, M.R., Ling-Mountford, N., Weston, P., Carter, K., Askey, K., Dallal, G.E.
et al., Adjunctive daily supplementation with encapsulated fruit, vegetable and berry juice powder con-
centrates and clinical periodontal outcomes: A double-blind RCT. J. Clin. Periodontol., 39, 62–72,
2012.
52. Manganese in bone development. Nutr. Rev., 3, 174–175, 1945.
53. Orlando, L., The Amazing Pineapple, Hawaii’s Natural Health Booster, 2006. Published online at:
http://www.buzzle.com/ editorials/4–2–2006–92514.asp (accessed October 20, 2013).
54. Efros, M., Bromberg, W., Cossu, L., Nakeleski, E., and Katz, A.E., Novel concentrated cranberry liquid
blend, UTI-STAT with proantinox, might help prevent recurrent urinary tract infections in women.
Urology, 76, 841–845, 2010.
55. Daher, C.F., Abou-Khalil, J., and Baroody, G.M., Effect of acute and chronic grapefruit, orange, and
pineapple juice intake on blood lipid profile in normolipidemic rat. Med. Sci. Monit., 11, 465–472, 2005.
56. Bhowmik, D., Chiranjib Chandira, R.M., Jayakar, B., and Sampath Kumar, K.P., Recent trends of drug
used treatment of tuberculosis. J. Chem. Pharm. Res., 2, 113–133, 2009.
57. Arthey, D., Food Industries Manual, Chapman & Hall, London, UK, 1995.
58. Jan, A. and Masih, D.E., Development and quality evaluation of pineapple juice blended with carrot and
orange juice. Int. J. Sci. Res. Publ., 2, 1–8, 2012.
59. McLellan, M.R. and Padilla-Zakour, O.I., Juice processing, in Processing Fruits: Science and
Technology, Barrett, D.M., Somogyi, L.P., and Ramaswamy, H.S., Eds., CRC Press, Boca Raton, FL,
2005, pp. 73–96.
60. Sreenath, H.K., Sudarshanakrishna, K.R., and Santhanam, K., Improvement of juice recovery from
pineapple pulp/residue using cellulases and pectinases. J. Ferment. Bioeng., 78, 486–488, 1994.
61. Nguyen, T.P. and Le, V.V.M., Application of ultrasound to pineapple mash treatment in juice processing.
Int. Food Res. J., 19, 547–552, 2012.
62. Tran, P.P.T. and Le, V.V.M., Effects of ultrasound on catalytic efficiency of pectinase preparation during
the treatment of pineapple mash in juice processing. Int. Food Res. J., 18, 347–354, 2011.
63. Li, B., Zhang, Z., and Canhui, M., Comparison of effects of ultra-high pressure and heat sterilization on
qualifies of freshly-squeezed pineapple juice. Trans. Chinese Soc. Agric. Eng., 26, 359–364, 2010.
64. Hepton, A. and Hodgson, A., Processing, in The Pineapple: Botany, Production and Uses, Bartholomew,
D.P., Paul, R.E., and Rohrbach, K.G., Eds., CABI Publishing, New York, 2003, pp. 281–289.
65. Ramaswamy, H., Chen, C., and Marcotte, M., Novel processing technologies for food preservation, in
Processing Fruits: Science and Technology, Barrett, D.M., Somogyi, L.P., and Ramaswamy, H.S., Eds.,
CRC Press/Taylor & Francis Group, Boca Raton, FL, 2005, pp. 201–219.
66. Jaeger de Carvalho, L.M., Bento da Silva, C.A., and Pierucci, A.P.T.R., Clarification of pineapple juice
(Ananas comosus L. Merryl) by ultrafiltration and microfiltration: Physicochemical evaluation of clari-
fied juices, soft drink formulation, and sensorial evaluation. J. Agric. Food Chem., 46, 2185–2189, 1998.
41
Plum, Prune, and Ume Juices
Kent Fanning, Roger Stanley, Bruce Topp, Dougal Russell, and Michael Netzel
CONTENTS
41.1 Introduction................................................................................................................................... 501
41.4 Health Effects................................................................................................................................ 507
41.4.1 Plum and Prune Juices..................................................................................................... 507
41.4.2 Ume Juice......................................................................................................................... 508
41.6 Conclusion..................................................................................................................................... 509
References............................................................................................................................................... 509
41.1 Introduction
The two plum species of worldwide economic significance are the Japanese plum (Prunus salicina
Lindl.) and European plum (P. domestica L.) (Figure 41.1). The Japanese plum has a wide range of adap-
tation from temperate to the subtropical regions, whereas the European plum is grown in cool and tem-
perate climates [1]. Fruit of both species is used to produce juice. A third species, P. mume Sieb. et Zucc.,
produces fruit that is referred to in Asia as plum, although it is more closely related to apricot [2]. The
fruit is commonly called “ume” and is also used to produce a range of juices and alcoholic beverages [2].
In this contribution, the term “plum” refers to fresh the European plum and Japanese plum, “prune”
refers to the dried European plum, and “ume” refers to ume fruit. The focus in this chapter is exclusively
on juice and juice concentrates made from plum, prune, and ume. Information pertaining to whole fruit
and extracts of fruit and canned, dried, salted, or fiber products has not generally been included unless it
can represent the nutritional or bioactive properties of juice products.
Plum juice is derived from whole fruit by separating the juice from the flesh and seed using pressing,
slicing, chopping, or homogenization. A range of processing factors influence the properties of the final
product, namely, enzyme type, enzyme content, and preheating of fruit, all of which impact juice yield
and quality [3–6]. Enzymatic treatment is used commercially and can lower viscosity and pectin content
but increase yield, clarity, soluble solids, titratable acidity, anthocyanin concentration, and antioxidant
activity [3–6].
Prune juice has been developed as an easy form of consumption of prunes. It is made by cooking
prunes in water until soluble solid of 18.5–21°Brix is reached [7,8]. This is often followed by treatment
with cellulolytic and pectinolytic enzymes to dissolve insoluble cellular material, prevent gelling, and
aid filtration [4,7,8].
Several juice and juice concentrate products are made from ume, including umeshu (plum wine), ume
with green tea, ume juice, misatol, and Bainiku-ekisu [2]. Juice or nectar is prepared from the ripe fruits
by adding browning inhibitors, enzymes, and cane sugar to reduce bitterness [9]. The basic method for
making fruit juice concentrate (Bainiku-ekisu) is mincing fruit, filtering, and boiling down the filtrate
until the mass has decreased up to 50 times [10]. A sucrose osmotic extraction can also be used to extract
501
(a) (b)
(c)
FIGURE 41.1 Fresh Japanese plum (a), European plum (b), and ume (c).
juice from fruit [11]. Dilution with water is used to prepare these juice/juice concentrate products for
consumption.
No current figures of global plum, prune, or ume juice production could be sourced. Plum and prune
juice are produced in relatively small to medium quantities in the United States and Europe, and ume
juice products are produced commercially in countries including Japan, China, South Korea, and Taiwan.
This chapter highlights the differences in nutrient and bioactive composition of plum, prune, and ume
products, the health effects of these products, and how nutrient and/or health claims are, and potentially
will be, used to promote and position commercial products.

A compilation of nutritional data for plum, prune, and ume juices is given in Table 41.1. In general, plum
and prune juices have an acidic pH ranging from 2.7 to 3.9 [12–15], titratable acidity of 0.6–2.0 [12–15],
and relatively highly soluble solids with °Brix values ranging from 13 to 21 [3,12,13,15,16]. The absence
or very low sucrose content of prune juice appears to be a distinguishing factor to plum juice [14,17].
The major characteristic of ume juice is its high level of citric and malic acids [10,11], with consequent
low pH and sharp taste, with commercial concentrate diluted 1:5 (v/v) with water still having a pH
of 2.3 [18].
The fiber content of both plum and prune juices is the primary nutrient content claim that is
marketed. Thus, 100% prune juices produced by Sunsweet (Table 41.2) and Del Monte have 3 g
fiber/240 mL. However, some prune juice products have much lower amounts of intrinsic fiber and
Plum, Prune, and Ume Juices 503
TABLE 41.1
Compositional and Nutritional Characteristics of Plum, Prune, and Ume Juices (per 100 mL or 100 g)
Nutrient Unit Plum Juice Prune Juice Ume Juice References
Energy kcal 57 71 49 [19–21]
Water g 81.2 87.6 [20,21]
Protein g 0.61 tr [20,21]
Lipid (fat) g 0.03 tr [20,21]
Fructose g 1.0–6.0 4.6–6.9 8.0 [3,6,11,15,17]
Glucose g 2.0–10.1 7.0–10.9 8.0 [3,6,11,15,17]
Sucrose g 0.0–8.0 0.0–0.1 400 [3,6,11,15,17]
Sorbitol g 0.01–6.4 4.4–6.9 [3,6,15,17]
Total dietary fiber g 0.01–1.0 0.1 [20–22]
Minerals
Boron mg — 0.6 — [22]
Calcium mg 9.1–52 12–51 1.0 [14,15,19–21]
Copper mg 0.04–0.05 0.06–0.07 0.01 [14,15,20,21]
Fluoride mg — 0.06 — [20]
Iron mg 0.11–0.17 1.18–3.7 0.2 [14,15,19–21]
Magnesium mg 9.0–15.2 14–58 2.0 [14,15,19–21]
Manganese mg 0.15–1.3 0.01 [14,20,21]
Phosphorous mg 256–285 25–78 2.0 [14,15,19–21]
Potassium mg — 276 30 [20,21]
Selenium µg — 0.6 — [20]
Sodium mg 1.0–9.0 4.0–32 35 [14,15,19–21]
Zinc mg 0.08–0.2 0.21–0.62 tr [14,15,20,21]
Vitamins
β-carotene µg — 2.0 tr [20,21]
Folate µg — 0.0 0.0 [20,21]
Niacin mg — 0.79 0.0 [20,21]
Riboflavin mg — 0.07 0.0 [20,21]
Thiamin mg — 0.02 0.0 [20,21]
Vitamin B5 mg — 0.11 — [20]
Vitamin B6 mg — 0.22 0.01 [20,21]
Vitamin C mg 4.0 4.1 0.0 [19–21]
Vitamin E mg — 0.12 0.2 [20,21]
Vitamin K1 µg — 3.4 0.0 [20,21]
Abbreviation: tr, trace.
fiber is sometimes added. Several plum juice products, including Sunsweet’s PlumSmart®, also have
added fiber to reach levels between 3 g/200 mL and 3 g/240 mL. Citric acid content of ume products
is used on labeling of commercial products (Table 41.2). This can be used to connect with consumer
perception that the high acid content may be beneficial in preventing conditions such as urinary tract
infection [18].
Prune juice is generally more vitamin and mineral dense than plum or ume juice products, but
further analysis, particularly of plum juice, is warranted to allow a better comparison (Table 41.1).
The nutrition label for Sunsweet’s 100% prune juice for the United States claims that it has 10%
or greater of the daily value, per 240 mL serve, for potassium (430 mg, 12% daily value), niacin
(10% daily value), riboflavin (15% daily value), vitamin B6 (15% daily value), and copper (15% daily
value).
TABLE 41.2
Summary of Commercial Products with Health/Nutrient Claims or Associations
Health and Nutrition
Product Name Manufacturer Claims or Associations Composition
Prune juice Sunraysia, Australia Naturally rich in 100% prune juice from concentrate.
antioxidants
Prune juice with Sunsweet Growers, Contains 3 g fiber/240 mL 100% prune juice, with pulp.
pulp United States serving
NATURlax Naturfresh Most natural way to regulate Plum (98%) and vegetable fiber (2%).
Ciruela: Plum Productos your body, with fiber
fruit drink Naturales, Spain (3 g/200 g)
Sunsweet Sunsweet Growers, Lets you stay fit on the Plum juice from concentrate, grape juice
PlumSmart Juice United States inside; helps keep the from concentrate, dextrin, carrot, and
digestive system balanced blueberry juice for color, vitamin C,
and happy; a good source chamomile extract, ginger juice, and
of fiber (3 g/240 mL) citric acid.
Queensberry Kiviks Marknad, Red fruits (strawberry, cherry, and
Smoothie Red Brazil raspberry), concentrated juice (apple,
Fruits plum, cranberry, and pomegranate),
oligofructose and
fructooligosaccharides, and fruit pectin.
Naked Purple Naked Juice, Aims to promote healthy Purple machine blend (açai, puree,
Machine United States aging and sound memory purple plum puree, concord grape juice
Smoothie, all from concentrate, and soy lecithin),
natural superfood apple juice, banana puree, elderberry
100% fruit extract, beet powder, vitamin E,
smoothie blackcurrant powder, vitamin C, and
natural flavors.
Meiraku, Ume Tokyo Meiraku With citric acid Fructose corn syrup, plum juice, plum
Kuensan vitamin Chiba Sa, Japan pulp extract, acidifier, flavor, vitamin C,
C plum juice: chloridation, color (caramel), and
Plum juice sweetening.
Sapporo Ume no Sapporo, Japan With citric acid Plum juice, fructose corn syrup,
Tennen Kuensan: maltooligosaccharide,
Low-calorie plum β-glucooligosaccharide component
juice with citric syrup, trehalose, acidifier, flavoring, and
acid sweetener (acesulfame K).

Table 41.3 shows the bioactive content and antioxidant capacity of a range of plum, prune, and ume
juice products. 5-Hydroxymethyl furfural (Figure 41.2) is naturally generated in acidic sugar-containing
foods, like juices, during drying and heat treatment but is also slowly formed during storage [23]. It is
present in both prune and ume juice products and is a major component in some [10,24,25]. However, it
is absent or present in only very small amounts in plum juice [16,17]. In ume juice concentrate, mume-
fural (1-[5-(2-formylfuryl)methyl]-dihydrogen 2-hydroxypropane-1,2,3-tricarboxylate), a derivative of
5-hydroxymethyl furfural, has also been quantified [10] and three other similar compounds have been
identified [26]. The metabolism of 5-hydroxymethyl furfural, following prune juice consumption, has
been studied in six healthy female subjects [24]. A rapid metabolism to glycine conjugates and sev-
eral other metabolites that were excreted in the urine was observed [24]. A major difference between
plum juice, made from red plum, and prune juice is that plum juice contains anthocyanins that are lost
in prune juice through the drying and juicing processes [14,16,17]. The dark brown color of prune is
TABLE 41.3
Bioactive Composition and Antioxidant Activity of Plum, Prune, and Ume Juices (per 100 mL or 100 g)
Compound/Activity Unit Plum Juice Prune Juice Ume Juice References
5-Hydroxymethyl-2- mg nd–0.8 53–158 1.3–402 [10,16,17,24,25]
furfural
Mumefural mg 346 [10]
Catechin mg 2.8–41 15.9 [15,17]
Chlorogenic acid mg 2.0–38 5.7–34 0.5–3.5 [11,15–17,24]
4-O-Caffeoylquinic acid mg 22.4 [24]
Caffeic acid mg 4.4 [17]
Coumaric acid mg 0.4 [16]
Epicatechin mg 2.2–5.4 [15]
Flavan-3-ols mg EC Monomers, 4–5 [12]
Dimers, 3
Trimers, 1
Oligomers, 1
Neochlorogenic acid mg 126–222 22.5 [12,15,16]
3′-Coumaroylquinic mg 0.4 [16]
acid
Coumaroylglucose mg 4.6–10.3 [15]
Rutin mg 4.0 [16]
Quercetin-3-O- mg 0.5–3.2 [12,15]
rutinsoide
Quercetin-3-O- mg 0.3–0.6 [12]
glucoside
Procyanidin B1 mg 6.3–8.1 [15]
Anthocyanin mg 0.18–279 nd [3,6,12,14–16,29,31]
Carotenoid content mg 9–11 tr [13,21]
Total phenolics Units 294–367 (mg CE) 350 (mg GAE) [3,6,12,13,15,24,31]
indicated 43–254 (mg GAE)
in brackets 25–430 (mg TAE)
470–520 (mg CAE)
Antioxidant capacity Antioxidant TEAC (1.9–2.7 mg TE) ORAC [12,13,15,24,29,31]
assay and ORAC (3.1 mmol TE) (2130 µmol TE)
units ORAC (360–490 mg TE)
indicated FRAP/ORAC (8.2/
in brackets 3.9 mmol Fe2+/mmol TE)
DPPH/ORAC (0.2–0.3/
1.9–2.3, mmol TE)
nd, not detected; tr, trace; EC, epicatechin equivalents; CE, catechin equivalents; GAE, gallic acid
Abbreviations:
equivalents; TAE, tannic acid equivalents; CAE, chlorogenic acid equivalents; TEAC, trolox equivalents
antioxidant capacity; TE, trolox equivalents; ORAC, oxygen radical absorbance capacity; FRAP, ferric-
reducing antioxidant power; DPPH, 2,2-diphenyl-1-picrylhydrazyl.
attributed to melanoidins created during heating and drying [27]. Melanoidins are a major contributor to
antioxidant capacity in prunes [28]. The main anthocyanins in plum juice are cyanidin-3-O-rutinoside,
cyanidin-3-O-glucoside (Figure 41.2), peonidin-3-O-glucoside, and peonidin-3-O-rutinoside [3,15,29].
Both the method of juicing and the plum variety may significantly impact the content of anthocyanins
[6,15], with a high content of 279 mg/100 mL present in juice from the anthocyanin-rich plum vari-
ety Queen Garnet [29]. Ume juice products do not contain anthocyanins. Preliminary results with
Queen Garnet plum juice and healthy male subjects indicate an extensive metabolism of the native
OH
OH OH
OH O+
HO
O + HO OH
HO OH
O
O
O OH
HO O
OH
O O
OH H3C
OH OH
HO HO OH
Cyanidin 3-O-glucoside Cyanidin 3-O-rutinoside
O
OH
O O
OH O
HO OH
HO O
HO OH
Chlorogenic acid 5-Hydroxymethyl furfural
FIGURE 41.2 Chemical structures of bioactive compounds found in plum juice (cyanidin 3-O-glucoside, cyanidin
3-O-rutinoside, and chlorogenic acid) and prune and ume juices (chlorogenic acid and 5-hydroxymethyl furfural).
plum anthocyanins, cyanidin-3-O-glucoside, and cyanidin-3-O-rutinoside, mainly to glucuronidated and

methylated compounds potentially affecting their in vivo bioactivity [29,30].
Anthocyanin-rich plum juice appears to hold promise as a commercial colorant. A comparison of
plum juice with commercial grape colorant showed there was less color loss in plum juice at both pH
1 and 3 after 138 days of storage at 25°C (21% and 23% [plum], 30% and 31% [grape]), and after 2 h at
99°C at pH 1 (17% [plum], 69% [grape]) [31]. Furthermore, juice from plum peel had a significantly lower
anthocyanin degradation rate than strawberry, bilberry, and red raspberry juices, when stored at a pH of
4.5 at 20°C for 17 weeks [32]. The plum peel juice also exhibited very stable color properties (lightness,
hue, and chroma) during storage [32].
Chlorogenic acid (Figure 41.2) is a common phenolic compound in all three juice types. However,
differences between fruit varieties and processing methods can lead to large variations in its content
between products [25,33]. The content of other phytochemicals (including cinnamates, flavonols, procy-
anidins, and carotenoids) presumably also varies between different juice products. However, there is cur-
rently insufficient data to comment on clear specific differences between plum, prune, and ume juices.
Detailed comparative analysis of juices is warranted. Furthermore, future studies, which investigate
in vitro and in vivo effects and/or bioavailability and metabolism, must define the source, composition,
and phytochemical content of the studied products.
In a pilot study with Queen Garnet plum juice, the antioxidant capacity of male volunteers’ urine was
increased, after consumption. This was relative to the antioxidant capacity after consumption of water as
an antioxidant-free control [29].
Both plum and prune juice concentrates, when included in brine solution, which was injected into
roast beef, have shown the ability to significantly reduce lipid oxidation [34], and prune puree signifi-
cantly reduced lipid oxidation of beef patties [35]. Prune juice showed 62% and 97% inhibition of Cu2+-
catalyzed oxidation of low-density lipoprotein (LDL) at 10 and 20 µM [16]. However, this inhibition was
significantly lower (P < 0.05) than that seen for prune (whole dried fruit) extract, neochlorogenic acid,
or chlorogenic acid, at 5 and 10 µM. 5-Hydroxymethyl furfural showed no inhibition between 5 and
20 µM [16]. Ume juice concentrate (Bainiku-ekisu) showed a dose-dependent reduction of microsomal
lipid peroxidation [36].
41.4 Health Effects

41.4.1 Plum and Prune Juices
Both raw and heat-treated plum juices showed inhibition of mutagenicity induced by 2-amino-
3-methyllimidazo[4,5-f]quinoline in Salmonella typhimurium strains TA98 and TA100, suggesting
moderate antimutagenic potency [37].
The ingestion of 400 mL Queen Garnet plum juice (containing 2.66 g total phenolics and 1.11 g antho-
cyanins, respectively) by two healthy male subjects resulted in a decreased (no P-value was calculated due
to only n = 2) urinary malondialdehyde excretion, a biomarker for oxidative stress, within 24 h as com-
pared with the polyphenol-/antioxidant-free control [29]. In another human study with 10 healthy male
subjects, the effect of nine fruit juices on the antioxidant activity in blood plasma was investigated [38].
After consuming 150 mL of pear, apple, orange, grape, peach, plum, kiwi, melon, and watermelon juices,
blood samples were collected at 0, 30, 60, 90, and 120 min postconsumption. Eight fruit juices, including
plum juice, significantly suppressed (P < 0.05) the generation of reactive oxygen species in the plasma
samples [38].
Shukitt-Hale et al. [39] showed that a diet consisting of a diluted commercial plum juice concentrate,
with the only source of fluids being either plum juice or water (control) for 8–9 weeks, significantly
reduced (P < 0.05) the time taken and distance traveled for rats to find the hidden platform in the Morris
water maze. This significant decrease was not seen in rats receiving water [39]. These promising results
indicate potential efficacy of polyphenol-rich plum juice for mitigating age-related decline in brain func-
tion and cognition, and human studies are warranted.
Both plum juice concentrates [40] and prune juices [41] have shown promise in the area of bone health.
A plum juice concentrate was fed to nonpregnant female mice for 30 days (8 mL/kg/day) and significantly
increased (P < 0.05) both blood calcium levels and bone calcium content compared to mice receiving
water [40]. The supplementation of ovariectomized rats with prune juice (7.5%) and fructooligosac-
charides (2%) reversed ovariectomy-induced bone mineral density loss in the right femoral bone [41].
Trabecular number and trabecular separation were also significantly increased and decreased (P < 0.05),
respectively, following consumption of prune juice and fructooligosaccharides [41]. The mechanisms
behind the role of plum and prune juices in affecting bone structure and mineral content are not defined
but may include the content of calcium, phosphorous, magnesium, boron, and polyphenols [40,41].
However, no compositional data for the tested juices were presented in either study. Human stud-
ies are needed to investigate the potential benefits of plum and prune juice products. These proposed
studies need to include clear definition of the source and composition (both nutrient and phytochemical)
of tested products.
Both plum and prune juices have shown an ability to reduce symptoms of constipation and defeca-
tion difficulty [42,43]. A commercially formulated plum juice (PlumSmart®, Table 41.2), with a fiber
content of 3 g/240 mL, was compared with apple juice (placebo) and apple juice containing psyllium
(Metamucil®) in 36 adults reporting chronic constipation symptoms [42]. Consumption of 240 mL of
plum juice formulation per day for 14 days resulted in softer stools than apple juice and apple juice
containing psyllium, and plum juice provided relief of symptoms similar to apple juice containing psyl-
lium [42]. Prune juice (prepared from plum juice concentrate, prune puree, water, and fructose), when
consumed at 125 mL twice daily for 2 weeks by 54 subjects who had minor gastrointestinal symptoms,
significantly reduced (P < 0.05) days with defecation difficulty [43]. Prune juice has also been used in
bowel care management of hospital patients [44]. The mechanism of action for the laxative effect of
plum and prune juices is not clearly defined [43,45]. The hypothesis is that the effects are a combination
of fiber, sorbitol, xylitol, and polyphenolics but further studies investigating different juice products as
well as these individual components are warranted [43,45,46]. There was a lack of compositional data
presented for tested products in previous studies [42,43] and, as mentioned previously, product composi-
tion needs to be well defined in future studies.
Maintenance of digestive health is the primary health positioning for both prune and plum juices,
which is often linked to fiber content in product marketing messages (Table 41.2). The European
Food Safety Authority has accepted a claim that consumption of prunes maintains normal bowel
function, with the scientific evidence considered as reflecting “prunes can contribute to normal
bowel function” [46]. However, prune juice can only use this health claim if it contains at least 3 g
fiber/100 g serving, with a note given that prune juice only contains fiber if prune pulp or puree is
present in the product [47]. General well-being claims are marketed using red/purple color of plum
and prune juice products by established associations between color and other fruits that have a high
health profile including berry, grape, pomegranate, and acai (Table 41.2).
41.4.2 Ume Juice

Ume concentrate, when added to whole blood, showed an increase in in vitro passage time, indicating
increased ability of red blood cells to deform to pass through vasculature [10]. Fractions containing
5-hydroxymethyl furfural and mumefural showed the strongest activity, with the presence of organic
acids thought to exert a synergistic effect [10]. Condensed ume fruit juice (misatol) has inhibited the
growth of HL-60 promyelocytic leukemia cell line and Kato III cells [48]. Bainiku-ekisu showed inhibi-
tion of angiotensin II–induced epidermal growth factor transactivation in vascular smooth muscle cells
prepared from rat aorta [37]. Ume fruit juice concentrate has shown in vitro antihuman influenza A
activity [49] with mumefural and derivatives showing antisalidase activity and dose-dependent growth
inhibition of virus [26]. Commercial ume concentrate, reconstituted with water (pH 2.3), has shown
dose-dependent in vitro inhibition of urease-producing bacteria, inhibition of urease activity, inhibition
of crystal formation, and crystal weight induced by Proteus mirabilis [18]. Juice that was adjusted to a
pH of 7.2 showed some but less activity [18].
Ume juice concentrate displayed an ability to reduce gastric damage from Helicobacter pylori in ani-
mals [50]. Gerbils were given H. pylori inoculum intragastrically and then ume fruit juice concentrate
dissolved at 0%, 1%, and 3% in drinking water, which was freely consumed for 9 weeks [50]. After
9 weeks, dose-dependent effects of juice concentrate were seen with significantly lower (P < 0.05) micro-
scopic scores (less severe gastritis and inflammation) in groups receiving 1 and 3% and significantly
lower (P < 0.05) mucosal hyperplasia in the 3% group. Both juice concentrate–receiving groups had
significantly lower (P < 0.05) urease A gene expression in glandular stomach. However, ume concen-
trate (Bainiku-ekisu) did not significantly reduce urea breath test values in 14 H. pylori–positive human
subjects following 2 or 12 weeks of ingesting 130 mL of 1% Bainiku-ekisu (in water) twice a day [51].
Oral consumption of ekisu (concentrated ume juice) for 2 weeks has been shown to significantly
decrease (P < 0.05) blood glucose and plasma triacylglycerols (TAG) and significantly increase (P <
0.05) plasma adiponectin concentration in adipose tissue, in insulin-resistant obese Wistar fatty rats [52].
Condensed ume fruit juice (misatol) has been tested as a daily dose in 1 dog with fibrosarcoma (1–2 g
diluted in 50–100 mL of liquid) and one human patient (2 g diluted in 50 mL water) with pancreatic
cancer [48]. Shrinking of both tumors was reported to a stage of remission.

The use of emerging processing technologies, including nonthermal processing, may better preserve the
fresh flavor, color, and nutritional content of fruit to achieve premium juice products [53,54]. However, a
plum juice made using high-pressure processing had no difference in total phenolic or carotenoid content
when compared with conventional thermally treated juice [13]. Another new technology approach, using
microwave hydrodiffusion to enhance the efficiency of extraction of phytochemicals, has also been tried
in plums [3,55]. However, juice yield was significantly lower (P < 0.05) as well as the content of phyto-
chemicals when compared with juice made using the standard technology [3,55]. Neither emerging tech-
nology has yet been reported as being commercially applied to the production of plum juice. Improved
processing technologies have been tried with ume such as freezing of fruit prior to osmotic extraction,
which has shown lowering of browning index and increasing the content of chlorogenic acid, glucose,
fructose, and citric, tartaric, and malic acids [11].
No specific phytochemicals are currently used for the promotion or differentiation of commercial
plum, prune, or ume juices. However, the use of plum juice as a source of red or purple color (anthocy-
anin) and the coloration of prune juice (using grape, blueberry, etc.) are a positioning tool (Table 41.2).
The presence of red/purple color allows products that are predominantly plum juice to associate with the
higher health profile of berry, pomegranate, and açai that is linked to the color of the product. Secondly,
plum or prune juices are commonly mixed with these red/purple fruits to leverage the higher health
profile. Although the use of antioxidant content in food product labeling and positioning is widespread
and generally nondistinguishing, some prune juice products use antioxidant content as a positioning tool
with labeling such as “naturally rich in antioxidant” and include the oxygen radical absorbance capacity
(ORAC) value on nutritional table (Table 41.2).
Enhanced antioxidant and polyphenolic rich products from plums could be used as superior health
products to target the functional food markets. The optimization of cultivars, horticultural practices, and
processing methods can be utilized to produce products with high levels of phytochemicals, with plum
juice production being advantageous due to higher fruit and juice yield than many competing fruits [56].
The inclusion of skin extracts is a trend being tried to raise the phytochemical and antioxidant content
and, thus by implication, deliver superior health benefit [12,29]. The development, testing, and position-
ing of plum and prune juice products into the bone health market is expected in the near future due to the
promising results in animal trials [40,41].
41.6 Conclusion
Plum, prune, and ume juices are acidic products with ume juice having high contents of citric and malic
acids. Fiber content is the primary nutrient content claim used for commercial prune and plum juices,
with fiber being added as an ingredient in many products. All juice types appear to have chlorogenic
acid present as a common phenolic compound, but plum juice, made from red plum, has anthocyanins,
while prune and ume juices do not. Alternatively, 5-hydroxymethyl furfural is present in prune and ume
juices, but generally absent in plum juice. Plum and prune juices are primarily promoted in terms of their
role in maintaining digestive health although the mechanism of action is not fully defined. Animal stud-
ies indicate potential benefits to bone health with clinical trials warranted. However, any future in vitro
or in vivo studies require detailed analysis of nutrient and phytochemical content of the products and
investigation of their metabolic fate as well as the biological activity of their corresponding metabolites.
The benefits of ume juice product consumption in humans are unclear and further animal and human
studies are required. Although not as studied as some other fruit juices, there appears to be significant
scope to develop the evidence and profile for positioning plum, prune, and ume juices in terms of their
health benefits.
REFERENCES
1. Topp, B.L., Russell, D.M., Neumuller, M., Dalbo, M.A., and Liu, W., Plum, in Handbook of Plant
Breeding, 1, Badenes, M.L. and Byrne, D.H., Eds., Springer, Berlin, Germany, 2012, pp. 571–621.
2. Topp, B., Noller, J., and Russell, D., Development of Prunus Mume: A New Tree Crop for Australia,
RIRDC, Canberra, Australian Capital Territory, Australia, 2007.
3. Cendres, A.l., Chemat, F., Page, D., Le Bourvellec, C., Markowski, J., Zbrzezniak, M., Renard,
C.M.G.C., and Plocharski, W., Comparison between microwave hydrodiffusion and pressing for plum
juice extraction. LWT-Food Sci. Technol., 49, 229–237, 2012.
4. Shalom, P., Gur-Arie, A.R., and Levi, A.H., Extraction of plum juice after liquefaction with commercial
enzymes. Ital. J. Food Sci., 11, 29–38, 1999.
5. Chang, T.S., Siddiq, M., Sinha, N.K., and Cash, J.N., Commercial pectinases and the yield and quality
of Stanley plum juice. J. Food Process. Pres., 19, 89–101, 1995.
6. Chang, T., Siddiq, M., Sinha, N., and Cash, J., Plum juice quality affected by enzyme treatment and
fining. J. Food Sci., 59, 1065–1069, 1994.
7. Siddiq, M. and Sultan, M., Plums and prunes, in Handbook of Fruits and Fruit Processing, Hui, Y.H.,
Barta, J., Canor, M.P., Gusek, T.W., Sidhu, J.S., and Sinha, N., Eds., Wiley-Blackwell, Oxford, U.K.,
2012, pp. 551–564.
8. Somogyi, L.P., Plums and prunes, in Processing Fruits: Science and Technology, 2nd edn., Barrett, D.,
Somogyi, L.P., and Ramasway, H., Eds., CRC Press/Taylor & Francis Group, Boca Raton, FL, 2005,
pp. 513–529.
9. Fang, T.T., Wang, D.Y., and Liaw, Y.M., Studies on preparation of Japanese apricot fruit juice. I. Effect
of different ripening stages on juice quality. Mem. Coll. Agric. Natl. Taiwan Univ., 28, 26–38, 1988.
10. Chuda, Y., Ono, H., Ohnishi-Kameyama, M., Matsumoto, K., Nagata, T., and Kikuchi, Y., Mumefural,
citric acid derivative improving blood fluidity from fruit-juice concentrate of Japanese apricot (Prunus
mume Sieb. et Zucc). J. Agric. Food Chem., 47, 828–831, 1999.
11. Chung, H.-S., Kim, D.-S., Kim, H.-S., Lee, Y.-G., and Seong, J.-H., Effect of freezing pretreatment
on the quality of juice extracted from Prunus mume fruit by osmosis with sucrose. LWT-Food Sci.
Technol., 54, 30–34, 2013.
12. de Beer, D., Steyn, N., Joubert, E., and Muller, N., Enhancing the polyphenol content of a red-fleshed
Japanese plum (Prunus salicina Lindl.) nectar by incorporating a polyphenol-rich extract from the
skins. J. Sci. Food Agric., 92, 2741–2750, 2012.
13. Gonzalez-Cebrino, F., Garcia-Parra, J., Contador, R., Tabla, R., and Ramirez, R., Effect of high-pressure
processing and thermal treatment on quality attributes and nutritional compounds of “Songold” plum
puree. J. Food Sci., 77, C866–C873, 2012.
14. Pilando, L.S. and Wrolstad, R.E., Compositional profiles of fruit juice concentrates and sweeteners.
Food Chem., 44, 19–27, 1992.
15. Will, F. and Dietrich, H., Optimised processing technique for colour- and cloud-stable plum juices and
stability of bioactive substances. Eur. Food Res. Technol., 223, 419–425, 2006.
16. Donovan, J.L., Meyer, A.S., and Waterhouse, A.L., Phenolic composition and antioxidant activity of
prunes and prune juice (Prunus domestica). J. Agric. Food Chem., 46, 1247–1252, 1998.
17. van Gorsel, H., Li, C., Kerbel, E.L., Smits, M., and Kader, A.A., Compositional characterization of
prune juice. J. Agric. Food Chem., 40, 784–789, 1992.
18. Zhu, H., Sun, X., Lu, J., Wang, M., Fang, Y., and Ge, W., The effect of plum juice on the prevention of
struvite calculus formation in vitro. Bju. Int., 110, E362–E367, 2012.
19. Kessler, T., Jansen, B., and Hesse, A., Effect of blackcurrant-, cranberry-, and plum juice consumption
on risk factors associated with kidney stone formation. Eur. J. Clin. Nutr., 56, 1020–1023, 2002.
21. Kagawa, Y., Fruits, in Standard Tables of Food Composition in Japan 2003, Kagawa Nutrition
Publishing Division, Tokyo, Japan, 2003, pp. 104–121.
22. Gebhardt, S., Cutrufelli, R., and Matthews, R., Composition of Food: Fruits and Fruit Juices, USDA,
Washington, DC, 1982.
23. Arribas-Lorenzo, G. and Morales, F.J., Estimation of dietary intake of 5-hydroxymethylfurfural and
related substances from coffee to Spanish population. Food Chem. Toxicol., 48, 644–649, 2010.
24. Prior, R.L., Wu, X.L., and Gu, L.W., Identification and urinary excretion of metabolites of
5-(hydroxymethyl)-2-furfural in human subjects following consumption of dried plums or dried plum
juice. J. Agric. Food Chem., 54, 3744–3749, 2006.
25. Wu, S.F., Chen, Y.H., Lin, C.L., Yang, J.C., Du, Y.C., Lee, J.J., Wu, Y.C., and Chang, F.R., Qualitative
and quantitative analyses of the anti-allergic constituent of commercial Prunus mume products in
Taiwan. J. Food Drug Anal., 19, 66–72, 2011.
26. Sriwilaijaroen, N., Kadowaki, A., Onishi, Y., Gato, N., Ujike, M., Odagiri, T., Tashiro, M., and Suzuki,
Y., Mumefural and related HMF derivatives from Japanese apricot fruit juice concentrate show multiple
inhibitory effects on pandemic influenza A (H1N1) virus. Food Chem., 127, 1–9, 2011.
27. Madrau, M.A., Sanguinetti, A.M., Del Caro, A., Fadda, C., and Piga, A., Contribution of melanoidins to
the antioxidant activity of prunes. J. Food Qual., 33, 155–170, 2010.
28. Posadino, A.M., Cossu, A., Piga, A., Madrau, M.A., Del Caro, A., Colombino, M., Paglietti, B.
et al., Prune melanoidins protect against oxidative stress and endothelial cell death. Front. Biosci., 3,
1034–1041, 2011.
29. Netzel, M., Fanning, K., Netzel, G., Zabaras, D., Karagianis, G., Treloar, T., Russell, D., and Stanley, R.,
Urinary excretion of antioxidants in healthy humans following Queen Garnet plum juice ingestion:
A new plum variety rich in antioxidant compounds. J. Food Biochem., 36, 159–170, 2012.
30. Netzel, M., Fanning, K., Netzel, G., Frank, T., Zabaras, D., Russell, D., and Stanley, R., Urinary phar-
macokinetics of Queen Garnet plum anthocyanins in healthy human subjects, in Emerging Trends in
Dietary Components for Preventing and Combating Disease, Patil, B.S., Jayaprakasha, G.K., Murthy,
K.N.C., and Seeram, N.P., Eds., ACS Symposium Series 1093, American Chemical Society, Washington,
DC, 2012, pp. 375–392.
31. Cevallos-Casals, B.A., Byrne, D., Okie, W.R., and Cisneros-Zevallos, L., Selecting new peach and plum
genotypes rich in phenolic compounds and enhanced functional properties. Food Chem., 96, 273–280,
2006.
32. Hernandez-Herrero, J.A. and Frutos, M.J., Degradation kinetics of pigment, colour and stability of the
antioxidant capacity in juice model systems from six anthocyanin sources. Int. J. Food Sci. Technol., 46,
2550–2557, 2012.
33. Mubarak, A., Swinny, E.E., Ching, S.Y.L., Jacob, S.R., Lacey, K., Hodgson, J.M., Croft, K.D., and
Considine, M.J., Polyphenol composition of plum selections in relation to total antioxidant capacity.
J. Agric. Food Chem., 60, 10256–10262, 2012.
34. Nunez de Gonzalez, M.T., Hafley, B.S., Boleman, R.M., Miller, R.K., Rhee, K.S., and Keeton, J.T.,
Antioxidant properties of plum concentrates and powder in precooked roast beef to reduce lipid oxida-
tion. Meat Sci., 80, 997–1004, 2008.
35. Yildiz-Turp, G. and Serdaroglu, M., Effects of using plum puree on some properties of low fat beef pat-
ties. Meat Sci., 86, 896–900, 2010.
36. Utsunomiya, H., Takekoshi, S., Gato, N., Utatsu, H., Motley, E.D., Eguchi, K., Fitzgerald, T.G., Mifune,
M., Frank, G.D., and Eguchi, S., Fruit-juice concentrate of Asian plum inhibits growth signals of vascu-
lar smooth muscle cells induced by angiotensin II. Life Sci., 72, 659–667, 2002.
37. Edenharder, R., Kurz, P., John, K., Burgard, S., and Seeger, K., In vitro effect of vegetable and fruit juices
on the mutagenicity of 2-amino-3-methylimidazo[4,5-f]quinoline, 2-amino-3,4-dimethylimidazo[4,5-
f]quinoline and 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline. Food Chem. Toxicol., 32, 443–459,
1994.
38. Ko, S.H., Choi, S.W., Ye, S.K., Cho, B.L., Kim, H.S., and Chung, M.H., Comparison of the antioxidant
activities of nine different fruits in human plasma. J. Med. Food, 8, 41–46, 2005.
39. Shukitt-Hale, B., Kalt, W., Carey, A.N., Vinqvist-Tymchuk, M., McDonald, J., and Joseph, J.A., Plum
juice, but not dried plum powder, is effective in mitigating cognitive deficits in aged rats. Nutrition, 25,
567–573, 2009.
40. Monsefi, M., Parvin, F., and Farzaneh, M., Effects of plum extract on skeletal system of fetal and new-
born mice. Med. Prin. Pract., 22, 351–356, 2013.
41. Arjmandi, B.H., Johnson, C.D., Campbell, S.C., Hooshmand, S., Chai, S.C., and Akhter, M.P.,
Combining fructooligosaccharide and dried plum has the greatest effect on restoring bone mineral
density among selected functional foods and bioactive compounds. J. Med. Food, 13, 312–319, 2010.
42. Cheskin, L., Mitola, A., Ridore, M., Kolge, S., Hwang, K., and Clark, B., A naturalistic, controlled,
crossover trial of plum juice versus Psyllium versus control for improving bowel function. Internet J.
Nutr. Wellness, 7, 2008.
43. Piirainen, L., Peuhkuri, K., Backstrom, K., Korpela, R., and Salminen, S., Prune juice has a mild laxa-
tive effect in adults with certain gastrointestinal symptoms. Nutr. Res., 27, 511–513, 2007.
44. Ring, M., Implementation of a bowel care protocol within ICU. Aust. Crit. Care, 24, 73–74, 2011.
45. Stacewicz-Sapuntzakis, M., Bowen, P.E., Hussain, E.A., Damayanti-Wood, B.I., and Farnsworth, N.R.,
Chemical composition and potential health effects of prunes: A functional food ? Crit. Rev. Food Sci.,
41, 251–286, 2001.
46. ESFA, Scientific opinion on the substantiation of health claims related to dried plums of “prune” cul-
tivars (Prunus domestica L.) and maintenance of normal bowel function (ID 1164, further assessment)
pursuant to Article 13(1) of Regulation (EC) No 1924/2006. EFSA J., 10, 2712–2729, 2012.
47. EFSA, Scientific opinion on the substantiation of health claims related to dried plums of “prune”
cultivars (Prunus domestica L.) and maintenance of normal bowel function (ID 1164) pursuant to
Article 13(1) of Regulation (EC) No 1924/2006. EFSA J., 8, 1486 [14pp.]-1486 [14pp.], 2010.
48. Adachi, M., Suzuki, Y., Mizuta, T., Osawa, T., Adachi, T., Osaka, K., Suzuki, K. et al., The “Prunus
mume Sieb. et Zucc” (Ume) is a rich natural source of novel anti-cancer substance. Int. J. Food Prop.,
10, 375–384, 2007.
49. Yingsakmongkon, S., Miyamoto, D., Sriwilaijaroen, N., Fujita, K., Matsumoto, K., Jampangern, W.,
Hiramatsu, H. et al., In vitro inhibition of human influenza a virus infection by fruit-juice concentrate of
Japanese plum (Prunus mume SIEB. et ZUCC). Biol. Pharm. Bull., 31, 511–515, 2008.
50. Otsuka, T., Tsukamoto, T., Tanaka, H., Inada, K., Utsunomiya, H., Mizoshita, T., Kumagai, T.,
Katsuyama, T., Miki, K., and Tatematsu, M., Suppressive effects of fruit-juice concentrate of Prunus
mume Sieb. et Zucc. (Japanese apricot, Ume) on Helicobacter pylori induced glandular stomach lesions
in Mongolian gerbils. Asian Pac. J. Cancer Prev., 6, 337–341, 2005.
51. Nakajima, S., Fujita, K., Inoue, Y., Nishio, M., and Seto, Y., Effect of the folk remedy, Bainiku-ekisu,
a concentrate of Prunus mume juice, on Helicobacter pylori infection in humans. Helicobacter,
11, 589–591, 2006.
52. Utsunomiya, H., Yamakawa, T., Kamei, J., Kadonosono, K., and Tanaka, S., Anti-hyperglycemic effects
of plum in a rat model of obesity and type 2 diabetes, Wistar fatty rat. Biomed. Res. (Tokyo, Japan),
26, 193–200, 2005.
53. Oey, I., Lille, M., Van Loey, A., and Hendrickx, M., Effect of high-pressure processing on colour,
texture and flavour of fruit- and vegetable-based food products: A review. Trends Food Sci. Technol.,
19, 320–328, 2008.
54. Deliza, R., Rosenthal, A., Abadio, F.B.D., Silva, C.H.O., and Castillo, C., Application of high pressure
technology in the fruit juice processing: Benefits perceived by consumers. J. Food Eng., 67, 241–246,
2005.
55. Cendres, A.L., Chemat, F., Maingonnat, J.-F.O., and Renard, C.M.G.C., An innovative process for
extraction of fruit juice using microwave heating. LWT-Food Sci. Technol., 44, 1035–1041, 2011.
56. Fanning, K., Edwards, D., Netzel, M., Stanley, R., Netzel, G., Russel, D., and Topp, B., Increasing
anthocyanin content in Queen Garnet plum and correlations with in-field measures. Acta Horticult.,
985, 97–104, 2013.
42
Pomegranate Juice
CONTENTS
42.1 Introduction.....................................................................................................................................513
42.3 Bioactives and Antioxidant Efficacy...............................................................................................515
42.3.1 Anthocyanins......................................................................................................................515
42.3.2 Ellagitannins.......................................................................................................................515
42.3.3 Other Phenolics..................................................................................................................516
42.3.4 Phenolics and Antioxidant Capacity..................................................................................517
42.3.5 Bioavailability and Antioxidant Capacity..........................................................................521
42.4 Health Effects..................................................................................................................................521
42.4.1 Cancer............................................................................................................................... 522
42.4.2 Cardiovascular Disease..................................................................................................... 522
42.4.3 Diabetes............................................................................................................................. 522
42.4.4 Erectile Dysfunction.......................................................................................................... 522
42.4.5 Potential Drug Interactions............................................................................................... 523
42.6 Conclusion..................................................................................................................................... 524
References............................................................................................................................................... 524
42.1 Introduction
The pomegranate (Punica granatum L.) fruit is linked closely to culture, health, and medicine throughout
the history of man and is mentioned in many ancient texts [1,2]. The ripe pomegranate fruit varies in size
and can exceed 12 cm in diameter with a leathery skin (also referred to as the peel or husk) and crowned
by a pointed calyx. The fruit contains many arils (the seed casings that constitute a succulent fleshy coat
containing small amounts of red juice over a hard pit/seed) separated by a white membranous pith and
pericarp. With regard to the consumed part of the pomegranate, the culinary uses of this fruit are usually
restricted to the aril, which is the edible portion. Thus, while the entire seed can be consumed, the tasty
aril is usually the desired part although the dried seeds are used as a spice in Indian and Pakistani cuisine.
In addition, a thick grenadine syrup formed from concentrated juice is also popularly consumed in various
Middle Eastern cuisines. Interestingly, while the tough leathery peel of the fruit is inedible, it is important
to note that during the large-scale commercial process of squeezing the whole fruit to produce pomegran-
ate juice, numerous chemical constituents present in the pith, membranes, and the peel of the fruit are
extracted into pomegranate juice, thus imparting biological properties to the beverage.
The biological effects of pomegranate juice are extensive and there are several subclasses of bioactive
compounds found in pomegranate juice, which are discussed herein. The bioavailability and metabolism
of pomegranate juice bioactives have been well studied in humans, and the physiological effects of pome-
granate juice constituents include their preventive potential against several chronic human diseases.
513
Thus, given the wide array of published scientific studies on pomegranate juice, it is not surprising that
this beverage has attracted immense public and research interest for its functional beverage applications,
which is the overall focus of this chapter.

The nutritional characteristics of bottled pomegranate juice (Table 42.1) are available from the U.S.
Department of Agriculture (USDA) National Nutrient Database [3]. In terms of proximate composition,
the nutritional value based on 100 g of pomegranate juice consists of 13.1 g of carbohydrate (of which
12.7 g is the total sugars, primarily as glucose and fructose, and 0.1 g is the total dietary fiber), 0.3 g of
lipid, and 0.2 g of protein. Notably, 100 g of pomegranate juice provides 54 kcal and constitutes ~86 g
water. The major minerals present in 100 g of pomegranate juice include potassium (214 mg), followed
by phosphorous and calcium (~11 mg each), sodium (9 mg), and magnesium (7 mg). In addition, folate,
vitamin C, and B vitamins (niacin, riboflavin, and thiamine) are also present in pomegranate juice.
While the flavor and sensory quality of the whole pomegranate fruit has been attributed to a combination
of sugars (glucose and fructose), organic acids (citric and malic acids), and volatiles [4], similar data are
not available for bottled commercial pomegranate juice.
TABLE 42.1
Compositional and Nutritional Characteristics of Pomegranate Juice (per 100 g)
Nutrient Unit Pomegranate Juice
Water g 85.95
Energy kcal 54
Protein g 0.15
Lipid (fat) g 0.29
Minerals
Calcium mg 11
Iron mg 0.10
Magnesium mg 7
Phosphorus mg 11
Potassium mg 214
Sodium mg 9
Zinc mg 0.09
Vitamins
Folate (DFE) µg 24
Niacin mg 0.233
Riboflavin mg 0.015
Thiamin mg 0.015
Vitamin B6 mg 0.040
Vitamin C mg 0.1
Vitamin E mg 0.38
Vitamin K µg 10.4
Database for Standard Reference, Release 26, 2013, Published online at: http://ndb.nal.
usda.gov/ndb/foods/show/2516 (accessed April 28, 2014).
Pomegranate Juice 515

There is a wide structural diversity of phytochemicals present in various parts of the whole pomegranate
fruit (peel/husk, pith, membranes, arils, and seeds), which include phenolics, steroids, and fatty acids [5].
However, in commercially produced pomegranate juice, it is mostly the phenolics that are present in the
aril (mainly anthocyanins responsible for the red color of pomegranate juice) and in the peel (mainly
ellagitannins, a subclass of hydrolyzable tannins) that are extracted into the beverage when the whole
fruit is hydrostatically pressed. Therefore, while other phytochemicals have been reported from the
whole pomegranate fruit, this chapter will only focus on those compounds that have been reported in
pomegranate juice alone, and not the whole fruit, per se.
A vast number of published studies are available regarding the identification of bioactives present
in pomegranate juice, but only those phytochemicals that have been unequivocally identified in pome-
granate juice are discussed herein. These methods of “unequivocal identification” include isolation and
structure elucidation by nuclear magnetic resonance or by methods of high-performance liquid chroma-
tography (HPLC) ultraviolet detection and liquid chromatography mass spectroscopy (LC-MS) analyses.
To date, over 40 compounds have been unequivocally identified in pomegranate juice and these are listed
in Tables 42.2 and 42.3 with their corresponding chemical structures shown in Figures 42.1 through 42.3.
For the ease of discussion, the compounds that have been reported in pomegranate juice are categorized
into the following phenolic subclasses described in the following.
42.3.1 Anthocyanins
Among the numerous phytochemicals found in pomegranate juice, the anthocyanins are probably the most
well known since these are the water-soluble pigments that impart the bright red color to the beverage.
Similar to many other plant foods, anthocyanins occur naturally in pomegranate juice as their glycosyl-
ated forms, but their deglycosylated aglycon forms, namely, anthocyanidins, have also been reported in
pomegranate juice. Notably, all of the anthocyanins in pomegranate juice are either found as mono- or
diglucosides of three common anthocyanidins: cyanidin, delphinidin, and pelargonidin. The anthocyanins
and anthocyanidins found in pomegranate juice are pelargonidin-3-glucoside, cyanidin-3-glucoside,
delphinidin-3-glucoside, pelargonidin 3,5-diglucoside, cyanidin 3,5-diglucoside, delphinidin 3,5-digluco-
side, delphinidin, and cyanidin (Figure 42.1) [6,7]. The levels of these compounds in pomegranate juice vary
based on pomegranate fruit cultivar and maturity stage, processing, and storage conditions of the beverage.
42.3.2 Ellagitannins
Ellagitannins are hydrolyzable tannins composed of esters of hexahydroxydiphenic acid (HHDP,
6,6′-dicarbonyl-2,2′,3,3′,4,4′-hexahydroxybiphenyl moiety) that hydrolyze to release ellagic acid. It is
known that ellagitannin monomers can be further oxidized to form dimers, trimers, and tetramers with
molecular weights of up to 4000 Da.
The peel of the pomegranate fruit is a rich source of ellagitannins, and therefore during the commercial
processing of pomegranate juice wherein the whole fruit is pressed, these compounds are extracted in
large quantities into the juice. The major ellagitannin present in pomegranate is punicalagin (2,3-hexa
hydroxydiphenoyl-4,6-gallagylglucose; MW = 1084 g/mol), commonly also referred to as punicalagins
(plural) since the molecule occurs as a pair of anomers, α- and β-punicalagins. Consequently, given its
high abundance in the peel of the pomegranate fruit, punicalagin is considered as a reliable chemical
marker compound for the authentication and standardization of commercially squeezed pomegranate
juice and pomegranate extracts. Moreover, the potent antioxidant properties of pomegranate juice have
been attributed to its high content of punicalagin that can reach levels exceeding 2 g/L juice [8].
In addition to punicalagin, other ellagitannins have also been reported in pomegranate juice including
punicalin, pedunculagin, lagerstannin C, sanguiin-H10, punigluconin, and granatin B (Figure 42.2)
[7,9]. In addition, ellagic acid, which is formed from the hydrolysis of ellagitannins, is also found in
pomegranate juice.
TABLE 42.2
Phytochemicals Identified in Pomegranate Juice
Molecular Molecular
Compound Number Compound Identity Formula Weight (g/mol) References
Anthocyanins
1 Delphinidin-3,5-diglucoside C27H31O17 627 [7,9]
2 Cyanidin-3,5-diglucoside C27H31O16 611 [7,9]
3 Pelargonidin-3,5-diglucoside C27H31O15 595 [7,9]
4 Delphinidin-3-glucoside C21H21O12 465 [7,9]
5 Cyanidin-3-glucoside C21H21O11 449 [7,9]
6 Pelargonidin-3-glucoside C21H21O10 433 [7,9]
7 Cyanidin-3-rutinoside C27H31O15 595 [7,9]
8 Delphinidin C15H11O7 303 [7,9]
9 Cyanidin C15H11O6 287 [7,9]
Ellagitannins and Ellagic Acid
10 Ellagic acid C14H6O8 302 [7,9]
11 Punicalagin C48H28O30 1084 [7,9]
12 Punicalin C34H22O22 782 [7,9]
13 Pedunculagin I C34H24O22 784 [7,9]
14 Lagerstannin C C27H22O19 650 [9]
15 Sanguiin H-10 C68H48O44 1568 [9]
16 Punigluconin C34H26O23 802 [7]
17 Granatin B C41H28O27 952 [7]
Flavonoids
18 (+)-Catechin C15H14O6 290 [9]
19 (+)-Gallocatechin C15H14O7 306 [9]
20 (−)-Epicatechin C15H14O6 290 [9]
21 Phloretin C15H14O5 274 [9]
22 Phlorizin C21H24O10 436 [9]
23 Kaempferol rutinoside C27H30O15 594 [9]
24 Pinocembrin C15H12O4 256 [9]
25 Phellatin C26H30O12 534 [9]
26 Amurensin C26H30O12 534 [9]
27 Procyanidin B1 C30H26O12 578 [9]
28 Procyanidin B2 C30H26O12 578 [9]
Lignans
29 Pinoresinol C20H22O6 358 [9]
30 Secoisolariciresinol C20H26O6 362 [9]
31 Cyclolariciresinol C20H24O6 360 [9]
Other Phenolics
32 Syringaldehyde C9H10O4 182 [9]
33 Feruloyl coniferin C26H30O11 518 [9]
42.3.3 Other Phenolics

Apart from the two major subclasses of phenolic compounds discussed earlier, several other subclasses
including phenolic acids, flavonols, flavanols, lignans, and condensed tannins (proanthocyanidins) have
been identified in pomegranate juice. These compounds include kaempferol rutinoside, phellatin, amu-
rensin, catechin, epicatechin, and gallocatechin, procyanidin B1, procyanidin B2, phloretin, phloridzin,
pinocembrin, vanillic acid, syringaldehyde, pinoresinol, secoisolariciresinol, cyclolariciresinol, feruloyl
coniferin, and gallic acid (Figure 42.3) [7,9].
TABLE 42.3
Organic and Phenolic Acids Identified in Pomegranate Juice
Compound Number Compound Identity Molecular Formula Molecular Weight (g/mol) References
Organic and Phenolic Acids
34 Vanillic acid C8H8O4 168 [9]
35 Gallic acid C7H6O5 170 [5,9]
36 Caffeic acid C9H8O4 180 [5]
37 Chlorogenic acid C16H18O9 345 [5]
38 Cinnamic acid C9H8O2 148 [5]
39 Citric acid C6H8O7 192 [5]
40 o-Coumaric acid C9H8O3 164 [5]
41 p-Coumaric acid C9H8O3 164 [5]
42 Ferulic acid C10H10O4 194 [5]
43 l-Malic acid C4H6O5 134 [5]
44 Oxalic acid C2H2O4 90 [5]
45 Protocatechuic acid C7H6O4 154 [5]
46 Quinic acid C7H12O6 192 [5]
47 Succinic acid C4H6O4 118 [5]
48 Tartaric acid C4H6O6 150 [5]
R1
R2 R1
+
HO O R2
R3
+
HO OH HO O R3
O OH
OH O HO
HO O O HO OH
O OH
HO O
OH OH HO
1 R1—
— OH, R2—
— OH, R3—
—OH 4 R1—
— OH, R2—
— OH, R3—
—OH
1— OH, R2—OH, R3— H
2R — — — — OH, R2—
5 R1— —OH, R3——H
1 2
— H, R —— OH, R3—
—H
3 R1— H, R2— OH, R3—H
— — — 6R —
OH
OH R1
+
HO O R2
+
HO HO O
OH R3
O OH
O
OH O OH
O OH
HO
HO OH 8 R1—
— OH, R2—— OH, R3—
—OH
7 1— 2— 3—
9 R — OH, R —OH, R — H
FIGURE 42.1 Anthocyanins identified in pomegranate juice.
42.3.4 Phenolics and Antioxidant Capacity

Pomegranate juice contains a diverse group of phenolic compounds and there is immense variabil-
ity in phenolic content of various commercial brands of the beverage as shown in Table 42.4 [7,10].
For instance, pomegranate juice can vary in total phenolic content from 574 to 2520 mg gallic acid
equivalents (GAE)/L, and among these compounds, anthocyanins varied between 20% and 82% of the
total phenolic content, while phenolic acids and ellagitannins can vary from 16.4% to 65.8% [7].
HO OH
HO
O HO OH
HO
O HO
HO
HO O
O
O O
O HO
OH
O O OH O
O O
HO OH O O
O OH
O O O
HO O OH
O HO OH HO
O
HO OH
HO OH HO
O
HO OH HO OH HO
Punicalagin Punicalin
OH OH
HO HO
O
HO HO O
O
OH O O
HO O OH
O O
COOH HO O
O O
O O O
HO OH O
O HO O
OH
OH
OH
HO OH
HO OH OH
OH HO OH HO OH
Punigluconin Pedunculagin I
HO OH HO OH
HO OH
O
O O O OH
OH O
O
HO OH
O OH
O
OH
HO O OH
O O
O O O
O OH O
HO HO
COOH
OH HO O
O
HO
HO O OH
OH HO HO
Lagerstannin C Granatin B
FIGURE 42.2 Ellagitannins identified in pomegranate juice. (Continued)

HO OH
HO HO
OH
OH
HO O
HO
O O OH
HO O O
O
HO O
O O O
O O HO O O
O O O O
HO HO
OH HO OH
O
HO
OH HO
OH OH HO OH
OH
Sanguiin H10
O
O OH
HO OH
HO O
O
Ellagic acid
FIGURE 42.2 (Continued ) Ellagitannins identified in pomegranate juice.
Among the diverse subclasses of phenolic compounds found in pomegranate juice, the major con-
tributors to its antioxidant activity are its ellagitannins. In fact, it has been reported that ellagitannins
account for more than 92% of the antioxidant activity of pomegranate juice with punicalagin levels alone
exceeding 2 g/L of juice [8]. This is because of the natural abundance of ellagitannins present in the peel,
membranes, and pith of the fruit. Therefore, when pomegranate juice is obtained by hydrostatic press-
ing of the whole pomegranate fruit, which is commonly used for the large-scale commercial production
of the beverage, the phenolic composition, and hence the antioxidant capacity of pomegranate juice,
R
OH OH OH
HO O HO O HO OH
OH OH
OH OH
OH OH OR O
18 R—H 20 21 R—H
19 R—OH 22 R— Glu
OH
HO O
HO
O OH
OH HO O
O O
OH O
O
HO
HO OH OH O
23 24
FIGURE 42.3 Other phenolic compounds identified in pomegranate juice (refer to Tables 42.2 and 42.3 for compound
numbers). (Continued)
OH
OH R2
HO O O O OH
HO
OH O
R1 OH H3CO OCH3
OH O OH O
25 R1— X, R2— H HO
26 R1—H, R2 — X X 29
H3CO
OH H3CO
OH
HO
HO
OH
OH
OCH3
OCH3
OH
OH
30
31
H3CO OH
H3CO OH
HO O O
HO O
OH
O
33
O OH O H O OH
OCH3 H3CO OCH3 HO OH

OH OH OH
34 32 35
OH OH
HO O HO O
OH OH
OH
HO OH OH OH
OH OH
O OH HO O
OH
HO OH
OH OH
27 28
FIGURE 42.3 (Continued ) Other phenolic compounds identified in pomegranate juice (refer to Tables 42.2 and 42.3 for
compound numbers).
increases, compared to juice obtained from the arils alone [8,11]. However, the levels of ellagitannins in
pomegranate juice are highly variable depending on fruit maturity and cultivar and processing and stor-
age conditions [12]. All of these factors influence the overall total antioxidant capacity of pomegranate
juice and could explain the wide variation in reported antioxidant values of different commercial brands
of the pomegranate juice beverage [10]. Nevertheless, despite all of these factors, reputable commer-
cial brands of authentic pomegranate juice have been shown to contain a higher phenolic content and
TABLE 42.4
Contents of Individual Phenolic Compounds in Pomegranate Juice
Unit Concentration (Range) References
Anthocyanins
Delphinidin-3,5-diglucoside mg/L 15.5–1120 [7]
Cyanidin-3,5-diglucoside mg/L 3.0–603 [7]
Pelargonidin-3,5- mg/L 1.32–700 [7]
diglucoside
Delphinidin-3-glucoside mg/L 2.49–370 [7]
Cyanidin-3-glucoside mg/L 2.2–221 [7]
Pelargonidin-3-glucoside mg/L 1.4–4.2 [7]
Cyanidin-3-rutinoside mg/L 0.2–8.2 [7]
Total Anthocyanins mg CCE/L 117–1956 [7]
Ellagitannins and Ellagic Acid
Ellagic acid mg/L 140–490 [7]
Punicalagins mg/L 22.8–1353 [8]
Pedunculagin I mg/L 6.2–115 [7]
Punigluconin mg/L 5.9–55.5 [7]
Granatin B mg/L 4.6–34.5 [7]
Flavonoids
Phlorizin mg/L 3.4–34 [7]
Total Flavonoids mg CE/L 35–319 [7]
Total Phenolics mg GAE/L 574–2520 [7]
Abbreviations: CCE, cyaniding chloride equivalents; CE, catechin equivalents; GAE, gallic acid equivalents.
antioxidant capacity compared to other commonly consumed polyphenol-rich ready-to-drink beverages

in the United States [13]. Moreover, the antioxidant capacity of pomegranate juice obtained from whole
fruit was shown to be threefold higher compared to red wine and green tea based on the evaluation of
the free-radical scavenging and iron-reducing capacity of the beverages [8]. Pomegranate juice has also
been shown to have significantly higher antioxidant level in comparison to other commonly consumed
fruit juices such as grape, cranberry, grapefruit, and orange [14].
42.3.5 Bioavailability and Antioxidant Capacity

Bioavailability studies have been conducted in human subjects with pomegranate phenolics [15–17].
Notably, the native pomegranate ellagitannins, including punicalagin, are not detected intact in circulation
but instead hydrolyze to release ellagic acid in plasma and are then further converted by gut microflora into
metabolites known as urolithins. These urolithins persist in circulation but do not show a high antioxidant
capacity in vivo [15]. Interestingly, a study with dietary supplementation of 800 mg of a pomegranate extract
in 11 healthy volunteers increased the plasma antioxidant capacity significantly by 32% at 0.5 h postpran-
dial, but no positive effects were observed on the generation of reactive oxygen species and biomarkers of
inflammation [17]. On the basis of the published human studies conducted thus far with pomegranate juice,
the estimation of the bioavailability of its polyphenols is affected by several factors including interindi-
vidual variability. However, it is still unclear whether the in vivo derived metabolites of pomegranate juice
compounds enhance plasma antioxidant status significantly beyond the body’s endogenous antioxidants.
42.4 Health Effects

In the past decade, numerous studies have been published on the biological properties of pomegranate
constituents including the prevention and treatment of certain cancers, cardiovascular disease (CVD),
type 2 diabetes, metabolic syndrome, obesity, dental conditions, erectile dysfunction (ED), bacterial
infections and antibiotic resistance, ultraviolet radiation–induced skin damage, infant brain ischemia,
male infertility, Alzheimer’s disease, and arthritis, among others. Consequently, there is a plethora of
review articles available on these various subject areas [1,2,18–26], and this extensive body of literature
covers laboratory, preclinical, and human clinical studies with not only pomegranate juice but also pome-
granate extracts and purified constituents found in the whole fruit. Therefore, this chapter will only focus
on human clinical trials conducted with pomegranate juice alone. Indeed, it is impressive that the pome-
granate juice beverage has human clinical data to support its biological effects and health benefits that has
largely contributed toward its increased consumption, worldwide popularity, and functional applications.
42.4.1 Cancer
Among several types of cancers, prostate cancer has been the most well studied form of cancer in human
subjects with pomegranate juice. A single-arm phase II clinical trial in men with recurrent prostate
cancer and rising prostate-specific antigen (PSA) level showed delayed PSA doubling time after daily
consumption of pomegranate juice (240 mL containing 570 mg polyphenol GAE) [27]. However, a
phase IIb, double-blind placebo-controlled randomized study with pomegranate juice (500 mL contain-
ing 1147 mg polyphenol GAE) in men with advanced prostate cancer did not show any effects on PSA
levels [28]. Whether the differences in the findings of these two human clinical studies are because of
the levels of bioactives present in the different pomegranate juice study materials or due to differences
in bioavailability and metabolism among subjects is not known. However, it is noteworthy that the ella-
gitannin microbial metabolites, namely, the urolithins (discussed in Section 42.3.5), have been detected
in the prostate tissues of subjects after pomegranate juice (200 mL/day) consumption [29]. The tissue
disposition of pomegranate juice metabolites in the prostate gland has also been recently confirmed in
a separate study after the delivery of a pomegranate extract in prostate cancer patients before radical
prostatectomy [30]. Overall, based on the available human clinical data and the fact that pomegranate
juice–derived metabolites localize to the prostate gland, it is apparent that this beverage holds great
promise against prostate cancer, but further studies are still necessary to confirm this.
42.4.2 Cardiovascular Disease

Several studies have evaluated the effects of pomegranate juice on CVD endpoints in human subjects
[31–34]. For instance, pomegranate juice consumption (50 mL/day containing 1979 mg/L ellagitannins)
by 10 patients with carotid artery stenosis for 1 year led to a significant reduction in intima-media thick-
ness, systolic blood pressure, and serum lipid peroxidation [32]. Similarly, consumption of pomegranate
juice (50 mL/day) for 2 weeks reduced systolic blood pressure and serum angiotensin-converting enzyme
activity in 10 hypertensive patients [33]. While there have not been many double-armed clinical studies
with pomegranate juice, consumption of 240 mL of the beverage in a 3-month randomized, double-
blind, placebo-controlled study with 45 patients with incident myocardial ischemia resulted in a signifi-
cant decrease in stress-induced ischemia in comparison to placebo [34]. Therefore, the current published
data in human subjects support the positive effects of pomegranate juice against CVD.
42.4.3 Diabetes
Pomegranate juice has been shown to improve cholesterol and lipid profiles in type 2 diabetic patients
with hyperlipidemia after consumption of concentrated pomegranate juice (40 g) for 8 weeks. The patients
showed statistically significant decreases in total/low-density lipoprotein (LDL) cholesterol ratio, total/
high-density lipoprotein (HDL) cholesterol ratio, and LDL/HDL ratio [35,36]. These studies suggest
that pomegranate juice consumption could modify heart disease risk factors in hyperlipidemic patients.
42.4.4 Erectile Dysfunction

While there are a few available animal studies that have evaluated the effects of pomegranate juice on
ED, thus far, there has been only one randomized double-blind, placebo-controlled human clinical trial
to evaluate the effects of pomegranate juice on ED [37]. In this 10-week crossover study, 53 men (mean
age 46) received either pomegranate juice or placebo and were assessed via the International Index of
Erectile Function, and while statistical significance was not reached, there was a trend toward improve-
ment in ED in this cohort.
42.4.5 Potential Drug Interactions

Given the wide consumption of pomegranate juice, whether potential food–drug interaction exists has
attracted research attention. To date, there are two animal studies suggesting that there could be potential
interactions between pomegranate juice and drugs metabolized by human cytochrome P450 3A enzymes
that would affect the pharmacokinetics of such drugs including tolbutamide and carbamazepine [38,39].
However, a randomized single-dose crossover study with pomegranate juice and grapefruit juice in 13
healthy human volunteers demonstrated that pomegranate juice did not affect elimination half-life or dis-
tribution of intravenous midazolam (a benzodiazepine derivative), nor did it affect the Cmax or clearance
of the drug [40]. However, in that study grapefruit juice impaired clearance and elevated plasma levels
of the drug. Therefore, while there are observed interactions between drugs and some beverages such as
grapefruit juice, this human study contradicts the published animal studies and suggests that there are no
observed drug interactions with pomegranate juice.

While the edible part of the pomegranate fruit (arils) is widely consumed, the juice of this fruit can be
obtained either by squeezing the edible arils alone or by squeezing the whole fruit (peel included). While
it may be a common household practice to obtain juice from the arils, on an industrial commercial scale,
pomegranate juice is obtained from squeezing the whole fruit whereby bioactive ellagitannin constituents
naturally present in the peel are extracted into juice. The effects of the peel constituents on taste (result-
ing in increased astringency) and increased phenolic and antioxidant levels have been shown by different
groups [8,11]. Overall, based on scientific studies, squeezing of the whole fruit to yield pomegranate juice
would be more desirable from a human health perspective given the potency of the bioactives naturally
present in the peel of the fruit. However, given the premium price of pomegranate juice, “counterfeiting”
or adulteration of this beverage with other less expensive juices including apple juice, or with natural
grape pigments to imitate color, has been observed, but there are published methods to authenticate pure
pomegranate juice [41].
Apart from the two types of pomegranate juice discussed earlier, other formulations that are available
are pomegranate wine [11], which has been shown to have more protective effects against the oxidation
of LDL compared to pure red wine alone [42]. Notably, in a recent double-blind, randomized, placebo-
controlled study, a pomegranate vinegar beverage was shown to exert beneficial effects on adiposity by
reducing visceral fat accumulation in overweight women [43]. Another novel pomegranate formulation is
dealcoholized red wine containing pomegranate extracts [44]. Interestingly, the addition of pomegranate
extracts to dealcoholized red wine resulted in a product with more intense yeast odor and flavor, acidity,
and astringency and with a less intense berry flavor, which was well received by consumers [44].
A recent study explored the functional applications of organic pomegranate juice subjected to lactic
acid fermentation by Lactobacillus plantarum [45]. The authors found increased phenolic content and
antioxidant activity in the fermented pomegranate juice and proposed this method as a novel technology
option for the utilization of this beverage. Similarly, pomegranate juice subjected to alcoholic and acetic
acid fermentation did not greatly affect the total phenolic content of the juice, but led to some variability
in the contents of total anthocyanins and individual phenolic acid derivatives [46].
Apart from the pomegranate juice beverages described earlier, arguably, the greatest future trend of
pomegranate lies in the commercial potential of botanical extracts derived from the pressed fruit cake
remaining after pomegranate juice production [47]. Moreover, the utilization of pomegranate extracts
as dietary supplements is rapidly increasing in popularity among consumers. While these products are
beyond the scope of this chapter, it is noteworthy that there is a large and growing body of studies
supporting the biological effects and health benefits of pomegranate extracts and its purified constitu-
ents, including punicalagin and ellagic acid. Therefore, it is possible that these studies could also be
extrapolated to pomegranate juice.
42.6 Conclusion
Pomegranate juice is a highly regarded functional beverage due to its bioactive compounds including
anthocyanins and ellagitannins. Overall, the multitude of phytochemicals found in pomegranate juice act
together with greater potency than any single purified constituent alone. With regard to the production
of pomegranate juice, it is important to note that due to the natural abundance of ellagitannins present
in the fruit peel, pomegranate juice obtained from squeezing the whole fruit will have greater phenolic
content and antioxidant capacity than pomegranate juice obtained from the arils alone. Impressively,
compared to other functional beverages, pomegranate juice has a significant number of human clinical
studies to support its health benefits in various areas including cancer, diabetes, and heart disease. Apart
from these studies, there is also a large and growing body of published data with pomegranate extracts
and purified constituents in laboratory and preclinical animal models in various health areas including
neurodegenerative diseases and inflammation. The bioavailability and metabolism of pomegranate juice
bioactives, namely, ellagitannins, has been well studied and established in human subjects with consider-
able observed interindividual variability. Overall, it is apparent that pomegranate juice will continue on
its positive trajectory and high regard among consumers as a niche functional beverage.
REFERENCES
1. Schulman, R.N., Summary, in Pomegranates: Ancient Roots to Modern Medicine, Seeram, N.P.,
Schulman, R.N., and Heber, D., Eds., CRC Press/Taylor & Francis Group, New York, 2006, pp. 223–226.
2. Jurenka, J.S., Therapeutic applications of pomegranate (Punica granatum L.): A review. Altern. Med.
Rev., 13, 128–144, 2008.
3. U.S. Department of Agriculture (USDA), USDA National Nutrient Database for Standard Reference, Release
26, 2013. Published online at: http://ndb.nal.usda.gov/ndb/foods/show/2516 (accessed April 28, 2014).
4. Mayuoni-Kirshinbaum, L. and Porat, R., The flavor of pomegranate fruit: A review. J. Sci. Food Agric.,
94, 21–27, 2014.
5. Seeram, N.P., Zhang, Y., Reed, J.D., and Vaya, J., Pomegranate phytochemicals, in Pomegranates:
Ancient Roots to Modern Medicine, Seeram, N.P., Schulman, R.N., and Heber, D., Eds., CRC Press/
Taylor & Francis Group, New York, 2006, pp. 3–29.
6. Hernández, F., Melgarejo, P., Tomás-Barberán, F.A., and Artés, F., Evolution of juice anthocyanins
during ripening of new selected pomegranate (Punica granatum) clones. Eur. Food Res. Technol., 210,
39–42, 1999.
7. Gómez-Caravaca, A.M., Verardo, V., Toselli, M., Segura-Carretero, A., Fernández-Gutiérrez, A., and
Caboni, M.F., Determination of the major phenolic compounds in pomegranate juices by HPLC-DAD-
ESI- MS. J. Agric. Food Chem., 61, 5328–5337, 2013.
8. Gil, M.I., Tomás-Barberán, F.A., Hess-Pierce, B., Holcroft, D.M., and Kader, A.A., Antioxidant activ-
ity of pomegranate juice and its relationship with phenolic composition and processing. J. Agric. Food
Chem., 48, 4581–4589, 2000.
9. Mena, P., Calani, L., Dall’Asta, C., Galaverna, G., García-Viguera, C., Bruni, R., Crozier, A., and
Rio, D.D., Rapid and comprehensive evaluation of (poly)phenolic compounds in pomegranate (Punica
granatum L.) juice by UHPLC-MSn. Molecules, 17, 14821–14840, 2012.
10. Borges, G., Mullen, W., and Crozier, A., Comparison of the polyphenolic composition and antioxidant
activity of European commercial fruit juices. Food Funct., 1, 73–83, 2010.
11. Wasila, H., Li, X., Liu, L., Ahmad, I., and Ahmad, S., Peel effects on phenolic composition, antioxidant
activity, and making of pomegranate juice and wine. J. Food Sci., 78, C1166–C1172, 2013.
12. El Kar, C., Ferchichi, A., Attia, F., and Bouajila, J., Pomegranate (Punica granatum) juices: Chemical
composition, micronutrient cations, and antioxidant capacity. J. Food Sci., 76, C795–C800, 2011.
13. Seeram, N.P., Aviram, M., Zhang, Y., Henning, S.M., Feng, L., Dreher, M., and Heber, D., Comparison
of antioxidant potency of commonly consumed polyphenol-rich beverages in the United States. J. Agric.
Food Chem., 56, 1415–1422, 2008.
14. Rosenblat, M. and Aviram, M., Antioxidative properties of pomegranate: In vitro studies, in
Pomegranates: Ancient Roots to Modern Medicine, Seeram, N.P., Schulman, R.N., and Heber, D., Eds.,
CRC Press/Taylor & Francis Group, New York, 2006, pp. 31–43.
15. Cerdá, B., Espín, J.C., Parra, S., Martínez, P., and Tomás-Barberán, F.A., The potent in vitro anti-
oxidant ellagitannins from pomegranate juice are metabolized into bioavailable but poor antioxidant
hydroxy-6H-dibenzopyran-6-one derivatives by the colonic microflora of healthy humans. Eur. J. Nutr.,
43, 205–220, 2004.
16. Seeram, N.P., Henning, S.M., Zhang, Y., Suchard, M., Li, Z., and Heber, D., Pomegranate juice ellagi-
tannin metabolites are present in human plasma and some persist in urine for up to 48 hours. J. Nutr.,
136, 2481–2485, 2006.
17. Mertens-Talcott, S.U., Jilma-Stohlawetz, P., Rios, J., Hingorani, L., and Derendorf, H., Absorption,
metabolism, and antioxidant effects of pomegranate (Punica granatum L.) polyphenols after ingestion
of a standardized extract in healthy human volunteers. J. Agric. Food Chem., 54, 8956–8961, 2006.
18. Basu, A. and Penugonda, K., Pomegranate juice: A heart-healthy fruit juice. Nutr. Rev., 67, 49–56,
2009.
19. Banihani, S., Swedan, S., and Alguraan, Z., Pomegranate and type 2 diabetes. Nutr. Res., 33, 341–348,
2013.
20. Aviram, M. and Rosenblat, M., Pomegranate protection against cardiovascular diseases. Evid. Based
Complement. Alternat. Med., doi: 10.1155/2012/382763, 2012.
21. Stowe, C.B., The effects of pomegranate juice consumption on blood pressure and cardiovascular health.
Complement. Ther. Clin. Pract., 17, 113–115, 2011.
22. Medjakovic, S. and Jungbauer, A., Pomegranate: A fruit that ameliorates metabolic syndrome. Food
Funct., 4, 19–39, 2013.
23. Zarfeshany, A., Asgary, S., and Javanmard, S.H., Potent health effects of pomegranate. Adv. Biomed.
Res., 3, 100, 2014.
24. Al-Muammar, M.N. and Khan, F., Obesity: The preventive role of the pomegranate (Punica granatum).
Nutrition, 28, 595–604, 2012.
25. Adhami, V.M., Khan, N., and Mukhtar, H., Cancer chemoprevention by pomegranate: Laboratory and
clinical evidence. Nutr. Cancer, 61, 811–815, 2009.
26. Lansky, E.P. and Newman, R.A., Punica granatum (pomegranate) and its potential for prevention and
treatment of inflammation and cancer. J. Ethnopharmacol., 109, 177–206, 2007.
27. Pantuck, A.J., Leppert, J.T., Zomorodian, N., Aronson, W., Hong, J., Barnard, R.J., Seeram, N.P. et al.,
Phase II study of pomegranate juice for men with rising prostate-specific antigen following surgery or
radiation for prostate cancer. Clin. Cancer Res., 12, 4018–4026, 2006.
28. Stenner-Liewen, F., Liewen, H., Cathomas, R., Renner, C., Petrausch, U., Sulser, T., Spanaus, K. et al.,
Daily pomegranate intake has no impact on PSA levels in patients with advanced prostate cancer-
Results of a phase IIb randomized controlled trial. J. Cancer, 4, 597–605, 2013.
29. González-Sarrías, A., Giménez-Bastida, J.A., García-Conesa, M.T., Gómez-Sánchez, M.B., García-
Talavera, N.V., Gil-Izquierdo, A., Sánchez-Alvarez, C. et al., Occurrence of urolithins, gut microbiota
ellagic acid metabolites and proliferation markers expression response in the human prostate gland upon
consumption of walnuts and pomegranate juice. Mol. Nutr. Food Res., 54, 311–322, 2010.
30. Freedland, S.J., Carducci, M., Kroeger, N., Partin, A., Rao, J.Y., Jin, Y., Kerkoutian, S. et al., A double-
blind, randomized, neoadjuvant study of the tissue effects of POMx pills in men with prostate cancer
before radical prostatectomy. Cancer Prev. Res. (Phila), 6, 1120–1127, 2013.
31. Aviram, M., Dornfeld, L., Rosenblat, M., Volkova, N., Kaplan, M., Coleman, R., Hayek, T., Presser, D.,
and Fuhrman, B., Pomegranate juice consumption reduces oxidative stress, atherogenic modifications
to LDL, and platelet aggregations: Studies in humans and in atherosclerotic apolipoprotein E-deficient
mice. Am. J. Clin. Nutr., 71, 1062–1076, 2000.
32. Aviram, M., Rosenblat, M., Gaitini, D., Nitecki, S., Hoffman, A., Dornfeld, L., Volkova, N. et al.,
Pomegranate juice consumption for 3 years by patients with carotid artery stenosis reduces common
carotid intima-media thickness, blood pressure and LDL oxidation. Clin. Nutr., 23, 423–433, 2004.
33. Aviram, M. and Dornfeld, L., Pomegranate juice consumption inhibits serum angiotensin converting
enzyme activity and reduces systolic blood pressure. Atherosclerosis, 158, 195–198, 2001.
34. Sumner, M.D., Elliott-Eller, M., Weidner, G., Daubenmier, J.J., Chew, M.H., Marlin, R., Raisin, C.J.,
and Ornish, D., Effects of pomegranate juice consumption on myocardial perfusion in patients with
coronary heart disease. Am. J. Cardiol., 96, 810–814, 2005.
35. Esmaillzadeh, A., Tahbaz, F., Gaieni, I., Alavi-Majd, H., and Azadbakht, L., Cholesterol-lowering effect
of concentrated pomegranate juice consumption in type II diabetic patients with hyperlipidemia. Int. J.
Vitam. Nutr. Res., 76, 147–151, 2006.
36. Esmaillzadeh, A., Tahbaz, F., Gaieni, I., Alavi-Majd, H., and Azadbakht, L., Concentrated pomegranate
juice improves lipid profiles in diabetic patients with hyperlipidemia. J. Med. Food, 7, 305–308, 2004.
37. Forest, C.P., Padma-Nathan, H., and Liker, H.R., Efficacy and safety of pomegranate juice on improve-
ment of erectile dysfunction in male patients with mild to moderate erectile dysfunction: A randomized,
placebo-controlled, double-blind, crossover study. Int. J. Impot. Res., 19, 564–567, 2007.
38. Nagata, M., Hidaka, M., Sekiya, H., Kawano, Y., Yamasaki, K., Okumura, M., and Arimori, K., Effects
of pomegranate juice on human cytochrome P450 2C9 and tolbutamide pharmacokinetics in rats. Drug
Metab. Dispos., 35, 302–305, 2007.
39. Hidaka, M., Okumura, M., Fujita, K., Ogikubo, T., Yamasaki, K., Iwakiri, T., Setoguchi, N., and
Arimori, K., Effects of pomegranate juice on human cytochrome p450 3A (CYP3A) and carbamazepine
pharmacokinetics in rats. Drug Metab. Dispos., 33, 644–648, 2005.
40. Farkas, D., Oleson, L.E., Zhao, Y., Harmatz, J.S., Zinny, M.A., Court, M.H., and Greenblatt, D.J.,
Pomegranate juice does not impair clearance of oral or intravenous midazolam, a probe for cytochrome
P450–3A activity: Comparison with grapefruit juice. J. Clin. Pharmacol., 47, 286–294, 2007.
41. Zhang, Y., Krueger, D., Durst, R., Lee, R., Wang, D., Seeram, N.P., and Heber, D., International multidi-
mensional authenticity specification (IMAS) algorithm for detection of commercial pomegranate juice
adulteration. J. Agric. Food Chem., 57, 2550–2557, 2009.
42. Sezer, E.D., Akçay, Y.D., Ilanbey, B., Yildirim, H.K., and Sözmen, E.Y., Pomegranate wine has greater
protection capacity than red wine on low-density lipoprotein oxidation. J. Med. Food, 10, 371–374,
2007.
43. Park, J.E., Kim, J.Y., Kim, J., Kim, Y.J., Kim, M.J., Kwon, S.W., and Kwon, O., Pomegranate vinegar
beverage reduces visceral fat accumulation in association with AMPK activation in overweight women:
A double-blind, randomized, and placebo-controlled trial. J. Funct. Foods, 8, 274–281, 2014.
44. Tárrega, M.A., Varela, P., Fromentin, E., Feuillère, N., Issaly, N., Roller, M., Sanz-Buenhombre, M.
et al., Specific phenolic compounds and sensory properties of a new dealcoholized red wine with pome-
granate (Punica granatum L.) extract. Food Sci. Technol. Int., doi: 10.1177/1082013213489128, 2013.
45. Filannino, P., Azzi, L., Cavoski, I., Vincentini, O., Rizzello, C.G., Gobbetti, M., and Di Cagno, R.,
Exploitation of the health-promoting and sensory properties of organic pomegranate (Punica granatum
L.) juice through lactic acid fermentation. Int. J. Food Microbiol., 163, 184–192, 2013.
46. Ordoudi, S.A., Mantzouridou, F., Daftsiou, E., Malo, C., Hatzidimitriou, E., Nenadis, N., and Tsimidou,
M.Z., Pomegranate juice functional constituents after alcoholic and acetic acid fermentation. J. Funct.
Foods, 8, 161–168, 2014.
47. Seeram, N.P., Zhang, Y., and Heber, D., Commercialization of pomegranates: Fresh fruit, beverages
and botanical extracts, in Pomegranates: Ancient Roots to Modern Medicine, Seeram, N.P., Schulman,
R.N., and Heber, D., Eds., CRC Press/Taylor & Francis Group, New York, 2006, pp. 187–196.
43
Raspberry Juice
Bradley W. Bolling, Diana M. DiMarco, Katherine Lainas, and Sarah Kranz
CONTENTS
43.1 Introduction................................................................................................................................... 527
43.3.1 Polyphenol Content Reported in the Literature............................................................... 529
43.3.2 Polyphenol Databank Values.............................................................................................531
43.3.3 Antioxidants and Bioactives Responsible for Antioxidant Activity of Raspberry Juice..........531
43.4 Health Effects.................................................................................................................................533
43.4.1 Bioavailability of Bioactives from Raspberry Juice..........................................................533
43.4.2 Results of Human Intervention Studies with Raspberry Juice..........................................533
43.4.3 Results of Animal Studies with Raspberry Juice............................................................. 534
43.6 Conclusion..................................................................................................................................... 536
References............................................................................................................................................... 536
43.1 Introduction
The raspberry belongs to the Rubus genus, which includes a variety of edible berries. This chapter will focus
on juice from the red raspberry (Rubus idaeus L.) and black raspberry (R. occidentalis L.), which are culti-
vated and consumed in many parts of the world. Boysenberry, loganberry, and tayberry which are hybridiza-
tions of blackberry and red raspberry cultivars, belong to the Rubus genus and are reviewed elsewhere [1].
Worldwide raspberry production averaged 582,000 tons from 2010 to 2012 [2]. Europe produces 75%
of the world raspberry supply, while the Americas produce 22% [2]. The red raspberry is more widely
cultivated and consumed than the black raspberry [3]. The cultivated raspberry is consumed fresh, fur-
ther processed for juicing or jam production, or individually quick frozen. In the United States, per capita
consumption of frozen raspberry was 168 g/year, a third behind the strawberry and blueberry, and in
2011, U.S. consumers used 141 g per capita of fresh raspberries [4].
Due to its rich polyphenol content and associated health benefits, raspberry juice is often incorporated
into functional foods and beverages. In Europe, the estimated anthocyanin intake is 20–65 mg/day [5].
Thus, even modest incorporation of raspberry juice into the diet could significantly increase estimated
anthocyanin consumption. In addition to anthocyanins, raspberry is a source of ellagitannins, vitamin
C, and other nutrients. This chapter highlights the nutritional characteristics, antioxidant and polyphenol
content, health effects, and novel formulations of raspberry juice.

Due to its tart flavor, raspberry juice is most often consumed as a component of juice blends [3]. In the
United States, raspberry juice is available in blends of apple, grape, or berry juice. The nutrient content
of raw raspberry and representative raspberry juice blends is listed in Table 43.1.
527
TABLE 43.1
Compositional and Nutritional Characteristics of Raspberry Juice Blends and Raw Raspberry
in the United States
Raspberry Juice Blends Fresh Red Raspberry Juice Raw Red Raspberry
Nutrient Unit (per 237 mL, 8 fl oz) [6,7] (per 237 mL, 8 fl oz) [7–9] (per 1 Cup, ~123 g) [10]
Energy kcal 110–150 53 64
Protein g 0 0.05 1
Lipid (fat) g 0 0.58 1
Carbohydrate g 28–38 13.4 15
Total sugars g 27–37 nr 5
Total dietary fiber g 0 4.3 8
Minerals
Iron mg 0.36 0.8 0.85
Magnesium mg nr nr 27
Potassium mg 25–280 219 186
Sodium mg 5–70 0 1
Vitamins
Folate DFE nr nr 26
Vitamin A IU 0 187 41
Vitamin C mg 60–72 36 32
Vitamin E mg nr nr 1
Abbreviations: DFE, dietary folate equivalents; nr, not reported; IU, international unit.
Nutrient databanks do not differentiate the nutrient content of red and black raspberries. However, studies
involving these berries have found only minor differences in most nutrients examined [11–14]. Differences
in nutrient content between berry juices could be related to juice yields. Fresh red and black raspberry yield
approximately 50% juice, compared with currant and bilberry, which yield 31%–67% juice [13].
Raspberry juice contains a variety of minerals of importance. Black raspberry juice has 0.19 mg
iron/100 g (dry weight [DW] taken prior to juicing) and 0.14 mg copper/100 g [13]. These values are
marginally higher than red raspberry juice. Magnesium is also slightly higher in black raspberry than
red raspberry juice, with a content of 13 mg/100 g berry versus 10 mg in red raspberry. Red and black
raspberry has 25–35 mg calcium and 146–178 mg potassium/100 g berry, which is similar to that found
in other Rubus berries. Rubus berries are also an excellent source of manganese; red raspberry juice has
0.49–0.67 mg/100 g fresh weight (FW) [13]. Fresh raspberry also contains zinc, strontium, and boron,
at 0.760, 0.150, and 0.025 mg/kg, respectively [11]. Compared to green and black teas, red chicory, cran-
berry, and mushroom, raspberry is the richest source of strontium and boron [11].
Red and black raspberry is an excellent source of vitamin C, having 32 mg/100 g berry (Table 43.1).
A study of raspberry cultivars grown in Italy reported a range of 12–29 mg/100 mg berry, with an
average value of 18 mg/100 g [15]. Vitamin C is lost during juice processing; raspberry juice blends are
typically fortified with ascorbic acid.
Raspberry has 15 g carbohydrate/100 g edible fruit (Table 43.1). Raspberry sugar content is predomi-
nately fructose and glucose, with a lesser amount of sucrose [11,12]. Raspberry has 5.6–7.9 mg total
sugars/100 g berry, of which fructose ranges from 2.5 to 3.5 mg, glucose from 2.1 to 2.8 mg, and sucrose
from 0.8 to 1.3 mg [12]. A similar glucose/fructose/sucrose ratio was also observed in fresh raspberry
juice [14]. Notably, raspberry juice blends contain sugars from other fruits or in the form of added fruc-
tose (Table 43.1). Although raspberry has 8 g fiber/100 g berry, most is not retained in the juicing process.
However, trace amounts of inositols are present in raspberry juice, with myo-inositol and chiro-inositol
comprising the main fractions [14].
Raspberry has low protein and lipid content, with 0.84 g and 0.20 g/100 g edible fruit, respectively [11].
A study of seven cultivars of red raspberry grown in Turkey examined fatty acids, which are primarily
present in seeds [12]. Raspberry purees may contain fatty acids, but few remain in processed juices.
Raspberry Juice 529
In summary, raspberry juices and juice blends supply significant content of vitamin C, potassium,
magnesium, manganese, zinc, and copper. The raspberry appears unique from other fruits and berries
due to its abundance of strontium and boron, accompanied by its low carbohydrate content.

43.3.1 Polyphenol Content Reported in the Literature
The raspberry is a rich source of antioxidant polyphenols. The major polyphenol classes in raspberry are pro-
anthocyanidins, hydrolyzable tannins, anthocyanins, flavonols, and flavan-3-ols (Table 43.2 and Figure 43.1).
Reports of the polyphenol content of raspberry juice vary due to differences in analytical methodology.
Nonspecific methods such as total phenols and anthocyanins by pH differential, and s pecific methods such
as high-performance liquid chromatography (HPLC), ultraviolet-visible spectroscopy (UV-VIS), and mass
spectrometry (MS) have been used to characterize polyphenols in raspberry. Black raspberry has signifi-
cantly greater polyphenol content than red raspberry, mainly due to its increased anthocyanin content.
Anthocyanins are the most abundant flavonoids in black and red raspberry juices [18]. Raspberry
anthocyanins are predominately cyanidin and pelargonidin derivatives (Figure 43.1). However, malvidin,
delphinidin, peonidin, and petunidin derivatives have also been identified in raspberry [19]. The three most
abundant red raspberry juice anthocyanins include cyanidin-3-O-sophoroside, cyanidin-3-O-glucoside,
and pelargonidin-3-O-sophoroside, which compose 79.8%, 14.2%, and 6.0% of the total anthocyanins,
respectively [20]. The most abundant anthocyanins in black raspberry are cyanidin-3-xylosylrutinoside
and cyanidin-3-rutinoside [21]. Black raspberry juice contains nearly double the anthocyanins of red
raspberry juice, but few reports have extensively characterized black raspberry juice [13,22]. The profile
of anthocyanins in red raspberry juice, however, varies by cultivar, as discussed in further detail in the
succeeding text [23].
Red raspberry and its juice also contain flavonols and flavan-3-ols. The more abundant flavan-3-ols
present in red raspberry are (–)-epicatechin and (+)-catechin, with lesser amounts of gallocatechins
[24]. Quercetin and kaempferol are the primary flavonols in red raspberry juice, mainly present as
glycosides or glucuronides [25]. Raspberry juice also contains phenolic acids; caffeic, p-coumaric,
ferulic, p-hydroxybenzoic, and protocatechuic acids were detected in acid-hydrolyzed juices [25]. Gallic
acid was the primary phenolic acid in red raspberry juice [26], although Gülçin et al. [27] reported high
levels of p-coumaric acid (40–279 mg/100 g DW aqueous extract of red raspberry cultivars). In black
raspberry, protocatechuic acid was the most abundant phenolic acid, at 8.35 mg/100 g lyophilized black
raspberry powder [28].
Raspberry also contains ellagitannins and gallotannins. Red raspberry ellagitannins mainly consist of
sanguiin H-6 and lambertianin C, but also include a number of less abundant species [19,29]. The ellagi-
tannin content of red raspberry juice is less than that of whole raspberry, but processing can be tailored
TABLE 43.2
Reported Polyphenol Content of Red and Black Raspberries and Their Juices
Red Raspberry Black Raspberry
Juice (mg/100 mL) Fruit (mg/100 g) Juice (mg/100 mL) Fruit (mg/100 g)
Polyphenol [16,20,23,31,32] [19,24,26,30,32] [21] [17,28,30]
Anthocyanins 0.4–51 48.6 244–541 687
Flavan-3-ols nr 5.8 nr nr
Flavonols 9.5 1.1 nr 19
Phenolic acids nr 29 nr nr
Ellagitannins/ellagic acid 22–80 120–310 nr 324
Proanthocyanidins 770 505 nr nr
Total phenols 123 148.1 206–330 980
R1 Anthocyanins R1 R2
OH Cyanidin 3-O-sophoroside OH O-sophoroside
O+ Pelargonidin 3-O-sophoroside H O-sophoroside

HO
Cyanidin 3-O-glucoside OH O-glucoside
R2 Cyanidin 3-O-xylosylrutinoside OH O-xylosylrutinoside
OH Cyanidin 3-O-rutinoside OH O-rutinoside
R3
Flavonols R1 R2 R3
OH
Quercetin 3-O-glucuronide O-glucuronide H OH
HO O Quercetin 3-O-sophoroside O-sophoroside H OH
R2
Quercetin 3-O-rutinoside O-rutinoside H OH
R1
Kaempferol 3-O-glucuronide O-glucuronide H H
OH O
OH HO
OH HO O
OHO HO
O-sophoroside O
HO O R
O O
HO OH HO O
Anthocyanin and flavonol O R
HO OH HO
substitutions O
OH HO O HO HO
O HO O R O
HO O HO O
O R HO O OH
HO O R HO O
OH HO OH OH
O-glucoside O-rutinoside O-glucuronide O-xylosylrutinoside
HO
Ellagitannins O
HO
HO O O OH
HO O
HO O
HO OH
O O O O O
HO
HO O OH
HO O O O O
Sanguiin H-6 HO O O
HO OH
O O O
O OH HO OH OH OH
HO HO O OH OH
O
HO OH OH
OH OH OH
HO
O
HO
OH
HO O O
HO HO O O
O HO OH
HO OO O O
HO O O OH
HO O O O
HO O O
HO
HO OH
O
Lambertianin C HO O O O O
OH HO OHOH OH
OH
O HO O O O
HO O OH
HO O O
HO OH
O O O O HO OH
OH OH OH
HO HO O O OH OH
HO OH
OH
OH OH OH
FIGURE 43.1 Structures of major red and black raspberry polyphenols.

Raspberry Juice 531
to recover greater amounts of ellagitannins, as discussed in the succeeding text [30,31]. “Bristol” black
raspberry has ellagic acid and ellagitannin content comparable to that of red raspberry cultivars [30].
Red raspberry and its juice contain considerable amounts of proanthocyanidins. Hosseinian et al. [32]
reported 505 mg proanthocyanidins/100 g raspberry harvested in Manitoba, while Gu et al. [33] reported
30.2 mg/100 g. Gu et al. [33] did not detect polymeric proanthocyanidins (degrees of polymerization
>10), whereas Hosseinian et al. [32] reported 143 mg/100 g. Thus, further work is needed to characterize
raspberry proanthocyanidins in order to determine variation in content due to analytical method, sample
preparation, or cultivar. While quantitative reports on black raspberry juice proanthocyanidins are lack-
ing, proanthocyanidin dimers and trimers have been found in black raspberry seed, which is suggestive
of their presence in fresh black raspberry and its juice [34].
43.3.2 Polyphenol Databank Values

The polyphenol content of many foods is documented in nutrient databanks such as the Phenol-Explorer
and USDA Flavonoid and Proanthocyanidin Databases. These values are useful references for comparative
purposes and for epidemiological studies that investigate the association between consumption of foods
and health. Red raspberry juice is indexed in both the USDA Flavonoid Database and Phenol-Explorer
[19,24]. The USDA reports 19 mg anthocyanins/100 g berry based on research by Jakobek et al. [20,24].
In contrast, Phenol-Explorer does not report red raspberry juice anthocyanins but does report flavonols and
ellagic acid. Entries for red and black raspberry are consistent with literature reports where data are pro-
vided. Phenol-Explorer reports 980 mg total phenols/100 g black raspberry and 148 mg total phenols/100 g
red raspberry, the latter based on seven studies [19]. Thus, based on databank values and literature reports,
more work is needed to quantitate proanthocyanidins, phenolic acids, and flavonols in raspberry juice.
43.3.3 Antioxidants and Bioactives Responsible for Antioxidant Activity

of Raspberry Juice
As described earlier, raspberry is rich in antioxidant polyphenols. In vitro, polyphenols can act as radical
scavengers, metal chelators, and chain-breaking antioxidants. Raspberry polyphenols are extensively
metabolized and have limited bioavailability in vivo [35]. Although their low intracellular concentra-
tions may preclude them from having a large role in direct radical scavenging activity, many antioxidant
polyphenols induce antioxidant defenses by improving cellular antioxidant mechanisms. Furthermore,
bioavailable polyphenols and their metabolites may inhibit known pro-oxidant enzymes. Polyphenols
that traverse the intestine may also alter gut microflora, which can exert both positive and negative
systemic effects [36]. Therefore, it is useful to characterize sources and mechanisms responsible for the
polyphenol antioxidant activity of raspberry.
A variety of methods have been used to determine the antioxidant capacity of raspberry and raspberry
juice. These include ferric-reducing antioxidant power (FRAP), total oxyradical scavenging capacity (TOCS),
oxygen radical absorbance capacity (ORAC), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS)
scavenging, 2,2-diphenyl-1-picrylhydrazyl (DPPH), and cellular antioxidant activity (CAA) [13,37–40].
The ORAC capacity and DPPH scavenging activity of laboratory-prepared raspberry juices have been
previously reported (Table 43.3). The DPPH scavenging capacity of both red and black raspberry juices
was reported by Konić-Ristić et al. [13]. The IC50 value for black raspberry juice was 1.28 mg/mL, while
TABLE 43.3
Reported Antioxidant Capacities of Raspberry Juices
Antioxidant Assay Red Raspberry Juice Black Raspberry Juice References
DPPH IC50: 2.4 mg juice/mL IC50: 1.8 mg juice/mL [13]
8.2 µmol TE/mL [20]
ORAC 18.5–21.2 µmol TE/g 91–119 µmol TE/g [22,41]
179–603 mg TE/100 g [32]
Abbreviations: DPPH, 2,2-diphenyl-1-picrylhydrazyl; ORAC, oxygen radical absorbance capacity; TE, trolox equivalents.
that for red raspberry juice was greater, at 2.40 mg/mL [13]. The study determined total phenols and
anthocyanins of raspberry, currant, and bilberry juices, and among these berry juices, total phenols
and juice IC50 values were significantly correlated (r 2 = −0.980; P < 0.01) [13]. These data suggest that
anthocyanins alone are not primarily responsible for berry juice DPPH antioxidant values.
Snyder et al. [41] reported ORAC values of juices from red raspberry cultivars grown in Utah. Red
raspberry juice prepared from late-harvest berries had the highest ORAC capacity, and pulp and seed
ORAC values were higher than in juice alone. Similar to DPPH scavenging activity, total anthocyanin
content was not correlated to raspberry juice ORAC values [41]. In juices prepared from black raspberry,
~50% of the original berry ORAC value was retained prior to pasteurization [22]. Subsequent pasteuriza-
tion increased black nonclarified and clarified juice ORAC values by ~25%, and these values remained
stable over 6 months of storage at 25°C [22]. The loss of monomeric anthocyanins during juice storage
over the same time period implies that polymeric anthocyanins or other polyphenol components contrib-
ute to black raspberry juice ORAC values [22].
Data from whole raspberry complement these studies of raspberry juice, suggesting that raspberry
has high antioxidant capacity relative to other foods, and further define the contribution of individual
components to the antioxidant capacity of raspberry.
Based on a comprehensive evaluation of FRAP of foods in Western diets, red raspberry has the seventh
highest antioxidant capacity per serving size [42]. FRAP is a direct measurement of the reducing potential
of extracts and was used to define the total antioxidant capacity (TAC) of wild and cultivated red raspberry
[42]. Wild raspberry had an electron-donating TAC of 3.97 mmol/100 g compared to that of cultivated
raspberry, which had 3.06 mmol/100 g [38]. Using reverse-phase HPLC analysis coupled to the FRAP
assay, Borges et al. [43] determined that ellagitannins contributed to 58% of the FRAP values, and antho-
cyanins and vitamin C contributed 16% and 11%, respectively, from methanol extracts of red raspberry.
The major antioxidant components of red raspberry were determined by reverse-phase HPLC separa-
tion coupled to an ABTS radical scavenging assay [37]. Polar and water-soluble antioxidants such as
vitamin C provided ~20% of antioxidant activity of extracts, anthocyanins contributed ~25%, and ellagi-
tannins contributed ~40% of total activity [37]. Among the 14 red raspberry cultivars tested, ellagitan-
nins contributed 30%–60% of total ABTS radical scavenging activity, highlighting their importance as
antioxidants. Notably, this analysis did not resolve highly polymerized proanthocyanidins, which could
further contribute to antioxidant activity [37].
Isolated raspberry ellagitannins inhibited human low-density lipoprotein (LDL) oxidation ex vivo
[44,45]. Similarly, isolated raspberry ellagitannins inhibited lipid and protein oxidation in model lipo-
some systems and were particularly effective at inhibiting the formation of conjugated diene hydroperox-
ides at 1.4 µg ellagitannins/mL [46]. In contrast, raspberry ellagitannins were less effective at inhibiting
bulk oil oxidation [44].
As discussed earlier, black raspberry is richer in anthocyanins relative to red raspberry. Cyanidin-3-
rutinoside was the most potent black raspberry anthocyanin in the FRAP, DPPH, and ABTS assays [47].
In the same study, cyanidin-3-xylosylrutinoside was also found to contribute to the antioxidant activity
of methanolic black raspberry extracts [47]. Notably, other phenolics from fractionated black raspberry
extracts did not exhibit strong antioxidant activity, likely due to their relatively lower concentrations [47].
This was confirmed in another study of 19 black raspberry homogenates from Ohio that assessed the
cumulative contribution of polyphenols to FRAP and DPPH values by coupling to nuclear magnetic
resonance analysis of extracts [48]. Cyandin-3-xylosylrutinoside content contributed the most to FRAP
and DPPH variability between black raspberry samples [48]. Thus, anthocyanins appear to be the major
contributors to black raspberry antioxidant capacity.
Aqueous acetone extracts of red raspberry also exhibited antioxidant activity in the CAA assay, which
utilizes an intracellular probe in a human cell line challenged with peroxyl radicals [40]. By measur-
ing antioxidant activity in human cells, this method allows for determination of bioavailability. Wolfe
et al. [40] used both a “no phosphate-buffered saline (PBS) wash” protocol and a “PBS wash” protocol
to account for the direct interactions of extract constituents with the cell membrane [40]. Raspberry had
the third lowest EC50 value among the fruits tested, with 6.52 mg/mL in the no-wash protocol, and had an
EC50 value of 14.2 mg/mL when the cells were washed between treatments [40]. This difference indicates
the lack of cellular bioavailability of phenolics from raspberry extracts [40].
Raspberry Juice 533
43.4 Health Effects

43.4.1 Bioavailability of Bioactives from Raspberry Juice
Black and red raspberry polyphenol bioavailability has been determined from fresh berries but not their
juices. Red raspberry ellagitannins are metabolized to urolithins in the colon and excreted in the urine
as urolithin glucuronides [35,49]. Notably, some individuals lack the capacity to produce urolithins upon
raspberry consumption [35]. A small amount of red raspberry anthocyanins (<0.1%) are absorbed intact
in the upper gastrointestinal tract and recovered in urine [35]. Unabsorbed raspberry anthocyanins are
further transformed to phenolic acids and other metabolites by gut microbiota [49]. A similarly low
amount of intact anthocyanins and ellagic acid was absorbed intact and excreted in urine (<1%) after
consumption of lyophilized black raspberry powder [50]. Black raspberry cyanidin-3-xylosylrutinoside
may be more bioavailable than cyanidin 3-rutinoside from black raspberry [51].
The bioavailability of red raspberry juice anthocyanins and ellagitannins was determined in Sprague
Dawley rats [52]. Maximum plasma anthocyanin concentrations were 24 nM, with 16 nm of cyanidin-
3-sophoroside at 1 h after ingestion from a dose equivalent to a 70 kg adult consuming 0.7 L of juice [52].
After 24 h, 1.2% of intact anthocyanins were recovered in urine, indicating their low bioavailability and
extensive metabolism by gut microbiota [52]. Anthocyanins were below the limit of detection in liver
and brain tissue; ellagitannins were not absorbed intact and were not detected in the stomach or intes-
tine, suggesting they were degraded in the stomach [52]. Furthermore, urolithins were not detected in
rat plasma, despite the capacity of this strain to produce urolithins from ellagitannins. Thus, this study
confirms the low polyphenol bioavailability of red raspberry polyphenols and highlights the importance
of metabolism to its bioavailability and bioactivity.
43.4.2 Results of Human Intervention Studies with Raspberry Juice

Human interventions examining the role of pure raspberry juice in health promotion are lacking, to
the best of our knowledge, despite its abundance of antioxidant polyphenols (Table 43.4). A blend of
raspberry, grape, and currant juice improved markers of oxidative stress in cyclists relative to those
consuming a placebo beverage [54]. The supplemental antioxidant beverage contained 93 g raspberry
concentrate/L in addition to grape and currant concentrates and whey protein [54]. For individuals
consuming the placebo beverage, urinary 8-hydroxy-2′-deoxyguanosine, a marker of oxidative DNA
damage, was increased after a 90 min bout of exercise; consuming the antioxidant beverage prevented
this increase [54]. Furthermore, following exercise, protein carbonyls were lower in the antioxidant-
supplemented group than the placebo group [54].
TABLE 43.4
List of Human Intervention Studies with Raspberry Products and Their Key Results
Outcome Key Results References
Exercise-induced oxidative Cyclists consuming a beverage with raspberry, black grape, and red currant [54]
stress juice concentrates had improved markers of oxidative stress relative to
those consuming a placebo beverage.
Glycemic response Concurrent consumption of pancakes with 100 g raspberries did not change [53]
the postprandial glycemic response in n = 12 individuals.
Biomarkers relevant to Drinking 60 g/day of lyophilized black raspberry powder for 1–9 weeks [56]
colorectal cancer decreased colonic tissue markers of apoptosis and proliferation in n = 24
colorectal cancer patients.
Progression of premalignant Topical application of a black raspberry gel benefited the loss of [57]
oral lesions heterozygosity index in some individuals with oral lesions.
Biomarkers of colorectal Consumption of 60 g/day of black raspberry powder for at least 4 weeks [58]
cancer improved genetic biomarkers relevant to colon cancer risk in colorectal
tissues.
A limited number of human intervention studies have used whole black or red raspberries (Table 43.4).
Notably, lyophilized black raspberry interventions may have therapeutic and/or chemopreventive poten-
tial for colon and oral cancer [55–58]. Further studies are needed to differentiate the effects of the whole
raspberry from raspberry juice and extracts.
43.4.3 Results of Animal Studies with Raspberry Juice

Black raspberry juice powder and whole black raspberry were fed to C57/BL6 mice fed high-fat,
obesogenic diets but these did not inhibit weight gain [59]. Red raspberry juice was tested in two ham-
ster models of early atherosclerosis [60,61]. Rouanet et al. [60] provided raspberry juice for 12 weeks
to hamsters fed an atherosclerotic diet. Raspberry juice was among the most effective polyphenol-rich
beverage treatments at inhibiting aortic fatty streak area [60]. Similar to other treatments, rasp-
berry also reduced hepatic superoxide dismutase (SOD) and glutathione peroxidase (GPx) activity
relative to the control diet and did not reduce plasma cholesterol [60]. A subsequent study by the
same group provided hamsters the juice equivalent to a dose of 275 mL for a 70 kg human daily for
12 weeks [61]. Juices were from the named red raspberry cultivars “Cardinal,” “Glen Ample,” and
“Tulameen” [61]. “Cardinal” and “Tulameen” juices lowered plasma total cholesterol and triacylglycer-
ols (TAG), whereas “Glen Ample” juice increased plasma high-density lipoprotein (LDL) by ~11% and
hepatic paraoxonase (PON) activity two-fold from the control group [61]. Thus, preclinical evidence
suggests that raspberry cultivars may have unique actions on modifying cardiovascular disease (CVD)
risk. Further work is necessary to clarify the polyphenols or other bioactives in red raspberry juice
responsible for these changes.
Most animal studies have employed raspberry extracts or whole berries in their design
(Table 43.5). Aiyer et al. [62,63] found that supplementation of 2.5% lyophilized black raspberry
powder for 2 weeks was insufficient to prevent reflux-induced esophageal adenocarcinomas or

esophagitis. In contrast, raspberry has demonstrated promise in other rodent models of cancer chemo-
prevention. Red raspberry extract inhibited diethylnitrosamine-induced cell proliferation and induced
apoptosis in hepatic lesions [64]. Diets supplemented with 5% black or red raspberry fed to F344 rats
treated with N-nitrosomethylbenzylamine (NMBA) inhibited tumor instances by 60%–75% and fur-
ther reduced tumor multiplicity from the control diet [65]. An ellagitannin-rich fraction from black
raspberry also reduced tumorigenesis in the NMBA rodent model [66]. Thus, preclinical animal mod-
els suggest whole black and red raspberry has cancer-chemopreventive actions. If ellagitannins are the
active components from whole raspberry, it is plausible that raspberry juice would have similar efficacy
to these studies.
TABLE 43.5
Recent Animal Studies Utilizing Black and Red Raspberry Juice, Extracts, or Whole Berries
Animal Model References
Red Raspberry
Early atherosclerosis in hamsters (juice) [60,61]
Arthritis-induced inflammation in rats (extract) [67]
Chemical-induced hepatic lesions and serum proteomics (extract) [64]
Black Raspberry
Diet-induced obesity in C57BL/6J mice (juice) [59]
Esophageal adenocarcinomas in SD rats (lyophilized berries) [62]
Surgery-induced esophagitis in SD rats (lyophilized berries) [63]
DSS-induced colitis in C57BL/6J mice (lyophilized berries) [68]
Chemical-induced esophageal tumors (lyophilized berries, extract) [65,66]
Abbreviations: SD, Sprague-Dawley; DSS, dextran sulfate sodium.
Raspberry Juice 535

Black and red raspberry juices have unique polyphenol profiles and high antioxidant capacity. Therefore,
these juices are highly desirable to include in functional juice formulations. Strategies to improve the
sensory qualities, polyphenol composition, and formulation of raspberry juice have been described in
the literature.
Red raspberry juice is sold as 100% juice, as a nectar, or blended with other juices such as apple,
pomegranate, cranberry, and grape. In a study of pomegranate–berry juice blends, a ratio of 10%
red raspberry/90% pomegranate juice blend was the most favorable among red raspberry blends and
scored similarly to pomegranate–blueberry and pomegranate–blackberry blends [69]. Red raspberry
(cv. “Tulameen”)-pineapple juice blends were more highly rated by Italian adults than orange, pome-
granate, apple, or blood orange based juice (20% raspberry juice) [70]. Furthermore, this flavor was
more highly preferred than strawberry, blackberry, red currant, and blueberry blends [70]. Red raspberry
juice concentrate was blended with black or green tea infusions that were rated highly for taste and smell
[71]. However, black tea was rated as less favorable than green tea for its appearance in juice blends [71].
Raspberry juice can also be effectively incorporated into whey-stabilized emulsions. Red raspberry
juice stabilized whey protein oil-in-water emulsions prepared with 1% or 14% rapeseed oil [72]. A 5%
concentration of red raspberry juice prevented formation of protein and lipid carbonyls in both emulsion
systems [72]. Thus, whey and raspberry juice-based smoothies, sports drinks, and spreads are potential
functional products.
Black and red raspberry juices can also be fermented into wines. Duarte et al. [73] reported that
Saccharomyces cerevisiae strain UFLA FW produced a more favorable wine than other yeasts from
“Meeker” red raspberry juice, containing desirable ethyl ester, alcohol, and acetate volatiles, with rela-
tively lower acid concentrations. Kim et al. [74] reported a strategy to enhance γ-aminobutyric acid
(GABA) content of black raspberry wine by fermenting raspberry juice with Lactobacillus brevis GABA
100. After 12 days of fermentation, up to 25 mg GABA/mL was produced in black raspberry juice [74].
Research has also focused on processing strategies to increase raspberry juice yields and retention of
polyphenols. Anthocyanins are localized in the raspberry exocarp, and ~20% of red raspberry antho-
cyanins are delivered in raspberry juice [13]. Verbeyst et al. [75] examined high-pressure processing and
pectinase treatment of red raspberry juice as a strategy to increase anthocyanin and vitamin C content
in juice from the raspberry cultivar “Sugana.” Enzyme-treated berries yielded ~20% more juice than
untreated berries and had similar anthocyanin and total phenolic concentrations [75]. Application of
400–800 MPa did not affect anthocyanin recovery from red raspberry [75]. Raspberry can also be effec-
tively pressed with apple pomace to aid juice processing [76].
Microfiltration or ultrafiltration can also be used as an alternative method of raspberry juice clari-
fication. Vladisavljević et al. [77] reported that a 0.2 µm pore size multichannel microfilter that was
backwashed with compressed air every 6 min produced a highly clarified red raspberry juice following
pectinase treatment. Pasteurization of juice led to ~6% loss of anthocyanins, and classical clarification
caused a 36%–44% decrease in anthocyanins [77]. Ultrafiltration losses were greater but varied by mem-
brane choice, whereas microfiltered juice lost only 14% anthocyanins after pasteurization [77].
Hager et al. [22] described anthocyanin and antioxidant retention during black raspberry production
and storage. Maceration, blanching, and enzymatic clarification resulted in a loss of 45% of the original
berry anthocyanins [22]. Pasteurization caused another 19% loss of anthocyanins in clarified juices [22].
During subsequent storage, polymeric anthocyanins were formed as monomeric anthocyanin content
decreased. Over the same period, juice ORAC values remained constant [22]. Thus, the initial steps of
black raspberry juice processing as well as its long-term storage appear useful targets to enhance reten-
tion of polyphenols.
Red raspberry puree was treated with ultrasound frequencies, and 490 kHz treatment increased puree
anthocyanins and DPPH antioxidant activity [78]. Yousefi et al. [79] described conditions to prepare red
raspberry juice concentrates by microwave heating, which resulted in greater retention of anthocyanin,
phenolic, and DPPH antioxidant activity.
A number of novel products containing raspberry juice or raspberry powder have been developed to
enhance the delivery of raspberry polyphenols. Lyophilized black raspberry powder was processed into
nectars and pectin-based confections that were well liked [80]. Black raspberry nectars were formulated
with 4% or 8% black raspberry powder and stabilized with pectin [80]. At least 100% of ellagic acid
was retained in these products, while up to 31% of anthocyanins were lost in nectar processing [80].
Furthermore, ellagic acid was stable through 2 months of storage, although ellagitannin content decreased
in the 4% powder nectar [80].
Juice from red “Meeker” raspberry (10% w/w) was incorporated into muffins as a strategy to increase
dietary bioactives in bakery products [81]. Lyophilizing muffin batter increased the recovery of mono-
meric anthocyanins after baking [81].
Red raspberry juice has also been used as a vehicle for delivering probiotics. Probiotic L. rhamnosus
and L. acidophilus strains were spray dried with maltodextrin and red raspberry juice, with up to 84%
survival of each strain [82]. Inclusion of maltodextrin reduced the accumulation of raspberry solids on
the spray drying chamber and improved product yields [82].
Raspberry juice‒based products could also be designed based on synergy between functional com-
ponents. Raspberry and adzuki bean extracts exhibited synergy in FRAP, ORAC, and DPPH assays and
protected cardiomyocytes from peroxide-induced cell damage [83]. More research is needed to deter-
mine synergies between raspberry polyphenols and other functional dietary ingredients.
43.6 Conclusion
Black and red raspberry juices are rich in antioxidant polyphenols and vitamin C. Although pure rasp-
berry juice is not typically consumed due to its sourness and astringency, raspberry juice blends and
nectars are highly desirable. Furthermore, raspberry juice is suitable for use in whey-based emulsion
beverages. Black and red raspberry juices have different polyphenol profiles and sensory qualities.
Both are notable for their antioxidant capacity, anthocyanin content, and abundance of ellagitannins.
Further work is needed to translate preclinical studies of raspberry juice into human intervention studies.
Particularly promising areas of future research appear to be the ability of red raspberry juice to improve
biomarkers of CVD risk and the use of black raspberry juice in cancer chemoprevention.
REFERENCES
1. Lee, J., Dossett, M., and Finn, C.E. Rubus fruit phenolic research: The good, the bad, and the confusing.
Food Chem., 130, 785–796, 2012.
2. Food and Agriculture Organization (FAO), FAOSTAT, Food and Agriculture Organization of the United
Nations, 2014. Published online at: http://faostat3.fao.org (accessed January 24, 2014).
3. Celik, F. and Ercisli, S., Lipid and fatty acid composition of wild and cultivated red raspberry fruits
(Rubus idaeus L.). J. Med. Plants Res., 3, 583–585, 2009.
4. U.S. Department of Agriculture (USDA), Economic Research Service. Fruit: Per Capita Availability
(fresh weight equivalent), 2014. Published online at: http://www.ers.usda.gov (accessed January 24, 2014).
5. Zamora-Ros, R., Knaze, V., Luján-Barroso, L., Slimani, N., Romieu, I., Touillaud, M., Kaaks, R.
et al., Estimation of the intake of anthocyanidins and their food sources in the European Prospective
Investigation into Cancer and Nutrition (EPIC) study. Br. J. Nutr., 106, 1090–1099, 2011.
6. Welch’s. Juice Concentrates, 100% Juices, 2014. Published online at: http://www.welchs.com/products
7. Old Orchard Juices, Red Raspberry 100% Juice Blend, 2014. Published online at: http:// oldorchard.
com/products/100-juice-64oz/64oz-100-juice-red-raspberry (accessed April 7, 2014).
8. Walnut Acres, Raspberry, 2014. Published online at: http:// www.walnutacres.com/product_view.
php?id=36&cat=juice (accessed April 7, 2014).
9. Brownwood Acres Foods, Inc., Red Raspberry Juice Concentrate, 2011. Published online at: http://
www.brownwoodacres.com/red-raspberry-juice-concentrate-quart-32-oz.html (accessed April 7, 2014).
Raspberry Juice 537
11. Daglia, M., Papetti, A., Mascherpa, D., Grisoli, P., Giusto, G., Lingström, P., Pratten, J. et al., Plant
and fungal food components with potential activity on the development of microbial oral diseases.
J. Biomed. Biotechnol., 2011, 1–9, 2011.
12. Kafkas, E., Ozgen, M., Ozogul, Y., and Turemis, N., Phytochemical and fatty acid profile of selected red
raspberry cultivars: A comparative study. J. Food Qual., 31, 67–78, 2008.
13. Konić-Ristić, A., Šavikin, K., Zdunić, G., Janković, T., Juranic, Z., Menković, N., and Stanković, I.,
Biological activity and chemical composition of different berry juices. Food Chem., 125, 1412–1417, 2011.
14. Sanz, M.L., Villamiel, M., and Martinez-Castro, I., Inositols and carbohydrates in different fresh fruit
juices. Food Chem., 87, 325–328, 2004.
15. Rotundo, A., Bounous, G., Benvenuti, S., Vampa, G., Melegari, M., and Soragni, F., Quality and
yield of Ribes and Rubus cultivars grown in southern Italy hilly locations. Phytother. Res., 12(Suppl.),
S135–S137, 1998.
16. Rommel, A. and Wrolstad, R.E., Composition of flavonols in red raspberry juice as influenced by culti-
var, processing, and environmental factors. J. Agric. Food Chem., 41, 1941–1950, 1993.
17. Wada, L. and Ou, B.X., Antioxidant activity and phenolic content of oregon caneberries. J. Agric. Food
Chem., 50, 3495–3500, 2002.
18. Wu, X.L., Beecher, G.R., Holden, J.M., Haytowitz, D.B., Gebhardt, S.E., and Prior, R.L., Concentrations
of anthocyanins in common foods in the United States and estimation of normal consumption. J. Agric.
Food Chem., 54, 4069–4075, 2006.
metabolism and pharmacokinetics in humans and experimental animals. Database, bas031, 1–8, 2012.
dant activity of various red fruit juices. Deut. Lebensm-Rundsch., 103, 58–64, 2007.
21. Dossett, M., Lee, J., and Finn, C.E., Variation in anthocyanins and total phenolics of black raspberry
populations. J. Funct. Foods, 2, 292–297, 2010.
22. Hager, A., Howard, L.R., Prior, R.L., and Brownmiller, C., Processing and storage effects on monomeric
anthocyanins, percent polymeric color, and antioxidant capacity of processed black raspberry products.
J. Food Sci., 73, H134–H140, 2008.
23. Boyles, M.J. and Wrolstad, R.E., Anthocyanin composition of red raspberry juice—Influences of culti-
var, processing, and environmental-factors. J. Food Sci., 58, 1135–1141, 1993.
Foods, Release 3.1, USDA, Springfield, VA, 2013.
25. Rommel, A., Wrolstad, R.E., and Durst, R.W., Red raspberry phenolic: Influences of processing,
variety, and environmental factors, in Phenolic Compounds in Food and Their Effects on Health I,
Ho, C.-T., Lee, C.Y., and Huang, M.-T., Eds., ACS Symposium Series 506, American Chemical Society,
Washington, DC, pp. 259–286, 1992.
26. Mattila, P., Hellstrom, J., and Torronen, R., Phenolic acids in berries, fruits, and beverages. J. Agric.
Food Chem., 54, 7193–7199, 2006.
27. Gülçin, İ., Topal, F., Çakmakçı, R., Bilsel, M., Gören, A.C., and Erdogan, U., Pomological features,
nutritional quality, polyphenol content analysis, and antioxidant properties of domesticated and 3 wild
ecotype forms of raspberries (Rubus idaeus L.). J. Food Sci., 76, C585–C593, 2011.
28. Wu, X., Pittman, I.H.E., Hager, T., Hager, A., Howard, L., and Prior, R.L., Phenolic acids in black rasp-
berry and in the gastrointestinal tract of pigs following ingestion of black raspberry. Mol. Nutr. Food
Res., 53, 76–84, 2009.
29. Mullen, W., Yokota, T., Lean, M.E.J., and Crozier, A., Analysis of ellagitannins and conjugates of ellagic
acid and quercetin in raspberry fruits by LC-MSn. Phytochem., 64, 617–624, 2003.
30. Bobinaitė, R., Viškelis, P., and Venskutonis, P.R., Variation of total phenolics, anthocyanins, ellagic
acid and radical scavenging capacity in various raspberry (Rubus spp.) cultivars. Food Chem., 132,
1495–1501, 2012.
31. Rommel, A. and Wrolstad, R.E., Ellagic acid content of red raspberry juice as influenced by cultivar,
processing, and environmental-factors. J. Agric. Food Chem., 41, 1951–1960, 1993.
32. Hosseinian, F.S., Li, W., Hydamaka, A.W., Tsopmo, A., Lowry, L., Friel, J., and Beta, T., Proanthocyanidin
profile and ORAC values of Manitoba berries, chokecherries, and seabuckthorn. J. Agric. Food Chem.,
55, 6970–6976, 2007.
33. Gu, L.W., Kelm, M.A., Hammerstone, J.F., Beecher, G., Holden, J., Haytowitz, D., Gebhardt, S., and
Prior, R.L., Concentrations of proanthocyanidins in common foods and estimations of normal consump-
tion. J. Nutr., 134, 613–617, 2004.
34. Xu, Y., Zhang, Y., Chen, M., and Tu, P., Fatty acids, tocopherols and proanthocyanidins in bramble
seeds. Food Chem., 99, 586–590, 2006.
35. González-Barrio, R., Borges, G., Mullen, W., and Crozier, A., Bioavailability of anthocyanins and ella-
gitannins following consumption of raspberries by healthy humans and subjects with an ileostomy.
J. Agric. Food Chem., 58, 3933–3939, 2010.
36. Etxeberria, U., Fernandez-Quintela, A., Milagro, F.I., Aguirre, L., Martinez, J.A., and Portillo, M.P.,
Impact of polyphenols and polyphenol-rich dietary sources on gut microbiota composition. J. Agric.
Food Chem., 61, 9517–9533, 2013.
37. Beekwilder, J., Jonker, H., Meesters, P., Hall, R.D., van der Meer, I.M., and de Vos, C.H.R., Antioxidants
in raspberry: Online analysis links antioxidant activity to a diversity of individual metabolites. J. Agric.
Food Chem., 53, 3313–3320, 2005.
38. Halvorsen, B.L., Holte, K., Myhrstad, M.C., Barikmo, I., Hvattum, E., Remberg, S.F., Wold, A.B. et al.,
A systematic screening of total antioxidants in dietary plants. J. Nutr., 132, 461–471, 2002.
39. Liu, M., Li, X.Q., Weber, C., Lee, C.Y., Brown, J., and Liu, R.H., Antioxidant and antiproliferative
activities of raspberries. J. Agric. Food Chem., 50, 2926–2930, 2002.
40. Wolfe, K.L., Kang, X.M., He, X.J., Dong, M., Zhang, Q.Y., and Liu, R.H., Cellular antioxidant activity
of common fruits. J. Agric. Food Chem., 56, 8418–8426, 2008.
41. Snyder, S.M., Low, R.M., Stocks, J.C., Eggett, D.L., and Parker, T.L., Juice, pulp and seeds fractionated
from dry climate primocane raspberry cultivars (Rubus idaeus) have significantly different antioxidant
capacity, anthocyanin content and color. Plant Food Hum. Nutr., 67, 358–364, 2012.
42. Halvorsen, B.L., Carlsen, M.H., Phillips, K.M., Bohn, S.K., Holte, K., Jacobs, D.R., and Blomhoff, R.,
Content of redox-active compounds (i.e., antioxidants) in foods consumed in the United States. Am.
J. Clin. Nutr., 84, 95–135, 2006.
43. Borges, G., Degeneve, A., Mullen, W., and Crozier, A., Identification of flavonoid and phenolic antioxi-
dants in black currants, blueberries, raspberries, red currants, and cranberries. J. Agric. Food Chem., 58,
3901–3909, 2010.
44. Kähkönen, M., Kylli, P., Ollilainen, V., Salminen, J.-P., and Heinonen, M., Antioxidant activity of iso-
lated ellagitannins from red raspberries and cloudberries. J. Agric. Food Chem., 60, 1167–1174, 2012.
45. Vuorela, S., Kreander, K., Karonen, M., Nieminen, R., Hamalainen, M., Galkin, A., Laitinen, L. et al.,
Preclinical evaluation of rapeseed, raspberry, and pine bark phenolics for health-related effects. J. Agric.
Food Chem., 53, 5922–5931, 2005.
46. Viljanen, K., Kylli, P., Kivikari, R., and Heinonen, M., Inhibition of protein and lipid oxidation in
liposomes by berry phenolics. J. Agric. Food Chem., 52, 7419–7424, 2004.
47. Tulio, A.Z., Reese, R.N., Wyzgoski, F.J., Rinaldi, P.L., Fu, R., Scheerens, J.C., and Miller, A.R.,
Cyanidin 3-rutinoside and cyanidin 3-xylosylrutinoside as primary phenolic antioxidants in black
raspberry. J. Agric. Food Chem., 56, 1880–1888, 2008.
48. Wyzgoski, F.J., Paudel, L., Rinaldi, P.L., Reese, R.N., Ozgen, M., Tulio, A.Z., Miller, A.R., Scheerens,
J.C., and Jardy, J.K., Modeling relationships among active components in black raspberry (Rubus occi-
dentalis L.) fruit extracts using high-resolution1h nuclear magnetic resonance (NMR) spectroscopy and
multivariate statistical analysis. J. Agric. Food Chem., 58, 3407–3414, 2010.
49. Gonzalez-Barrio, R., Edwards, C.A., and Crozier, A., Colonic catabolism of ellagitannins, ellagic acid,
and raspberry anthocyanins: In vivo and in vitro studies. Drug Metab. Dispos., 39, 1680–1688, 2011.
50. Stoner, G.D., Sardo, C., Apseloff, G., Mullet, D., Wargo, W., Pound, V., Singh, A. et al., Pharmacokinetics
of anthocyanins and ellagic acid in healthy volunteers fed freeze-dried black raspberries daily for 7 days.
J. Clin. Pharmacol., 45, 1153–1164, 2005.
51. Tian, Q.G., Giusti, M.M., Stoner, G.D., and Schwartz, S.J., Urinary excretion of black raspberry (Rubus
occidentalis) anthocyanins and their metabolites. J. Agric. Food Chem., 54, 1467–1472, 2006.
52. Borges, G., Roowi, S., Rouanet, J.-M., Duthie, G.G., Lean, M.E.J., and Crozier, A., The bioavailability
of raspberry anthocyanins and ellagitannins in rats. Mol. Nutr. Food Res., 51, 714–725, 2007.
Raspberry Juice 539
53. Clegg, M.E., Pratt, M., Meade, C.M., and Henry, C.J.K., The addition of raspberries and blueberries to
a starch-based food does not alter the glycaemic response. Br. J. Nutr., 106, 335–338, 2011.
54. Morillas-Ruiz, J., Zafrilla, P., Almar, M., Cuevas, M.J., López, F.J., Abellán, P., Villegas, J.A., and
González-Gallego, J., The effects of an antioxidant-supplemented beverage on exercise-induced oxi-
dative stress: Results from a placebo-controlled double-blind study in cyclists. Eur. J. Appl. Physiol.,
95, 543–549, 2005.
55. Kresty, L.A., Frankel, W.L., Hammond, C.D., Baird, M.E., Mele, J.M., Stoner, G.D., and Fromkes, J.J.,
Transitioning from preclinical to clinical chemopreventive assessments of lyophilized black raspberries:
Interim results show berries modulate markers of oxidative stress in barrett’s esophagus patients. Nutr.
Cancer, 54, 148–156, 2006.
56. Mentor-Marcel, R.A., Bobe, G., Sardo, C., Wang, L.-S., Kuo, C.-T., Stoner, G., and Colburn, N.H.,
Plasma cytokines as potential response indicators to dietary freeze-dried black raspberries in colorectal
cancer patients. Nutr. Cancer, 64, 820–825, 2012.
57. Shumway, B.S., Kresty, L.A., Larsen, P.E., Zwick, J.C., Lu, B., Fields, H.W., Mumper, R.J., Stoner, G.D.,
and Mallery, S.R., Effects of a topically applied bioadhesive berry gel on loss of heterozygosity indices
in premalignant oral lesions. Clin. Cancer Res., 14, 2421–2430, 2008.
58. Wang, L.S., Arnold, M., Huang, Y.W., Sardo, C., Seguin, C., Martin, E., Huang, T.H. et al., Modulation
of genetic and epigenetic biomarkers of colorectal cancer in humans by black raspberries: A phase I
pilot study. Clin. Cancer Res., 17, 598–610, 2010.
59. Prior, R.L., Wilkes, S., Rogers, T., Khanal, R.C., Wu, X., Hager, T.J., and Howard, L., Dietary black
raspberry anthocyanins do not alter development of obesity in mice fed an obesogenic high-fat diet.
J. Agric. Food Chem., 58, 3977–3983, 2010.
60. Rouanet, J.-M., Décordé, K., Rio, D.D., Auger, C., Borges, G., and Cristol, J.-P., Berry juices, teas, anti-
oxidants and the prevention of atherosclerosis in hamsters. Food Chem., 118, 266–271, 2010.
61. Suh, J.-H., Romain, C., González-Barrio, R., Cristol, J.-P., Teissèdre, P.-L., Crozier, A., and Rouanet,
J.-M., Raspberry juice consumption, oxidative stress and reduction of atherosclerosis risk factors in
hypercholesterolemic golden syrian hamsters. Food Funct., 2, 400–405, 2011.
62. Aiyer, H.S., Li, Y., Losso, J.N., Gao, C., Schiffman, S.C., Slone, S.P., and Martin, R.C., Effect of freeze-
dried berries on the development of reflux-induced esophageal adenocarcinoma. Nutr. Cancer, 63,
1256–1262, 2011.
63. Aiyer, H.S., Li, Y., Liu, Q.H., Reuter, N., and Martin, R.C.G., Dietary freeze-dried black raspberry’s
effect on cellular antioxidant status during reflux-induced esophagitis in rats. Nutr., 27, 182–187,
2011.
64. Chen, H.-S., Liu, M., Shi, L.-J., Zhao, J.-L., Zhang, C.-P., Lin, L.-Q., Liu, Y. et al., Effects of raspberry
phytochemical extract on cell proliferation, apoptosis, and serum proteomics in a rat model. J. Food
Sci., 76, T192–T198, 2011.
65. Stoner, G.D., Wang, L.-S., Seguin, C., Rocha, C., Stoner, K., Chiu, S., and Kinghorn, A.D., Multiple
berry types prevent N-nitrosomethylbenzylamine-induced esophageal cancer in rats. Pharm. Res.,
27, 1138–1145, 2010.
66. Wang, L.-S., Hecht, S., Carmella, S., Seguin, C., Rocha, C., Yu, N., Stoner, K., Chiu, S., and Stoner, G.,
Berry ellagitannins may not be sufficient for prevention of tumors in the rodent esophagus. J. Agric.
Food Chem., 58, 3992–3995, 2010.
67. Jean-Gilles, D., Li, L., Ma, H., Yuan, T., Chichester, C.O., and Seeram, N.P., Anti-inflammatory effects
of polyphenolic-enriched red raspberry extract in an antigen-induced arthritis rat model. J. Agric. Food
Chem., 60, 5755–5762, 2012.
68. Montrose, D.C., Horelik, N.A., Madigan, J.P., Stoner, G.D., Wang, L.S., Bruno, R.S., Park, H.J.,
Giardina, C., and Rosenberg, D.W., Anti-inflammatory effects of freeze-dried black raspberry powder
in ulcerative colitis. Carcinogenesis, 32, 343–350, 2010.
69. Vázquez-Araújo, L., Chambers, I.E., Adhikari, K., and Carbonell-Barrachina, Á.A., Sensory and physi-
cochemical characterization of juices made with pomegranate and blueberries, blackberries, or raspber-
ries. J. Food Sci., 75, S398–S404, 2010.
70. Endrizzi, I., Pirretti, G., Calò, D.G., and Gasperi, F., A consumer study of fresh juices containing berry
fruits. J. Sci. Food Agric., 89, 1227–1235, 2009.
71. Owczarek, L., Juices and beverages with a controlled phenolic content and antioxidant capacity. Polish
J. Food Nutr. Sci., 13, 261–268, 2004.
72. Viljanen, K., Halmos, A.L., Sinclair, A., and Heinonen, M., Effect of blackberry and raspberry juice on
whey protein emulsion stability. Eur. Food Res. Techol., 221, 602–609, 2005.
73. Duarte, W.F., Dragone, G., Dias, D.R., Oliveira, J.M., Teixeira, J.A., Silva, J.B.A., and Schwan, R.F.,
Fermentative behavior of saccharomyces strains during microvinification of raspberry juice (Rubus
idaeus L.). Int. J. Food Microbiol., 143, 173–182, 2010.
74. Kim, J.Y., Lee, M.Y., Ji, G.E., Lee, Y.S., and Hwang, K.T., Production of γ-aminobutyric acid in black
raspberry juice during fermentation by Lactobacillus brevis GABA100. Int. J. Food Microbiol., 130,
12–16, 2009.
75. Verbeyst, L., Hendrickx, M., and Loey, A., Characterisation and screening of the process stability of
bioactive compounds in red fruit paste and red fruit juice. Eur. Food Res. Techol., 234, 593–605, 2012.
76. Roberts, J.S., Gentry, T.S., and Bates, A.W., Utilization of dried apple pomace as a press aid to improve
the quality of strawberry, raspberry, and blueberry juices. J. Food Sci., 69, S181–S190, 2004.
77. Vladisavljević, G.T., Vukosavljević, P., and Veljović, M.S., Clarification of red raspberry juice using
microfiltration with gas backwashing: A viable strategy to maximize permeate flux and minimize a loss
of anthocyanins. Food Bioprod. Proc., 91, 473–480, 2013.
78. Golmohamadi, A., Möller, G., Powers, J., and Nindo, C., Effect of ultrasound frequency on antioxidant
activity, total phenolic and anthocyanin content of red raspberry puree. Ultrason. Sonochem.,
20, 1316–1323, 2013.
79. Yousefi, G., Yousefi, S., and Emam-Djomeh, Z., A comparative study on different concentration meth-
ods of extracts obtained from two raspberries (Rubus idaeus L.) cultivars: Evaluation of anthocyanins
and phenolics contents and antioxidant activity. Int. J. Food Sci. Techol., 48, 1179–1186, 2013.
80. Gu, J., Ahn-Jarvis, J.H., Riedl, K.M., Schwartz, S.J., Clinton, S.K., and Vodovotz, Y., Characterization
of black raspberry functional food products for cancer prevention human clinical trials. J. Agric. Food
Chem., 62, 3997–4006, 2014.
81. Rosales-Soto, M.U., Powers, J.R., and Alldredge, J.R., Effect of mixing time, freeze-drying and baking
on phenolics, anthocyanins and antioxidant capacity of raspberry juice during processing of muffins.
J. Sci. Food Agric., 92, 1511–1518, 2012.
82. Anekella, K. and Orsat, V., Optimization of microencapsulation of probiotics in raspberry juice by spray
drying. LWT—Food Sci. Technol., 50, 17–24, 2013.
83. Wang, S., Meckling, K.A., Marcone, M.F., Kakuda, Y., and Tsao, R., Synergistic, additive, and antago-
nistic effects of food mixtures on total antioxidant capacities. J. Agric. Food Chem., 59, 960–968, 2011.
44
Strawberry Juice
Rong Tsao and Hongyan Li
CONTENTS
44.1 Introduction....................................................................................................................................541
44.3.1 Anthocyanins.................................................................................................................... 546
44.3.2 Flavonols........................................................................................................................... 546
44.3.3 Hydroxybenzoic and Hydroxycinnamic Acids................................................................. 546
44.4 Health Effects................................................................................................................................ 547
44.4.1 Antioxidant Activity......................................................................................................... 547
44.4.2 Anti-Inflammatory Activity............................................................................................. 547
44.4.3 Reduced Cardiovascular Disease..................................................................................... 547
44.4.4 Against Diabetes.............................................................................................................. 548
44.4.5 Anticancer Effects............................................................................................................ 548
44.6 Conclusion..................................................................................................................................... 549
References............................................................................................................................................... 549
44.1 Introduction
Strawberry (Fragaria x ananassa), the only fruit with its seeds on the outside, is a widely appreciated fruit
with a bright red color, characteristic aroma, and sweet and sour flavor. Based on a rough estimate, 4.3
million metric tons (MT) of strawberry is produced every year worldwide. The potential health benefits of
strawberry include preventing heart disease, diabetes, cancers, and other chronic diseases [1]. The prefer-
ence for fresh strawberry is challenging because it has an extremely short shelf life at its best quality, due to
its susceptibility to fungal spoilage, mechanical injury, and rapid dehydration. For this reason, strawberries
are processed into jams, juice, concentrates, and candied fruits and added in dairy products such as yogurt.
Strawberry juice, a virtually fat-free and nutrient-dense beverage, is one of the most popular fruit
juices due to its attractive color, aroma, mouthfeel, and functional properties. It is also relatively low in
calories and often blended with other fruit juices and cocktails. Strawberry juice is rich in fiber; potas-
sium; vitamins A, B, and C; polyphenols such as anthocyanins (mainly pelargonidin-3-O-glucoside),
flavonols, and hydroxybenzoic and hydroxycinnamic acids; and amino acids. Vitamin C, total phenolic,
and total anthocyanin contents as well as total antioxidant activity in different processed strawberry
products such as strawberry juice, nectar, wine, and puree were found to be significantly lower than in
raw strawberry fruits. Among the processed products, however, strawberry juices were found to be rela-
tively less affected in terms of the total vitamin C and phenolic contents, particularly anthocyanins and
the antioxidant activities [2].
Strawberry juice is an excellent choice for defending against inflammation, cardiovascular disease
(CVD), diabetes, and cancer due to the aforementioned phytochemicals and vitamin C that are strong
antioxidants with relevant in vitro and in vivo activities. The quality of strawberry juice is dictated by
541
factors such as cultivar and maturity at the time of processing. This chapter provides a comprehensive
and critical review on functional compounds of strawberry juice and their beneficial role in human
health. These include effects on antioxidant, inflammation, CVD, diabetes, and cancers.

Strawberry is a good source of amino acids, organic acids, soluble sugars, dietary fiber, and important
minerals and vitamins (Table 44.1). The contents of these functional compounds differ with cultivar
and degree of ripeness. These essential and nonessential nutrients are particularly important in helping
maintaining good health beyond basic nutritional functions.
Free amino acids in strawberry juice are related to sensory quality loss due to their reaction with
reducing sugars in browning reactions. The total free amino acid content in fresh strawberry juice
(7.5°Brix) was 445 mg/L [3]. Among free amino acids, alanine (134.8 mg/L), serine (94.3 mg/L), aspar-
tic acid (60.1 mg/L), glutamic acid (56.7 mg/L), and threonine (40.6 mg/L) were the most abundant in
the juice. Minor free amino acids in strawberry juice include valine, tyrosine, phenylalanine, glycine,
TABLE 44.1
Nutritional Characteristics of Strawberry Juice
Food Description Compound Unit Content References
7.5°Brix strawberry juice Free Amino Acids
Alanine mg/L 134.8 [3]
Serine mg/L 94.3 [3]
Aspartic acid mg/L 60.1 [3]
Glutamic acid mg/L 56.7 [3]
Threonine mg/L 40.6 [3]
Commercial strawberry juice (United Kingdom) Organic Acids
Citric acid g/L 6.7 [4]
Laboratory-made strawberry juice Malic acid g/L 1.74 [6]
Single-strength strawberry juice (Germany) Sugars
Sucrose g/L 0.49 [5]
Glucose g/L 5.84 [5]
Fructose g/L 7.23 [5]
Strawberry shake Minerals
Potassium mg/100 g 182 [7]
Magnesium mg/100 g 13 [7]
Phosphorus mg/100 g 100 [7]
Calcium mg/100 g 113 [7]
Zinc mg/100 g 0.36 [7]
Strawberry juice (Brazil) Iron µg/L 187 [8]
Manganese µg/L 42 [8]
Laboratory-made strawberry juice Vitamins
Ascorbic acid mg/100 mL 60 [10]
Strawberry shake Riboflavin mg/100 g 0.20 [7]
Vitamin B6 mg/100 g 0.04 [7]
Vitamin B12 µg/100 g 0.31 [7]
Folate (DFE) µg/100 g 3 [7]
Vitamin A (RAE) µg/100 g 26 [7]
Commercial yogurt with strawberry juice Thiamine ng/mL 122 [5]
Niacin ng/mL 262 [5]
Commercial strawberry juice (United Kingdom) Dietary Fiber g/L 8.0 [4]
Abbreviations: DFE, dietary folate equivalents; RAE, retinol activity equivalents.
Strawberry Juice 543
isoleucine, leucine, and lysine (2.4–17.2 mg/L) [3]. The total content of free amino acids was reduced
significantly from 445 to 418.7 mg/L in heat-treated (90°C for 60 s) strawberry juice, probably caused
by the Maillard reaction, but can be reserved and even slightly enhanced by nonthermal high-intensity
pulsed electric field (HIPEF) treatment of the juice [3]. HIPEF-treated strawberry juice had higher free
amino acid (alanine, leucine, valine, and glycine) contents than thermally treated juice, after storage at
4°C for 56 days [3].
The organic acids and soluble sugars contribute to the excellent flavor of strawberry juice. The citric
acid and total sugar (expressed as the sum of sucrose, glucose, and fructose concentrations) contents in
strawberry juice were reported to be 6.7 and 84.0 g/L, respectively. After 6 weeks storage at 4°C, the cit-
ric acid and total sugar showed small changes (7.0 and 83.1 g/L, respectively) [4]. The individual sucrose,
glucose, and fructose concentrations in concentrate and single-strength strawberry juice were reported to
be 2.28, 26.09, and 29.39, and 0.12, 5.82, and 7.34 g/L, respectively [5]. Citric acid (7.33 g/L) and malic
acid (1.74 g/L) were reported as the primary organic acids in strawberry juice [6]. The concentration of
dietary fiber in strawberry juice was 8 g/L, which was much higher than those in orange, grapefruit,
pineapple, black currant, and pomegranate juices (in all cases between 0.5 and 3.5 g/L) [4].
Strawberry juice is a healthy food choice due to its nutrient profile, including minerals. The potassium,
magnesium, phosphorus, calcium, and zinc contents of strawberry shake were 182, 13, 100, 113, and
0.36 mg/100 g, respectively [7]. Iron and manganese were present at 187 and 42 µg/L in a kind of straw-
berry juice in Brazil, respectively [8]. In addition, cinnamic acid (13.87 ng/mL) also existed in drinkable
yogurt with strawberry juice [9].
Strawberry juice is also a good source of ascorbic acid (Table 44.1) and present at 60 mg/100 mL,
although it may vary depending on cultivar and degree of ripening [10]. Riboflavin (0.20 mg/100 g), vita-
min B6 (0.04 mg/100 g), vitamin B12 (0.31 µg/100 g), folate (3 µg/100 g), and vitamin A (26 µg/100 g) are
reported in strawberry shake [7]. Small amounts of thiamine (121.56 ng/mL), niacin (262.41 ng/mL), and
pyridoxine (3.4 ng/mL) were also present in strawberry juice [9]. Ascorbic acid is the main vitamin in
strawberry and its related products. There is a strong inverse relationship between the intake of ascorbic
acid (particularly serum ascorbic acid) and chronic diseases (in vitro and animal carcinogenesis studies).
More details about the epidemiology of ascorbic acid and its role in human health can be found in the
literature [11]. Substantial epidemiological evidences have shown that consumption of fruits and vegeta-
bles, a reasonable measure of dietary vitamin C intake, reduces human health risks [12,13]. Ascorbic acid
is not very stable in strawberry juice under ambient conditions, and it degrades via a first-order kinetics
and leads to decreased nutritional values [14]. Nearly half of the ascorbic acid in strawberry juice was
lost after 15 days of storage at 5°C or 10°C, and totally lost at 25°C [10]. When exposed to fluctuating
temperatures in transit, significant decrease of shelf life (about 5 days) was observed in strawberry juice.

Strawberry juice is rich in antioxidant polyphenols including phenolic acids and flavonols (Figure 44.1).
The phytochemical antioxidants, mainly the polyphenols, have been characterized in recent years in detail
using advanced analytical tools such as high-performance liquid chromatography/ultrahigh-performance
liquid chromatography (HPLC/UPLC) coupled with ultraviolet–visible spectroscopy (UV-VIS), fluores-
cence detection, and/or mass or tandem mass spectrometry [15]. Table 44.2 summarizes the phytochemi-
cals identified in strawberry juice. Among the phytochemicals of strawberry juice, polyphenols are the
most important for their strong antioxidant activities. The total phenolic and flavonoid contents of the
lyophilized powders from strawberry juice (supernatants of fresh strawberry) have been reported to be
20.9 and 0.87 mg gallic acid equivalents (GAE)/g lyophilized powder, respectively [16].
The contents of some phytochemicals and the physicochemical properties of strawberry juice may be
affected by oxygen, light, temperature, and enzyme activities such as polyphenol oxidase and by microbial
contamination during storage. Fresh strawberry juice was found to have increased pH (from 3.2 to 4.0) and
alcohol content, and significantly altered color after being stored at room temperature for 2 weeks. Yeast
and mold were also observed [17]. Strawberry juice concentrate (68°Brix) stored at 20°C also developed to
be musty/moldy and to have astringent taste and pungent aromas and brown color after 6 days [18].
R1 O
O HO O
HO
OH HO OH
R2
—R2 —
4-Hydroxybenzoic acid, R1— —H; O OH
— H, R2—
3,4-Dihydroxybenzoic acid, R1— — OH; O
Gallic acid, R1— Ellagic acid
—R2—
— OH;
— R2—
Syringic acid, R1— — OCH3
Benzoic Acids
R1 O
R3 O–R4
R2
Cinnamic acid, R1——R2——R3 — —R4 —
—H;
p-Coumaric acid, R1—— R2——R4— —H, R3— —OH;
p-Coumaroyl glucose, R1— — R2—
— H, R3——OH, R4 —
— glucoside;
Ferulic acid, R1—R
— 4——H , R —
2— OCH , R
3 3—
—OH;
— R2—
Sinapic acid, R1— —OCH3, R3— — OH, R4 ——H;
Caffeic acid, R1—
— R4—
— H, R2——OH, R3— —OH;
—H, R2—
Chlorogenic acid, R1— —R3 —
—OH, R4 — —5-quinoyl
Cinnamic Acids
R1
OH
R1
HO O
OH R2
+
HO O OR3
R2 OH
O
OR3 Kaempferol-3-glucoside, R1— —R2 — —H, R3— —glucose
Quercetin-3-glucoside, R —
— H, R —
— OH, — glucose
R3—
OH 1 2
Quercetin-3-ruticoside, R —
— H, R —
— OH, R —
—rutinose
Cyanidin-3-glucoside: R1— — —
— OH, R2—H, R3 —glucose 1 2 3
— — —
Pelargonidin-3-glucoside: R1— H, R2 —H, R3 —glucose Quercetin-3-glucuronide, R —
1— H, R —
2— OH, R3 —
— glucuronic acid
— H, R2—
Pelargonidin-3-rutinoside: R1— — H, R3 —
—rutinose Myricetin-3-rutinoside, R1——R2— —OH, R3— — rutinose
Anthocyanins Flavonols
OH
OH
HO O
OH
OH OH OH
OH
OH OH
OH HO O R1
HO O HO O
OH
OH OH OH
OH (–)-Epicatechin, R1—
—H; OH
Procyanidin B1 —OH
(–)-Epigallocatechin, R1— (+)-Catechin
Monomeric and dimeric flavanols
FIGURE 44.1 Chemical structures of polyphenols found in strawberry juice.

TABLE 44.2
Phenolic Contents of Strawberry Juice (mg/100 mL)
Food Description Class Compound Content References
6.8°Brix strawberry Hydroxybenzoic acid Gallic acid 6.35 [25]
juice 4-Hydroxybenzoic acid 7.21 [25]
3,4-Dihydroxybenzoic acid 3.20 [25]
Syringic acid 3.79 [25]
Ellagic acid 5.33 [25]
Monomeric flavan-3-ol (−)-Epigallocatechin 0.62 [25]
(−)-Epicatechin 10.87 [25]
(+)-Catechin 6.56 [25]
Hydroxycinnamic acid p-Coumaroyl glucose 64.24 [25]
Chlorogenic acid 3.44 [25]
p-Coumaric acid 4.46 [25]
Cinnamic acid 0.51 [25]
Anthocyanin Cyanidin-3-O-glucoside 0.23 [25]
Pelargonidin-3-O-glucoside 15.13 [25]
Pelargonidin-3-O-rutinoside 1.68 [25]
Flavonol Myricetin-3-O-rutinoside 1.03 [25]
Quercetin-3-O-rutinoside 0.44 [25]
Quercetin-3-O-glucoside 1.36 [25]
Quercetin-3-O-glucuronide 0.44 [25]
Kaempferol-3-O-glucoside 0.45 [25]
Clear strawberry juice Hydroxybenzoic acid Ellagic acid 3.3 [26]
Monomeric flavan-3-ol (+)-Catechin 20.3 [26]
Hydroxycinnamic acid p-Coumaric acid 29.4 [26]
Pelargonidin-3-O-malonylglucoside 14.9 [26]
Flavonol Quercetin 1.9 [26]
Kaempferol 1.8 [26]
Cloudy strawberry juice Hydroxybenzoic acid Ellagic acid 2.4 [26]
Pelargonidin-3-O-glucoside 228 [26]
Kaempferol 6.3 [26]
Puree strawberry juice Hydroxybenzoic acid Ellagic acid 15.8 [26]
Kaempferol 6.7 [26]
44.3.1 Anthocyanins
Pelargonidin-3-glucoside, cyanidin-3-glucoside, and pelargonidin-3-rutinoside are the predominant
anthocyanins found in strawberry juice [14,19] (Figure 44.1). Among them, pelargonidin-3-glucoside
(15.13 mg/100 mL, Table 44.2) is responsible for the appealing, bright, and red color of strawberry
juice. The total anthocyanin contents were at 176–445 mg/L in the supernatants of fresh strawberry
pulp, whereas in clear juice the main anthocyanin pelargonidin-3-glucoside alone was found at
414 mg/L [20,21]. Intake of anthocyanin-rich foods alleviates oxidative stress-related chronic dis-
eases [7,22,23].
The natural pH of strawberry fruits ranges from 3.3 to 3.8, in which anthocyanins (red pigment) should be
fairly stable. The color parameters in strawberry juice (CIE L*a*b*) of lightness (L*), redness (a*), and yel-
lowness (b*) were 28, 44.8, and 33.7, respectively [21]. However, the color of strawberry juice, an indicator of
quality and degree of deterioration, is highly unstable and very susceptible to degradation. The rate of color
degradation of strawberry concentrate (65°Brix) is higher as compared to strawberry juice (8°Brix) [14].
The color stability is greatly affected by processing conditions including light, pH, oxygen, enzymes, ther-
mal treatment, metal ions, and other components such as degradation products of anthocyanins. The high
amount of ascorbic acid in strawberry juice may also contribute to the degradation of anthocyanins [20].
Anthocyanin degradation in strawberry occurs during juice processing and storage. Therefore, factors that
might affect the stability of anthocyanins of these two processes must be studied to ensure good color stabil-
ity. Metallic salts have been found to stabilize strawberry purée color [24]. Phenolic acids such as sinapic
acid enhance the color intensity of strawberry juice, thus improving color quality during storage, and rose
petal polyphenolic extract significantly reduced the thermal degradation of strawberry anthocyanins [24,25].
44.3.2 Flavonols
Procyanidin B1, catechin, quercetin-3-glucuronide, quercetin-3-glucoside, and quercetin-3-rutenoside
were identified in strawberry juices [26] (Figure 44.1). Additional flavonoids including quercetin, kaemp-
ferol, myricetin-3-O-rutinoside, and kaempferol 3-O-glucoside were also found in strawberry purees and
cloudy juices in recent studies [27,28]. Concentrations of individual flavonols along with other phenolic
compounds in strawberry juice are listed in Table 44.2. p-Coumaric acid, rutin, quercetin, isorhamne-
tin, and morin have been reported in strawberry juice [29]. Quercetin, kaempferol, and myricetin are
the three main flavonol aglycones of strawberry juice. These flavonols are strong antioxidants and have
potential antiangiogenic effects on human umbilical vein endothelial cells (HUVECs), by suppress-
ing the vascular endothelial growth factor–stimulated HUVEC tubular structure formation, which are
important processes in the development of cancer and atherosclerosis [30].
44.3.3 Hydroxybenzoic and Hydroxycinnamic Acids

Strawberry juice contains hydroxybenzoic acids, including gallic, 4-hydroxybenzoic, 3,4-dihydroxyben-
zoic, syringic, and ellagic acids (Table 44.2 and Figure 44.1). Ellagic acid and its derivatives in particular
are a diverse group of hydroxybenzoic acids often exist in glycosylated, methylated, methoxylated, and
acylated forms in strawberry juice [31]. Hydroxycinnamic acids in berries are present mainly in the form
of simple esters with hydroxycarboxylic acids or glucose. p-Coumaroyl glucose (64.24 mg/100 mL)
was the main hydroxycinnamic acid derivative in strawberry juice (6.8°Brix), followed by caffeic acid
(7.53 mg/100 mL), chlorogenic acid (3.44 mg/100 mL), and cinnamic acid (0.51 mg/100 mL) [27].
Hydroxybenzoic and hydroxycinnamic acids could scavenge free radicals significantly and have
been consistently associated with reduced risk of cardiovascular and chronic diseases [32,33]. Ellagic
acid and its derivatives have been found to demonstrate certain health effects due to the antiprolifera-
tive and antioxidant properties [34,35]. It inhibited the expression of oxidized low-density lipoprotein
(ox-LDL)-mediated lectin-like ox-LDL (LOX-1) receptor in human endothelial cells as well as the
generation of reactive oxygen species (ROS), which therefore could be used to reduce the development
of atherosclerosis [36]. Kampa et al. [37] studied the antiproliferative effects of several phenolic acids
and a time- and dose-dependent inhibitory effect on T47D human breast cancer cell growth was found
in the order of caffeic acid > ferulic acid = protocatechuic acid = 3,4-dihydroxyphenylacetic acid >
sinapic acid = syringic acid.
Total phenolic content and antioxidant activity of strawberry juice may also be affected during the
storage. When refrigerated for 2 days, the total phenolic content increased significantly from 1302 to
1671 mg GAE/L, and the increase also led to 5.3% higher antioxidant activity. Extended storage of
strawberry juice at the same temperature continued to show increased total phenolic content (6.2%), but
drastically lost its antioxidant activity by 47.4% after 29 days [38]. Longer storage (3 months) of straw-
berry juice at 4°C eventually reduced both the total phenolic content and antioxidant activity, but the
degradation rates were most intense during the first month [39]. The spoilage of fresh strawberry juice
can be controlled by processing and storage under aseptic and/or sanitation conditions; however, an obvi-
ous compromise between sensorial and nutritional values must be considered [40].
44.4 Health Effects

44.4.1 Antioxidant Activity
Total phenolic content of strawberry juice (supernatant of fresh strawberry juice) is 1.98 mg GAE/g [41],
and the antioxidant activities as measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric-reducing
antioxidant potential (FRAP), photochemiluminescence, and oxygen radical absorbance capacity
(ORAC) assays were 10.17, 10.58, 8.48, and 190 μmol trolox equivalents (TE)/g, respectively [42].
The L-929 cell-based assay, using 2′,7′-dichlorofluorescin diacetate as a useful indicator of ROS,
showed that the cellular antioxidant activity of strawberry juice (IC50 values: 52 μg/mL) was superior to
those of highbush blueberry juice (116 μg/mL), kiwi juice (214 μg/mL), and peach juice (1119 μg/mL),
respectively [42].
It was reported that cyanidin-3-glucoside, one of the main anthocyanins in strawberry juice, could act
as an antioxidant in vivo. It suppressed oxidative injure to the liver of rats (2 g/kg diet for 14 days) caused
by hepatic ischemia–reperfusion by lowering the serum thiobarbituric acid–reactive substance (TBARS)
concentration and increasing the oxidation resistance of the serum to lipid peroxidation [43].
44.4.2 Anti-Inflammatory Activity

The effects of strawberry juice administrations on the cytokine secretions by peritoneal macrophages
under different lipopolysaccharide (LPS)-stimulated macrophage models were studied [16]. It was
found that strawberry juice (10–500 μg/mL) administration may exhibit a prophylactic effect against
LPS-induced inflammation of primary peritoneal macrophages via increasing the secretion of anti-
inflammatory cytokine interleukin (IL)-10 (613%). Simultaneously, the inhibitions of pro-inflammatory
cytokine secretion of IL-1β and IL-6 were 38% and 21%, respectively. The same trends of decreased
IL-1β and IL-6 and increased IL-10 were also reported by others, suggesting that strawberry juice has
anti-inflammation potential via modulating pro-/anti-inflammatory cytokine secretion profiles [44]. The
same authors also found that strawberry juice could protect LPS-stimulated macrophages from apoptotic
cell death by modulating Bak and Bcl-2 protein levels [44].
It was suggested that phenolics such as ellagic acid, anthocyanin, catechin, quercetin, and kaempferol
in strawberry juice might contribute to anti-inflammatory activity [16]. The strong immunomodula-
tory effects of strawberry juice and its major phenolic compounds were expressed via decreasing CD4+
T-helper type 1/CD4+ T-helper type 2 ([IFN-γ + IL-2 + IL-12]/IL-10) and pro-/anti-inflammatory or
tumor necrosis factor-alpha/IL-10 cytokine secretion ratios in dose-dependent manners [29].
44.4.3 Reduced Cardiovascular Disease

High fiber content, folates, and antioxidants such as ellagic acid and anthocyanin in strawberry juice
together could strengthen the cardiac function and help lower the risk of CVD. Strawberry juice (at the
concentration of 0.01%) showed complete inhibition of copper-induced oxidation of human low-density
lipoprotein (LDL) in vitro [45]. Its effect in minimizing oxidative damage to LDL lipids was also shown
in a cholesterol-lowering dietary portfolio [46]. Participants were fed a hyperlipidemic diet (soy, viscous
fiber, plant sterol, and nuts) for 2.5 years (body mass index: 19.8–32.3) and then received supplements of
strawberry (454 g/d, 112 kcal) for 1 month. Strawberry-supplemented group showed greater reduction
in oxidative damage to LDL cholesterol (13.4%) and the ratio of total to high-density lipoprotein (HDL)
cholesterol (15.2%) [46]. Long-term strawberry juice consumption may lead to lower levels of circulating
ox-LDL and therefore help reduce the risk of CVD.
Moderate consumption of strawberry juice was also beneficial for preventing early atherosclerosis.
It was found that a strawberry beverage containing 10 g freeze-dried fruit showed lower triacylglycerols
(TAG) and ox-LDL after high-fat meals, in a study involving 24 hyperlipidemic men and women (14
women, 10 men; mean age 50.9 ± 15 years) [47]. After 12 weeks of an atherogenic diet and a daily dose
corresponding to the consumption of 275 mL by a 70 kg human (0.22 g strawberries/mL) in hamsters,
the aortic lipid deposition in hamsters was inhibited by 97%, and the activities of hepatic antioxidant
enzymes superoxide dismutase and glutathione peroxidase were reduced [48].
44.4.4 Against Diabetes

Strawberry juice contains phytochemicals such as ellagic acid derivatives and anthocyanins that are
considered to be beneficial to diabetics. In addition, both the fruit and seeds offer a proper dosage of
dietary fiber, which is also very important for diabetes sufferers. The polyphenols in strawberry juice
were proven as effective α-amylase and α-glucosidase inhibitors [49]. Strawberry juice was reported to
have potential to reduce symptoms of the hyperglycemia-linked type 2 diabetes and related complication
of hypertension by inhibiting the α-amylase (50%), α-glucosidase (80%), and angiotensin-1-converting
enzyme activity (38%) [50].
Cyanidin-3-glucoside in strawberry juice was reported to be an effective insulin secretagogue by stim-
ulating insulin secretion from rodent pancreatic β-cells (INS-1 832/13) in vitro [51]. Moreover, cyanidin-
3-glucoside (0.2% diet for 5 weeks) also showed significant potency in antidiabetic effect by ameliorating
hyperglycemia and insulin sensitivity via downregulation of the retinol binding protein 4 expression and
upregulation of the glucose transporter in type 2 diabetic mice [52]. Pelargonidin-3-glucoside (injection
3 mg/kg body weight) was also observed to increase insulin secretion in streptozotocin-induced diabetic
Wistar rats [53].
44.4.5 Anticancer Effects

Recent evidence suggests that strawberry juice is effective in modulating enzyme activities and inhibiting
cancer cell proliferation. Strawberry juice at concentrations ranging from 25 to 200 μg/mL inhibited the
growth and stimulated apoptosis of human cancer cells including oral (KB, CAL-27), breast (MCF-7), colon
(HT-29, HCT116), and prostate (LNCaP) tumor cell lines in a dose-dependent manner [54,55]. Similar
results have been obtained in stomach adenocarcinoma AGS, prostatic adenocarcinoma PC-3, colorectal
adenocarcinoma Caco-2, and other cancer cell lines (50 μg/mL supernatant of fresh strawberry) [56].

Strawberry (200 g/kg) and orange (800 g/kg) mixed juice was a popular juice and showed good accept-
ability with sensory characteristics and nutritional advantages that support the development of innova-
tive fresh juice [57]. Frozen aerated fruit juice including strawberry juice dessert has been patented to aid
the sweetening and freezing of juices [58]. To obtain a high solid fruit product, a method was reported
to produce ready-to-pour frozen concentrated clarified fruit juice [59]. Strawberry juice was also used
to coat a ready-to-eat cereal [60]. A method of process for curing meat with fruit juice has also been
developed [61].
Microbial contamination is a key concern for product development of strawberry juice and related
products. Strawberry juice is easy to be spoiled by many microorganisms such as yeasts and molds.
Spoilage and metabolism produce off flavor and cause a very short shelf life due to endogenous enzyme
activities. The contents of some phytochemicals and the physicochemical properties of strawberry juice
may be affected by oxygen, light, temperature, and enzyme activities such as polyphenol oxidase and
by microbial contamination. Not only the phenolic content but also the antioxidant activities are signifi-
cantly affected by storage and processing conditions [38–40].
Heat is an effective and perhaps the most widely used technique for preserving and extending the
shelf life of strawberry juice. However, thermal processing causes nonenzymatic browning, leading
to changes in color and formation of undesirable products. Thermal processing can seriously affect
the nutritional quality of strawberry juice resulting not only from the loss of anthocyanins, free amino
acid, and vitamin C but also from changes of color, flavor, texture, and antioxidant properties [3,62].
Therefore, to develop strawberry-based functional foods or beverages, conditions during processing and
storage must be optimized to minimize the loss of nutritionally important bioactives and to retain the
characteristic flavor and color of strawberry juice. Novel nonthermal technologies such as high hydro-
static pressure (HHP), HIPEF, and ultrasound treatment have, therefore, been developed to replace or
complement conventional heat technologies. HIPEF-treated strawberry juice had higher free amino acid
(alanine, leucine, valine, and glycine) concentrations than thermally treated juice, after storage at 4°C for
56 days [3]. HHP combined with low-temperature storage was more effective in retaining the nutritional
value in terms of the ascorbic acid, anthocyanins, and total phenols [63]. Ultrasound treatment was less
destructive to anthocyanins compared to thermal pasteurization [64]. Even so, during both storage and
processing, the strawberry juice still has concomitant loss of flavor, color, and sensory as well as lower
nutritional value due to the loss of antioxidant compounds. Developing novel products based on straw-
berry juice with promising health benefits is still a challenge.
44.6 Conclusion
Strawberry juice is a rich source of functional compounds including ascorbic acid and polyphenols such
as anthocyanins, flavonols, and hydroxybenzoic and hydroxycinnamic acids, in addition to amino acids,
dietary fiber, and unique flavor compounds. A number of in vitro and in vivo studies suggest that straw-
berry juice is a good source of antioxidants and may possess beneficial effects on human health includ-
ing improving cardiovascular function and heart health, reducing inflammation and risk of diabetes and
cancers. Further research is needed to generate sufficient evidence to substantiate these health benefits
of strawberry juice.
REFERENCES
1. Hannum, S.M., Potential impact of strawberries on human health: A review of the science. Crit. Rev.
Food Sci. Nutr., 44, 1–17, 2004.
2. Klopotek, Y., Otto, K., and Bohm, V., Processing strawberries to different products alters contents of
Vitamin C, total phenolics, total anthocyanins, and antioxidant capacity. J. Agric. Food Chem., 53,
5640–5646, 2005.
3. Odriozola-Serrano, I., Garde-Cerdán, T., Soliva-Fortuny, R., and Martín-Belloso, O., Differences in
free amino acid profile of non-thermally treated tomato and strawberry juices. J. Food Compos. Anal.,
32, 51–58, 2013.
4. Nualkaekul, S., Salmeron, I., and Charalampopoulos, D., Investigation of the factors influencing
the survival of Bifidobacterium longum in model acidic solutions and fruit juices. Food Chem., 129,
1037–1044, 2011.
5. Sadilova, E., Stintzing, F.C., Kammerer, D.R., and Carle, R., Matrix dependent impact of sugar and
ascorbic acid addition on color and anthocyanin stability of black carrot, elderberry and strawberry
single strength and from concentrate juices upon thermal treatment. Food Res. Int., 42, 1023–1033, 2009.
6. del Campo, G., Berregi, I., Caracena, R., and Santos, J.I., Quantitative analysis of malic and citric acids
in fruit juices using proton nuclear magnetic resonance spectroscopy. Anal. Chim. Acta, 556, 462–468,
2006.
7. U.S. Department of Agriculture (USDA), USDA National Nutrient for Standard References, Release 23.
National Technical Information Service, USDA, Springfield, VA, 2010.
8. Froes, R.E.S., Neto, W.B., Silva, N.O.C.E., Naveira, R.L.P., Nascentes, C.C., and da Silva, J.B.B.,
Multivariate optimization by exploratory analysis applied to the determination of microelements in fruit
juice by inductively coupled plasma optical emission spectrometry. Spectrochim. Acta B, 64, 619–622,
2009.
9. Mendiola, J.A., Marin, F.R., Señoráns, F.J., Reglero, G., Martín, P.J., Cifuentes, A., and Ibáñez, E.,
Profiling of different bioactive compounds in functional drinks by high-performance liquid chromatog-
raphy. J. Agric. Food Chem., 1188, 234–241, 2008.
10. Derossi, A., De Pilli, T., and Fiore, A.G., Vitamin C kinetic degradation of strawberry juice stored under
non-isothermal conditions. LWT—Food Sci. Technol., 43, 590–595, 2010.
11. Enstrom, J., The epidemiology of vitamin C, in International Encyclopedia of Public Health,
Heggenhougen, H.K., Ed., Academic Press, Oxford, U.K., 2008, pp. 413–424.
12. Wolf, A. and Elmadfa, I., Fruit and vegetable intake of mothers in Europe: Risks/benefits, in Bioactive
Foods in Promoting Health, Watson, R.R. and Preedy, V.R., Eds., Academic Press, San Diego, CA,
2009, pp. 161–172.
13. Thompson, M.D. and Thompson, H.J., Botanical diversity in vegetable and fruit intake: Potential health
benefits, in Bioactive Foods in Promoting Health, Watson, R.R. and Preedy, V.R., Eds., Academic
Press, San Diego, CA, 2019, pp. 1–17.
14. Garzón, G.A. and Wrolstad, R.E., Comparison of the stability of pelargonidin-based anthocyanins in
strawberry juice and concentrate. J. Food Sci., 67, 1288–1299, 2002.
15. Tsao, R. and Yang, R., Optimization of a new mobile phase to know the complex and real polyphenolic
composition: Towards a total phenolic index using high-performance liquid chromatography. J. Agric.
Food Chem., 1018, 29–40, 2003.
16. Lin, J.-Y. and Tang, C.-Y., Strawberry, loquat, mulberry, and bitter melon juices exhibit prophylac-
tic effects on LPS-induced inflammation using murine peritoneal macrophages. Food Chem., 107,
1587–1596, 2008.
17. Saddozai, A.A., Raza, S., and Saleem, S.A., Microbial count and shelf life of strawberry juice. Pak.
J. Agric. Res., 25, 218–223, 2012.
18. Lundahl, D.S., McDaniel, M.R., and Wrolstad, R.E., Flavor, aroma, and compositional changes in
strawberry juice concentrate stored at 20°C. J. Food Sci., 54, 1255–1258, 1989.
19. Wang, S.Y. and Zheng, W., Effect of plant growth temperature on antioxidant capacity in strawberry.
J. Agric. Food Chem., 49, 4977–4982, 2001.
20. Özkan, M., Yemenicioğlu, A., and Cemeroğlu, B., Degradation of various fruit juice anthocyanins by
hydrogen peroxide. Food Res. Int., 38, 1015–1021, 2005.
21. Tiwari, B.K., O’Donnell, C.P., Patras, A., Brunton, N., and Cullen, P.J., Effect of ozone processing on
anthocyanins and ascorbic acid degradation of strawberry juice. Food Chem., 113, 1119–1126, 2009.
22. Hassan, H.A. and Abdel-Aziz, A.F., Evaluation of free radical-scavenging and anti-oxidant properties
of black berry against fluoride toxicity in rats. Food Chem. Toxicol., 48, 1999–2004, 2010.
23. Morimitsu, Y., Kubota, K., Tashiro, T., Hashizume, E., Kamiya, T., and Osawa, T., Inhibitory effect of
anthocyanins and colored rice on diabetic cataract formation in the rat lenses. Int. Congr. Ser., 1245,
503–508, 2002.
24. Rein, M.J. and Heinonen, M., Stability and enhancement of berry juice color. J. Agric. Food Chem.,
52, 3106–3114, 2004.
25. Mollov, P., Mihalev, K., Shikov, V., Yoncheva, N., and Karagyozov, V., Colour stability improvement of
strawberry beverage by fortification with polyphenolic copigments naturally occurring in rose petals.
Innov. Food Sci. Emerg., 8, 318–321, 2007.
26. Abad-García, B., Berrueta, L.A., López-Márquez, D.M., Crespo-Ferrer, I., Gallo, B., and Vicente, F.,
Optimization and validation of a methodology based on solvent extraction and liquid chromatogra-
phy for the simultaneous determination of several polyphenolic families in fruit juices. J. Agric. Food
Chem., 1154, 87–96, 2007.
27. Díaz-García, M.C., Obón, J.M., Castellar, M.R., Collado, J., and Alacid, M., Quantification by UHPLC
of total individual polyphenols in fruit juices. Food Chem., 138, 938–949, 2013.
28. Oszmiański, J. and Wojdyło, A., Comparative study of phenolic content and antioxidant activity of
strawberry puree, clear, and cloudy juices. Eur. Food Res. Technol., 228, 623–631, 2009.
29. Liu, C.J. and Lin, J.Y., Anti-inflammatory effects of phenolic extracts from strawberry and mulberry
fruits on cytokine secretion profiles using mouse primary splenocytes and peritoneal macrophages.
Int. Immunopharmacol., 16, 165–170, 2013.
30. Kim, J.-D., Liu, L., Guo, W., and Meydani, M., Chemical structure of flavonols in relation to modulation
of angiogenesis and immune-endothelial cell adhesion. J. Nutr. Biochem., 17, 165–176, 2006.
31. Versari, A., Biesenbruch, S., Barbanti, D., Farnell, P.J., and Galassi, S., Effects of pectolytic enzymes on
selected phenolic compounds in strawberry and raspberry juices. Food Res. Int., 30, 811–817, 1997.
32. Puangpraphant, S., Berhow, M.A., Vermillion, K., Potts, G., and Gonzalez de Mejia, E., Dicaffeoylquinic
acids in Yerba mate (Ilex paraguariensis St. Hilaire) inhibit NF-kB nucleus translocation in macro-
phages and induce apoptosis by activating caspases-8 and -3 in human colon cancer cells. Mol. Nutr.
Food Res., 55, 1509–1522, 2011.
33. Herrmann, K. and Nagel, C.W., Occurrence and content of hydroxycinnamic and hydroxybenzoic acid
compounds in foods. Crit. Rev. Food Sci., 28, 315–347, 1989.
34. Larrosa, M., García-Conesa, M.T., Espín, J.C., and Tomás-Barberán, F.A., Ellagitannins, ellagic acid
and vascular health. Mol. Asp. Med., 31, 513–539, 2010.
35. Landete, J.M., Ellagitannins, ellagic acid and their derived metabolites: A review about source, metabo-
lism, functions and health. Food Res. Int., 44, 1150–1160, 2011.
36. Lee, W.-J., Ou, H.-C., Hsu, W.-C., Chou, M.-M., Tseng, J.-J., Hsu, S.-L., Tsai, K.-L., and Sheu, W.H.-H.,
Ellagic acid inhibits oxidized LDL-mediated LOX-1 expression, ROS generation, and inflammation in
human endothelial cells. J. Vasc. Surg., 52, 1290–1300, 2010.
37. Kampa, M., Alexaki, V.-I., Notas, G., Nifli, A.-P., Nistikaki, A., Hatzoglou, A., Bakogeorgou, E. et al.,
Antiproliferative and apoptotic effects of selective phenolic acids on T47D human breast cancer cells:
Potential mechanisms of action. Breast Cancer Res., 6, R63–R74, 2004.
38. Piljac-Žegarac, J., Valek, L., Martinez, S., and Belščak, A., Fluctuations in the phenolic content and
antioxidant capacity of dark fruit juices in refrigerated storage. Food Chem., 113, 394–400, 2009.
39. Kalisz, S., Effect of production method of strawberry juices on the content of anthocyanins and colour.
ZYWN.-Nauk Technol. Jakosc, 15, 149–160, 2008.
40. Cordenunsi, B.R., Genovese, M.I., Oliveira do Nascimento, J.R., Aymoto Hassimotto, N.M., José dos
Santos, R., and Lajolo, F.M., Effects of temperature on the chemical composition and antioxidant activ-
ity of three strawberry cultivars. Food Chem., 91, 113–121, 2005.
41. Holasova, M. and Fiedlerova, V., Application and comparison of methods of antioxidant activity deter-
mination in fruit and vegetable juices. Chem. Listy, 105, 766–772, 2011.
42. Girard-Lalancette, K., Pichette, A., and Legault, J., Sensitive cell-based assay using DCFH oxidation for
the determination of pro- and antioxidant properties of compounds and mixtures: Analysis of fruit and
vegetable juices. Food Chem., 115, 720–726, 2009.
43. Tsuda, T., Horio, F., Kitoh, J., and Osawa, T., Protective effects of dietary cyanidin 3-O-β-d-glucoside
on liver ischemia-reperfusion injury in rats. Arch. Biochem. Biophys., 368, 361–366, 1999.
44. Liu, C.-J. and Lin, J.-Y., Anti-inflammatory and anti-apoptotic effects of strawberry and mulberry
fruit polysaccharides on lipopolysaccharide-stimulated macrophages through modulating pro-/anti-
inflammatory cytokines secretion and Bcl-2/Bak protein ratio. Food Chem. Toxicol., 50, 3032–3039,
2012.
45. Kulisic-Bilusic, T., Schnäbele, K., Schmöller, I., Dragovic-Uzelac, V., Krisko, A., Dejanovic, B., Milos,
M., and Pifat, G., Antioxidant activity versus cytotoxic and nuclear factor kappa B regulatory activities
on HT-29 cells by natural fruit juices. Eur. Food Res. Technol., 228, 417–424, 2009.
46. Jenkins, D.J., Nguyen, T.H., Kendall, C.W., Faulkner, D.A., Bashyam, B., Kim, I.J., Ireland, C. et al.,
The effect of strawberries in a cholesterol-lowering dietary portfolio. Metabolism, 57, 1636–1644,
2008.
47. Burton-Freeman, B., Linares, A., Hyson, D., and Kappagoda, T., Strawberry modulates LDL oxidation
and postprandial lipemia in response to high-fat meal in overweight hyperlipidemic men and women.
J. Am. Coll. Nutr., 29, 46–54, 2010.
48. Rouanet, J.M., Décordé, K., Rio, D.D., Auger, C., Borges, G., Cristol, J.P., Lean, M.E.J., and Crozier,
A., Berry juices, teas, antioxidants and the prevention of atherosclerosis in hamsters. Food Chem., 118,
266–271, 2010.
49. McDougall, G.J., Shpiro, F., Dobson, P., Smith, P., Blake, A., and Stewart, D., Different polyphenolic
components of soft fruits inhibit α-amylase and α-glucosidase. J. Agric. Food Chem., 53, 2760–2766,
2005.
50. Cheplick, S., Kwon, Y.-I., Bhowmik, P., and Shetty, K., Phenolic-linked variation in strawberry cul-
tivars for potential dietary management of hyperglycemia and related complications of hypertension.
Bioresour. Technol., 101, 404–413, 2010.
51. Jayaprakasam, B., Vareed, S.K., Olson, L.K., and Nair, M.G., Insulin secretion by bioactive anthocya-
nins and anthocyanidins present in fruits. J. Agric. Food Chem., 53, 28–31, 2005.
52. Sasaki, R., Nishimura, N., Hoshino, H., Isa, Y., Kadowaki, M., Ichi, T., Tanaka, A. et al., Cyanidin
3-glucoside ameliorates hyperglycemia and insulin sensitivity due to downregulation of retinol binding
protein 4 expression in diabetic mice. Biochem. Pharmacol., 74, 1619–1627, 2007.
53. Roy, M., Sen, S., and Chakraborti, A.S., Action of pelargonidin on hyperglycemia and oxidative damage
in diabetic rats: Implication for glycation-induced hemoglobin modification. Life Sci., 82, 1102–1110,
2008.
54. Seeram, N.P., Adams, L.S., Zhang, Y., Lee, R., Sand, D., Scheuller, H.S., and Heber, D., Blackberry,
black raspberry, blueberry, cranberry, red raspberry, and strawberry extracts inhibit growth and stimu-
late apoptosis of human cancer cells in vitro. J. Agric. Food Chem., 54, 9329–9339, 2006.
55. Zhang, Y., Seeram, N.P., Lee, R., Feng, L., and Heber, D., Isolation and identification of strawberry
phenolics with antioxidant and human cancer cell antiproliferative properties. J. Agric. Food Chem.,
56, 670–675, 2008.
56. Boivin, D., Blanchette, M., Barrette, S., Mograbi, A., and Béliveau, R., Inhibition of cancer cell pro-
liferation and suppression of TNF-induced activation of NFκB by edible berry juice. Anticancer Res.,
27, 937–948, 2007.
57. Endrizzi, I., Pirretti, G., Calò, D.G., and Gasperi, F., A consumer study of fresh juices containing berry
fruits. J. Sci. Food Agric., 89, 1227–1235, 2009.
58. Wade, B.R. and Wade, T.L., Frozen aerated fruit juice dessert, US Patent # US4609561 A, Original
Assinee: Olympus Industries, Inc., 1986.
59. Chen, C.S. and Chen, W.A., Method for producing ready to pour frozen concentrated clarified fruit
juice, fruit juice produced therefrom, and high solids fruit product, US Patent # US5756141 A, Original
Assignee: Chin Shu Chen, William Apollo Chen, 1998.
60. Carpenter, T.L., Fisher, W., and Smith, T.A., Coating cereal with fruit juice, US Patent # US4880645 A,
General Foods Corp., 1989.
61. Stumpf, R.E. and Stumpf, R.W., Process for curing meat with fruit juice, US Patent # US4806373 A,
Original Assignee: Robert W. Stumpf, Rebecca Fitch, 1989.
62. Odriozola-Serrano, I., Soliva-Fortuny, R., and Martín-Belloso, O., Impact of high-intensity pulsed
electric fields variables on vitamin C, anthocyanins and antioxidant capacity of strawberry juice.
LWT—Food Sci. Technol., 42, 93–100, 2009.
63. Cao, X., Bi, X., Huang, W., Wu, J., Hu, X., and Liao, X., Changes of quality of high hydrostatic pres-
sure processed cloudy and clear strawberry juices during storage. Innov. Food Sci. Emerg., 16, 181–190,
2012.
64. Dubrovic, I., Herceg, Z., Jambrak, A.R., Badanjak, M., and Dragovic-Uzelac, V., Effect of high intensity
ultrasound and pasteurization on anthocyanin content in strawberry juice. Food Technol. Biotechnol.,
49, 196–204, 2011.
45
Watermelon Juice
Beraat Ozcelik and Merve Yavuz
CONTENTS
45.1 Introduction....................................................................................................................................553
45.4 Health Effects................................................................................................................................ 556
45.6 Conclusion......................................................................................................................................559
References................................................................................................................................................559
45.1 Introduction
Watermelon (Citrullus lanatus) belongs to the Cucurbitaceae family and is cultivated in almost all
warm regions of the world. It can exist in different colors such as red, orange, and yellow depending
on the lycopene and β-carotene content. To date, watermelon has been viewed as a nonnutritional crop,
but in recent years, several bioactive compounds have been determined and the beneficial effects have
been demonstrated by in vivo and in vitro studies [1]. Watermelon contains phenolics, which are mainly
hydroxycinnamic acid derivatives and a large amount of lycopene giving its characteristic red color and
powerful antioxidant activity [2,3]. Watermelon juice is gaining popularity in recent years due to its
sensorial, physical, and nutritional characteristics [4].
Watermelon juice is produced commercially with thermal treatment to inactivate microorganisms and
enzymes. However, this may lead to some undesirable changes in its color, flavor, or other attributes.
Nonthermal treatments are used in obtaining more acceptable watermelon juice [3–9]. Due to its pleasant
flavor, watermelon juice is also used in alcoholic cocktail beverages.
Lucier and Lin [10] indicated that 85.3% of watermelon is consumed in the home, but there is no avail-
able consumption data for watermelon juice or other processed watermelon products such as roasted
watermelon seeds. This chapter highlights the nutritional properties of watermelon juice and its bioac-
tive components, antioxidant activity, and effects on human health as well as novel formulations and
processes, which are studied to produce more acceptable watermelon juice.

Crandall and Kesterson [11] determined that watermelon is composed of 41% juice, 8% pulp, 1% seeds,
and 50% rind on fresh weight (FW). In addition, Sogi et al. [12] reported that watermelon is composed
of 42% juice, 33.6% rind, and 23.6% pomace (including seeds) on FW. The composition varies among
cultivars and seasons [13,14]. Watermelon juice contains a high level of potassium, which is the respon-
sible constituent of diuretic property of watermelon [15]. Watermelon juice is also a good source of
vitamin C. A cup (237 mL) of watermelon juice contains 20% of the daily value for vitamin C in addition
553
TABLE 45.1
Compositional and Nutritional Characteristics of Watermelon and Watermelon Juice
(per 100 g or 100 mL)
Unit Watermelon [17] Watermelon Juice [18,19,32]
Water g 91.45 91.8
Energy kcal 30 36
Protein g 0.61 0.44
Lipid (fat) g 0.15 0.04
Reducing sugar g 4.99 4.5
Sucrose g 1.21 2.71
Ash g 0.25 0.29
Minerals
Calcium mg 7 na
Magnesium mg 10 na
Phosphorus mg 11 na
Potassium mg 112 80
Sodium mg 1 20
Vitamins
Niacin mg 1.78 na
Riboflavin mg 0.021 na
Thiamine mg 0.033 na
Vitamin A (RAE) mg 0.028 na
Vitamin B6 mg 0.045 na
Abbreviations: na, not available; RAE, retinol activity equivalents.
to potassium and vitamin A [16]. USDA [17] Recommended Dietary Allowances (RDA) of vitamin C as
60 mg/day. It is an essential nutrient for normal metabolic functions of the human body. Compositional
and nutritional characteristics of watermelon and watermelon juice are given in Table 45.1.

Numerous studies have demonstrated that watermelon is a rich source of bioactive compounds and
possesses high antioxidant activity [20–28]. It includes a variety of bioactive compounds, such that
71 phytochemicals, among which phenolic compounds, namely, phenolic acids (hydroxybenzoic and
hydroxycinnamic acids), flavonoids derivatives, lignans, iridoids, coumarins, and stilbenoids were
identified. Protocatechuic acid glucoside, p-coumaric acid glucoside, 3-O-feruloylsucrose, sinapic acid
glucoside, O-caffeoylshikimic acid, rutin, quercetin rhamnoside, vanillyl catalpol (picroside), ajugol,
catalposide, (+)-aviprin, saligenin glucopyranoside, and hydroquinone glucuronide are among the char-
acterized phenolic compounds in watermelon [29]. Chemical structures of some common phenolic com-
pounds found in watermelon are shown in Figure 45.1.
Oms-Oliu et al. [7] analyzed untreated watermelon juice for its antioxidant capacity and found
4.92% of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical inhibition. Antioxidant capacity was found
to be 42.70 mg ascorbic acid equivalents (AAE)/kg in fresh watermelon juice, 36.80 mg AAE/kg in
pasteurized noncentrifuged juice, and 33.80 mg AAE/kg in centrifuged pasteurized juice [30]. Naz et al.
[21] determined DPPH and ferric-reducing antioxidant power (FRAP) in watermelon juice at levels of
29.11% radical inhibition and 21.67 mM α-tocopherol equivalents (α-TE)/g, respectively. Furthermore,
lycopene extract showed DPPH and FRAP as 57% radical inhibition and 37.6 mM α-TE/g, respectively.
Antioxidant activity of watermelon juice is also found to be 49% inhibition of β-carotene [21].
Watermelon Juice 555
HO
HO CO2H
O
HO O
OH OH
Protocatechuic acid glucoside
HO
HO CO2H
O
HO O
OH
p-Coumaric acid glucoside
OH
OH OH
O O
HO O OH
O
HO OH
O
OH
OMe
3-O-Feruloylsucrose
FIGURE 45.1 Chemical structures of common phenolic compounds found in watermelon.
Polyphenol contents of watermelon juice, centrifuged–pasteurized watermelon juice, and noncentri-

fuged–pasteurized juice have been reported to be 341, 234, and 255 mg chlorogenic acid equivalents
(CAE)/kg, respectively [30]. Naz et al. [21] determined total phenolic content of watermelon juice and
lycopene extract at the level of 23.63 and 97.15 mg gallic acid equivalents (GAE)/100 g, respectively.
Freezing of watermelon juice did not induce significant changes (P > 0.05) in total phenolic and flavonoid
content of watermelon juice [22].
Watermelon is rich in carotenoids particularly lycopene, β-cryptoxanthin, and β-carotene as well as
α- and γ-tocopherols at the lower level [24]. According to the study conducted by Sharma et al. [31]
total carotenoid content in fresh watermelon juice was reduced from 4.57 to 0.93 mg/100 g and lyco-
pene level from 4.40 to 0.82 mg/100 g after a severe heat treatment (50°C–90°C up to 5 h). Tarazona-
Diaz and Aguayo [30] determined the lycopene levels in acidified–pasteurized–noncentrifuged juice and
acidified–pasteurized–centrifuged juice at the levels of 10.99 and 6.99 mg/kg, respectively. Lycopene
content found in the literature for watermelon juice is given in Table 45.2.
There are several contradictory results in the literature for ascorbic acid level in both watermelon and
watermelon juice. Watermelon juice has higher ascorbic acid than that of numerous fruit juices such as
apple, carrot, grape, peach, and pear [25]. Levels of some bioactive compounds in watermelon juice are
represented in Table 45.2.
Hayoglu and Fenercioglu [32] observed a 50% reduction of ascorbic acid levels after pasteurization
in watermelon juice, and the same levels of it was noted at pasteurization temperatures of 70°C–100°C.
Doodnath and Badrie [33] applied pasteurization at 85°C for 20 min and indicated that ascorbic acid
level was decreased, but the reduction rate was not provided. Sharma et al. [31] indicated that lycopene
and ascorbic acid values showed a significant reduction by exposing the watermelon juice to thermal pas-
teurization at 90°C for 5 h. However, conditions used in the study were more severe than that of thermal
TABLE 45.2
Bioactive Compounds Found in Watermelon Juice
TSS Total Phenolics Total Flavonoids Total Carotenoids Lycopene Ascorbic Acid
(°Brix) (mg GAE/100 mL) (µg QE/mL) (mg/100 mL) (mg/L) (mg/100 mL) References
na 13.89 na na 52.9 4.02 [3]
9.47 1694 na na 36.76 na [8]
na 249 9.84 na na na [22]
9.55 1.62 na na 62.56 na [5]
na na na na 62 2.6 [7]
na na na 4.57a 44.03a na [31]
na na na na na 9.5 [25]
8.5 na na na na 6.4a [32]
8.5 na na na na 3.2b [32]
9.03 na na na 14.97a na [28]
na 23.63 na na 45.3 na [21]
a Fresh juice.
b Thermally treated juice.
Abbreviations: TSS, total soluble solids; GAE, gallic acid equivalents; QE, quercetin equivalents; na, not available.
O O
H2N N OH
H
NH2
FIGURE 45.2 Chemical structure of l-citrulline.
treatments used in the studies on thermally treated watermelon juice. Moreover, Tarazona-Diaz and
Aguayo [30] observed approximately 50% reduction of lycopene levels in watermelon juice after pasteur-
ization at 87.7°C for 20 s. They also studied effect of centrifugation on watermelon juice. Lycopene level
of pasteurized–centrifuged watermelon juice was 6.99 mg/L, while that of fresh one was 14.97 mg/L
(Table 45.2). For commercial production of watermelon juice, strict thermal processing is needed in order
to have a pH > 5, which led to significant reduction in lycopene and visual color of the juice [31].
Watermelon has high content of l-citrulline, which is almost absent in other natural foods. l-citrulline
is the compound that was first isolated from watermelon and therefore named l-citrulline (Figure 45.2).
Watermelon flesh contained 2.33 g/kg FW l-citrulline [34]. Wu et al. [34] found total free amino acids at
the level of 4.5 g/L and l-citrulline plus arginine accounted for 71% of this level in the watermelon juice.
Watermelon is a natural and rich source of amino acid. It was proven that l-citrulline has much higher
bioavailability in a matrix of watermelon juice when it was not exposed to heat treatment by in vitro study
conducted on Caco-2 cells. The highest absorption percentage (18.87%) was achieved for unpasteurized
watermelon juice, while l-citrulline in the control (l-citrulline + water) sample had absorption at 12%.
Pasteurized (80°C for 40 s) watermelon juice gave a lower rate of bioavailability of l-citrulline (~9%) [28].
45.4 Health Effects

Watermelon juice is reported to decrease the risk of chronic diseases such as asthma, atherosclerosis,
diabetes, cancers, arthritis, and age-related degenerative pathologies on account of being a powerful
antioxidant and a rich source of bioactives [26,27]. The beneficial effects of watermelon juice are related
to its possible antioxidant, anti-inflammatory, and vasodilatory properties [35].
l-citrulline is a nonprotein amino acid, which is an intermediate product of a reaction in ureagenesis.
In recent years, research has focused on its crucial role in nitric oxide (NO) metabolism and regulation.
During nitric acid cycle, it acts as a precursor of arginine, and arginine is recycled from citrulline [36].
Collins et al. [37] stated that l-citrulline from watermelon was converted into arginine to a large extent.
l-citrulline has also hydroxyl radical scavenging activity that can protect DNA [38]. Because of its long-
side-chain structure, it has multiple sites to scavenge hydroxyl radicals. In addition, l-citrulline has an
ability to generate NO apart from its antioxidant properties. These two features of l-citrulline make
it a preventative constituent for hypertension, heart failure, atherosclerosis, sickle cell disease, sexual
stamina, and erectile functions by decreasing oxidative stress and increasing arginine availability [28].
Watermelon juice as a rich source of l-citrulline can also decrease hyperglycemia, hyperlipidemia, and
oxidative stress [39].
Asita and Molise [40] studied antimutagenic effects of watermelon juice on cyclophosphamide (CP)-
induced genotoxicity, which was assessed by proportion of polychromatic erythrocyte (PCE) and fre-
quency of micronucleated polychromatic erythrocytes (MNPCE) in mice. Results of the proportion of
PCE showed no significant difference (P > 0.05) in any of the experimental groups. However, they found
that pretreatment with 50% and 25% doses of watermelon juice reduced the frequency of CP-induced
MNPCE. According to the results of the study, natural antigenotoxins are present in watermelon juice.
Watermelon juice also has a protective effect on liver, kidney, and brain tissues by preventing the
increase in lipid peroxide formation probably due to its antioxidant activity according the study con-
ducted on rats [22].
Wu et al. [34] asserted that watermelon juice can be a functional food because of its effects on reduc-
ing serum concentrations of cardiovascular risk factors, improving glycemic status, and ameliorating
vascular dysfunction in obese animals with type II diabetes. Dietary supplementation with watermelon
pomace juice of rats had higher serum concentrations of arginine, lower fat accumulation and serum
concentrations of glucose, free fatty acids, homocysteine, and dimethylarginine. Serum concentrations
of glucose, free fatty acids, triacylglycerols (TAG), and cholesterol in rats having noninsulin-dependent
diabetes mellitus, which received a 4-week dietary supplementation of watermelon pomace juice, were
17.8 1.26, 5.80, and 4.93 mmol/L, respectively, while those of the control group were 22.8, 1.53, 5.92,
and 5.04 mmol/L, respectively.
Watermelon has a high content of carotenoids and exerts a decreasing effect on the risk of myocardial
infarction, anticancer properties, and supports a healthy eye [41]. Lycopene is the major carotenoid of
red-fleshed watermelons having positive health effects. Prostate cancer, coronary heart disease (CHD),
and lung cancer may be prevented by intake of watermelon [42]. Edwards et al. [43] demonstrated that
a diet on fresh-frozen watermelon juice used by healthy, nonsmoking adults (36–69 years) for a 3-week
period showed lycopene bioavailability to be extremely high. The diet contained 20.1 mg/day lycopene
and 2.5 mg/day β-carotene from watermelon juice (W-20), doubled dose of that (W-40), and 18.4 mg/d
lycopene and 0.6 mg/d from canned tomato juice (T-20). Plasma lycopene concentrations for the W-20,
W-40, T-20, and control treatments were 1078, 118, 960, and 272 nmol/L, respectively. W-20 and W-40
treatments resulted in plasma concentrations of β-carotene levels at 574 and 694 nmol/L, respectively,
which are higher than that of the control treatment (313 nmol/L).

Numerous studies have been conducted on watermelon juice to investigate the effects of novel technolo-
gies; formulations consisting watermelon juice are also available. Shin et al. [44] produced watermelon
juice from watermelon fruit with an average yield of 56.2% of the FW applying a thermal treatment
at 100°C for 5 min. Sedimentation (within 24 h) was chosen to separate colloidal components from
watermelon juice. The addition of cane sugar to reach a total solid content of 11–13°Brix could give
organoleptically preferred watermelon juice [44]. In addition to that, Marks et al. [45] patented a com-
mercial packaged watermelon juice that included water, malic acid, polysaccharide, sweetener, and natu-
ral fl
avor. Watermelon juice having 57% and 74% juice content, titratable acidity in a range of 0.27–0.33,
and total solid content in a range of 10–13°Brix was subjected to heat treatment at 77°C for 15 s. It was
then cooled and bottled at refrigeration temperature.
To obtain watermelon juice having high nutritional and microbiological quality, Yingliang at al. [46]
introduced a new processing method, which included boiling at 65°C and 0.095 MPa for 7 min and
addition of N2 for sterilization of watermelon juice. Juice processing with this method had a shelf life
<12 months and retained its flavor.
Rawson et al. [3] subjected watermelon juice to thermosonication, which is an alternative method to
thermal treatment. However, to produce watermelon juice high in bioactive components, process param-
eters of thermosonication should be adjusted well.
High pressure is another treatment applied to thermosensitive watermelon juice. Lycopene content
and pH value did not change with pressure during concentration of watermelon juice [47]. Zhang et al.
[6] also determined higher lycopene levels in watermelon juice, which was subjected to high-pressure
treatment compared to that of ultraviolet (UV)-C and thermal treatment. The total lycopene and phenolic
contents remained unchanged upon UV-C treatment in Teflon® coil even at high UV-C exposure with
longer shelf stability observed for the resultant juice [8].
To prevent nonenzymatic browning in thermally treated watermelon juice, high-intensity pulsed elec-
tric field processing (HIPEF) was introduced to increase shelf life of the juice. During HIPEF process-
ing, low frequencies (<100 Hz) and short pulses (up to 2.5 ls) afforded watermelon juices with minor
change in color and no change in the initial amount of hydroxymethylfurfural (HMF) [4].
The high-pressure carbon dioxide (HPCD) and mild-heat treatment was combined and provided a
significant reduction on peroxidase and polyphenol oxidase residual activity with treatment time and
pressure during the first 5 min. Inactivation of pectin methylesterase at high levels required longer times
of processing. Lycopene content and pH value of HPCD-treated watermelon juice decreased slightly, but
total phenolic content and total soluble solids remained unchanged [5].
Spray drying was applied to watermelon juice using 5% maltodextrin as a drying agent and at 150°C
giving the best quality powder. Total soluble solids, acidity, and glucose content did not change signifi-
cantly, but lycopene content was reduced from 28.45 to 18.24 mg/mL. However, this was still a useable
method to produce reconstituted watermelon juice because of providing longer shelf life and also repre-
senting a sensorially acceptable juice [48].
Reverse osmosis is another treatment applied to watermelon juice to date. The results of a related study
showed that lycopene content and antioxidant capacity increased in concentrated watermelon juice by
reverse osmosis [49]. In addition, Das Gupta and Jayarama [50] reported that reconstituted watermelon
juice prepared by reverse osmosis reached a high level of sensory acceptability by panelists.
Gusina and Trostinskaya [15] introduced a new recipe by using low acidity of watermelon juice that
included a mixture of 85.4% watermelon juice with 8.5% apple paste, 6% sugar, and 0.1% citric acid.
Acidification using citric acid preserved its fresh sensory properties [30]. It is reported that watermelon
juice at 20 or 25°Brix and pH 3.75–3.87 consisting 0.20% xanthan gum and 0.15% citric acid was the
most acceptable in sensory analysis [33]. Bai-Hui and Cheng-Yu [51] recommended a composite sta-
bilizer formula for quick-freeze watermelon juice beverage consisting of xanthan gum (0.05%), pectin
(0.08%), and carboxymethyl cellulose (0.1%). In addition, Saini and Bains [52] introduced a new method
to produce vitamin-enriched watermelon juice. Low ascorbic acid content of fresh juice was increased by
fortification with different levels of ascorbic acid at 60, 80, or 100 mg/100 g juice before bottling. They
also increased the shelf life of watermelon juice due to the lower pH.
Qian et al. [53] introduced a clarifying agent, chitosamine, to watermelon juice production, which did
not cause a change in total acidity, soluble solid content, and ascorbic acid level and also provided a high
transparency.
Watermelon juice is not only used as a beverage but also used in formulations of different products
such as sorbet, yogurt, sauce, salad dressing, fruit salad, dessert, bakery filling, candy, or bar mix [54].
Furthermore, an alcoholic beverage was obtained from watermelon juice using Saccharomyces cere-
visiae, S. cerevisiae var. ellipsoideus, and Kluyveromyces fragilis. An alcoholic beverage containing
alcohol at 5.3% was produced from watermelon juice having a total reducing sugar at 3.2% naturally that
was fortified with 12% glucose [55].
Shin et al. [44] produced a novel product from watermelon juice pasteurized at 90°C for 5 min by
lactic acid fermentation, which was acceptable by 60% of panelists when considering commercial
fruit nectars on the market. Moreover, Mestry et al. [56] developed a non-dairy-based food formula-
tion containing a watermelon and carrot juice mixture in the ratio 70:30 (v/v), which was sensorially
most acceptable ratio by the panelists. The juice mixture was fermented at 37°C with Lactobacillus
acidophilus to obtain a probiotic juice and then spray dried at a temperature of 120°C–160°C and
flow rate of 2.0–5.0 mL/min using maltodextrin as a carrier at a concentration of 10%–15% by weight.
Lycopene and β-carotene content of the dried powder of fermented mixed juice of carrot and water-
melon showed a reduction at higher temperatures and longer exposure, while lower inlet air tempera-
tures provided a good retention [56].
Owing to its low acidic character and high fructose to glucose ratio, watermelon juice has a good
potential to be used in juice formulations. Organoleptic quality of several ratios was determined and vol-
ume ratios of watermelon/pineapple/orange of 80:10:10, watermelon/sour cherry/apple of 50:25:25, and
watermelon/pomegranate of 75:25 were highly acceptable by panelists in different studies [57]. Citrus
juices may also be blended with watermelon juices for producing nonbitter juices by adjusting the limo-
nin content [58].
Oludemi and Akanbi [59] blended 50% tomato, 25% watermelon, and 25% pineapple juice, which was
given the highest total antioxidant capacity of 3.69 mg catechin equivalents (CE)/100 mL. They recom-
mended blending the tomato juice with watermelon juice because of it has a lower viscosity and very
attractive flavor.
45.6 Conclusion
Watermelon juice is a nutritional beverage with reported health benefits such as antioxidant, anti-
inflammatory, and vasodilatory properties as a possible preventive beverage for cardiovascular disease
(CVD). It is a rich source of carotenoids, predominantly lycopene, and contains multiple minerals and
vitamins. Watermelon juice is also a rich source of l-citrulline improving NO release, which has sev-
eral positive health effects. There is, however, a lack of studies in the literature on the characteriza-
tion and quantification of the phytochemicals, antioxidant properties, and health benefits of watermelon
juice. Thermal treatments cause color and viscosity changes and also reduction of bioactive compounds.
Studies on the bioactive compounds of watermelon juice are mostly conducted on fresh juice; therefore,
more studies are needed on pasteurized commercial watermelon juice. Novel technologies represent
a good option for the processing of the watermelon juice. It is possible to mix watermelon juice with
other juices to reach a desired product with a pleasant aroma and flavor. Despite the nutritional benefits
of watermelon juice, there is still a limited number of such products available on the market. However,
commercial watermelon juice may be a healthy option for people living in metropolitan areas where
watermelon consumption is lower.
REFERENCES
1. Perkins-Veazie, P., Davis, A., and Collins, J.K., Watermelon: From dessert to functional food. Israel J.
Plant Sci., 60, 395–402, 2012.
2. Gil, M.I., Aguayo, E., and Kader, A.A., Quality changes and nutrient retention in fresh-cut versus whole
fruits during storage. J. Agric. Food Chem., 54, 4284–4296, 2006.
3. Rawson, A., Tiwari, B.K., Patras, A., Brunton, N., Brennan, C., Cullen, P.J., and O’Donnell, C., Effect
of thermosonication on bioactive compounds in watermelon juice. Food Res. Int., 44, 1168–1173, 2011.
4. Aguilo-Aguayo, I., Soliva-Fortuny, R., and Martín-Belloso, O., Avoiding non-enzymatic browning by
high-intensity pulsed electric fields in strawberry, tomato and watermelon juices. J. Food Eng., 92,
37–43, 2008.
5. Liu, Y., Hu, X., Zhao, X.Y., and Song, H., Combined effect of high pressure carbon dioxide and mild
heat treatment on overall quality parameters of watermelon juice. Innov. Food Sci. Emerg., 13, 112–119,
2011.
6. Zhang, C., Trierweiler, B., Li, W., Butz, P., Xu, Y., Rufer, C.E., and Zhao, X., Comparison of thermal,
ultraviolet-C, and high pressure treatments on quality parameters of watermelon juice. Food Chem.,
126, 254–260, 2010.
7. Oms-Oliu, G., Odriozola-Serrano, I., Soliva-Fortuny, R., and Martín-Belloso, O., Effects of high-
intensity pulsed electric field processing conditions on lycopene, vitamin C and antioxidant capacity of
watermelon juice. Food Chem., 115, 1312–1319, 2009.
8. Feng, M., Ghafoor, K., Seo, B., Yang, K., and Park, J., Effects of ultraviolet-C treatment in Teflon® -coil
on microbial populations and physico-chemical characteristics of watermelon juice. Innov. Food Sci.
Emerg., 19, 133–139, 2013.
9. Liu, Y., Hu, X.S., Zhao, X.Y., and Zhang, C., Inactivation of polyphenol oxidase from watermelon juice
by high pressure carbon dioxide treatment. J. Food Sci. Technol., 50, 317–324, 2013.
10. Lucier, B. and Lin, B.H., Factors affecting watermelon consumption in the United States, U.S.,
Department of Agriculture-Economic Research Service, USDA, Springfield, VA, 2001.
11. Crandall, P.G. and Kesterson, J.W., Components of processed watermelon fruit. J. Am. Soc. Hortic. Sci.,
106, 493–495, 1981.
12. Sogi, D.S., Oberoi, D.P.S., and Malik, S., Effect of particle size, temperature, and total soluble solids
on the rheological properties of watermelon juice: A response surface approach. Int. J. Food Prop., 13,
1207–1214, 2010.
13. Ghazizadeh, M., Razeghi, A., Valaee, N., Mirbagheri, E., Tahbaz, F., Motevallizadeh, H., Seyedahmadian,
F., and Mirzapour, H., Watermelon juice concentrate. Asia Pac. J. Clin. Nutr., 13, 162–162, 2004.
14. Arocho, Y.D., Bellmer, D., Maness, N., McGlynn, W., and Rayas-Duarte, P., Watermelon pomace com-
position and the effect of drying and storage on lycopene content and color. J. Food Qual., 35, 331–340,
2012.
15. Gusina, G.B. and Trostinskaya, L.O., Watermelon juice with pulp. Konserv. Iovoschesuch. Prom., 3,
17–18, 1974.
16. Cook, L.N., Comparisons of physicochemical properties of watermelon juice treated with pulsed electric
fields and thermal pasteurization, MSc thesis, Louisiana State University, Baton Rouge, Los Angeles,
CA, 2007.
18. Chan, S., Problems in production of canned and bottled watermelon juice, MSc thesis, Oregon State
College, Corvallis, OR, 1949.
19. Khan, G. and Uddin, F., Some important metallic constituents in juice of watermelon. Hamdard Med.,
46, 48–50, 2003.
20. Djuric, Z. and Powell, L.C., Antioxidant capacity of lycopene-containing foods. Int. J. Food Sci. Nutr.,
52, 143–149, 2001.
21. Naz, A., Butt, M.S., Pasha, I., and Nawaz, H., Antioxidant indices of watermelon juice and lycopene
extract. Pak. J. Nutr., 12, 255–260, 2013.
22. Altas, S., Kızıl, G., Kızıl, M., Ketani, A., and Haris, P.I., Protective effect of Diyarbakır watermelon
juice on carbon tetrachloride-induced toxicity in rats. Food Chem. Toxicol., 49, 2433–2438, 2011.
23. Leong, L.P. and Shui, G., An investigation of antioxidant capacity of fruits in Singapore markets.
Food Chem., 76, 69–75, 2002.
24. Isabelle, M., Lee, B.L., Lim, M.T., Koh, W.P., Huang, D., and Ong, C.N., Antioxidant activity and
profiles of common fruits in Singapore. Food Chem., 123, 77–84, 2010.
25. Leonard, S.S., Cutler, D., Ding, M., Vallyathan, V., Castranova, V., and Shi, X., Antioxidant properties
of fruit and vegetable juices: More to the story than ascorbic acid. Ann. Clin. Lab. Sci., 32, 193–200,
2002.
26. Choo, W.S. and Sin, W.Y., Ascorbic acid, lycopene and antioxidant activities of red-fleshed and yellow-
fleshed watermelons. Adv. Appl. Sci. Res., 3, 2779–2784, 2012.
27. Tlili, I., Hdider, C., Lenucci, M.S., Ilahy, R., Jebari, H., and Dalessandro, G., Bioactive compounds and anti-
oxidant activities during fruit ripening of watermelon cultivars. J. Food Comp. Anal., 24, 923–928, 2011.
28. Tarazona-Diaz, M.P., Alacid, F., Carrasco, M., Martinez, I., and Aguayo, E., Watermelon juice: Potential
functional drink for sore muscle relief in athletes. J. Agric. Food Chem., 61, 7522–7528, 2013.
29. Abu-Reidah, I.M., Arraez-Roman, D., Carretero, A.S., and Fernandez-Gutierrez, A., Profiling of
phenolic and other polar constituents from hydro-methanolic extract of watermelon (Citrullus lana-
tus) by means of accurate-mass spectrometry (HPLC-ESI-QTOF-MS). Food Res. Int., 51, 354–362,
2012.
30. Tarazona-Diaz, M.P. and Aguayo, E., Influence of acidification, pasteurization, centrifugation and
storage time and temperature on watermelon juice quality. J. Sci. Food Agric., 93, 3863–3869, 2013.
31. Sharma, R., Kaur, D., Oberoi, D.P.S., and Sogi, D.S., Thermal degradation kinetics of pigments and
visual color in watermelon juice. Int. J. Food Prop., 11, 439–449, 2008.
32. Hayoglu, I.A. and Fenercioglu, H.A., Research on the possibility of using watermelon in the fruit juice
industry. Gıda, 15, 329–332, 1990.
33. Doodnath, L. and Badrie, N., Processing and quality evaluation of ready-to-serve watermelon nectars.
J. Food Sci. Technol., 38, 495–498, 2001.
34. Wu, G., Collins, J.K., Perkins-Veazie, P., Siddiq, M., Dolan, K.D., Kelly, K.A., and Meininger, C.J.,
Dietary supplementation with watermelon pomace juice enhances arginine availability and ameliorates
the metabolic syndrome in Zucker diabetic fatty rats. J. Nutr., 137, 2680–2685, 2007.
35. Poduri, A., Rateri, D.L., Saha, S.K., Saha, S., and Daugherty, A., Citrullus lanatus ‘sentinel’(watermelon)
extract reduces atherosclerosis in LDL receptor-deficient mice. J. Nutr. Biochem., 24, 882–886, 2012.
36. Bahri, S., Zerrouk, N., Aussel, C., Moinard, C., Crenn, P., Curis, E., Chaumeil, J., Cynober, L., and Sfar,
S., Citrulline: From metabolism to therapeutic use. Nutrition, 29, 479–484, 2013.
37. Collins, J.K., Wu, G., Perkins-Veazie, P., Spears, K., Claypool, P.L., Baker, R.A., and Clevidence, B.A.,
Watermelon consumption increases plasma arginine concentrations in adults. Nutrition, 23, 261–266,
2007.
38. Akashi, K., Miyake, C., and Yokota, A., Citrulline, a novel compatible solute in drought-tolerant wild
watermelon leaves, is an efficient hydroxyl radical scavenger. Febs. Lett., 508, 438–442, 2001.
39. El-Razek, F.H.A. and Sadeek, E.A., Dietary supplementation with watermelon (Citrullus lanatus) juice
enhances arginine availability and modifies hyperglycemia, hyperlipidemia and oxidative stress in
diabetic rats. Aust. J. Basic Appl. Sci., 5, 1284–1295, 2011.
40. Asita, A.O. and Molise, T., Antimutagenic effects of red apple and watermelon juices on cyclophospha-
mide-induced genotoxicity in mice. Afr. J. Biotechnol., 10, 17763–17768, 2011.
41. Davis, A.R., Collins, J., Fish, W.W., Tadmor, Y., Webber, C.L., and Perkins-Veazie, P., Rapid method for
total carotenoid detection in canary yellow-fleshed watermelon. Food Sci., 72, 319–323, 2007.
42. Tadmor, Y., King, S., Levi, A., Davis, A., Meir, A., Wasserman, B., and Lewinsohn, E., Comparative
fruit colouration in watermelon and tomato. Food Res. Int., 38, 837–841, 2005.
43. Edwards, A.J., Vinyard, B.T., Wiley, E.R., Brown, E.D., Collins, J.K., Perkins-Veazie, P., and Clevidence,
B.A., Consumption of watermelon juice increases plasma concentrations of lycopene and β-carotene in
humans. J. Nutr., 133, 1043–1050, 2013.
44. Shin, D.H., Koo, Y.J., Kim, C.O., Min, B.Y., and Suh, K.B., Studies on the production of watermelon and
cantaloupe melon juice. Korean J. Food Sci. Technol., 10, 215–223, 1978.
45. Marks S., Lunt, M., and Mattson, P., Method of making a commercial packaged watermleon juice drink.
U.S. Patent No. 6,589,581, U.S. Patent and Trademark Office, Washington, DC, 2003.
46. Yingliang, G., Lingbo, M., Jiwei, W., and Quan, Z., Study on simple processing of pure watermelon
juice. Food Sci. Technol., 79, 74–76, 2005.
47. Alemi, A., Emam Djomeh, Z., and Mirzaei, H.A., Effect of pressure and temperature of concentration
on some of quality attributes of watermelon juice. Iranian J. Food Sci. Technol., 9, 37–44, 2012.
48. Jain, R. and Vir Singh, S., Spray drying of watermelon juice. Indian Food Packer, 67, 84–87, 2013.
49. Dos Santos Gomes, F., Albuquerque da Costa, P., Domingues de Campos, M.B., Couri, S., and Cabral,
L.M.C., Concentration of watermelon juice by reverse osmosis process. Desalin. Water Treat., 27,
120–122, 2011.
50. Das Gupta, D.K. and Jayarama, K.S., Studies on the membrane concentration of watermelon juice.
J. Sci. Ind. Res., 55, 966–970, 1996.
51. Bai-Hui, A. and Cheng-Yu, G., Processing technology and stability of quick-freeze watermelon juice
beverage. Food Sci. Technol., 38, 92–99, 2013.
52. Saini, S.P.S. and Bains, G.S., A new method for mechanized production of watermelon seeds and juice.
Indian Food Packer, 48, 55–57, 1994.
53. Qian, Y.J., Yu, W.X., Yang, X.J., and Fu, C.M., Effects of chitosamine on the composition of water-
melon juice. Food Ind., 2, 4–6, 1998.
54. DeJong, B.D., Johnson, E.A., Luther, D.L., and Taylor, G.R., Watermelon juice products and food prod-
ucts produced with the juice products. U.S. Patent No. 7,807,209, U.S. Patent and Trademark Office,
55. Kim, S.L., Kim, W.J., and Lee, S.Y., Alcohol fermentation of Korean R. watermelon juice. J. Korean
Agric. Chem. Soc., 27, 139–145, 1984.
56. Mestry, A.P., Mujumdar, A.S., and Thorat, B.N., Optimization of spray drying of an innovative func-
tional food: Fermented mixed juice of carrot and watermelon. Dry. Technol., 29, 1121–1131, 2011.
57. Hayaloglu, I. and Vardin, H., Sensory evaluation of fruit punch including watermelon and pomegranate
juices at various levels. Gıda, 26, 267–270, 2001.
58. Bhardwaj, R.L. and Pandey, S., Juice blends—A way of utilization of under-utilized fruits, vegetables,
and spices: A review. Crit. Rev. Food. Sci. Nutr., 51, 563–570, 2011.
59. Oludemi, F.O. and Akanbi, C.T., Chemical, antioxidant and sensory properties of tomato-watermelon-
pineapple blends, and changes in their total antioxidant capacity during storage. Int. J. Food Sci.
Technol., 48, 1416–1425, 2013.
Section III
Herbal and Vegetable Juices

46
Carrot Juice
Ralf Martin Schweiggert and Reinhold Carle
CONTENTS
46.1 Introduction................................................................................................................................... 565
46.3.1 Carotenoids....................................................................................................................... 569
46.3.2 Anthocyanins.................................................................................................................... 569
46.3.3 Nonanthocyanin Phenolic Compounds.............................................................................571
46.3.4 Polyacetylenes.................................................................................................................. 572
46.4 Health Effects................................................................................................................................ 572
46.4.1 Vitamin A Supply and Further Potential Health Benefits of Carotenoids....................... 572
46.4.2 Anticarcinogenic Effects.................................................................................................. 573
46.4.3 Dietary Fiber and Gastrointestinal Infections...................................................................574
46.4.4 Vitamin C Supply..............................................................................................................574
46.4.5 Legal Health Claims Regarding Carrot Juice and Related Foods....................................574
46.6 Conclusion..................................................................................................................................... 577
References............................................................................................................................................... 578
46.1 Introduction
The very first reports about the carrot (Daucus carota L.), as it is known today, date back to the tenth
century, although many other members of the Apiaceae family such as parsnips, parsley, and fennel were
already used by the ancient Greeks and Romans. Moreover, until the fifteenth century, orange-colored
cultivars were completely unknown, while rather whitish, yellow, and purple carrots were widely used
from East Asia to Europe. The first documentation of the orange carrot (cv. “Horn” and “Long Orange”)
was found in paintings of 1618–1621, while the origin of the orange storage root color still remains a scien-
tific mystery. However, orange carrots continuously gained popularity during the following centuries and,
to date, is the predominant type used for fresh and processing markets [1]. The total world production of
carrots (including turnips) was ~36.9 metric tons (MT) in 2012 [2], with the highest proportions being des-
ignated for fresh consumption. According to the U.S. Department of Agriculture (USDA), approximately
77% (1.0 MT) of total carrots harvested in the United States were destined for the fresh market in 2009 [3].
In China and Japan, the young foliage is used as a stir-fried herb and in salads [4]. In addition to using the
fresh or minimally processed vegetable, the carrot is canned and processed into purees, juice, smoothies,
and mixed pickles, among others. Carrot juice is the second most popular vegetable juice after tomato
juice, containing high concentrations of provitamin A carotenoids and other valuable phytochemicals
such as vitamin C, polyphenols, polyacetylenes, oligogalacturonic acids, and soluble dietary fiber. In par-
ticular, the juice and concentrate from the purple carrot is rich in highly stable acetylated anthocyanins,
being which is used as a coloring foodstuff or natural colorant by the food industry. An enormous body
of scientific literature regarding the nutritional characteristics, functionality, bioavailability, bioactivity,
565
and the consequent ultimate health effects of carrots, carrot juice, other carrot products, and their con-
tained phytochemicals has accumulated over the past decades. This chapter highlights the nutritional and
phytochemical content of carrot juice, its antioxidant capacity, the preclinical and clinical evidence for
supporting human health, and novel formulations that may improve this functionality.

Due to its economic importance, most nutritional information that is available is on raw carrots. Table
46.1 shows the overall nutrient composition of raw carrot, carrot juice, and other vegetable and fruit
juices for comparison. Fresh carrots and carrot juice do not contribute significant amounts of calories,
protein, fat, or other major nutrients to the human diet, but their nutritional value is clearly the supply
of various phytochemicals described as follows. The data depicted in Table 46.1 represents mean values
reported in the USDA Nutrient Database [3], although, noteworthy, the nutrient content of carrot was
shown to vary mostly with cultivar, and also with season, environmental conditions, and maturity [4,5].
A large variety of different colored carrots is available, having very high contents in phytochemicals
such as carotenoids, anthocyanins, and other phenolic compounds. The commercially most important
orange cultivars are good sources of the provitamin A carotenoids α- and β-carotene, mostly occurring
at a molar ratio of approximately 1:2 (α/β) in the roots [6]. In contrast, red and yellow carrots contain
considerable amounts of lycopene and lutein, respectively [7]. High concentrations of anthocyanins, a
class of highly antioxidative plant pigments, were found in purple carrot, sometimes called black
carrot [8]. Different cultivars with a purple cortex, but orange or white color in the inner core, are available
as well as carrots being purple colored throughout. Continuous breeding programs have led to novel
purple–orange–red cultivars, concurrently containing high amounts of lycopene, β-carotene, and
anthocyanins [9]. Separate sections were dedicated to these phytochemicals later.
Regarding further micronutrients, carrot juice contains appreciable amounts of B vitamins, namely,
thiamin, riboflavin, niacin, and pantothenic acid, as compared to other fruits and vegetables (Table 46.1).
Moreover, a 100 g serving of carrot juice accounts for 7% vitamin C, 5% folate, 4% vitamin E, 11% vita-
min K, and 7% potassium of the Recommended Dietary Allowances (RDA). Several trace elements such
as iron, manganese, selenium, and zinc are also present in minor amounts. Nitrate levels were reported
to be high in specific vegetable juices, particularly when derived from spinach, lettuce, and red beet,
which were conventionally grown under the application of nitrogen fertilizers [10]. As shown in Table
46.1, carrot juice contains an only minor amount (2.2 mg/100 g of fresh weight [FW]) when compared
to red beet (41.9 mg/100 g of FW) and to the acceptable daily intake limit of 222 mg for a 60 kg male of
19–30 years as stated by the European Food Safety Authority (EFSA) [11].
The chemical composition and, thus, the nutritional characteristics substantially change during
processing of carrots to juice (Figure 46.1). After several washing steps for removal of sand and stones,
carrots are sorted and peeled. The commonly applied steam peeling results in significant reduction of
the microbial load and inactivation of oxidative enzymes located in the outer layers. Insufficient inac-
tivation may lead to a loss in polyphenolic constituents and ultimately to a brown discoloration due to
the activity of polyphenol oxidases. During the subsequent blanching step, water-soluble constituents
such as sugars may be leached out. Comminution or milling of the carrots can either be carried out
prior to or after blanching. After blanching and milling, a treatment with pectinases, hemicellulases,
and cellulases can be included to enhance the yield in juice and phytochemicals [12]. Using 0.2% of a
pectinase preparation, Khandare et al. [13] increased the total anthocyanin content in a purple carrot
juice from 504 to 1005 mg/L. Total liquefaction, however, might result in separation problems during
the subsequent decanting step, then being mandatorily replaced by horizontal presses. To date, how-
ever, decanters are most commonly used for industrial carrot juice production [12,14]. High proportions
of insoluble fiber remain in the pomace after dejuicing, consequently implying the reduction of total
dietary fiber from 2.8% (raw carrot) to 0.8% (carrot juice, canned). For preservation, the obtained juice
(pH 5–6) is either sterilized or pasteurized after acidification to pH < 4.5, which is commonly achieved
by the addition of citric acid or via lactic acid fermentation. Thermal treatment has a negative impact on
valuable constituents such as degradation of carotenoids and isomerization [15]. Regardless, vitamin A
Carrot Juice 567
TABLE 46.1
Compositional and Nutritional Characteristics of Some Fruits and Vegetable Juices (per 100 g Fresh
Weight)
Carrot Carrot Tomato Apple Orange Celery Red Beet
(Raw) (Juice) (Juice) (Juice) (Juice) (Raw) (Raw) RDAa
Nutrient Unit [3] [3,68] [3] [3] [3] [3,68] [3,68] [69]
Energy kcal 41 40 17 46 47 16 43 —
Water g 88 89 94 88 88 95 88 —
Protein g 0.93 0.95 0.76 0.10 0.68 0.69 1.61 56
Lipid (fat) g 0.24 0.15 0.05 0.13 0.15 0.17 0.17 —
Ash g 0.97 0.75 1.05 0.23 0.43 0.75 1.08 —
Carbohydrate g 9.56 9.30 4.24 11.30 11.01 2.97 9.56 130
Total sugars g 4.74 3.90 3.56 9.62 8.76 1.83 6.76 —
Total dietary fiber g 2.8 0.8 0.4 0.2 0.3 1.6 2.8 38
Minerals
Calcium mg 33 24 10 8 10 40 16 1000
Iron mg 0.30 0.46 0.43 0.12 0.10 0.20 0.80 8
Magnesium mg 12 14 11 5 10 11 23 400
Manganese mg 0.143 0.13 0.070 0.074 0.021 0.103 0.329 2.3
Phosphorus mg 35 42 18 7 17 24 40 700
Potassium mg 320 292 229 101 184 260 325 4700
Selenium µg 0.1 0.6 0.3 0.1 0.1 0.4 0.7 55
Sodium mg 69 66 10 4 4 80 78 1500
Zinc mg 0.24 0.18 0.15 0.02 0.04 0.13 0.35 11
Vitamins
Niacin mg 0.983 0.386 0.673 0.073 0.201 0.320 0.334 16
Pantothenic acid mg 0.273 0.228 0.250 0.049 0.180 0.246 0.155 5
Riboflavin mg 0.058 0.055 0.031 0.017 0.021 0.057 0.040 1.3
Thiamin mg 0.066 0.092 0.047 0.021 0.039 0.021 0.031 1.2
Total folate µg 19 4 20 0 24 36 109 400
Vitamin A (RAE) µg 835 956 23 0 9 22 2 900
Vitamin B 6 mg 0.138 0.217 0.111 0.018 0.031 0.074 0.067 1.3
Vitamin C mg 5.9 8.5 18.3 0.9 30.1 3.1 4.9 90
Vitamin E mg 0.66 1.16 0.32 0 0.20 0 0.04 15
Vitamin K µg 13.2 15.5 2.3 0 0.1 29.3 0.2 120
Nitrate
Nitrate (N-NO3) mg nr 2.2 nr 4.3 1.4 9.7 41.9 <222b
[10,11,68]
a For adult males (19–30 years).
b Acceptable daily intake as proposed by EFSA, EFSA J., 689, 1, 2008.
Abbreviations: RAE, retinol activity equivalents; RDA, Recommended Dietary Allowances; nr, not reported.
values (retinol activity equivalents, RAE) calculated from the concentrations of α- and β-carotenes in
orange carrot juice still remain very high (Table 46.1), unless serious technological mistreatments were
made. For instance, a significant degradation of carotenoids was observed during harsh heat sterilization
treatments [16]. Although acidified carrot juice requires less intense heating, carotenoid degradation
reactions were also observed to some limited extent [17]. Anthocyanins present in purple carrot juice and
juice concentrate are similarly prone to degradation, despite their enhanced stability when compared to
anthocyanins from other sources such as black currant, elderberry, and blackberry [18].
Although final quality parameters and nutritional characteristics of carrot juice depend on the applied
technology, cultivar selection has a substantial impact, sometimes surpassing negative processing
Fresh carrots
Multistage washing
Sorting
Steam peeling
Comminution/milling Blanching
Eventual addition of carrot juice Comminution/milling

(better flowing of the mash)
Heat treatment of the mash
Optional: Enzymatic liquefaction
Dejuicing (decanter or press)
Lactic acid fermentation and

Sterilization/UHT treatment Acidification and pasteurization
pasteurization
Filling, packaging, cooling
Carrot juice
FIGURE 46.1 Carrot juice production scheme. (Adapted from Marx, M.N., Einfluss der Herstellungstechnologie auf
die trans-cis-Isomerisierung von β-Carotin in Karottensaft [in German], Dissertation Hohenheim University, Stuttgart,
Germany, 2003.)
effects. For instance, even higher contents of >4.3 mg of α-carotene and >9.3 mg of β-carotene per 100 g
of FW were reported for canned carrot juice when compared to 3.5 and 8.3 mg/100 g of FW for raw
carrot of a different cultivar, respectively [3]. Interestingly, some studies revealed significantly higher
levels of carotenoids in orange–purple cultivars as compared to pure orange ones [19]. Therefore, the
selection of raw materials is most important for producing high-quality carrot juices.

Owing to their deep yellow, orange, red, and purple colors, the most obvious bioactive constituents of
carrot and the derived juice are their potent antioxidant pigments, for example, the lipid-soluble carot-
enoids and water-soluble anthocyanins. In contrast to colored cultivars, plant pigments are virtually
absent in white cultivars, although trace amounts of carotenoids may sometimes be present [20]. Being
frequently overlooked, carrot juice contains further potential bioactives, such as polyacetylenes, phenolic
Carrot Juice 569
acids (mainly chlorogenic acid), and isocoumarins like 6-methoxymellein (6-MM), all of which are
known to be fundamental principles of carrot bitterness [21]. Other water-soluble vitamins and dietary
carrot fibers may also be of considerable importance. The bioactivity and antioxidant properties of these
compounds (Figure 46.2) are outlined in the following sections.
46.3.1 Carotenoids
Among over 750 different naturally occurring carotenoids, the nutritionally most important ones
are the so-called provitamin A carotenoids. Among them, α- and β-carotene are the most abundant
in nature (Figure 46.2), both also occurring at high levels in orange carrot juice (up to 1.1–4.3 and
2.8–9.3 mg/100 mL, respectively [3,16]). In addition to their potent antioxidant activity, the most promi-
nent bioactivity of these compounds is their role as a vitamin A precursor. The human organism possesses
efficient intestinal transporter proteins (SR-BI) for their absorption and a β-carotene-15,15′-oxygenase
(BCO) for their conversion to vitamin A. As shown in Figure 46.3, BCO catalyzes the oxidative cleavage
of one molecule of β-carotene into two molecules of retinal, which are subsequently dehydrogenated to
retinol (vitamin A). Although β-carotene is frequently occurring in plant foods, carrot, carrot juice, and
derived products contain exceptionally high amounts of α- and β-carotene. For instance, high β-carotene
cultivars were shown to contain more than 18 mg β-carotene per 100 g of FW [20]. For comparison,
other fruits and vegetables such as papaya (0.3 mg/100 g of FW), mango (0.6 mg/100 g of FW), pumpkin
(3.1 mg/100 g of FW), and spinach (5.6 mg/100 g of FW) have significantly lower β-carotene contents [3].
Although carotenoid levels are reduced during juice production, carrot juice still represents an excellent
dietary source of α- and β-carotene (1.1–4.3 and 2.8–9.3 mg/100 mL, respectively [3,16]). In addition,
cell wall degradation and thermal treatment during processing were shown to drastically enhance carot-
enoid bioavailability; that is, their liberation from the food matrix, absorption in the small intestine, and
release to the blood plasma. Thus, despite having lower carotenoid levels than the initial raw material,
the juice may represent a better dietary source. The poor bioavailability of carotenes from raw carrot
was related to the deposition form in large crystalline aggregates (up to 25 µm), which are broken down
during processing. In contrast, papaya and mango fruits were reported to contain carotenoids in a lipid-
dissolved and liquid-crystalline form within small nanoscale elements of globular-tubular chromoplasts
[22]. Schweiggert et al. [23] hypothesized that these differences in physical state of deposition are the
decisive factor for the observed three-fold enhanced carotenoid bioavailability from raw papaya com-
pared to raw carrot.
Yellow carrot contains the nutritionally important antioxidant lutein (Figure 46.2), in addition to
minor amounts of α- and β-carotene. Although relative lutein levels (~0.5 mg/100 g of FW) exceed
the β-carotene levels (<0.2 mg/100 g of FW) in yellow carrot and derived products, the absolute lutein
content as compared to other plant foods is considered to be only “moderate” according to the classifi-
cation of Britton and Khachik [7,20]. However, the bioavailability of lutein was previously shown to be
significantly higher than that of β-carotene [24]. Molldrem et al. [25] fed 1.7 mg lutein contained in 350
g of cooked yellow carrot daily for 7 days, providing evidence of its high bioavailability by significantly
increasing blood serum levels. Lutein shows a strong antioxidant capacity, as it effectively quenches
singlet oxygen and other oxidizing species and also inhibits lipid peroxidation [26].
Lycopene is a further potentially bioactive carotenoid of red carrot (Figure 46.2), containing levels
easily surpassing those of β-carotene. According to Britton and Khachik’s classification [7], “very high”
levels of 6–10 mg/100 g of FW were frequently reached in red carrot, being similar or even higher than
in common red tomatoes [19,20]. The antioxidant potential as measured by physical quenching rates was
highest for lycopene among many biological oxygen quenchers examined so far [27], thus representing a
potentially powerful bioactive. Since juices made from red and yellow carrots are commercially unavail-
able in most countries, the discussion about lycopene and lutein has been limited here.
46.3.2 Anthocyanins
Purple carrot juice and juice concentrate gained a high industrial relevance as natural food colorants due
to their high content in anthocyanins. Consumer demand for such natural colorants was recently boosted
β-Carotene
α-Carotene
(all-E)-Lycopene
OH
HO
Lutein
OH
OH
HO O+
OH
OH
Cyanidin
O
HO OH
O
O OH
HO OH
OH
5΄-Caffeoylquinic acid (chlorogenic acid)
OH O
6-Methoxymellein
HO R
Falcarinol, R=H
Falcarindiol, R=OH
FIGURE 46.2 Major bioactive phytochemicals in differently colored carrot cultivars. (Orange color is derived from α- and
β-carotene, red color from lycopene, yellow color from lutein, and purple color from cyanidin derivatives. Falcarindiol
represents the main bitter principle in fresh carrot, particularly in the outer cortex. 6-Methoxymellein is responsible for
stress-induced bitterness.)
Carrot Juice 571
15
15΄
β-Carotene
β-Carotene-15, 15΄-oxygenase (BCO)

Retinol dehydrogenase
OH
Retinol (vitamin A)
FIGURE 46.3 Simplified illustration of β-carotene conversion to vitamin A in humans.
by the so-called Southampton study. This human trial involved almost 300 children and related the
consumption of synthetic food dyes to an increased risk in hyperactivity [28]. Since then, the food indus-
try has undertaken enormous efforts to replace synthetic dyes with natural alternatives. Purple carrot
juice represents a promising and, meanwhile, widely used alternative to synthetic food colorants, since it
contains high amounts of several highly stable glycosylated and acylated anthocyanins (up to 17.4 g/kg
of dry weight) [8]. Kammerer et al. [8] reported cyanidin xylosylglucosylgalactoside, cyanidin xylosyl-
galactoside, and sinapoyl-, feruloyl-, and p-coumaroyl derivatives of the aforementioned triglycoside to
be the major pigments. The aglycone cyanidin of these pigments is depicted in Figure 46.2. Particularly,
the contained acylated anthocyanins reveal exceptional stability at acidic pH during heat treatment and
storage when compared to other natural colorants [18,29].
In addition to color, anthocyanins also possess a strong antioxidant activity, which was reported to be
comparable to other flavonoid compounds [30]. Moreover, anticancer [31] and anti-inflammatory [32]
bioactivities have been proposed based on in vitro studies, although clear-cut investigations on humans
are still lacking to support these findings. Several human in vivo studies dealt with the absorption,
metabolism, and urinary excretion of anthocyanins in humans. Recently, the bioavailability of anthocya-
nins from purple carrot juice was studied [33]. Acylated anthocyanins (76% of total anthocyanins) were
initially less bioavailable than the nonacylated ones (24% of total anthocyanins). However, absorption of
nonacylated anthocyanins was only more rapid than that of acylated ones, and, ultimately, total bioavail-
ability of both pigment subclasses was similar 8 h after consumption. Thus, acylation of anthocyanins
appeared to be a key factor for delaying their bioavailability [33]. The bioactivity in humans and its
implication on potential health effects are under current investigation by several groups.
46.3.3 Nonanthocyanin Phenolic Compounds

In addition to anthocyanins, a variety of phenolic compounds with considerable antioxidant activity was
described in carrot juice. So-called chlorogenic acids are the major compounds. These represent esters of
(−)-quinic acid with hydroxycinnamic acid derivatives such as caffeic, ferulic, and p-coumaric acids. In all
different colored carrots, 5′-caffeoylquinic acid (5′-CQA) is the most abundant phenolic acid (Figure 46.2).
Approximately, 68, 66, and 580 mg of 5′-CQA per 100 g of fresh purple carrots, per 100 mL of purple carrot
juice, and per 100 g of the derived concentrate were reported previously, respectively [34]. For comparison,
coffee beverages (2%, w/v brewing) contain approximately 25–75 mg chlorogenic acid per 100 mL, depend-
ing on the coffee bean variety and the roasting process. In apples and pears, 5′-CQA is the dominant pheno-
lic acid with levels ranging from 6 to 39 mg and 6 to 28 mg/100 g of FW, respectively [35]. Thus, the purple
carrot can be considered a rich source of chlorogenic acid. An important and widely accepted b ioactivity
of phenolic acids is their biological role as phytoalexins, that is, their formation is an essential part of plant
resistance against phytopathogens [36]. As observed in many fruits and vegetables, carrot roots also respond
to stress factors such as cold, injury, and ethylene exposure with a rapid accumulation of polyphenols,
mainly 5′-CQA and 6-MM (Figure 46.2) and some other phenolics [37]. Among these “stress-related” phe-
nolics, 6-MM is of particular interest for juice production due to its strong bitter taste. The accumulation of
6-MM during carrot root storage was shown to be enhanced by ethylene treatment and to be inhibited by
application of its antagonist 1-methylcyclopropene (1-MCP). Concomitant sensory trials revealed a strong
correlation of ethylene-induced bitter taste and the levels of 6-MM and other phenolics [37].
46.3.4 Polyacetylenes
The eventual bitter taste of carrot juice was previously thought to be related to the polyacetylenes fal-
carindiol, falcarinol, and falcarindiol-3-acetate. The importance of 6-MM for the bitter taste of fresh
carrot and carrot juice was doubted due to an insufficient ratio of its concentration and bitter detection
threshold [21]. However, while 6-MM appeared to be generally absent in unstressed carrot, Kramer et al.
[37] showed that the 6-MM concentration drastically increased after ethylene treatment, thus reaching
levels exceeding its bitter threshold. Thus, 6-MM is believed to be responsible for the ethylene-induced
or, more generally, stress-induced bitterness of carrot. In contrast, polyacetylenes are also present in
sound and freshly harvested carrots and may cause substantial bitterness even in the absence of 6-MM,
particularly in the presence of certain volatile terpenoids and concurrently low sugar levels. Specifically,
falcarindiol (Figure 46.2), which is mainly present in the peel and the outer parts of the root, seemed to
have a high impact on bitterness [21,37]. Due to an increased fungal resistance of the outer carrot layers
as compared to the inner xylem, antifungal properties of falcarindiol were proposed. A strong negative
correlation was found between falcarindiol levels and susceptibility to Mycocentrospora acerina for 14
of 16 tested cultivars [38]. Furthermore, falcarindiol exhibited cytotoxic [39], anti-inflammatory [40],
antimutagenic [41], and anticarcinogenic [42] activities. In contrast to the specific pericyclic localization
of falcarindiol, the polyacetylenes falcarinol and falcarindiol-3-acetate occur at similar concentrations
in the inner and outer parts. Also, the contribution of the latter two compounds to the bitterness of car-
rot juice appears to be less important due to their low concentrations being below the bitter threshold
[43]. Thus, the reduction of falcarindiol seems to be mostly relevant for the production of nonbitter juice.
Czepa and Hofmann [43] showed that bitterness in the product can be significantly reduced by peeling
and removal of dark and green parts (Figure 46.1).
Although not contributing significantly to the bitterness of carrot juice, falcarinol and falcarin-
diol-3-acetate might still possess considerable bioactivity. While detailed and reliable information on
falcarindiol-3-acetate is scarce, in vitro and in vivo anticancer effects were proposed for falcarinol.
Moreover, falcarinol exhibited an anti-inflammatory, antiplatelet-aggregatory, antifungal, and anti-
mycobacterial bioactivity. A detailed overview of these bioactivities was compiled by Christensen
[44]. In order to express these bioactivities, falcarinol-type phytochemicals need to be released from
the food matrix and absorbed in the human gut. Recently, the bioavailability of falcarinol from carrot
juice (900 mL containing ~12 mg falcarinol) was demonstrated. Maximum concentrations of about
2.3 and 2.0 ng/mL blood serum were reached after 2 and 5 h after consumption, respectively [44,45].
However, whether such concentrations suffice to exert any significant bioactivity in humans remains
unknown.
46.4 Health Effects

46.4.1 Vitamin A Supply and Further Potential Health Benefits of Carotenoids
Due to the high content of provitamin A carotenoids, only 100 g of carrot juice contain 956 µg RAE
(Table 46.1) and, thus, would suffice to meet the RDA for vitamin A as stated by the U.S. Institute of
Medicine (700–900 µg RAE, [46]). While the bioavailability of carotenoids from raw carrot is rather low
Carrot Juice 573
[4,23], many studies suggest particularly processed carrot to be a readily bioavailable source for dietary
α- and β-carotenes. Therefore, carrot juice represents a most valuable source of vitamin A for deficient
populations [4]. According to several strategies of the World Health Organization, the consumption of
foods with high levels of provitamin A carotenoids should be strongly encouraged for diminishing vita-
min A deficiency and its severe implications for human health [47].
In order to exert any health effects or other bioactivities, carotenoids need to be released from the
food matrix and absorbed by the human enterocyte. In order to enhance the low bioavailability from
raw carrot, thermal processing and the addition of lipid to the carrot meal revealed to be powerful
measures. For instance, Rock et al. [48] fed ~9.3 mg β-carotene from either raw or thermally processed
and pureed carrot daily over two 4-week periods in a crossover human study. The consumption of
processed carrot increased the plasma concentrations of β-carotene in the subjects about three-fold
more effectively than that of the raw vegetable. Although detailed human studies directly comparing
carotenoid bioavailability from carrot juice to raw carrot are scarce, an enhanced bioavailability from
juice is to be expected. Comparing the carrot puree used in the aforementioned study with commercial
carrot juice, the reduced dietary fiber content in the juice may further boost carotenoid bioavailability,
since the presence of fiber during digestion was shown to hinder carotenoid absorption [49]. Thürmann
et al. [50] showed a slightly lower bioavailability of β-carotene from carrot juice to that from an arti-
ficial drink containing a synthetic water-soluble formulation of β-carotene. Nevertheless, β-carotene
from the carrot juice was still highly bioavailable, and juice consumption resulted in a rapid increase in
β-carotene plasma concentration. After absorption and enzymatic cleavage, vitamin A plays a crucial
role in human vision and is of particular importance for seeing objects under dim light conditions.
Mechanistic knowledge on the bioactivity of vitamin A outside the visual system is rapidly expand-
ing and is mostly related to its specific binding to nuclear receptors activated by retinoic acid. These
receptors regulate the expression level of a large number of genes [47,51]. In addition to its function
as a vitamin A precursor, β-carotene represents a potent lipid-soluble antioxidant due to its efficient
radical-trapping property under physiological conditions. The daily consumption of 473 mL (= 16 oz)
orange carrot juice for 3 months was recently shown to significantly improve the antioxidant status and
to decrease lipid peroxidation in human subjects [52].
By analogy, the major carotenoid in yellow carrot, lutein, was described to possess a strong antioxidant
capacity, to effectively quench singlet oxygen and other oxidizing species, to inhibit lipid peroxidation,
and to prevent promotion and replication of tumor cells [26]. Beyond its antioxidative properties, lutein
consumption was associated with a lower incidence of age-related macular degeneration and improved
cognitive functions in the elderly, although the need for further study of the latter was claimed by other
researchers [53].
The main carotenoid in red carrot is lycopene, which, as described earlier, exerts an outstandingly high
antioxidant activity [27]. In a variety of epidemiological studies, a high intake of lycopene-rich foods was
associated with decreased cardiovascular disease (CVD) and prostate cancer risk in a variety of epide-
miological studies. Since lycopene-rich carrot juices are practically unavailable on commercial markets,
the discussion about further possible health benefits of this compound may be followed elsewhere [54,55].
46.4.2 Anticarcinogenic Effects

Mechanistic studies on the putative anticancer activity of carrot juice and fresh carrot are scarce.
Pool-Zobel et al. [56] investigated the effect of carotenoid intake from tomato juice, carrot juice, and
spinach powder in water and milk on DNA damage. While dietary supplementation with each of three
carotenoid-rich foods reduced endogenous levels of strand breaks in lymphocyte DNA, only carrot
juice supplementation reduced oxidative base damage. In contrast, Briviba et al. [57] reported that
supplementation of a diet low in carotenoids with tomato or carrot juice did not reduce lipid peroxida-
tion in plasma and feces. Due to the assumed associations of carotenoid intake and a reduced risk in
colon cancer, Schnäbele et al. [58] investigated the cytotoxic and antiproliferative properties of human
fecal water after a 2-week consumption of either carrot or tomato juice, respectively. Although carot-
enoid levels were significantly increased in the feces, the cytotoxic and antiproliferative properties
remained unchanged compared to the initial activity. The researchers concluded that the carotenoids in
feces are not directly involved in an anticarcinogenic mechanism in the colon. Although most research
on carrot-related anticancer effects focused on carotenoids, other compounds such as fiber, polyphe-
nols, and polyacetylenes might be of interest in future investigations. Despite limited in vivo research
to date, particularly falcarinol was associated with numerous biological activities as described ear-
lier. Furthermore, the epidemiologically observed protective effects of carrot consumption may be due
to a mere correlation with the actual cause, such as a high-vegetable dietary pattern and a generally
healthy lifestyle [4]. Thus, further research is urgently needed to corroborate the assumed b eneficial
health effects.
46.4.3 Dietary Fiber and Gastrointestinal Infections

Total dietary fiber content is significantly reduced during juice production. Large quantities of pomace
with very high contents of dietary fiber are generated and might be used as a dietary supplement due
to the interesting potential bioactivity of carrot fiber. By administration of the so-called Moro’s carrot
soup, significant progress in the therapy of diarrheal disease in infants was achieved in Germany at the
beginning of the twentieth century [59]. Much later, this bioactivity was associated with the occurrence
of oligosaccharides in Moro’s carrot soup, which are formed from fiber after the rigorous cooking for
2 h as described in the recipe. Such oligosaccharides were also obtained after enzymatic liquefaction
of carrot and carrot pomace, when long-chain pectins were cleaved to form lower-molecular-weight
fragments [60,61]. As shown by in vitro experiments, these oligosaccharides were able to block the
adherence of various enteropathogenic microorganisms to HEp-2 cells and human intestinal mucosa.
As a consequence, such aqueous carrot extracts were described to be “significantly superior to the
basic glucose-electrolyte-solution for oral rehydration in acute gastro-intestinal infections of children,”
as stated by researchers from Vienna [62]. With regard to carrot juice, the pomace as by-product
might represent a most valuable source for carrot fiber with medicinal application. A process for the
enzymatic liquefaction of carrot pomace was described earlier, resulting in a liquid carotene-rich food
ingredient containing high concentrations of several potentially bioactive oligosaccharides [61,63].
Nevertheless, the majority of the pomace is still underutilized being disposed as feed or fertilizer.
Examples of minor food applications are its addition to bread, cake, dressings, pickles, and functional
drinks, among others.
46.4.4 Vitamin C Supply

A serving of 100 g of carrot juice delivers about 8.5% vitamin C to the RDA. For comparison, 100 g of
orange juice supplies 30.1% vitamin C to the RDA (Table 46.1). Numerous important biological func-
tions of vitamin C were described, for example, its importance for collagen synthesis (bone, cartilage,
skin, gum, and teeth health), the functionality of the immune system, and the absorption of nonheme
iron. Regarding the aforementioned and some other functions, the EU authorized several health claims
for vitamin C, which can be used on food labels in the European Union [64].
46.4.5 Legal Health Claims Regarding Carrot Juice and Related Foods
In the EU, health-related properties of a food product can only be used for advertisement and market-
ing, if sufficient scientific evidence including well-powered human studies has unequivocally demon-
strated such so-called “health claims.” The EFSA carefully evaluates current knowledge regarding
the numerous requested health claims and decides whether the health claim may be granted, that is,
approved, or not. A consolidated list of authorized and nonauthorized health claims can be found on
the EFSA website [64]. For instance, approved health claims for vitamin A are “vitamin A contributes
to the maintenance of normal vision” (claim entry IDs 16, 4239, 4701) and “vitamin A helps the proper
functioning of the immune system” (claim entry IDs 14, 200, 1462). Examples for submitted health
claims directly related to carrot and associated products are shown in Table 46.2. However, at the time
of writing this chapter, none of the claims for carrot has been authorized due to the substantial lack of
scientific evidence.
Carrot Juice 575
TABLE 46.2
Selected Authorized and Nonauthorized Health Claims Listed by the European Food Safety Authority
European Food
Health Claim Health Safety Authority
Entry IDa Food/Nutrient Relationship Health Claim Status
16, 4239, 4701 Vitamin A Visual system Contributes to the maintenance of Authorized
normal vision.
14, 200, 1462 Vitamin A Immune system Contributes to the normal function Authorized
of the immune system.
15, 17, 4660, Vitamin A Skin health Contributes to the maintenance of Authorized
4702 normal skin.
2431 Carrot Eye health Promotes maintenance of vision Nonauthorized
apparatus functions, improves dark
adaption, strengthens eye
capillaries, and reduces eye
tiredness in case of vision exertion.
2522 Carrot root Antioxidant and Helps to keep the skin healthy, Nonauthorized
photoprotection helps to retard skin aging, helps to
protect your skin from excessive
UV radiation and sunburns, and
promotes healthy skin
pigmentation and tanning.
3075 Carrot juice, lactic Healthy Supports a healthy digestion. Nonauthorized
acid fermented digestion
3076 Carrot juice, lactic Intestinal flora Supports a healthy intestinal and Nonauthorized
acid fermented colon flora.
3077 Carrot juice, lactic Immune system Supports the immune system and Nonauthorized
acid fermented and antioxidant supports the natural antioxidant
properties system in the body.
Note: This table was created to the best of the authors’ knowledge in August 2013 to provide an overview of the current
situation. Since the legal situation may change at any time, the table, all related statements, and views do not
represent any form of legal advice.
a ID according to the EU Register on Nutrition and Health Claims [64], last accessed in March 2014.

Due to difficult supply of red and yellow carrots and the simultaneous availability of alternative
“noncarrot” lycopene- and lutein-rich coloring foodstuffs, a positive development for the economic
importance of such carrot juices cannot be predicted. Nevertheless, El-Abasy et al. [65] showed the
potential of juice blends from red and yellow carrot for the coloration of fermented dairy products. In
contrast, an increasing importance of orange and purple carrots and derived products such as juice and
juice concentrate can clearly be expected, as indicated by the significantly growing world production
during the last decade (Table 46.3).
Considering the per capita consumption of conventional juice from carrot and other vegetables, stable
annual values were observed during the last several years. Annual per capita consumption of vegetable
juice and nectar in Germany remained mostly constant at slightly variable levels from 1.20 to 1.35 L [66].
The European Fruit Juice Association reports an increasing demand for premium products, especially in
the chilled juice and “not from concentrate” segments. According to their annual market report in 2012,
many producers of fruit and vegetable juices have taken advantage of consumers’ willingness to purchase
high-quality products. They invested in innovations and, particularly, in value-added chilled and func-
tional juices. Overall, however, the EU juice market volume again declined by 1.9% in 2012, although the
rate of decline is expected to decelerate until a stabilization might be reached in the near future [66]. In
smoothies composed of fruits and vegetables, carrot juice has become the most popular ingredient; how-
ever, such blends made from juices and purees are still niche products having minor nutritional relevance.
TABLE 46.3
Top 20 Carrot- and Turnip-Producing Countries in 2001 and Their 10-Year Development
Production in MT
Rank 2011 Rank Development Country 2001 2011
1 China 6,111,984 16,219,573
2 Russian Federation 1,575,070 1,735,030
3 United States 1,714,996 1,298,800
4 Uzbekistan 320,900 1,220,000
5 Poland 921,911 887,374
6 Ukraine 462,500 864,200
7 United Kingdom 901,800 694,104
8 France 649,489 624,459
9 Japan 691,300 617,300
10 Turkey 230,000 602,078
11 Italy 599,500 542,691
12 Germany 444,448 533,717
13 Indonesia 300,648 526,917
14 India 383,793 514,889
15 The Netherlands 378,000 482,000
16 Morocco 198,370 427,734
17 Canada 279,050 421,444
18 Iran 150,000 418,325
19 Mexico 355,903 404,726
20 Kazakhstan 189,100 370,000
Source: Adapted from FAO, FAOSTAT Database of the Food and Agriculture Organization of the
United Nations, Production crops: Carrot and turnip, FAO, Rome, Italy, 2012.
Abbreviation: MT, metric tons.
Although fruit and vegetable consumption of children is alarmingly low, smoothies have been criticized
due to the reduced satiety resulting from the ingestion of fluid foods. Nevertheless, the propagation of
vegetable-based variants would at least contribute to better meet the “5 A Day” recommendation, in
particular for junior consumers. Carrot fiber may be added to juices to improve the satiety effect of bever-
ages, and also differently fermented carrot juice and puree, respectively, may receive nutritional interest,
as lactic and oligogalacturonic acids are known to have beneficial health properties.
Carrot Juice 577
Also, their use for direct consumption, exploiting the coloring properties of orange and purple car-
rots will be an opportunity of high demand. Synthetic dyes have been associated with hyperactivity in
children in the “Southampton study” [28], and, therefore, the replacement of synthetic dyes by natural
colorants such as carotenoids and anthocyanins has rigorously been carried out during the last decade.
Meanwhile, a high proportion of food products colored with natural dyes is available in many European
countries. A similar development and progress is ongoing and expected to be extended in the United
States and other parts of the world during the upcoming years. A strong trend to market products prom-
ising improved health (“functional food”) was recently shown in the United States [67]. However, the
vagueness of the term “functional food” and often unproven associations of such products and health
benefits are likely to provoke consumer confusion rather than confidence. Nevertheless, worldwide
consumers show an increasing trend toward buying and consuming foods having potential health
benefits [67]. Different colored fresh carrot types have been introduced to the consumer, for example,
in the United Kingdom by Tesco in 2011. A growing variety of foods containing purple carrot and its
juice and concentrate is available on the shelves of many big supermarket chains. Purple carrot juice,
concentrate, and powder is commercialized in large quantities by several industrial suppliers, suggesting
their application as colorant in beverages, ice creams, sweets, pastries, coatings, jellies, and even some
dairy products.
Since knowledge about the numerous possible health effects of carrot consumption can be
expected to spread among costumers, the food industry might be making more and more use of
carrot products as a functional ingredient, particularly using the highly techno- and biofunctional
purple cultivars. Driven by the same trend, breeders also aimed at the “biofortification” of carrot and
have intensified the development of colored breeding lines with exceptional levels of phytochemicals
in order to increase pigment levels, nutritional quality, and visual appeal [4]. This trend has already
led to the development of several functional beverages with extremely high concentrations of bioac-
tive phytochemicals such as polyphenols and anthocyanins. However, studies on the evaluation of
dose recommendations during regular intake of such extremely enriched foods and supplements
are missing. In addition, the effects of long-term supplementation of such “nutraceuticals” may be
completely unexpected and, to date, are still mostly unknown, but urgently needed for the sake of
consumer safety.
46.6 Conclusion
Differently colored carrot juices and concentrates are highly functional foodstuffs. In particular, the
potential of orange and purple carrot juices for coloration is being widely exploited in many countries
worldwide. While carotenoids, such as β-carotene, provide stable yellow and orange color hues, the acyl-
ated and comparably stable anthocyanins from purple carrot juice and concentrate are appreciated for
their red, purple, and bluish colors by the food industry. In addition to coloration, many health benefits
are associated with the consumption of carrot, carrot juice, and other carrot products. Readily bioavail-
able vitamin A precursors are provided in high amounts by carrot juice and processed products. While
some scientific evidence such as the beneficial effects of vitamins A and C have meanwhile been trans-
lated into legal health claims, numerous associations and cause–effect relationships are not yet unques-
tionably proven. Further epidemiological, pharmacological, and clinical research is urgently required to
better understand the contribution of carrots and their assumed bioactives to human health. Randomized
controlled trials are required for assured knowledge on potential protective effects against today’s most
challenging diseases, such as various types of cancer and obesity. Metabolic products and mechanisms
of action of less intensely studied carrot constituents such as anthocyanins, phenolic acids, and poly-
acetylenes need to be elucidated. However, even if many health benefits such as a reduced risk in cancer
could not yet be related to single fruits, vegetables, or even single compounds, a diet rich in fruits and
vegetables is clearly and consistently considered to be beneficial for human health in many ways. Carrot
and its products derived thereof are important components of human diets, and several further health
benefits may still need to be discovered.
REFERENCES
1. Simon, P.W., Domestication, historical development, and modern breeding of carrot. Plant Breed. Rev.,
19, 157–190, 2000.
2. FAO, FAOSTAT database of the Food and Agriculture Organization of the United Nations. Production
crops: Carrot and turnip, FAO, Rome, Italy, 2012.
3. U.S. Department of Agriculture (USDA), National nutrient database for standard reference, release 24,
National Technical Information Service, USDA, Springfield, VA, 2012.
4. Arscott, S.A. and Tanumihardjo, S.A., Carrots of many colors provide basic nutrition and bioavailable
phytochemicals acting as a functional food. Compr. Rev. Food Sci. Food Saf., 9, 223–239, 2010.
5. Sharma, K.D., Karki, S., Thakur, N.S., and Attri, S., Chemical composition, functional properties and
processing of carrot—A review. J. Food Sci. Technol., 49, 22–32, 2012.
6. Marx, M., Schieber, A., and Carle, R., Quantitative determination of carotene stereoisomers in carrot
juices and vitamin supplemented (ATBC) drinks. Food Chem., 70, 403–408, 2000.
7. Britton, G. and Khachik, F., Carotenoids in food, in Carotenoids Volume 5: Nutrition and Health,
Britton, G., Liaaen-Jensen, S., and Pfander, H., Eds., Birkhäuser Verlag, Berlin, Germany, 2009,
pp. 45–66.
8. Kammerer, D., Carle, R., and Schieber, A., Quantification of anthocyanins in black carrot extracts
(Daucus carota ssp. sativus var. atrorubens Alef.) and evaluation of their color properties. Eur. Food
Res. Technol., 219, 479–486, 2004.
9. Mills, J.P., Simon, P.W., and Tanumihardjo, S.A., Biofortified carrot intake enhances liver antioxidant
capacity and vitamin A status in Mongolian gerbils. J. Nutr., 138, 1692–1698, 2008.
10. Smiechowska, M., The content of nitrates and vitamin C in juices obtained from organic and conven-
tional raw materials. Pol. J. Food Nutr. Sci., 12, 57–61, 2003.
11. EFSA, Nitrate in vegetables—Scientific opinion of the panel on contaminants in the food chain. EFSA
J., 689, 1–79, 2008.
12. Marx, M.N., Einfluss der Herstellungstechnologie auf die trans-cis-Isomerisierung von β-Carotin in
Karottensaft (in German), Dissertation, Hohenheim University, Stuttgart, Germany, 2003.
13. Khandare, V., Walia, S., Singh, M., and Kaur, C., Black carrot (Daucus carota ssp. sativus) juice:
Processing effects on antioxidant composition and color. Food Bioprod. Process., 89, 482–486,
2011.
14. Reiter, M., Stuparic, M., Neidhart, S., and Carle, R., The role of process technology in carrot juice cloud
stability. LWT Food Sci. Technol., 36, 165–172, 2003.
15. Marx, M., Stuparic, M., Schieber, A., and Carle, R., Effects of thermal processing on trans–
cis-isomerization of β-carotene in carrot juices and carotene-containing preparations. Food Chem., 83,
609–617, 2003.
16. Chen, B.H., Peng, H.Y., and Chen, H.E., Changes of carotenoids, color, and vitamin A contents during
processing of carrot juice. J. Agric. Food Chem., 43, 1912–1918, 1995.
17. Wu, J.S.B. and Shen, S.C., Processing of vegetable juice and blends, in Handbook of Vegetables and
Vegetable Processing, Sinha, N.K., Ed., Blackwell Publishing Ltd., Ames, IA, 2011, pp. 335–350.
18. Zozio, S., Pallet, D., and Dornier, M., Evaluation of anthocyanin stability during storage of a coloured
drink made from extracts of the Andean blackberry (Rubus glaucus Benth.), açai (Euterpe oleracea
Mart.) and black carrot (Daucus carota L.). Fruits, 66, 203–215, 2011.
19. Grassmann, J., Schnitzler, W.H., and Habegger, R., Evaluation of different coloured carrot cultivars on
antioxidative capacity based on their carotenoid and phenolic contents. Int. J. Food Sci. Nutr., 58, 603–611,
2007.
20. Surles, R.L., Weng, N., Simon, P.W., and Tanumihardjo, S.A., Carotenoid profiles and consumer sen-
sory evaluation of specialty carrots (Daucus carota L.) of various colors. J. Agric. Food Chem., 52,
3417–3421, 2004.
21. Czepa, A. and Hofmann, T., Structural and sensory characterization of compounds contributing to the
bitter off-taste of carrots (Daucus carota L.) and carrot puree. J. Agric. Food Chem., 51, 3865–3873,
2003.
22. Schweiggert, R.M., Mezger, D., Schimpf, F., Steingass, C.B., and Carle, R., Influence of chromoplast
morphology on carotenoid bioaccessibility of carrot, mango, papaya, and tomato. Food Chem., 60,
2577–2585, 2012.
Carrot Juice 579
23. Schweiggert, R.M., Kopec, R.E., Villalobos-Gutierrez, M.G., Högel, J., Quesada, S., Esquivel, P.,
Schwartz, S.J., and Carle, R., Carotenoids are more bioavailable from papaya than from tomato and
carrot in humans: A randomised cross-over study. Br. J. Nutr., 111, 490–498, 2014.
24. Van Het Hof, K.H., Brouwer, I.A., West, C.E., Haddeman, E., Steegers-Theunissen, R.P.M., Van
Dusseldorp, M., Weststrate, J.A., Eskes, T.K.A.B., and Hautvast, J.G.A.J., Bioavailability of lutein from
vegetables is 5 times higher than that of β-carotene. Am. J. Clin. Nutr., 70, 261–268, 1999.
25. Molldrem, K.L., Li, J., Simon, P.W., and Tanumihardjo, S.A., Lutein and β-carotene from lutein-con-
taining yellow carrots are bioavailable in humans. Am. J. Clin. Nutr., 80, 131–136, 2004.
26. Khachik, F., Beecher, G.R., and Smith Jr., J.C., Lutein, lycopene, and their oxidative metabolites in
chemoprevention of cancer. J. Cell. Biochem., 58, 236–246, 1995.
27. Di Mascio, P., Kaiser, S., and Sies, H., Lycopene as the most efficient biological carotenoid singlet oxy-
gen quencher. Arch. Biochem. Biophys., 274, 532–538, 1989.
28. McCann, D., Barrett, A., Cooper, A., Crumpler, D., Dalen, L., Grimshaw, K., Kitchin, E. et al., Food
additives and hyperactive behaviour in 3-year-old and 8/9-year-old children in the community: A ran-
domised, double-blinded, placebo-controlled trial. Lancet, 370, 1560–1567, 2007.
29. Sadilova, E., Stintzing, F.C., and Carle, R., Thermal degradation of acylated and nonacylated anthocya-
nins. J. Food Sci., 71, C504–C512, 2006.
30. Wang, H., Cao, G., and Prior, R.L., Oxygen radical absorbing capacity of anthocyanins. J. Agric. Food
Chem., 45, 304–309, 1997.
31. Kamei, H., Kojima, T., Hasegawa, M., Koide, T., Umeda, T., Yukawa, T., and Terabe, K., Suppression of
tumor cell growth by anthocyanins in vitro. Cancer Invest., 13, 590–594, 1995.
32. Wang, H., Nair, M.G., Strasburg, G.M., Chang, Y., Booren, A.M., Gray, J.I., and DeWitt, D.L.,
Antioxidant and antiinflammatory activities of anthocyanins and their aglycone, cyanidin, from tart
cherries. J. Nat. Prod., 62, 294–296, 1999.
33. Charron, C.S., Kurilich, A.C., Clevidence, B.A., Simon, P.W., Harrison, D.J., Britz, S.J., Baer, D.J., and
Novotny, J.A., Bioavailability of anthocyanins from purple carrot juice: Effects of acylation and plant
matrix. J. Agric. Food Chem., 57, 1226–1230, 2009.
34. Kammerer, D., Carle, R., and Schieber, A., Characterization of phenolic acids in black carrots (Daucus
carota ssp. sativus var. atrorubens Alef.) by high-performance liquid chromatography/electrospray
ionization mass spectrometry. Rapid Commun. Mass Spectrom., 18, 1331–1340, 2004.
35. Clifford, M.N., Chlorogenic acids and other cinnamates—Nature, occurrence and dietary burden.
J. Sci. Food Agric., 79, 362–372, 1999.
36. Carle, R., Phytoalexins and their significance for the resistance of higher plants to noxious organisms.
Pharm. Unserer Zeit, 21, 99–104, 1992.
37. Kramer, M., Bufler, G., Ulrich, D., Leitenberger, M., Conrad, J., Carle, R., and Kammerer, D.R., Effect
of ethylene and 1-methylcyclopropene on bitter compounds in carrots (Daucus carota L.). Postharvest
Biol. Technol., 73, 28–36, 2012.
38. Olsson, K. and Svensson, R., The influence of polyacetylenes on the susceptibility of carrots to storage
diseases. J. Phytopathol., 144, 441–447, 1996.
39. Bernart, M.W., Cardellina II, J.H., Balaschak, M.S., Alexander, M.R., Shoemaker, R.H., and Boyd,
M.R., Cytotoxic falcarinol oxylipins from Dendropanax arboreus. J. Nat. Prod., 59, 748–753, 1996.
40. Metzger, B.T., Barnes, D.M., and Reed, J.D., Purple carrot (Daucus carota L.) polyacetylenes decrease
lipopolysaccharide-induced expression of inflammatory proteins in macrophage and endothelial cells.
J. Agric. Food Chem., 56, 3554–3560, 2008.
41. Miyazawa, M., Shimamura, H., Bhuva, R.C., Nakamura, S., and Kameoka, H., Antimutagenic activity
of falcarindiol from Peucedanum praeruptorum. J. Agric. Food Chem., 44, 3444–3448, 1996.
42. Jin, H.R., Zhao, J., Zhang, Z., Liao, Y., Wang, C., Huang, W., Li, S.., He, T., Yuan, C., and Du, W., The
antitumor natural compound falcarindiol promotes cancer cell death by inducing endoplasmic reticulum
stress. Cell Death Dis., 3, 1–9, 2012.
43. Czepa, A. and Hofmann, T., Quantitative studies and sensory analyses on the influence of cultivar,
spatial tissue distribution, and industrial processing on the bitter off-taste of carrots (Daucus carota L.)
and carrot products. J. Agric. Food Chem., 52, 4508–4514, 2004.
44. Christensen, L.P., Aliphatic C17-polyacetylenes of the falcarinol type as potential health promoting
compounds in food plants of the apiaceae family. Recent Pat. Food Nutr. Agric., 3, 64–77, 2011.
45. Christensen, L.P. and Brandt, K., Bioactive polyacetylenes in food plants of the Apiaceae family:
Occurrence, bioactivity and analysis. J. Pharm. Biomed. Anal., 41, 683–693, 2006.
46. Institute of Medicine, Vitamin A, in Dietary Reference Intakes for Vitamin A, Vitamin K, Arsenic,
Boron, Chromium, Copper, Iodine, Iron, Manganese, Molybdenum, Nickel, Silicon, Vanadium, and
Zinc, Institute of Medicine, Food and Nutrition Board, Ed., National Academic Press, Washington, DC,
2001, pp. 82–161.
47. World Health Organization (WHO), Global Prevalence of Vitamin A Deficiency in Populations at Risk
1995–2005, WHO Global Database on Vitamin A Deficiency, WHO, Geneva, Switzerland, 2009.
48. Rock, C.L., Lovalvo, J.L., Emenhiser, C., Ruffin, M.T., Flatt, S.W., and Schwartz, S.J., Bioavailability of
β-carotene is lower in raw than in processed carrots and spinach in women. J. Nutr., 128, 913–916, 1998.
49. Riedl, J., Linseisen, J., Hoffmann, J., and Wolfram, G., Some dietary fibers reduce the absorption of
carotenoids in women. J. Nutr., 129, 2170–2176, 1999.
50. Thürmann, P.A., Steffen, J., Zwernemann, C., Aebischer, C., Cohn, W., Wendt, G., and Schalch, W.,
Plasma concentration response to drinks containing ß-carotene as carrot juice or formulated as a water
dispersible powder. Eur. J. Nutr., 41, 228–235, 2002.
51. Clagett-Dame, M. and Knutson, D., Vitamin A in reproduction and development. Nutrients, 3, 385–428,
2011.
52. Potter, A.S., Foroudi, S., Stamatikos, A., Patil, B.S., and Deyhim, F., Drinking carrot juice increases
total antioxidant status and decreases lipid peroxidation in adults. Nutr. J., 10, 96, 2011.
53. Johnson, E.J., A possible role for lutein and zeaxanthin in cognitive function in the elderly. Am. J. Clin.
Nutr., 96, 1161–1165, 2012.
54. Voutilainen, S., Nurmi, T., Mursu, J., and Rissanen, T.H., Carotenoids and cardiovascular health. Am. J.
Clin. Nutr., 83, 1265–1271, 2006.
55. Wei, M.Y. and Giovannucci, E.L., Lycopene, tomato products, and prostate cancer incidence: A review
and reassessment in the PSA screening era. J. Oncol., 2012, 1–7, 2012.
56. Pool-Zobel, B.L., Bub, A., Müller, H., Wollowski, I., and Rechkemmer, G., Consumption of vegetables
reduces genetic damage in humans: First results of a human intervention trial with carotenoid-rich
foods. Carcinogenesis, 18, 1847–1850, 1997.
57. Briviba, K., Schnäbele, K., Rechkemmer, G., and Bub, A., Supplementation of a diet low in carot-
enoids with tomato or carrot juice does not affect lipid peroxidation in plasma and feces of healthy men.
J. Nutr., 134, 1081–1083, 2004.
58. Schnäbele, K., Briviba, K., Bub, A., Roser, S., Pool-Zobel, B.L., and Rechkemmer, G., Effects of car-
rot and tomato juice consumption on faecal markers relevant to colon carcinogenesis in humans. Br. J.
Nutr., 99, 606–613, 2008.
59. Moro, E., Karottensuppe bei Ernährungsstörungen der Säuglinge. Munchen. Med. Wochen., 31,
1637–1640, 1908.
60. Fanaro, S., Jelinek, J., Stahl, B., Boehm, G., Kock, R., and Vigi, V., Acidic oligosaccharides from pectin
hydrolysate as new component for infant formulae: Effect on intestinal flora, stool characteristics, and
pH. J. Pediatr. Gastroenterol. Nutr., 41, 186–190, 2005.
61. Stoll, T., Schieber, A., and Carle, R., Quantitative determination of saturated oligogalacturonic acids
in enzymatic digests of polygalacturonic acid, pectin and carrot pomace by online LC-ESI-MS. Anal.
Bioanal. Chem., 377, 655–659, 2003.
62. Kastner, U., Glasl, S., Follrich, B., Guggenbichler, J.P., and Jurenitsch, J., Acidic oligosaccharides as
active principle of aqueous carrot extracts in the prophylaxis and therapy of gastrointestinal infections.
Wien. Med. Wochenschr., 152, 379–381, 2002.
63. Stoll, T., Schweiggert, U., Schieber, A., and Carle, R., Process for the recovery of a carotene-rich func-
tional food ingredient from carrot pomace by enzymatic liquefaction. Innov. Food Sci. Emerg. Technol.,
4, 415–423, 2003.
64. European Commission, EU Register of Nutrition and Health claims, 2013, Published online at: http://
ec.europa.eu/nuhclaims/ (accessed in March 2014).
65. El-Abasy, A.E., Abou-Gharbia, H.A., Mousa, H.M., and Youssef, M.M., Mixes of carrot juice and some
fermented dairy products: Potentiality as novel functional beverages. Food Nutr. Sci., 3, 233–239, 2012.
66. European Fruit Juice Association (AIJN), Liquid fruit market report, European Fruit Juice Association,
Rue de la Loi, Brussels, Belgium, 2013.
Carrot Juice 581
67. Urala, N., Schutz, H., and Spinks, J., Consumer perceptions of “functional food” in the United States.
J. Food Prod. Market., 17, 407–419, 2011.
68. Domagala-Swiatkiewicz, I. and Gastol, M., Comparative study on mineral content of organic and
conventional carrot, celery and red beet juices. Acta Sci. Pol. Hort. Cult. 11, 173–183, 2012.
69. Food and Nutrition Board of the Institute of Medicine, Dietary reference intakes (DRIs), Published
online at: http://iom.edu/ (accessed January 28, 2014).
47
Chinese Medicinal Herbs and Root-Based Beverages
Chin-Lin Hsu, Chi-Cheng Lu, and Gow-Chin Yen
CONTENTS
47.1 Introduction................................................................................................................................... 583
47.4 Health Effects................................................................................................................................ 586
47.4.2 Preventing DNA Damage................................................................................................. 587
47.4.3 Antimutagenic Action...................................................................................................... 587
47.4.4 Hepatoprotective Action................................................................................................... 588
47.4.5 Preventing Liver Fibrosis.................................................................................................. 588
47.4.6 Renal Protective Activity................................................................................................. 588
47.4.7 Protection of Myocardium................................................................................................ 588
47.4.8 Antihypertensive Action................................................................................................... 589
47.4.9 Anti-Inflammatory Activity............................................................................................. 589
47.6 Conclusion..................................................................................................................................... 590
References............................................................................................................................................... 590
47.1 Introduction
Chinese herbal or medicinal tea is essentially an herbal mixture made from leaves, seeds, and/or roots of
various plants [1]. Herbal tea is a medicinal drink and a traditional beverage that originated from China
over 2000 years ago. The herbal tea is usually caffeine free and is may be divided into single compo-
nent (e.g., mulberry tea, Tochu tea [Eucommia ulmoides leaves], ginseng tea, and Hsian-tsao [Mesona
procumbens Hemsl.] tea) or mixtures of several ingredients (e.g., Guangdong herbal tea [well-known as
Wanglaoji-Liang-Cha]). Liu et al. [2] demonstrated that herbal drinks consist of different plant parts,
such as leaf, root, fruit, flower, branch, bud, pollen, stigma, pith, bulb, tuber, kernel, stem, peel, aerial
part, seed, bark, and rhizome from herbaceous and woody plants. The root, whole plant, fruit, flower,
seed, aerial part, and leaf were the common plant parts used for making herbal teas. In single-component
herbal tea, mulberry tea inhibits postprandial hyperglycemia in type 2 diabetic patients [3]. Tochu tea has
a potential antimutagenicity due to (1) the suppressing effect on the urine mutagenicity after ingestion
of raw fish and cooked beef and (2) the clastogen-suppressing effects as evidenced in Chinese hamster
ovary (CHO) cells and mice [4,5]. It has also been indicated that ginseng tea has a cerebroprotective
action against global and focal ischemic injury [6]. Many studies have shown that Hsian-tsao herbal tea
has several health benefits (e.g., antioxidant, antimutagenic, hepatoprotective, antifibrosis, renal protec-
tive, myocardium protective, antihypertensive, and anti-inflammatory activities) and is a good choice for
a beverage [7–14]. Furthermore, He et al. [15] revealed that Guangdong tea consists of multiple Chinese
herbs, including Ilex asprella (Hook. Et Arn.) Champ. ex Benth., Lophatherum gracile Brongn.,
Vitex negundo L., Microcos paniculata L., Oroxylum indicum (L.) Vent., Rosa laevigata Michx.,
583
Lygodium japonicum (Thumb.) Sw., Desmodium styracifolium (Osb.) Merr., Polygonum chinense L.,
and Helicteres angustifolia L. This tea has been used as a health-promoting beverage for prevention of
infectious diseases, relieving fever, alleviating pain, and restoring strength in Hong Kong and China
for nearly 200 years [1]. Recently, Chinese medicinal materials used in functional food or beverage are
scoring an increasingly positive trend. Hence, we summarize some common Chinese medicinal materi-
als and point out the part(s) of plant used in beverage and herbal tea [16], as summarized in Table 47.1.
Importantly, Hsian-tsao is usually used as one of the major ingredients in Chinese traditional herbal tea
such as Wanglaoji-Liang-Cha, which is a popular herbal beverage in China, and in other herbal teas. The
whole plant of Hsian-tsao can be used for food and beverage. It is also a widespread drink and a healthy
beverage in Taiwan.
Hsian-tsao is a traditional herb in Asian countries such as Taiwan, China, and Hong Kong. In Taiwan,
it is usually harvested after 6 months from planting (average planting date: March–April), and the main
growing regions are in northern and eastern Taiwan, such as Hsinchu, Miaoli, Taoyuan, and Hualien.
This tea is consumed as a functional beverage in Taiwan and China. Moreover, Lai and Lin [17] indi-
cated that Hsian-tsao leaf contains polysaccharide gum, called Hsian-tsao leaf gum. It is used in mak-
ing Hsian-tsao jello-type dessert in the orient, and hot-/cold-food products are very popular in the four
seasons of Taiwan. Several studies demonstrate that the herb of Hsian-tsao contains several compounds
TABLE 47.1
List of Some Chinese Medicinal Materials Used in Beverages and Herbal Teas
Scientific Name Common Name Part(s) Used in Beverage
Mesona procumbens Hemsl. Hsian-tsao Root, whole plant, and leaf
Morus alba Linn. Mulberry Fruit
Eucommia ulmoides Tochu Leaf
Rhodiola rosea Rhodiola Root
Benincasa hispida White gourd Fruit
Chaenomeles speciosa Papaya Fruit
Crataegus pinnatifida/C. cuneata Chinese hawthorn fruit Fruit
Ziziphus jujuba Jujube fruit Fruit
Lycium barbarum Wolfberry Fruit
Momordica grosvenori Luo han guo/Grosvenor momordica fruit Fruit
Prunus mume Japanese apricot fruit Fruit
Ganoderma lucidum Lucid ganoderma/lingzhi Sporophore (fruiting body)
Mentha piperita/M. haplocalyx Wild mint herb Leaf
Acanthopanax senticosus Siberian ginseng Radix and rhizome (root)
Codonopsis pilosula Tangshen root Radix (root)
Glycyrrhiza uralensis Liquorice Radix (root)
Panax ginseng Ginseng root Radix and rhizome (root)
Panax quinquefolius American ginseng root Radix (root)
Polygonum multiflorum Tuber fleeceflower root Radix (root)
Pueraria lobata Lobed kudzuvine root Radix (root)
Zingiber officinale Ginger Rhizome (root)
Prunus armeniaca Almond Seed
Cassia tora Sickle senna seed Seed
Nelumbo nucifera Lotus seed Seed
Sterculia lychnophora Boat-fruited sterculia seed Seed
Prunella vulgaris Common self-heal spike Fruit spike
Tremella fuciformis Tremella Sporophore (fruiting body)
Auricularia auricula-judea Jew’s ear (black fungus) Sporophore (fruiting body)
Source: Adapted and modified from Agri-Food & Veterinary Authority of Singapore, 2014, Published online
at: http://www.ava.gov.sg/ (accessed January 20, 2015).
Chinese Medicinal Herbs and Root-Based Beverages 585
including macrocomponents (e.g., fat, protein, fiber, and ash), as well as minerals (e.g., Na, K, Ca, Mg,
Fe, and Zn), and bioactives (e.g., stigmasterol, β-sitosterol, oleanolic acid, ursolic acid, protocatechuic
acid, p-hydroxybenzoic acid, vanillic acid, syringic acid, and caffeic acid) [18,19]. The herb of Hsian-
tsao has been reported to exhibit antioxidant activity, preventing DNA damage, antimutagenic action,
hepatoprotective effect, liver fibrosis, renal-protective activity, myocardium, antihypertensive action,
and anti-inflammatory activity [7–14,18,20]. This chapter highlights the nutritional characteristics, anti-
oxidant efficacy, health effects, and novel products of Hsian-tsao, where possible comparison is made
with medicinal herbs and root-based beverages.

Proximate compositions, mainly fat, protein, ash, fiber, and nitrogen-free extracts (NFE), of Hsian-tsao
dried leaves have been reported to be 2.65, 9.55, 9.55, 31.42, and 46.83 g/100 g, respectively. Moreover,
the contents of Na, K, Ca, Mg, Fe, and Zn in Hsian-tsao dried leaves were 502, 566, 660, 75, 101, and
15 mg/100 g, respectively [18,19]. Nutritional characteristics of commercial Hsian-tsao products are
given in Table 47.2 [21].

Hung and Yen [18] reported the antioxidative components that are isolated from Hsian-tsao. This study
indicated that the antioxidant activity (an inhibition of peroxidation of linoleic acid) of the ethyl acetate
extracts of Hsian-tsao was greater than those of butylated hydroxyanisole, α-tocopherol, and other sol-
vent extracts of Hsian-tsao. The ethyl acetate extracts of Hsian-tsao was separated into nine fractions by
silica gel chromatography. The yield and antioxidant activity of fraction II was higher than that of other
fractions. Moreover, fraction II was further separated into five subfractions (A, B, C, D, and E) using
silica gel chromatography. In the subfraction IIC, four compounds including stigmasterol, β-sitosterol,
oleanolic acid, and ursolic acid were isolated, purified, and identified by ultraviolet (UV), electron
impact–mass spectroscopy (EI-MS), proton nuclear magnetic resonance (1H NMR), and carbon nuclear
magnetic resonance (13C NMR). Chemical structures of isolated compounds (stigmasterol, β-sitosterol,
oleanolic acid, and ursolic acid) from Hsian-tsao are shown in Figure 47.1. Oleanolic acid and ursolic
acid have antioxidant activities in hypertension cases [22]. This acid also plays a role in the antioxidant
action, including quenching reactive oxygen species, inhibiting lipid peroxidation, and modulating cel-
lular antioxidant defenses system [23]. do Nascimento et al. [24] indicated that ursolic acid and its deriva-
tives have strong 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity.
Hung and Yen [19] also reported the antioxidant activity of phenolic compounds isolated from Hsian-
tsao, in which the content of total phenolic and β-tocopherol was 185 and 0.33 mg/g of Hsian-tsao extract,
TABLE 47.2
Nutritional Characteristics of Hsian-Tsao Products
Hsian-Tsao Tea (Serving Size: 960 mL) Instant Hsian-Tsao (Serving Size: 3 g)
Nutrient Unit (Quantity per 100 mL) (Quantity per 3 g)
Energy kcal 13 7.3
Protein g 0.0 0.2
Fat (lipid) g 0.0 0.1
Saturated fat g 0.0 0.0
Sodium mg 0 266
Source: Adapted from Guanxi Farmers’ Associations, 2014, Published online at: http://kusi.myweb.hinet.net/
page05.html (accessed January 20, 2015).
CH3 CH3
H3C
H3C CH3 CH3
CH3 CH3
CH3 CH3
CH3 CH3
HO HO
Stigmasterol β-Sitosterol
CH3
H3C CH3
H3C
OH OH
CH3 CH3 CH3 CH3
O O
CH3 CH3
HO HO
H3C CH3 H3C CH3
Oleanolic acid Ursolic acid
O OH O OH O OH
OH O CH3
OH OH OH
Protocatechuic acid p-Hydroxybenzoic acid Vanillic acid
O OH
O
HO OH
H3C O O CH3
OH HO
Syringic acid Caffeic acid
FIGURE 47.1 Chemical structures of isolated compounds from Hsian-tsao.
respectively. However, other tocopherols (α, γ, and δ forms) and β-carotene were not detected in the
Hsian-tsao extracts. Additionally, this study indicated that acidic ethyl acetate (pH 2) fraction, isolated
from Hsian-tsao extracts, contained large amounts of phenolic compounds and a strong antioxidant
activity on peroxidation of linoleic acid. The acidic ethyl acetate fraction of Hsian-tsao extracts was sub-
sequently separated into four subfractions (A–D), among which subfraction B had a high yield and strong
antioxidant activity. Therefore, five phenolic compounds (including protocatechuic acid, p-hydroxyben-
zoic acid, vanillic acid, syringic acid, and caffeic acid) were isolated, purified, and identified (using UV,
EI-MS, 1H NMR, and 13C NMR) from acidic ethyl acetate subfraction B of Hsian-tsao extracts. Chemical
structures of isolated compounds (including protocatechuic acid, p-hydroxybenzoic acid, vanillic acid,
syringic acid, and caffeic acid) from Hsian-tsao are shown in Figure 47.1.
47.4 Health Effects

The biological functions of different extracts from Hsian-tsao are shown in Table 47.3. Health effects of
Hsian-tsao are detailed in the succeeding text.
TABLE 47.3
Biological Functions of Different Extracts from Hsian-Tsao
Extracts Dose (Duration) Functions References
Water extract np Preventing DNA damage: reduced DNA damage in human [7]
lymphocytes induced by UV-C and/or H2O2.
Water extract np Antimutagenic action: inhibited IQ-induced mutagenesis [8]
in Salmonella tester strains TA98 and TA100.
Water extract 0.1, 0.5, and 1.0 g/kg Hepatoprotective effect: inhibited t-BHP-induced acute [9]
rat (13 days) hepatic damage.
Water extract 1.2 g/kg rat (8 weeks) Preventing liver fibrosis: inhibited carbon tetrachloride [10]
(CCl4)-induced liver injury.
Water extract 1.5 g/kg rat (4 weeks) Renal protective activity: prevented renal pathological [11]
changes and decreased the TSP-1 expression in
streptozotocin-induced diabetic rats.
Water extract 1.5 g/kg rat (4 weeks) Protection of myocardium: prevented pathological changes [12]
of myocardium and decreased TSP-1 expression in
streptozotocin-induced diabetic rats.
Water extract 1.0 g/kg rat (6 weeks) Antihypertensive action: decreased the levels of systolic [13]
pressure and arterial pressure.
Water extract 250 and 500 mg/kg Anti-inflammatory activity: inhibited carrageenan-induced [14]
mouse (2 h) mouse paw edema.
0.1%–0.3% np Antioxidant activity: scavenging capability on DPPH and [20]
alkaline solution superoxide anion radicals.
(NaHCO3) extract
np, not provided/in vitro model; DPPH, 1,1-diphenyl-2-picryl-hydrazyl; UV-C, ultraviolet–C; IQ, 2-amino-
Abbreviations:
3-methylimidazo(4,5-f)quinolone; t-BHP, tert-butyl hydroperoxide; TSP-1, thrombospondin-1.

Hsian-tsao extracts scavenge DPPH and superoxide anion radicals and inhibit lipid peroxidation [18,20].
Sun-dried Hsian-tsao sample was extracted with Na2CO3 or NaHCO3 at concentrations of 0%–1.5% upon
heating for 0.5–3 h. The antioxidant properties of Hsian-tsao extracts were evaluated by using different
in vitro antioxidant assays, such as antioxidant activity by thiocyanate method, DPPH radical scaveng-
ing, superoxide radical anion scavenging, and total phenolic content. The results indicated a higher
amount of total phenolics and stronger antioxidant activity in the Hsian-tsao extracts upon alkaline
(0.1%–0.3% Na2CO3) and heat treatments (2 h) [20].
47.4.2 Preventing DNA Damage

Hsian-tsao extracts reduce DNA damage in human lymphocytes induced by ultraviolet-C (UV-C) and/or
H2O2 [7]. In this study, dried Hsian-tsao powder was extracted with boiling water for 30 min, and the water
extract solutions were filtered and freeze dried (sample called water extract of Hsian-tsao). The protective
effect of water extracts of Hsian-tsao on DNA damage in UV-C- and/or H2O2-induced human lymphocytes
was evaluated using single-cell electrophoresis (comet assay). The comet assay is a sensitive and rapid
technique for quantifying and analyzing DNA damage that can be applied both in vitro and in vivo models
[25–27]. Water extracts of Hsian-tsao could attenuate UV-C- and H2O2-induced DNA damage in human
lymphocytes, which might mainly be attributed to their polyphenolic compounds. Therefore, this study
suggests that water extract of Hsian-tsao has a potential role in the chemoprevention of photooxidation.
47.4.3 Antimutagenic Action

Water extracts of Hsian-tsao have been found to inhibit benzo[a]pyrene (B[a]P)- and 2-amino-
3-methylimidazo(4,5-f)quinoline (IQ)-induced mutagenesis in Salmonella strains TA98 and TA100 [8].
These antimutagenic studies are good in vitro model to identify natural substances with antigenotoxic
and antimutagenic potential [28–30]. The dried Hsian-tsao powder, prepared via extraction with boiling
water for 30 min, filtration, and freeze drying (sample called water extract of Hsian-tsao), had inhibi-
tory effect [8]. This was found to be effective in a dose-dependent manner at 0.2–0.6 mg/plate on the
mutagenicity of benzo[a]pyrene (B(a)P) and IQ toward Salmonella typhimurium TA98 and TA100 [8].
This study suggests that the antimutagenic activity of water extract of Hsian-tsao might be due to its
polyphenolic compounds and ascorbic acid.
47.4.4 Hepatoprotective Action

Water extracts of Hsian-tsao suppress tert-butyl hydroperoxide (t-BHP)-induced acute hepatic damage
in rats [9]. t-BHP is an organic peroxide and widely applied to investigate the cell injury initiated by
oxidative stress [31,32]. Yen et al. [9] showed the protective effect of water extracts of Hsian-tsao against
t-BHP-induced acute hepatic damage in male Sprague Dawley rats. These rats were orally pretreated
with water extracts of Hsian-tsao (0.1, 0.5, and 1 g/kg of body weight) or its active compound, caffeic
acid (0.1 g/kg of body weight), for 13 days, before a single dose of t-BHP (0.2 mmol/kg of body weight)
was intraperitoneally injected into each animal, and 18 h later, the rats were sacrificed by decapitated.
These results demonstrated that intake of water extracts of Hsian-tsao and its active compound, caffeic
acid, can be beneficial for the suppression of t-BHP-induced acute hepatic damage (lowered the serum
levels of the hepatic enzyme markers [including alanine aminotransferase, aspartate aminotransferase,
and lactate dehydrogenase], reduced hepatic oxidative stress [including lower levels of malondialdehyde,
glutathione disulfide, and 8-hydroxy-2′-deoxyguanosine, and higher levels of glutathione, glutathione
peroxidase, and glutathione reductase], and incidence of liver lesions [including cloudy swelling, pykno-
sis, and cytolysis]) in rats. Caffeic acid was isolated, purified, and identified from Hsian-tsao extract [19].
Yang et al. [33] indicated that caffeic acid isolated from fermented field water dropwort (Oenanthe
javanica) extract ameliorates t-BHP-induced hepatotoxicity in HepG2 cells and carbon tetrachloride
(CCl4)-induced liver damage in rats.
47.4.5 Preventing Liver Fibrosis

Shyu et al. [10] showed that water extracts of Hsian-tsao inhibited CCl4-induced liver injury in rats.
Hsian-tsao extracts prevented rat liver fibrosis induced by CCl4 via inhibition of hepatic stellate cell acti-
vation. In preparation of Hsian-tsao extracts, sample powder was extracted with boiling water for 2 h,
and then the liquid extracts were filtered and freeze dried to produce a powder, named as water extracts
of Hsian-tsao. These results demonstrated that intake of water extracts of Hsian-tsao can be beneficial
for the suppression of CCl4-induced liver fibrosis in rats via the modulations of redox state, smooth
muscle α-actin (α-SMA) protein expression, and activations of matrix metalloproteinase-2 (MMP-2) and
MMP-9. This study also indicated that their active compounds (including oleanolic acid and ursolic acid)
decreased the levels of pro-MMP-2 activity and MMP-2 activation, and α-SMA protein expression in
cultured rat liver stellate HSC-T6 cells.
47.4.6 Renal Protective Activity

It is reported that water extracts of Hsian-tsao prevented renal pathological changes and decreased throm-
bospondin-1 (TSP-1) expression in streptozotocin-induced diabetic rats [11]. Dried Hsian-tsao leaves
were extracted with boiling water for 2 h, filtrated, and evaporated under reduced pressure (34–36 kPa)
using a rotary vacuum evaporator at 40°C, and the contents were freeze dried. The intake of these water
extracts at 1.5 g/kg for 4 weeks can be beneficial for renal protection in diabetic rats. They are mediated
by reducing TSP-1 expression and minimizing kidney’s ultrastructural damages.
47.4.7 Protection of Myocardium

Water extracts of Hsian-tsao have been found to prevent pathological changes of myocardium and
to decrease TSP-1 expression in streptozotocin-induced diabetic rats [12]. The Hsian-tsao extracts
(1.5 g/kg rat for 4 weeks) prevented myocardium damages in streptozotocin-induced diabetic rats via
decreased expression of TSP-1 [12]. Therefore, Yang’s study team [11,12] demonstrated that water
extracts of Hsian-tsao prevented the renal and myocardium damages in streptozotocin-induced diabetic
rats through a key modulator of TSP-1.
47.4.8 Antihypertensive Action

Water extracts of Hsian-tsao decreased the levels of systolic pressure and arterial pressure in spontane-
ously hypertensive rats. A supplement of water extracts of Hsian-tsao (1 g/kg rat for 6 weeks) may sup-
press the development of increased blood pressure in spontaneously hypertensive rats [13]. This study
also demonstrated that its active component, caffeic acid, had potential blood pressure‒lowering effect in
spontaneously hypertensive rats. Intakes of water extracts of Hsian-tsao and its active component, caffeic
acid, decreased the level of malondialdehyde (in liver and plasma) and increased the levels of glutathione
(in liver), total superoxide dismutase (in liver and kidney), glutathione peroxidase (in liver and kidney),
and catalase (in liver and kidney) in spontaneously hypertensive rats.

Water extracts of Hsian-tsao possess an inhibition ability in carrageenan-induced mouse paw edema in
mice. Water extracts of Hsian-tsao (250 and 500 mg/kg for 2 h) suppressed carrageenan-induced paw
edema in mice through decreasing the levels of serum nitric oxide (NO), tissue malondialdehyde, serum
tumor necrosis factor-alpha (TNF-α), and serum interleukin-1 beta (IL-1β) [14]. High-performance liq-
uid chromatography (HPLC) profile demonstrated protocatechuic acid, chlorogenic acid, vanillic acid,
and caffeic acid as the active components in water extracts of Hsian-tsao [14]. Protocatechuic acid
inhibited carrageenan-induced paw edema in rats, and this might be attributed to its anti-inflammatory
activity, antioxidant, and membrane-stabilizing property [34]. Methanol extract from the bark of Odina
wodier and its major constituent chlorogenic acid ameliorated carrageenan-induced paw edema in rats
[35]. In addition, vanillic acid from wild and cultivated Amburana cearensis inhibited the paw edema
in rat peritoneal cavity induced by carrageenan [36]. Finally, caffeic acid derivatives (e.g., butyl caffe-
ate, octyl caffeate, and caffeic acid phenethyl ester) inhibited carrageenan-induced paw edema and
prevented the increase level of IL-1β in the mouse paw [37].

A number of novel Hsian-tsao’s food products (e.g., Hsian-tsao tea bag, instant Hsian-tsao powder, Hsian-
tsao cake, Hsian-tsao noodles, and Hsian-tsao sugar) have been developed in Taiwan.
Instant Hsian-tsao powder: Dried Hsian-tsao plants were washed, extracted (0.1% Na2CO3 solu-
tion), filtrated, and spray dried. In general, selecting suitable carriers play an important role
in spray-drying process. Some carriers (e.g., maltodextrin, cyclodextrin, potato starch, wheat
starch, green bean starch, and carboxyl methyl cellulose) were used in the instant Hsian-tsao
spray-drying procedure [38]. This product looks like coffee and designated as a convenience
commodity that is easy to prepare and easy to drink. Nutritional characteristics of Hsian-tsao
products are shown in Table 47.2.
Hsian-tsao cake: Hsian-tsao cake is a green-colored steam rice cake. Steam rice cake is a tradi-
tional Chinese dessert and a ready-to-eat food item. In the process, the raw materials of Hsian-
tsao cake (including water extracts of Hsian-tsao, rice, flour, sugar, baking powder, and water)
are mixed, fermented, and steamed for 1–2 h to produce the final product.
In previous studies, Hsian-tsao herbs were reported to have multiple functions, such as antioxidant,
antimutagenic, hepatoprotective, renal protective, antihypertensive, and anti-inflammatory activities
[8–11,13,14,20]. The polyphenol-rich extract of Hsian-tsao can be used as functional food components.
Many studies have indicated that polyphenolic compounds (including phenolic acids and flavonoids)
have pharmacological properties such as antioxidant activity, antimutagenicity, anti-inflammation, anti-
obesity, and antifatigue activity [39–44]. Therefore, the water extracts of Hsian-tsao or combination for-
mulas have potential applications in the development of functional foods with associated health effects
related to antihypertension, hepatoprotection, antiobesity, and antifatigue.
47.6 Conclusion
Hsian-tsao is one of the edible plants and traditional herbs, containing bioactive components, minerals,
and polysaccharide gum. It is used in making Hsian-tsao tea, Hsian-tsao jello-type dessert, and steam
cake in the Orient and these products are very popular in Taiwan. This herb has been reported to have
many biological functions, such as antioxidant activity, preventing DNA damage, antimutagenic action,
hepatoprotective action, preventing liver fibrosis, renal protective activity, protection of myocardium,
antihypertensive action, and anti-inflammatory activity. Moreover, the water extracts of Hsian-tsao or
combination formulas have potential applications in the development of multiple functional foods.
REFERENCES
1. Hu, S.Y., Herbal teas and populace health care in tropical China. Am. J. Chin. Med., 25, 103–134, 1997.
2. Liu, Y., Ahmed, S., and Long, C., Ethnobotanical survey of cooling herbal drinks from southern China.
J. Ethnobiol. Ethnomed., 9, 82, 2013.
3. Banu, S., Jabir, N.R., Manjunath, N.C., Khan, M.S., Ashraf, G.M., Kamal, M.A., and Tabrez, S.,
Reduction of post-prandial hyperglycemia by mulberry tea in type-2 diabetes patients. Saudi J. Biol.
Sci., 22, 32–36, 2015.
4. Sasaki, Y.F., Chiba, A., Murakami, M., Sekihashi, K., Tanaka, M., Takahoko, M., Moribayashi, S. et al.,
Antimutagenicity of Tochu tea (an aqueous extract of Eucommia ulmoides leaves): 2. Suppressing effect
of Tochu tea on the urine mutagenicity after ingestion of raw fish and cooked beef. Mutat. Res., 371,
203–214, 1996.
5. Nakamura, T., Nakazawa, Y., Onizuka, S., Satoh, S., Chiba, A., Sekihashi, K., Miura, A., Yasugahira,
N., and Sasaki, Y.F., Antimutagenicity of Tochu tea (an aqueous extract of Eucommia ulmoides leaves):
1. The clastogen-suppressing effects of Tochu tea in CHO cells and mice. Mutat. Res., 388, 7–20, 1997.
6. Shah, Z.A., Gilani, R.A., Sharma, P., and Vohora, S.B., Cerebroprotective effect of Korean ginseng tea
against global and focal models of ischemia in rats. J. Ethnopharmacol., 101, 299–307, 2005.
7. Yen, G.C., Hung, Y.L., and Hsieh, C.L., Protective effect of extracts of Mesona procumbens Hemsl. on
DNA damage in human lymphocytes exposed to hydrogen peroxide and UV irradiation. Food Chem.
Toxicol., 38, 747–754, 2000.
8. Yen, G.C., Duh, P.D., and Hung, Y.L., Contributions of major components to the antimutagenic effect of
Hsian-tsao (Mesona procumbens Hemsl.). J. Agric. Food Chem., 49, 5000–5004, 2001.
9. Yen, G.C., Yeh, C.T., and Chen, Y.J., Protective effect of Mesona procumbens against tert-butyl hydro-
peroxide-induced acute hepatic damage in rats. J. Agric. Food Chem., 52, 4121–4127, 2004.
10. Shyu, M.H., Kao, T.C., and Yen, G.C., Hsian-tsao (Mesona procumbens Heml.) prevents against rat
liver fibrosis induced by CCl4 via inhibition of hepatic stellate cells activation. Food Chem. Toxicol., 46,
3707–3713, 2008.
11. Yang, M., Xu, Z.P., Xu, C.J., Meng, J., Ding, G.Q., Zhang, X.M., and Weng, Y., Renal protective activity
of Hsian-tsao extracts in diabetic rats. Biomed. Environ. Sci., 21, 222–227, 2008.
12. Yang, M., Xu, Z., Zhang, R., Zhang, P., Weng, Y., Shen, Y., and Zhang, X., Protection of myocardium
in streptozotocin-induced diabetic rats by water extracts of Hsian-tsao (Mesona procumbens Hemsl.).
Asia Pac. J. Clin. Nutr., 17, 23–29, 2008.
13. Yeh, C.T., Huang, W.H., and Yen, G.C., Antihypertensive effects of Hsian-tsao and its active compound
in spontaneously hypertensive rats. J. Nutr. Biochem., 20, 866–875, 2009.
14. Huang, G.J., Liao, J.C., Chiu, C.S., Huang, S.S., Lin, T.H., and Deng, J.S., Anti-inflammatory activities
of aqueous extract of Mesona procumbens in experimental mice. J. Sci. Food Agric., 92, 1186–1193,
2012.
15. He, R.R., Tsoi, B., Li, Y.F., Yao, X.S., and Kurihara, H., The anti-stress effects of Guangdong herbal tea
on immunocompromise in mice loaded with restraint stress. J. Heal. Sci., 57, 255–263, 2011.
16. Agri-Food & Veterinary Authority of Singapore, 2014. Published online at: http://www.ava.gov.sg/
17. Lai, L.S. and Lin, T.P.S., Solution properties of hsian-tsao (Mesona procumbens Hemsl) leaf gum. Food
Hydrocolloid., 14, 287–294, 2000.
18. Hung, C.Y. and Yen, G.C., Extraction and identification of antioxidative components of Hsian-tsao
(Mesona procumbens Hemsl.). LWT-Food Sci. Technol., 34, 306–311, 2001.
19. Hung, C.Y. and Yen, G.C., Antioxidant activity of phenolic compounds isolated from Mesona procum-
bens Hemsl. J. Agric. Food Chem., 50, 2993–2997, 2002.
20. Yen, G.C. and Hung, C.Y., Effects of alkaline and heat treatment on antioxidative activity and total
phenolics of extracts from Hsian-tsao (Mesona procumbens Hemsl.). Food Res. Int., 33, 487–492, 2000.
21. Guanxi Farmers’ Association, 2014. Published online at: http://kusi.myweb.hinet.net/page05.html
22. Somova, L.O., Nadar, A., Rammanan, P., and Shode, F.O., Cardiovascular, antihyperlipidemic and anti-
oxidant effects of oleanolic and ursolic acids in experimental hypertension. Phytomedicine, 10, 115–121,
2003.
23. Wang, X., Ye, X.L., Liu, R., Chen, H.L., Bai, H., Liang, X., Zhang, X.D., Wang, Z., Li, W.L., and Hai,
C.X., Antioxidant activities of oleanolic acid in vitro: Possible role of Nrf2 and MAP kinases. Chem.
Biol. Interact., 184, 328–337, 2010.
24. do Nascimento, P.G., Lemos, T.L., Bizerra, A.M., Arriaga, A.M., Ferreira, D.A., Santiago, G.M.,
Braz-Filho, R., and Costa, J.G., Antibacterial and antioxidant activities of ursolic acid and derivatives.
Molecules, 19, 1317–1327, 2013.
25. Kirkland, D. and Speit, G., Evaluation of the ability of a battery of three in vitro genotoxicity tests to
discriminate rodent carcinogens and non-carcinogens III. Appropriate follow-up testing in vivo. Mutat.
Res., 654, 114–132, 2008.
26. Dusinska, M. and Collins, A.R., The comet assay in human biomonitoring: Gene–environment interac-
tions. Mutagenesis, 23, 191–205, 2008.
27. Collins, A.R., Azqueta, A., and Langie, S.A., Effects of micronutrients on DNA repair. Eur. J. Nutr., 51,
261–279, 2012.
28. Nieva Moreno, M.I., Zampini, I.C., Ordonez, R.M., Jaime, G.S., Vattuone, M.A., and Isla, M.I.,
Evaluation of the cytotoxicity, genotoxicity, mutagenicity, and antimutagenicity of propolis from
Tucuman, Argentina. J. Agric. Food Chem., 53, 8957–8962, 2005.
29. Arriaga-Alba, M., Blasco, J.L., Ruiz-Perez, N.J., Sanchez-Navarrete, J., Rivera-Sanchez, R., and
Gonzalez-Avila, M., Antimutagenicity mechanisms of the Rhoeo discolor ethanolic extract. Exp. Toxicol.
Pathol., 63, 243–248, 2011.
30. Del-Toro-Sanchez, C.L., Bautista-Bautista, N., Blasco-Cabal, J.L., Gonzalez-Avila, M., Gutierrez-
Lomeli, M., and Arriaga-Alba, M., Antimutagenicity of methanolic extracts from Anemopsis califor-
nica in relation to their antioxidant activity. Evid. Based Complement. Alternat. Med., 2014, 273878,
2014.
31. Rush, G.F., Gorski, J.R., Ripple, M.G., Sowinski, J., Bugelski, P., and Hewitt, W.R., Organic hydroper-
oxide-induced lipid peroxidation and cell death in isolated hepatocytes. Toxicol. Appl. Pharmacol., 78,
473–483, 1985.
32. Martin, C., Martinez, R., Navarro, R., Ruiz-Sanz, J.I., Lacort, M., and Ruiz-Larrea, M.B., tert-Butyl
hydroperoxide-induced lipid signaling in hepatocytes: Involvement of glutathione and free radicals.
Biochem. Pharmacol., 62, 705–712, 2001.
33. Yang, S.A., Jung, Y.S., Lee, S.J., Park, S.C., Kim, M.J., Lee, E.J., Byun, H.J., Jhee, K.H., and Lee, S.P.,
Hepatoprotective effects of fermented field water-dropwort (Oenanthe javanica) extract and its major
constituents. Food Chem. Toxicol., 67, 154–160, 2014.
34. Lende, A.B., Kshirsagar, A.D., Deshpande, A.D., Muley, M.M., Patil, R.R., Bafna, P.A., and Naik, S.R.,
Anti-inflammatory and analgesic activity of protocatechuic acid in rats and mice. Inflammopharmacology,
19, 255–263, 2011.
35. Ojha, D., Mukherjee, H., Mondal, S., Jena, A., Dwivedi, V.P., Mondal, K.C., Malhotra, B., Samanta, A.,
and Chattopadhyay, D., Anti-inflammatory activity of Odina wodier Roxb, an Indian folk remedy,
through inhibition of toll-like receptor 4 signaling pathway. PLoS One, 9, e104939, 2014.
36. Leal, L.K., Pierdona, T.M., Goes, J.G., Fonseca, K.S., Canuto, K.M., Silveira, E.R., Bezerra, A.M., and
Viana, G.S., A comparative chemical and pharmacological study of standardized extracts and vanillic
acid from wild and cultivated Amburana cearensis A.C. Smith. Phytomedicine, 18, 230–233, 2011.
37. da Cunha, F.M., Duma, D., Assreuy, J., Buzzi, F.C., Niero, R., Campos, M.M., and Calixto, J.B., Caffeic
acid derivatives: In vitro and in vivo anti-inflammatory properties. Free Radic. Res., 38, 1241–1253,
2004.
38. Shih, H.T., Study on the techniques of spray dry instant Hsian-tsao. Bulletin of Taoyuan District
Agricultural Research and Extension Station, 9, 11–19, 1992.
39. Son, S. and Lewis, B.A., Free radical scavenging and antioxidative activity of caffeic acid amide and
ester analogues: Structure–activity relationship. J. Agric. Food Chem., 50, 468–472, 2002.
40. Hsu, C.L. and Yen, G.C., Effect of gallic acid on high fat diet-induced dyslipidaemia, hepatosteatosis
and oxidative stress in rats. Br. J. Nutr., 98, 727–735, 2007.
41. Hsu, C.L., Wu, C.H., Huang, S.L., and Yen, G.C., Phenolic compounds rutin and o-coumaric acid ame-
liorate obesity induced by high-fat diet in rats. J. Agric. Food Chem., 57, 425–431, 2009.
42. Wu, R.E., Huang, W.C., Liao, C.C., Chang, Y.K., Kan, N.W., and Huang, C.C., Resveratrol protects
against physical fatigue and improves exercise performance in mice. Molecules, 18, 4689–4702, 2013.
43. Michel, P., Dobrowolska, A., Kicel, A., Owczarek, A., Bazylko, A., Granica, S., Piwowarski, J.P., and
Olszewska, M.A., Polyphenolic profile, antioxidant and anti-inflammatory activity of eastern teaberry
(Gaultheria procumbens L.) leaf extracts. Molecules, 19, 20498–20520, 2014.
44. Wang, X., Yang, L., Yang, X., and Tian, Y., In vitro and in vivo antioxidant and antimutagenic activities
of polyphenols extracted from hops (Humulus lupulus L.). J. Sci. Food Agric., 94, 1693–1700, 2014.
48
Tomato Juice
María Jesús Periago and Francisco-Javier García-Alonso
CONTENTS
48.1 Introduction................................................................................................................................... 593
48.3.1 Lycopene........................................................................................................................... 596
48.3.2 Phenolic Compounds........................................................................................................ 596
48.3.3 Tomatine........................................................................................................................... 599
48.4 Health Effects................................................................................................................................ 600
48.4.1 Prevention of Cardiovascular Disease.............................................................................. 600
48.4.2 Prevention of Cancer........................................................................................................ 600
48.4.3 Nonalcoholic Fatty Liver Disease.................................................................................... 601
48.4.4 Reduction of Oxidative Stress and Inflammation Biomarkers......................................... 601
48.5.1 Industrial Processing........................................................................................................ 603
48.5.2 Added Ingredients............................................................................................................ 604
48.6 Conclusion..................................................................................................................................... 604
References............................................................................................................................................... 604
48.1 Introduction
Tomato is the second plant food most produced and consumed in the world. According to the data of the
World Processors Tomato Council in 2013, 35 million metric tons of tomatoes were processed into differ-
ent industrial products such as tomato puree, tomato paste, canned tomato, tomato sauce, diced tomato,
and tomato juice [1]. In Spain, it is estimated that around 80% of the tomato production is destined to
the tomato paste, whereas the other 20% is processed toward other tomato products [2]. Tomato juice
is a product obtained from the fermentable but unfermented edible part of tomato fruit (Lycopersicum
esculentum) having the characteristic color, flavor, and taste typical of the fresh fruit [3]. Tomato juice
is commercially available after applying different processes for preserving its nutritional properties and
criteria of food safety. In contrast to other fruit juices, tomato juice is mainly consumed as beverage
as an appetizer, and other ingredients are often added, such as salt, spices, olive oil, and vegetable oil.
In addition, tomato juice is the main ingredient of different cold beverages, cocktails, and soups, which
are made by mixing the tomato juice with other vegetables (onion, pepper, cucumber, garlic, celery, etc.),
beef broth, clam broth, and alcoholic drinks.
From a nutritional point of view, tomato juice has the same nutritional properties as fresh tomato
fruit, and hence it is characterized by a low fat and protein content, soluble sugar, and a low glycemic
index. It also provides a wide range of minerals and vitamins, being low in sodium, high in potassium,
and containing small amounts of vitamins C, E, and folate [4]. Other bioactive compounds of tomato
593
fruit, with antioxidant properties, such as phenolic compounds (hydroxycinnamic acids and flavonoids)
and carotenoids (mainly lycopene and β-carotene), remain intact in the tomato juice after industrial pro-
cesses [5–7]. Moreover, their contents increase during processing due to the extraction, concentration,
and thermal processing, leading to a reduction of water content and hence concentrating the bioactive
compounds. In addition, industrial processing enhances the availability of these compounds as a result of
the breakdown of cell walls and other cellular organelles, increasing the extractability of the compounds
and facilitating their absorption during digestion in the human large bowel [8]. Taking into consideration
these premises, tomato juice preserves the nutritional properties and the bioactive compounds with anti-
oxidant properties, with the exception of vitamin C, and hence it can be considered a good dietary source
of antioxidants, especially lycopene. This chapter highlights the composition of tomato juice in relation
to nutrients, antioxidant activity, phytochemicals, and potential health benefits of tomato juice consump-
tion, and future perspectives of tomato juice industry.

Numerous factors contribute to the nutrient composition of the tomato juice. In addition, tomato variet-
ies contain varying amounts of the macronutrients, micronutrients, and phytochemicals. The process-
ing method and the type of juice can also determine the final nutritional characteristics of the juice.
According to the International and European Laws [3], tomato juice is a fermentable but unfermented
product obtained from the edible part of tomato fruit that is sound, appropriately mature, and fresh or
preserved by chilling, freezing, or other authorized postharvest treatments, with the characteristic color,
flavor, and taste typical of tomato fruit. Tomato juice is processed from fruits with pips, seeds, and peels,
but parts of them are not incorporated into the final product. As with other fruit juices, flavor, pulp, and
cells obtained by suitable physical means from tomatoes may be restored to the juice. It can also be
obtained by reconstituting concentrated tomato juice with water of suitable quality for human consump-
tion. In this case, concentrated tomato juice is obtained after removal of at least 50% of the water content.
Tomato juice from concentrate is prepared by suitable processes that maintain the essential physical,
chemical, organoleptic, and nutritional characteristics of the average type of juice made from the product
[3,9]. Several ingredients are also authorized to enhance the nutritional composition of tomato juice, so
minerals, vitamins, salt, spices, and aromatic herbs can be added to the juice during the manufacturing
process [3,9].
The compositional and nutritional characteristics of the tomato juice are shown in Table 48.1. From the
nutritional point of view, the main component of tomato juice is water, which ranges from 87.40% to 95%
[10,11], showing reduced macronutrient content. The protein and lipid contents are low, less than 1.0%
and 0.1%, respectively, while carbohydrate is the most abundant macronutrient, mainly formed by rap-
idly absorbed carbohydrate or naturally occurring free sugars [10,11]. According to Directive 2012/12/
EU [3], the simple sugar content in tomato juice obtained from fresh fruit must be higher than 5% sucrose
(°Brix), since tomato fruits used are completely ripened before processing. Even when tomato juice is
prepared from a tomato concentrate, the reconstituted juice must show the same minimum °Brix level of
5%. The content of total dietary fiber ranges from 0.4 to 0.8 g/100 mL [10,11], of which 80% water insol-
uble (mainly composed of cellulose, hemicelluloses, and lignins) and 20% water soluble (represented
by pectins) [12]. The caloric value of tomato juice ranges from 16 to 48 kcal/100 mL [10,11]. In general,
plain tomato juice is considered a product with very low caloric value, due to its high water content and
low concentration of macronutrients, but when it is mixed with beef or clam broth, the energy value
increases up to 37 and 48 kcal/100 mL, respectively.
An issue related to the micronutrient composition of tomato juice that deserves special attention
is its potassium content (89–236 mg/100 mL). The natural content of sodium in a squeezed tomato
juice varies between 3.3 and 10 mg/100 mL. However, in the commercial tomato juice, salt is usually
added as an ingredient, increasing the content of sodium to around 300 mg/100 mL [10,11,13]. The
ratio of potassium to sodium in tomato fruits and in different tomato products has been considered a
beneficial factor for the prevention of cardiovascular disease (CVD) [14]. The content of other miner-
als ranges between 8 and 19 mg/100 mL for calcium, 3 and 11 mg/100 mL for magnesium, and 11 and
Tomato Juice 595
TABLE 48.1
Compositional and Nutritional Characteristics of Tomato Juice (per 100 mL)
Nutrient Unit Tomato Juice References
Energy kcal 16–48 [10,11]
Moisture g 87.40–95 [10,11]
Protein g 0.60–0.82 [10,11]
Lipid (fat) g 0.00–0.10 [10,11]
Carbohydrate g 2.6–10.95 [10,11]
Total sugars g 2.88–3.70 [10,11]
Total dietary fiber g 0.4–0.8 [10,11]
Minerals
Potassium mg 89–236 [10,11]
Sodium mg 3.3–300 [10,11,13]
Calcium mg 8–19 [10,11,13]
Magnesium mg 3–11 [10,11,13]
Phosphorus mg 11–18 [10,11]
Iron mg 0.15–0.9 [10,11,13]
Zinc mg 0.01–0.15 [10,11,13]
Cupper mg 0.05–0.2 [13]
Manganese mg 0.003–0.07 [13]
Vitamins
Vitamin A (RAE) µg 90–2058 [10,11]
Vitamin C mg 10–59.4 [10,11,15,17]
Vitamin E (ATE) mg 0.32–5.56 [10,11,22]
Total folate (DFE) µg 31.61–26.87 [19]
Abbreviations: RAE, retinol activity equivalents; ATE, alpha-tocopherol equivalents; DFE, dietary
folate equivalents.
18 mg/100 mL for p hosphorus; whereas the contents of iron, zinc, copper, and manganese are below
1 mg/100 mL (Table 48.1) [10,11,13].
Concerning the vitamin content, vitamin A content ranges from 90 to 2058 µg/100 mL, and it is mainly
represented by the carotenoids with vitamin A activity, such as β- and γ-carotenes [10,11]. Vitamin C
exists in the reduced ascorbic acid and oxidized dehydro-L-ascorbic acid forms, but its content is lower
than in raw tomato [15,16], since vitamin C is a thermolabile compound that could be completely elimi-
nated during industrial juice processing [17]. For this reason, some processors add ascorbic acid to tomato
juice. Hence in fortified tomato juice, this vitamin could reach value up to 60 mg/100 mL [10,11,15,17],
close to the Recommended Dietary Allowances (RDA) [18]. Despite the addition of ascorbic acid, the
storage conditions, time, and temperature have an impact on the stability of this vitamin reaching losses
around 50% after 12 months of storage at room temperature [17]. The content of vitamin E ranges
from 0.32 to 5.56 µg/100 mL [10,11], lower than tomato fruit since this vitamin is not extracted with
the mechanical extraction process, which removes the tomato seeds during the sieving step. Regarding
B-group vitamins, folate is present at various plant foods, including tomatoes and products derived from
them, as tetrahydrofolate (H4-folate) and 5-methyltetrahydrofolate (5-CH3-H4-folate) [16,19]. In tomato
juice, the two folate vitamins have been observed, with 5-CH3-H4-folate being the predominant natural
form, with content level of around 30 µg/100 g of fresh weight (FW), whereas the H4-folate content
represents less than 10% of the total folate content [19] (Table 48.1). Concerning the stability of folate
in tomato juice during 1 year commercial shelf life, Iniesta et al. [19] have reported final losses near to
50% and 80% in tetrapacks and glass-bottled tomato juices, respectively. Folate degradation was not
temperature dependent, and these data reveal that folate is the most labile bioactive in tomatoes and that
light exposure enhances folate degradation.

The nutritional interest in tomatoes and the products derived from them is mainly focused on carotenoids
and phenolics, which contribute to the tomato’s antioxidant capacity and beneficial effects on human
health [20]. Though these compounds are mainly concentrated in the tomato peel, the mechanical pro-
cess applied during industrial processing extracts a high proportion of them, and due to loss of water
during thermal treatment, there are important quantities of them in tomato juices [21,22]. In addition,
glycoalkaloids dehydrotomatine, α-tomatine, and esculoside have been reported to elicit several benefi-
cial effects in both in vivo and in vitro studies [23]. However, the content of these bioactives in the tomato
juice and their bioactivity can vary according to several factors, such as cultivar, agronomical conditions,
ripening stage, storage conditions, postharvest treatment, and industrial processing conditions.
48.3.1 Lycopene
Lycopene is the most predominant carotenoid of the tomato fruit and tomato products representing
around 60%–65% of total carotenoids, and it is responsible for the red color. Other carotenoids, β-, α-,
and γ-carotenes, phytoene, phytofluene, lutein, and xanthophyll are also found in lower concentrations
(Figure 48.1) [24]. The content of lycopene and other carotenoids in tomato juice is mainly determined
by the content of these compounds in the fresh tomatoes used as raw materials in the industry. In general,
tomato juice shows a higher content of lycopene than tomatoes for fresh or salad consumption, since tomato
cultivars for industrial processing have higher amounts of natural colorants [21,25]. There are many data in
the scientific literature about the content of carotenoids in tomato juice and as affected by industrial process-
ing [5,7,21,22,26]. However, much of them are related to tomato juices obtained experimentally in pilot plant
and not under industrial conditions, hence these data do not provide any real information about the content
of these compounds in commercially available tomato juices. The content of lycopene and other carotenoids
in commercial tomato juices is shown in Table 48.2. As can be seen, there is a great variability in the lyco-
pene isomers and β-carotene concentration according to the kind of juice, which is mainly due to the differ-
ences of raw materials, as have been mentioned earlier, since the content of these compounds increase and
are relatively stable during different industrial steps [5,7,22,26–29]. Moreover, lycopene content is relatively
stable during the shelf life of sterilized tomato juice, because its losses are below 20% after 12 months of
storage, regardless of the storage temperature or the packaging material used [17]. Therefore, in minimally
processed tomato juice, higher losses of lycopene could be observed due to the residual activity of lipoxy-
genase and hydroperoxide lyase enzymes [29,30]. In general, tomato juice can be considered as a dietary
source of lycopene, because it is more available in processed tomato products. However, whereas in tomato
juice, lycopene is present in its natural (all-E) isomeric, in the human body (serum and tissues), lycopene is
found mainly as (Z) isomers, accounting for more than 50% of total lycopene present [6,31].
48.3.2 Phenolic Compounds

The content of phenolic compounds in tomatoes varies widely according to the cultivars; hence, the pres-
ence of these bioactives in tomato juice is also variable. In addition, the contribution of tomato juice to the
daily intake of these bioactives is not relevant as there are only few data available in the scientific litera-
ture, compared to other fruit juices. The total phenolics content in tomatoes range from 16 to 50 mg gallic
acid equivalents (GAE)/100 g of FW, whereas total flavonoids represent under 10 mg catechin equivalents
(CE)/100 g [20,25]. Related to the individual compounds, hydroxycinnamic acid such as chlorogenic acid
and its derivatives, rutin, and kaempferol are the main phenolic compounds of fresh tomatoes and products
derived from them [25,32,33] (Figure 48.2). In addition, small quantities of naringenin can be detected,
mainly in the tomato peel, and can be obtained during the processing of tomato juice [34]. Regarding the
content of total phenolics in tomato juice measured by Folin–Ciocalteu’s test, the values ranged from 10 to
30 mg GAE/100 g [28,32,33,35,36], whereas total flavonoids was lower, with values around 9.8–11.0 mg
CE/100 g [17,28]. Despite variability of raw material, the different technological processes applied dur-
ing tomato juice manufacture modify the content of phenolic compounds [11,32,33,37] (Table 48.3).
Tomato Juice 597
Lycopene
β-Carotene
OH
HO
Lutein
Phytoene
Phytofluene
FIGURE 48.1 Chemical structure of common carotenoids found in tomato juice.
TABLE 48.2
Content of Lycopene Isomers and β-Carotene (mg/L) in Different Tomato Juices
Tomato Juices E-Lycopene Z-Lycopene β-Carotene References
Commercial traditional pasteurized 21.29–34.35 na 1.82–2.65 [15]
Commercial glass bottled 106–114 4–7 na [17]
Commercial tetrapacks 119–106 5–11 na [17]
High-pressure homogenized 100–145 8–16 4.5–6.7 [28]
Minimally processed 23.18–24.44 1.94–2.78 na [29]
HO CO2H
O
O O
HO O H3CO
OH OH OH
OH
HO OH HO
p-Coumaric acid Chlorogenic acid Ferulic acid
OH
HO O
OH OH
HO OH
O OH
O
O
OH O HO O
H3C O
HO
HO
OH
OH O
Rutin Naringenin
OH
HO O
OH
OH O
Kaempferol
FIGURE 48.2 Chemical structures of common polyphenolic compounds found in tomato juice.
TABLE 48.3
Content of Phenolic Compounds in Tomato Juices (per 100 g)
Phenolics Unit Tomato Juice References
Total Phenolics mg GAE 24–30 [17,28,32]
Total Flavonoids mg CE 9.8–11 [17,28]
Hydroxycinnamic Acid
Chlorogenic acid mg 2.19–4.44 [32,33,47]
Ferulic acid mg 0.5–0.9 [32,33,47]
p-Coumaric mg 0.30–0.64 [32,33,47]
Flavones
Apigenin mg 0.01 [11]
Luteolin mg 0.02 [11]
Flavanols
Kaempferol mg 0.01–0.55 [11,32,33,37]
Myricetin mg 0.03 [11]
Quercetin mg 0.50–2.8 [11,32,33,37]
Rutin mg 2.12 [47]
Flavanones
Naringenin mg 0.64 [33,47]
Abbreviations: GAE, gallic acid equivalents; CE, catechin equivalents.
Tomato Juice 599
H
N
O
HO OH
O
O O
O O
HO
O
HO HOHO
HO OH
Tomatine
FIGURE 48.3 Chemical structure of α-tomatine.
As mentioned for carotenoids, phenolic compounds remain stable after processing and during stor-
age of the tomato juice, preserving the nutritional quality in terms of their content of bioactive
compounds [17,28,32,33].
48.3.3 Tomatine
The glycoalkaloid referred to as tomatine consist of a mixture of α-tomatine (Figure 48.3) and dehy-
drotomatine. Contrary to lycopene, whereas tomato fruit ripens and change color, the increase of this
carotenoid is accompanied by a concurrent decrease in the concentration of tomatine. Green tomatoes,
therefore, have a high content of tomatine, as high as 500 mg/kg FW, whereas the highest content in
red tomatoes is only about 5 mg/kg [23]. Tomatine is relatively stable to different thermal processing.
In tomato juice, the mean content of 0.27 mg/100 g, hence a serving size of 200 mL, provides 0.54 mg
of this alkaloid [38]. A recent review has analyzed the beneficial effect of tomato products with special
attention to the beneficial effect of lycopene and tomatine. Although tomatine has been shown to exert
an anticarcinogenic effect in in vitro cell and in vivo studies through different mechanisms [23], its rel-
evance as bioactive compounds in tomato juice must be considered as minimum, since full ripe tomatoes
are used as raw material during the juice manufacture.

Antioxidants of tomato juice (mainly carotenoids and phenolic compounds), together with antioxidant
vitamins, can prevent the damage to macromolecules (carbohydrate, protein, lipid, and DNA) caused by
free radicals, mechanisms involved in carcinogenesis, aging, and the development of CVD. It is recog-
nized that the daily ingestion of tomato juice and tomato puree results in a significant increase of the
total antioxidant activity in plasma [31,39], which has been described as being efficient in ameliorating
the biomarkers of oxidative stress in lipids and DNA [31,39,40] and the prevention of several pathologies.
Therefore, the total antioxidant activity of tomato juice is due to the synergistic activity of natural lipo-
philic and hydrophilic antioxidants. However, analytical methods for antioxidant activity analysis gener-
ally determine only the bioactivity of the hydrophilic compounds, but not of the lipophilic ones, such as
lycopene. Although antioxidant activity of tomato juice has been widely reported in the literature, due to
the high mechanistic variability in the in vitro chemical methods, different results and values have been
published [15,17,26,28,33,35,36].
Phenolic compounds represent the main hydrophilic antioxidants of tomato juice in the absence of
vitamin C [16,17,41]. Different authors have reported an increase in the antioxidant activity after ther-
mal treatment of tomato purée destined to prepare tomato juice [28,35,36], which have been related to a
greater release of phenolic and flavonoid compounds from the cell matrix during processing. In addition,
it has been suggested that if heating time during processing is prolonged, the loss of naturally occurring
antioxidants can be minimized by a recovery or even an enhancement of the antioxidant activity due to
the formation of advanced Maillard reaction products, which may even induce an increase in the antioxi-
dant capacity [42]. On the other hand, lycopene is responsible for lipophilic antioxidant activity in tomato
juice, but its contribution to total antioxidant activity is small [17]. Therefore, it is reasonable to assume
that other antioxidants, most likely lipophilic phenolic compounds, may contribute to maintaining the
lipophilic antioxidant activity of the tomato juice during storage [15,17].
48.4 Health Effects

The beneficial effects on human health associated with the consumption of tomato juice are mainly
related to the high content of lycopene and its antioxidant properties, but other bioactive compounds
of tomato juice, such as vitamin C, phenolic compounds, and folate, can exhibit a synergistic effect in
quenching free radicals.
48.4.1 Prevention of Cardiovascular Disease

The systematic review of Mordente et al. [43] evaluated the relationship between tomato intake and
CVD. Although several epidemiological studies have investigated this association, controversial results
have been obtained. Since CVD is a multifactorial disease, a picture has emerged concerning the possi-
ble role of the intake of tomato products in the progression of CVD risk factors. This beneficial effect on
human health is related to various mechanisms, such as a reduction in plasmatic cholesterol, inhibition of
low-density lipoprotein (LDL) oxidation and lipid peroxidation, and decreasing platelet activation. The
regulation of all the cited parameters could, therefore, represent an important tool for the prevention of
CVD [31,43–49].
The intake of tomato juice for a minimum of 2 or 3 weeks, providing more than 20 mg of lycopene/
day, increased the level of lycopene in plasma, resulting in a reduction in both total and LDL choles-
terols [31,44]. Lycopene has been shown to act as a hypocholesterolemic agent by inhibiting 3-hydroxy-
3-methylglutaryl-coenzyme A (HMG-CoA) reductase, the rate-limiting enzyme in cholesterol synthesis.
This effect has been described by various authors in macrophage cell lines, showing a similar activity to
that of fluvastatin [45,46]. Recently, this effect has been evaluated in vivo in an intervention study with
Sprague Dawley rats, which were given tomato juice for 5 weeks. After this period, lycopene was found
to have accumulated in the liver, and inhibition of the HMG-CoA reductase activity was observed, also
decreasing total cholesterol in the plasma [47]. In addition, computational molecular modeling showed
that lycopene can bind to the active site of the enzyme and compete with the enzyme ligand HMG-CoA,
inhibiting mevalonate formation and consequently reducing cholesterol synthesis, acting similarly to the
statin cerivastatin [48].
After absorption, lycopene binds inside the hydrophobic core of lipoprotein particles and distributed
into very-low-density lipoprotein (VLDL) and LDL, decreasing LDL oxidation. Since an increase in
LDL oxidation is thought to be causally associated with the risk of atherosclerosis and coronary heart
disease (CHD), the reduction of LDL and lipid oxidation biomarkers is considered a factor to mini-
mize CVD risk. The consumption of tomato juice reduces the lipid oxidation biomarkers of plasma and
urine, measured as thiobarbituric acid–reactive substances (TBARS) [43]. In general, the consumption
of tomato juice reduces the oxidative stress biomarkers of macromolecules [26], an effect that could also
be enhanced by synergism of different antioxidants present in tomato and tomato products [44,48]. The
in vivo antithrombotic effect of lycopene has been described by O’Kennedy et al. [49] after the intake
of a single dose of tomato juice. Reduced platelet activation and aggregation is due to the inhibition of
glycoprotein IIb/IIIa and platelet secretory mechanisms.
48.4.2 Prevention of Cancer

The intake of tomato and processed tomato products has been found to inhibit the proliferation of several
types of cancer cells, mainly related to their lycopene. Several in vitro and in vivo studies have demon-
strated the positive effect of this bioactive in the prevention of prostate, lung, colon, and breast cancers [50].
Tomato Juice 601
However, different epidemiologic studies have produced inconsistent results, since in clinical trials, it is
difficult to control for disease stage, genetic background, diet, age, and other cancer risk factors.
Whether lycopene is the only compound responsible for this effect remains an open question, but it exhib-
its several biochemical mechanisms for cancer-prevention activity. The most frequently studied mecha-
nisms have been the antioxidant and the free radical scavenging activity, which reduce oxidative damage
to macromolecules, particularly to DNA. The intake of tomato juice during 5 weeks providing 15 mg of
lycopene per day significantly reduced the serum levels of 8-oxo-d-guanosine after an extensive physical
exercise, indicating a protective effect against reactive oxygen species DNA damage [51]. Moreover, the
addition of tomato juice to prostate carcinoma PC-3 cell could induce the breakage of DNA, inhibiting the
proliferation of this cell line [52]. Related to the colon cancer, it has been described in rats that the intake of
tomato juice prevents N-methylnitrosourea-induced carcinogenesis [53]. However, in an intervention trial
with humans, the intake of tomato juice during 2 weeks led only to minor changes in the luminal biomark-
ers relevant to colon carcinogenesis [54]. Despite of different studies have described an inverse relationship
between the intake of tomato juice and some biomarkers related to different cancer, no significant effect has
been found between the carotenoids plasma concentration and the risk of cancer [55,56].
48.4.3 Nonalcoholic Fatty Liver Disease

Nonalcoholic fatty liver disease (NAFLD), which is the most common cause of liver dysfunction, char-
acterized by fatty infiltration in the absence of an excess of alcohol consumption or any other liver
disease, is the most common liver disorder in western countries. In addition, it is currently considered
the hepatic manifestation of metabolic syndromes and other related metabolic derangements, such as
obesity, insulin resistance, hypertension, and dyslipidemia, being the major contributors to CVD and
overall obesity-related morbidity and mortality [57]. Recently, the effect of the consumption of tomato in
the dietary treatment of rats with induced NAFLD has been studied, showing that tomato juice admin-
istration partially modify the hepatocyte’s metabolic pattern from a high-fat diet to a normal diet, and
it even improved the redox balance of the liver [58]. This effect could be related to the accumulation of
lycopene in the liver, since dietary supplementation with 0.05% pure lycopene can regulate the hepatic
lipid metabolism in mice fed with a high-fat diet through the upregulation of microRNA-21, which mark-
edly blocked intracellular lipid accumulation by blocking the expression of fatty acid binding protein-7
(FABP7) [59]. Moreover, the consumption of tomato extract and lycopene can inhibit the progression of
NAFLD to nonalcoholic steatohepatitis-promoted hepatocarcinogenesis, mainly as a result of reduced
oxidative stress in the liver, which can be fulfilled through various mechanisms [60].
48.4.4 Reduction of Oxidative Stress and Inflammation Biomarkers

The effect of intake of tomato juice in the amelioration of different biomarkers of oxidative stress and
inflammation has been studied in several human intervention studies. In two intervention trials, the syner-
gistic activity of lycopene and vitamin C or n-3 fatty acids and the effect on some biomarkers of oxidative
stress and inflammation related to CVD have been studied. Volunteers were randomly assigned to ingest
tomato juice either with or without vitamin C fortification for 2 weeks (500 mL of tomato juice, with a
daily dose of 20.6 mg of lycopene, and 45.5 versus 435 mg of vitamin C). A reduction in total cholesterol
was correlated with lycopene intake, showing that tomato juice fortified with vitamin C slightly raised
the antioxidant capacity in urine and decreased lipid oxidation in plasma and urine, some risk factors
associated with CVD [31]. In a 2-week intervention study with healthy women, the daily intake of plain
and polyunsaturated fatty acids (PUFA)-enriched tomato juice was compared with respect to different
biomarkers also related to CVD. Each serving (500 mL) of both tomato juices provided natural phenolic
compounds (181 mg/day) and lycopene (26.5 mg/day), and the enriched juice with 250 mg of eicosa-
pentaenoic acid + docosahexaenoic acid. Although lipid profiles remained unchanged, a decrease in the
serum levels of homocysteine after the intake of n-3 PUFA-enriched juice was observed. A decrease in
vascular adhesion molecule 1 (VCAM-1) levels was also noted after the intake of both juices, whereas
intracellular adhesion molecule 1 (ICAM-1) only decreased following the intake of enriched juice, which
suggests a possible synergistic action between n-3-PUFA and tomato antioxidants [40] (Figure 48.4).
50
40
30
FRAP relative changes (%)

20
10
0
–10
–20
–30
–40
–50
(a)
50
40
HOMOCYSTEINE relative changes (%)
30
20
10
0
–10
–20
–30 *
–40
–50
(b)
50
40
30
VCAM-1relative changes (%)
20
10
0
–10
–20
–30
–40
–50
(c)
FIGURE 48.4 Relative percent changes in serum ferric-reducing antioxidant power (FRAP) (a), homocysteine (b), vascular
adhesion molecule 1 (VCAM-1) (c). (Adapted from García-Alonso, F.J. et al., Eur. J. Nutr., 5, 415, 2012. With permission.)
Note: Reference juice, plain tomato juice with lycopene (26.5 mg/day), and n-3 PUFA-enriched tomato juice (26.5 mg/day
with 250 mg of eicosapentaenoic plus docosahexaenoic acids). (Continued)
Tomato Juice 603
50
40
ICAM-1 relative changes (%)

30
20
10
0
–10
–20
–30
–40
–50
(d)
FIGURE 48.4 (Continued) Intracellular adhesion molecule 1 (ICAM-1) (d) concentrations in the study groups after plain
(black bars) and n-3 polyunsaturated fatty acids (PUFA)-enriched (white bars) tomato juice consumption. *Significantly
different compared with reference juice group (P < 0.01). (Adapted from García-Alonso, F.J. et al., Eur. J. Nutr., 5, 415,
2012. With permission.) Note: Reference juice, plain tomato juice with lycopene (26.5 mg/day), and n-3 P UFA-enriched
tomato juice (26.5 mg/day with 250 mg of eicosapentaenoic plus docosahexaenoic acids).

Novel products based on tomato juice can be developed with the aim of improving the bioactive
compounds and to enhance its bioactivity, as a result of the synergistic action between them. First,
considering the variability in the bioactive compounds of tomatoes, industrial processors should know
the chemical characteristics of the main tomato varieties to obtain a tomato juice with good bioactive
profiles, providing beneficial effects for human health [16,20,25]. Today, new strategies and technolo-
gies applied in agriculture allow for the development and growth of new tomato cultivars with higher
levels of these bioactive compounds. In addition, the postharvest treatments, such as the exposure to
ultraviolet-C (UV-C) light, improved the bioactive compounds of tomatoes, and hence it could be applied
as a t echnological process before tomato juice processing to obtain novel, value-added products [61–63].
Despite raw material selection and treatment, to design new tomato juice, other strategies can be applied
in giving special attention to the industrial processing and the addition of different ingredients.
48.5.1 Industrial Processing

The optimization of industrial processing and the different industrial steps can improve the extraction
of bioactive compounds from the cell organelles, yielding a higher concentration of these functional
compounds in tomato juice. Authorized treatments for obtaining fruit juice are mentioned in Directive
2012/12/EU [3]. In general, mechanical extraction processes such as the sieving, grinding, and milling
of the edible parts of whole or peeled tomato fruits are used and can be combined with the application of
physical processes, including in-line water extraction of the edible part of the tomato. One-step and two-
step homogenization processes for raw and hot break tomato purée before the thermal treatment of tomato
juice have been experimentally applied to evaluate the effect on the lycopene, β-carotene, total phenolic
compounds, vitamin C, and folate content. After applying the different pressure conditions, only an
increase in the extractability of methyltetrahydrofolate (5-CH3-H4-folate) content was observed, when the
homogenization was combined with the thermal treatment [28]. Other authorized treatments during fruit
juice manufacturing include the addition of enzymatic preparations, such as pectinases (to break down
pectin), cellulases and hemicellulases (to break down cellulose and hemicellulose), proteinases (to break
down proteins), and amylases (to break down starch). Their application, mixed or alone, has a significant
effect on the extraction of bioactive compounds, mainly due to the breakdown of complex carbohydrates
from the cell walls, liberating the phenolic compounds associated with the components of dietary fiber
[12]. Experimentally, the application of different commercially available enzymatic preparations (with
pectinases, hemicellulases, cellulases, and arabinases) can improve the extraction of total phenolics and
the aglycone quercetin during the manufacturing of a cold tomato soup, namely gazpacho [64].
48.5.2 Added Ingredients

A wide range of nutrients and other ingredients may be added in tomato juice during manufacturing,
including vitamins, minerals, amino acids, essential fatty acids, fiber, various plants, and herbal extracts.
New tomato juice products may be designed by the addition of different authorized ingredients and nutri-
ents, beyond salt and herbal extracts. One strategy that can be followed with the aim of improving the
nutritional quality and bioactivity of the tomato juice is to increase the vitamins that are found in tomato
juice in low levels, such as vitamin A (increasing the β-carotene content), vitamin C (which can be added
in the different authorized chemical forms such as ascorbic acid or different ascorbate salts), and folate
or vitamin B9, which can be added as pteroylmonoglutamic acid. These vitamins, once incorporated,
improve the content and also the bioactivity of the natural compounds of the juice, but other micronutri-
ents could also be added according to the law [65].
Ingredients other than vitamin and minerals, which are ingested in a low proportion in the diet and are
beneficial for human health, can be added to the juice as they are generally recognized as safe, such as
n-3 PUFA and dietary fiber. When adding n-3 PUFA to tomato juice, one of the most important problems
is the stabilization and the odor of this ingredient, hence an appropriate preparation should be selected to
obtain an acceptable product for consumers. The addition of tomato peel, which is a by-product derived
from the tomato industry, is considered a good strategy to increase the dietary fiber content in tomato
juice, since more than 85% is dietary fiber and, at the same time, its addition can even improve the natu-
ral lycopene and phenolic compounds of the juice [12]. In addition, the reutilization of tomato peel as
a source of dietary fiber to increase the content of fiber in tomato products and other foods is a way to
improve the added value use of tomato pomace and to reduce volume of the tomato waste.
In general, the enrichment of the tomato juice with all the aforementioned ingredients can improve
the bioactivity and functionality, because these compounds can act synergistically with lycopene and
phenolics, improving the beneficial effect associated with tomato juice intake.
48.6 Conclusion
Tomato juice can be considered as a natural source of bioactive compounds, especially lycopene.
In general, all bioactive compounds remain more or less stable after tomato juice processing and during
the commercial shelf life. The presence of different phytochemical compounds and small amounts of
some vitamins with antioxidant activity results in a vegetable juice that can exert a wide range of benefi-
cial effects in the prevention of diet-related chronic diseases. In addition, low density of macronutrients
and caloric value indicate that this juice could be consumed daily without increasing caloric intake,
which is especially recommended for persons subjected to a low-calorie diet. However, it is very impor-
tant to diversify traditional tomato juice and design new products that are more attractive to consumers,
as well to provide information about the chemical properties of this juice and its beneficial effects on
human health, with the aim of promoting their consumption in a healthy diet.
REFERENCES
1. World Producer Tomato Council (WPTC), World Production Estimate of Tomatoes for Processing,
2013. Published online at: http://www.wptc.to (accessed March 30, 2014).
2. Guerrero-Acosta, J., Perspectivas de futuro del tomate de industria. Vida Rural, 15, 10–14, 2008.
Tomato Juice 605
3. European Community (EC), Directive 2012/12/EU of the European Parliament and of the Council of
19 April 2012 amending Council Directive 2001/112/EC relating to fruit juices and certain similar
products intended for human consumption. Off. J. Eur. Union, 24/04/2012, L115/1, 2012.
4. Martínez-Álvarez, J.R. and Villariño Marín, A., Zumos de frutas y verduras, in Frutas, Verduras y
Salud, Aranceta-Bartrina, J. and Pérez-Rodrigo, C., Eds., Masson S.A. & Elsevier Group, Barcelona,
Spain, 2006, pp. 85–98 (in Spanish).
5. Gupta, R., Balasubramaniam, V.M., Schwartz, S.J., and Francis, D.M., Storage stability in tomato juice
subjected to combined pressure-heat treatments. J. Agric. Food Chem., 58, 8305–8313, 2010.
6. Jacob, K., García-Alonso, F.J., Ros, G., and Periago, M.J., Stability of carotenoids, phenolic compounds,
ascorbic acid and antioxidant capacity of tomatoes during thermal processing. Arch. Latinoam. Nutr.,
60, 192–198, 2010.
7. Gupta, R., Kopec, R.E., Schawartz, S.J., and Balasubramaniam, V.M., Combined pressure-temperature
effects on carotenoids retention and bioaccessibility in tomato juice. J. Agric. Food Chem., 59,
7808–7817, 2011.
8. Frölich, K., Kaufmann, K., Bitsch, R., and Böhm, V., Effects of ingestion of tomatoes, tomato juice and
tomato purée on contents of lycopene isomers, tocopherols and ascorbic acid in human plasma as well
as on lycopene isomer pattern. Br. J. Nutr., 95, 734–741, 2006.
9. Codex Alimentarius, Codex general standard for fruit juices and nectars (CODEX STAN 247–2005),
2005. Published online at: http://www.codexalimentarius.org/inputs/download/standards/ (accessed
March 25, 2014).
10. Agencia Española de Seguridad Alimentaria y Nutrición, Base Española de Composición de Alimentos
(BEDCA), 2009, Published online at: http://www.bedca.net (accessed April 28, 2014).
11. U.S. Department of Agriculture (USDA), USDA national nutrient database for standard reference,
Release 26, 2013. Published online at: http://ndb.nal.usda.gov/ (accessed March 29, 2014).
12. Navarro-Gónzalez, I., García-Valverde, V., García-Alonso, J., and Periago, M.J., Chemical profile, func-
tional and antioxidant properties of tomato peel fiber. Food Res. Int., 44, 1528–1535, 2011.
13. Cámara, M., Pérez, M.L., López, M., Martí, N., Saura, N.D., and Micol, V., Nutrición y salud, in El
Libro del Zumo, Agrícola Española, S.A., Ed., Asociación Española de Fabricantes de Zumos, Madrid,
Spain, 2011, pp. 117–137 (in Spanish).
14. Willcox, J.K., Catignani, G.L., and Lazarus, S., Tomatoes and cardiovascular health. CRC Crit. Rev.
Food Sci. Nutr., 43, 1–18, 2003.
15. Sánchez-Moreno, C., Plaza, L., de Ancos, B., and Cano, M.P., Nutritional characterization of com-
mercial traditional pasteurized tomato juices: Carotenoids, vitamin C and radical-scavenging capacity.
Food Chem., 98, 749–756, 2006.
16. Periago, M.J., García-Alonso, J., Jakob, K., Olivares, A.B., Bernal, M.J., Iniesta, M.D., Martínez, C.,
and Ros, G., Bioactive compounds, folates and antioxidant properties of tomatoes (Lycopersicum escu-
lentum) during vine ripening. Int. J. Food Sci. Nutr., 60, 694–708, 2009.
17. Garcia-Alonso, J., Bravo, S., Casas, J., Pérez-Conesa, D., Jacob, K., and Periago, M.J., Changes in anti-
oxidant compounds during the shelf-life of commercial tomato juices in different packaging materials.
J. Agric. Food Chem., 57, 6815–6822, 2009.
18. U.S. Department of Health and Human Services, National Institute of Health (NIH), 2013. Published
online at: http://ods.od.nih.gov/factsheets/VitaminC-HealthProfessional/ (accessed April 4, 2014).
19. Iniesta, M.D., Pérez-Conesa, D., García-Alonso, J., Ros, G., and Periago, M.J., Folate content in tomato
(Lycopersicum esculentum). Influence of cultivar, ripeness, year of harvest and pasteurization and stor-
age temperatures. J. Agric. Food Chem., 57, 4739–4745, 2009.
20. Martínez-Valverde, I., Periago, M.J., Provan, G., and Chesson, A., Phenolic compounds, lycopene and
antioxidant activity in commercial varieties of tomato (Lycopersicum esculentum). J. Sci. Food Agric.,
82, 323–330, 2002.
21. Abushita, A.A., Dado, H.G., and Biacs, P.A., Change in carotenoids and antioxidant vitamins in tomato
as a function of varietal and technological factors. J. Food Chem., 48, 2075–2081, 2000.
22. Seybold, C., Frölich, K., Bitsch, R., Otto, K., and Böhm, V., Changes in contents of carotenoids and
vitamin E during tomato processing. J. Agric. Food Chem., 52, 7005–7010, 2004.
23. Friedman, M., Anticarcinogenic, cardioprotective and other health benefits of tomato compounds:
Lycopene, α-tomatine, and tomatidine in pure form and in fresh and processed tomatoes. J. Agric. Food
Chem., 61, 9534–9550, 2013.
24. Periago, M.J., Martínez-Valverde, I., Ros, G., Martínez-Graciá, C., and López-Martínez, G., Propiedades
químicas, biológicas y valor nutritivo del licopeno. Revista Anales de Veterinaria, 17, 51–66, 2001.
25. García-Valverde, V., Navarro-González, I., García-Alonso, J., and Periago, M.J., Antioxidant bioactive
compounds in selected industrial processing and fresh consumption tomato cultivars. Food Bioprocess
Technol., 6, 391–402, 2013.
26. Hsu, K.C., Evaluation of processing qualities of tomato juice induced by thermal and pressure processing.
Lebensm. Wisse. Technol., 41, 450–459, 2008.
27. Lin, C.H. and Chen, B.H., Stability of carotenoids in tomato juice during storage. Food Chem., 90,
837–846, 2005.
28. Pérez-Conesa, D., García-Alonso, J., García-Valverde, V., Iniesta, M.D., Jacob, K., Sánchez-Siles,
L.M., Ros, G., and Periago, M.J., Changes in bioactive compounds and antioxidant activity during
homogenization and thermal processing of tomato puree. Innov. Food Sci. Emerg. Technol., 10,
179–188, 2009.
29. Bravo, S., Optimización del Contenido y Disponibilidad del Licopeno y otros Compuestos Bioactivos
en Tomates y Productos Elaborados con Tomate (in Spanish), PhD thesis, Universidad de Murcia,
Murcia, Spain, 2013.
30. Odriozola-Serrano, I., Soliva-Fortuny, R., and Martin-Belloso, O., Changes of health-related compounds
throughout cold storage of tomato juice stabilized by thermal or high intensity pulsed electric field treat-
ments. Innov. Food Sci. Emerg. Technol., 9, 272–279, 2008.
31. Jacob, K., Periago, M.J., Böhm, V., and Berruezo, G., Influence of lycopene and vitamin C from tomato
juice on biomarkers of oxidative stress and inflammation. Br. J. Nutr., 99, 137–146, 2008.
32. Odriozola-Serrano, I., Soliva-Fortuny, R., Hernñandez-Jover, T., and Martín-Belloso, O., Carotenoid
and phenolic profile of tomato juices processed by high intensity pulsed electric fields compared with
conventional thermal treatments. Food Chem., 112, 258–266, 2009.
33. Vallverdu-Queralt, A., Odriozola-Serrano, I., Oms-Oliu, G., Lamuela-Raventós, R.M., Elez- Martínez,
P., and Martín-Belloso, O., Changes in the polyphenol profile of tomato juices processed by pulsed elec-
tric fields. J. Agric. Food Chem., 60, 9667–9672, 2012.
34. Tomás-Barberán, F.A. and Clifford, M.N., Flavanones, chalcones and dihydrochalcones-nature, occur-
rence and dietary burden. J. Sci. Food. Agric., 80, 1073–1080, 2000.
35. Dewanto, V., Wu, X., Adom, K.K., and Liu, R.H., Thermal processing enhances the nutritional value of
tomatoes by increasing total antioxidant activity. J. Agric. Food Chem., 50, 3010–3014, 2002.
36. Gahler, S., Otto, K., and Böhm, V., Alterations of vitamin C, total phenolics, and antioxidant capac-
ity as affected by processing tomatoes to different products. J. Agric. Food Chem., 51, 7962–7968,
2003.
37. Stewart, A.J., Bozonnet, S., Mullen, W., Jenkins, G.I., Lean, M.E.J., and Crozier, A., Occurrence of
flavonols in tomato and tomato based products. J. Agric. Food Chem., 48, 2663–2669, 2000.
38. Friedman, M. and Levin, C.E., α-Tomatine content in tomato and tomato products determined by HPLC
with pulsed amperometric detection. J. Agric. Food Chem., 43, 1507–1511, 1995.
39. Porrini, M. and Riso, P., Lymphocyte lycopene concentration and DNA protection from oxidative dam-
age is increased in women after a short period of tomato consumption. J. Nutr., 130, 189–192, 2000.
40. García-Alonso, F.J., Jorge-Vidal, V., Ros, G., and Periago, M.J., Effect of consumption of tomato juice
enriched with ω-3 polyunsaturated fatty acids on the lipid profile, antioxidant biomarker status, and
cardiovascular disease risk in healthy women. Eur. J. Nutr., 5, 415–424, 2012.
41. Lavelli, V., Peri, C., and Rizzolo, A., Antioxidant activity of tomato products as studied by a model reac-
tions using xanthine oxidase, myeloperoxidase and copper-induced lipid peroxidation. J. Agric. Food
Chem., 48, 1442–1448, 2000.
42. Nicoli, M.C., Anse, M., Parpinel, M.T., Franceschi, S., and Lerici, C.R., Loss and/or formation of anti-
oxidants during food processing and storage. Cancer Lett., 114, 71–74, 1997.
43. Mordente, A., Guantario, B., Meucci, E., Silvestrini, A., Lombardi, E., Martorana, G.E., Giardina, B., and
Böhm, V., Lycopene and cardiovascular diseases: An update. Curr. Med. Chem., 18, 1146–1163, 2011.
44. Silaste, M.L., Alfthan, G., Aro, A., Kesäniemil, Y.A., and Hörkkö, S., Tomato juice decreased LDL
cholesterol levels and increased LDL resistance to oxidation. Br. J. Nutr., 98, 1251–1258, 2007.
Tomato Juice 607
45. Fuhrman, B., Elis, A., and Aviram, M., Hypocholesterolemic effect of lycopene and β-carotene is
related to suppression of cholesterol synthesis and augmentation of LDL receptor activity in macro-
phages. Biochem. Biophys. Res. Commun., 233, 658–662, 1997.
46. Palozza, P., Simone, R., Catalano, A., Parrone, N., Monego, G., and Ranelletti, F.O., Lycopene regula-
tion of cholesterol synthesis and efflux in human macrophages. J. Nutr. Biochem., 22, 971–978, 2011.
47. Navarro-Gónzález, I., Pérez-Sánchez, H., Martín-Pozuelo, G., García-Alonso, J., and Periago, M.J., The
inhibitory effects of bioactive compounds of tomato juice binding to hepatic HMGCR: In vivo study and
molecular modeling. PLoS One, 9, e83968.
48. Riso, P., Visioli, F., Erba, D., Testolin, G., and Porrini, M., Lycopene and vitamin C concentrations
increase in plasma and lymphocytes after tomato intake. Effects on cellular antioxidant protection. Eur.
J. Clin. Nutr., 58, 1350–1358, 2004.
49. O`Kennedy, N., Crosbie, L., van Lieshout, M., Broom, J.I., Webb, D.J., and Duttaroy, A.K., Effects of
antiplatelet components of tomato extract on platelet function in vitro and ex vivo: A time course can-
nulation study in healthy humans. Am. J. Clin. Nutr., 84, 570–579, 2006.
50. Giovannucci, E., Tomatoes, tomato-based products, lycopene and cancer: Review of the epidemiologic
literature. J. Natl. Cancer Inst., 91, 317–331, 1999.
51. Harms-Ringdahl, M., Jenssen, D., and Haghdoost, S., Tomato juice intake suppressed serum concentra-
tion of 8-oxo-dG after extensive physical activity. Nutr. J., 2, 11–29, 2012.
52. Luo, Q., Pang, Y.Q., Li, Z.N., Yan, J., Yang, M., and Zeng, X., Study on the inhibitory effects of juice of
tomato on the growth of human prostate carcinoma PC-3 cells. Wei Sheng Yan Jiu, 34, 336–337, 2005.
53. Narisawa, T., Fukaura, Y., Hasebe, M., Nomura, S., Oshima, S., Sakamoto, H., Inakuma, T., Ishiguro,
Y., Takayasu, J., and Nishino, H., Prevention of N-methylnistrosourea-induced colon carcinogenesis in
F344 rats by lycopene and tomato juice rich in lycopene. Jpn. J. Cancer Res., 89, 1003–1008.
54. Schnäbele, K., Briviba, K., Roser, S., Pool-Zobel, B.L., and Rechkemmer, G., Effects of carrot and
tomato juice consumption on fecal markers relevant to colon carcinogenesis in humans. Br. J. Nutr., 99,
606–613, 2008.
55. Sato, R., Helzlsouer, K.J., Alberg, A.J., Hoffman, S.C., Norkus, E.P., and Comstock, G.W., Prospective
study of carotenoids, tocopherols, and retinoid concentrations and the ris of breast cancer. Cancer
Epidemiol. Biomar. Prev., 11, 451–457, 2002.
56. Key, T.J., Appleby, P.N., Allen, N.E., Travis, R.C., Roddam, A.W., Jenab, M., Egevab, L. et al., Plasma
carotenoids, retinol, and tocopherols and the risk of prostate cancer in the European Prospective
Investigation into Cancer and Nutritional study. Am. J. Clin. Nutr., 86, 672–681, 2007.
57. Tessari, P., Coracina, A., Cosma, A., and Tiengo, A., Hepatic lipid metabolism and non-alcoholic fatty
liver disease. Nutr. Metab. Cardiovasc. Dis., 19, 291–302, 2009.
58. Bernal, C., Martín-Pozuelo, G., Lozano, A.B., Sevilla, A., García-Alonso, J., Cánovas, M., and Periago,
M.J., Lipid biomarkers and metabolic effects of lycopene from tomato juice on liver of rats with induced
hepatic steatosis. J. Nutr. Biochem., 24, 1870–1881, 2013.
59. Ahn, J., Lee, H., Jung, C.H., and Ha, T., Lycopene inhibits hepatic steatosis via microRNA-21-induced
downregulation of fatty acid-binding protein 7 in mice fed a high-fat diet. Mol. Nutr. Food Res., 56,
1665–1674, 2012.
60. Wang, Y., Ausman, L.M., Greenberg, A., Russell, R.M., and Wang, X.D., Dietary lycopene and tomato
extract supplementations inhibit non-alcoholic steatohepatitis-promoted hepatocarcinogenesis in rats.
Int. J. Cancer, 126, 1788–1796, 2010.
61. Jagadeesh, S.L., Charles, M.T., Gariepy, Y., Goyette, B., Raghavan, G.S.V., and Vigneault, C., Influence
of post-harvest UV-C hormesis on the bioactive components of tomato during post-treatment handling.
Food Bioprocess Technol., 4, 1463–1472, 2009.
62. Bravo, S., García-Alonso, J., Martín-Pozuelo, G., Gómez, V., Santaella, M., Navarro-González, I., and
Periago, M.J., The influence of post-harvest UV-C hormesis on lycopene, β-carotene, and phenolic
content and antioxidant activity of breaker tomatoes. Food Res. Int., 49, 296–302, 2012.
63. Bravo, S., García-Alonso, J., Martín-Pozuelo, G., Gómez, V., García-Valverde, V., Navarro-González, I.,
and Periago, M.J., Effects of postharvest UV-C treatment on carotenoids and phenolic compounds of
vine-ripe tomatoes. Int. J. Food Sci. Technol., 48, 1744–1749, 2013.
64. Martínez-Valverde, I., Evaluación nutricional y optimización de compuestos bioactivos en gazpacho

(in Spanish), PhD thesis, Universidad de Murcia, Murcia, Spain, 2001.
65. European Community (EC), Regulation (EC) no. 1925/2006 of the European Parliament and of the
Council of 20 December 2006, on the addition of vitamins and minerals and of certain other substances
to foods. Off. J. Eur. Union, 30/12/2006, L404/26, 2006.
49
Vegetable-Containing Juices
(Carrot, Kale, and Sprout)
Daniel A. Jacobo-Velázquez, Erika Ortega-Hernández, and Luis Cisneros-Zevallos
CONTENTS
49.1 Introduction................................................................................................................................... 609
49.4 Health Effects.................................................................................................................................612
49.4.1 Carrot Juice.......................................................................................................................617
49.4.2 Kale Juice..........................................................................................................................618
49.4.3 Sprout Juices (Wheat and Broccoli)..................................................................................621
49.5 Vegetable-Containing Juices: Relationship between Chronic Disease, Food, and Medicine...... 622
49.7 Conclusion..................................................................................................................................... 623
References............................................................................................................................................... 624
49.1 Introduction
The consumption of fresh fruits and vegetables has been associated with the prevention of different
chronic and degenerative diseases due to their high levels of antioxidants and other bioactive compounds.
Vegetable-containing juices possess most of the beneficial properties of fresh vegetables.
Among the different vegetables, carrot (Daucus carota) is one of the most common raw materials
utilized for the production of juices, and it contains high levels of pro–vitamin A carotenoids, vitamins,
and minerals [1–3]. Likewise, kale (Brassica oleracea var. sabellica) is also a common ingredient in veg-
etable juices, and it is an excellent source of glucosinolates and carotenoids, which are associated with
the prevention of cancer and eye diseases [4,5]. Similarly, sprouts, specifically broccoli (B. oleracea var.
italica) sprout, are utilized as an ingredient in vegetable juices. This represents an effective approach to
increase the intake of antioxidant, since bioactive compounds in sprouts are more concentrated than in
mature plants, and in the specific case of broccoli sprouts, it contains 20–50 times more phase II detoxi-
fication enzyme inducers than the mature plants [6–8].
This chapter reviews the nutritional characteristics of carrot, kale, and sprout juices. Likewise,
the nutritional characteristics of vegetable-containing juices as well as the main bioactive present in
their composition are described. The core part of the chapter reviews the health benefits of vegetable-
containing juices based on the results from human intervention studies. At the end, a diagram explaining
the relationship between vegetable juice consumption and their potential use as a preventive or interven-
tion/therapeutic treatment is shown.
609
TABLE 49.1
Compositional and Nutritional Characteristics of Vegetable-Containing Juices (per 100 g)
Carrot Juice Kale Juice Broccoli Sprout Juice Wheat Sprout
Nutrient Unit [1] [4,33,34] [1,6–8] Juice [1]
Water g 88.87 94.80 90.00 99.00
Protein g 0.95 1.39 3.19 1
Lipid (fat) g 0.15 0.3 0.09 0
Carbohydrate g 9.28 na 3.44 0
Total sugars g 3.91 na na na
Total dietary fiber g 0.8 1.03 0.90 0
Minerals
Calcium mg 24 146 0.89 28
Iron mg 0.46 0.32 na 2.14
Magnesium mg 14 23 0.47 82
Phosphorus mg 42 30 0.37 200
Potassium mg 292 289 0.48 169
Sodium mg 66 74.5 0.75 16
Vitamins
Folate (DFE) mg 0.004 0.52 — 0.04
Riboflavin mg 0.06 0.15 na 0.16
Thiamin mg 0.09 0.3 na 0.23
Vitamin A IU 19.12 580 na na
Vitamin C mg 8.5 184 na 2.6
Vitamin E (ATE) mg 1.16 0.4 na na
Carotenoids
α-Carotene mg 4342 na na na
β-Carotene mg 9303 2004 na na
Abbreviations: DFE, dietary folate equivalents; ATE, alpha-tocopherol equivalents; na, not available.

The compositional and nutritional characteristics of carrot, kale, and sprout (broccoli and wheat) juices
are summarized in Table 49.1. Carrot juice is one of the most popular vegetable juices and represents a
rich source of vitamins and minerals. Vitamin A is the most abundant compound in carrot. One serv-
ing size of carrot juice (236 g) can provide up to 250% of Recommended Dietary Allowances (RDA) of
vitamin A (as pro–vitamin A carotenoids), 33% of vitamin C, 40% of vitamin K, 20.70% of vitamin E,
14.14% of phosphorous, 14.65% of potassium, and less than 10% of the content of protein, fiber, calcium,
magnesium, iron, zinc, thiamin, and riboflavin [1,2]. Likewise, β-carotene represents the major portion
(60%–80%) of carotenoids in carrot juice, followed by α-carotene (10%–40%) and lutein (1%–5%) [3].
Detailed information about carrot juice is also provided in Chapter 46.
Kale juice is prepared from a plant akin to cabbage of high consumption in Japan [4]. Kale is a leafy
green vegetable belonging to the Brassicaceae family. In recent years, it has gained the attention of the
scientific community due to its high content of bioactive compounds such as vitamin C, pro–vitamin A,
glucosinolates, phenolic antioxidants, dietary fiber, micronutrients (iron, zinc, and manganese), and
macronutrients (calcium and magnesium) [5]. A serving size of kale juice (240 g) provides 306% of
RDA of vitamin C, 130% of folate, more than 100% of vitamin A, 25.5% of thiamin, 13% of riboflavin,
14.6% of calcium, and other compounds in minimal proportions.
Vegetable-Containing Juices (Carrot, Kale, and Sprout) 611
The use of sprout juices is an alternative approach for the prevention of metabolic diseases. Broccoli
sprout is of increasing interest in nutrition science as a consequence of the beneficial effects of different
phytochemicals, including glucosinolates, isothiocyanates such as sulforaphane, and phenolic acids. A cup
of broccoli sprout juice (244 g) contains around 876 mg of total polyphenols, 1.84 mg of flavonoids, and
12.98 mg of sulforaphane [6–8]. On the other hand, sprouts obtained from cereals also contain many
health-promoting compounds. For instance, a serving size of wheat sprout juice (244 g) provides 45.75%
of RDA of thiamin, 34.38% of riboflavin, 32.63% of protein, 23.18% of folate, 69.71% of phosphorous, and
64.54% of magnesium [1].

Carrot juice obtained from orange carrots provides high levels of hydroxycinnamic acids such as chlo-
rogenic acid [2,9]. On the other hand, carrot juice obtained from purple carrot contains high levels
of anthocyanins, which are the predominant pigment. These phytochemicals possess high antioxidant
activity and thus could be used on the treatment of different chronic and degenerative diseases, since
they have shown to inhibit low-density lipoprotein (LDL) oxidation, inhibit platelet aggregation and
adhesion, decrease total and LDL cholesterols, induce endothelium-dependent vasorelaxation, and
reduced oxidative DNA damage [10–13]. Anthocyanins in purple carrot are acylated with p-coumaric,
ferulic, p-hydroxybenzoic, and sinapic acids, and they are more stable and resistant to factors such as
pH and light. In unclarified purple carrot juice, the major anthocyanin is cyanidin-3-galactoside-
xyloside-glucoside-ferulic acid, followed by cyanidin-3-galactoside-xyloside-glucoside-coumaric acid
and cyanidin-3-galactoside-xyloside-glucoside [10,12,13].
In addition to phenolic compounds, carrot juice is an important dietary source of carotenoids. Carotenes
are the predominant carotenoids in orange carrot, whereas lycopene and lutein are the predominant
carotenoid species in red and yellow carrot cultivars, respectively [14]. However, purple carrot contains
2.2- and 2.3-fold more α- and β-carotenes than orange carrot, respectively [9]. Structures of chlorogenic
acid, cyanidin-3-galactoside-xyloside-glucoside-ferulic acid, cyanidin-3-galactoside-xylosideglucoside-
coumaric acid, cyanidin-3-galactoside-xyloside-glucoside, and α- and β-carotenes, which are the major
phytochemicals that can be found in carrot juice (depend on variety), are shown in Figure 49.1.
Juices obtained from cruciferous vegetables (e.g., kale and broccoli) contain high levels of phyto-
chemicals that have a direct and indirect effect as antioxidants. For instance, carotenoids present in
these vegetables are capable of acting directly as antioxidants by quenching singlet oxygen, whereas iso-
thiocyanates have an indirect effect as antioxidants by increasing the activity of phase II detoxification
enzymes. Kale and broccoli sprout juices are rich in numerous carotenoids such as lutein, zeaxanthin,
and β-carotene [2]. Lutein and zeaxanthin are well known to prevent cataract and macular degeneration,
since they can filter high-energy wavelengths of visible light, acting as antioxidants that protect against
the formation of free radicals [15].
Cruciferous vegetable juices are also an excellent source of indole compounds, mainly indole-3-
carbinol, which is known to suppress the proliferation of various cancer cell lines, including those of
breast, colon, prostate, and endometrium, by targeting a wide spectrum of signaling pathways governing
hormonal homeostasis, cell cycle progression, and cell proliferation [16–18].
In addition to carotenoids and indole 3-carbinol, juices obtained from cruciferous vegetables, espe-
cially from broccoli, contain high levels of glucosinolates, which release isothiocyanates (e.g., sulfora-
phane) when hydrolyzed by the enzyme myrosinase. The precursor of sulforaphane is the glucoraphanin,
which is abundant in broccoli and represents approximately 44%–56% of the total glucosinolate content.
Broccoli sprouts contain 20- to 50-fold more glucoraphanin than mature plants [19]. The mechanism of
the action of sulforaphane against different chronic diseases is related to the induction of phase II detoxi-
fication enzymes, inhibition of carcinogen-activating phase I enzymes, induction of apoptosis and cell
cycle arrest, and anti-inflammation [20,21]. Structures of lutein, zeaxanthin, glucoraphanin, and indole
3-carbinol, which are the main bioactive compounds present in kale and broccoli sprout juices, are given
in Figure 49.2.
Skeleton Compound R
Cyanidin-3-galactoside-xyloside- OH
HO glucoside
O+ OH O O
HO
Cyanidin-3-galactoside-xyloside-
OH O
O glucoside-ferulic acid
HO
O OH
O
HO HO O
HO O O
OH CH3 OH
HO OH
O
R
Cyanidin-3-galactoside- O
xylosideglucoside-coumaric acid HO
O
HO OH
O
HO
O OH
OH
HO
Chlorogenic acid
CH3 CH3 H3C

H3C CH3
CH3 CH3 H3C CH3

CH3
α-Carotene
CH3
H 3C CH3 H3C
H 3C
CH3
CH3 CH3 CH3
CH3
β-Carotene
FIGURE 49.1 Major bioactive compounds found in carrot juice. Anthocyanins are only found in purple carrot.
Cereals are also a rich source of phytochemicals; the most bioactive compounds studied in wheat
sprout juice are the phenolics and carotenoids, including β-carotene, ferulic acid, and vanillic acid
[22,23]. Structures of these bioactive molecules are shown in Figure 49.3.
49.4 Health Effects

In the following sections of this chapter, the health effects of carrot, kale, and sprout juices are described
based on the results from human intervention studies (Table 49.2).
H3C OH
H3C CH3 CH3 CH3
CH3 CH3 H3C CH3

HO CH3
Lutein
H3C OH
H3C CH3 CH3 CH3
CH3 H3C CH3

CH3
HO CH3
Zeaxanthin
O
O S N O S O–
HO
O
HO OH O
OH S
CH3
Glucoraphanin
HO
N
H
Indole 3-carbinol
FIGURE 49.2 Major bioactive compounds found in kale and broccoli sprouts juices.
CH3
H3C CH3 H3C
H3C
CH3
CH3 CH3 CH3
CH3
β-Carotene
H3C
O
HO
OH
O
Ferulic acid
H3C
O
HO
OH
O
Vanillic acid
FIGURE 49.3 Major bioactive compounds found in wheat sprout juice.

TABLE 49.2
614
Health Effects of Vegetable-Containing Juices in Clinical Studies

Vegetable-
Containing Juices Subjects Study Details Experimental Findings References
Carrot Juice
Daily freshly n = 17 subjects (8 men) with 16 oz (480 mL) carrot juice given daily for 3 months. Carrot juice consumption resulted in significant [13]
squeezed carrot elevated TC and TAG levels decrease (P < 0.05) in systolic pressure, while
juice diastolic pressure was unaffected. It also significantly
increased (P < 0.05) the plasma total capacity and
decreased the plasma MDA production. Levels of TC,
TAG, Apo A, Apo B, LDL, HLD, body fat
percentage, insulin, leptin, and IL-1α were unchanged
(P > 0.05).
Fresh Balero (orange) n = 69 women (breast cancer 8 oz (240 mL) carrot juice mixed with two teaspoons There was an increase of total plasma carotenoids, [27]
and BetaSweet survivors, aged 33–72) of olive oil daily for 3 weeks. which is related to significant changes in the oxidative
(purple) carrot juice stress and inflammation biomarker (8-iso-PGF2α).
Carrot juice n = 20 healthy (nonsmoking Crossover study of two 14-day treatments. Each Carrot juice related to increase in plasma carotenoid [28]
and non-supplement-taking treatment was preceded by a low-carotenoid period concentration (P = 0.0002). Significant changes in
men) of 2 weeks. 330 mL daily carrot juice. secretion of IL-2, proliferation of peripheral blood
mononuclear cells, and lytic activity of NK cells, with
maximum responses during the low-carotenoid
periods.
Carrot juice n = 22 healthy men Crossover study of two 14-day treatments. Each Significant increase of β- and α-carotenes in fecal [44]
treatment was preceded by depletion period. 330 mL contents. Significant dose-dependent inhibition of cell
daily carrot juice. growth (HT29) in fecal water after 5 days of
treatment.
Commercial 100% n = 48 smoking men, aged Randomized study and placebo-controlled design. Lymphocyte DNA damage was significantly decreased [26]
carrot juice 23–57 After a depletion period of 14 days, the subjects were in the carrot juice and the purified β-carotene groups.
divided into three groups and supplemented with However, no significant changes in DNA damage was
either 300 mL/day carrot juice (n = 18), 20.49 mg observed for the placebo group except tail moment
purified β-carotene (n = 16), or 150 mg lactose as (P = 0.016). Neither erythrocyte antioxidant enzyme
placebo (n = 14) for 8 weeks. or plasma lipid profiles were significantly changed
after supplementation.
Carrot juice plus n = 15 non-supplement- Study of two 3-week periods. First period: 375 mL Juice supplementation showed a significant decreased [24]
orange juice taking, smoking, and flavored water mineral daily. Second period: 250 mL in MDA (P < 0.01). However, the rate of LDL
normolipidemic men. orange juice plus 300 mL carrot juice. oxidations was not affected.
(Continued)
Vegetable-
Carrot juice, tomato n = 23 healthy (nonsmoking Week 1 and 2: depletion period. Significant decrease in endogenous levels of strand [25]
juice, and dried men, aged 27–40) Week 3 and 4: 330 mL/day tomato juice. breaks in lymphocyte DNA. Carrot juice consumption
spinach Week 5 and 6: 330 mL/day carrot juice. significantly reduced the oxidative base damage in
Week 7 and 8: 10 g dried spinach powder (in water or contrast with tomato juice or spinach.
milk).
Kale Juice
Kale juice n = 84 subclinical 300 mL/day for 6 weeks kale juice. Kale juice supplementations significantly decreased [35]
hypertensive men systolic and diastolic blood pressure in all patients
regardless of their GSTM1 or GSTT1polymorphisms.
However, blood glucose level decreased only in the
GSTM1 genotype.
TC and TAG levels of the participants were not
changed after kale juice supplementation regardless
Vegetable-Containing Juices (Carrot, Kale, and Sprout)
of GST polymorphisms
HDL and LDL level only showed significant increase
and decrease, respectively, in GSTT1 genotype.
Fresh filtered kale n = 32 men with 150 mL/day of kale juice for 12 weeks. Significant increase in glutathione peroxidase activity [33]
juice, soaked in hypercholesterolemia and (P = 0.0005) and serum concentrations of HDL and
malic acid solution normal triglyceridemia ratio HDL–LDL cholesterols (P < 0.0001). The LDL
(non-supplement-taking concentration was significantly reduced. There was
and nonalcoholic subjects) no difference in the concentration of MDA.
Kale and garlic juice n = 40 (27 men) healthy, The subjects were divided into four groups. Days 1–3 There was a significantly decrease in urinary NDMA [45]
nonsmoking, and of the study were control days. Day 4, the subjects excretion after strawberry, garlic, or kale juice
non-supplement-taking received the following: consumption.
subjects G1: (400 mg) nitrate
G2: (300 g) whole strawberry
G3: (200 g) garlic juice
G4: (200 g) kale juice
(Continued)
615
616

Vegetable-
Kale juice n = 32 men with 150 mL/day of kale juice for 12 weeks. Serum concentrations of HDL and the ratio of HDL/ [34]
hypercholesterolemia LDL were significantly increased, while LDL
concentration was reduced (P < 0.001) after consume
of kale juice.
Levels of 12:0, 14:0, 18:1ω9, 18:3ω6, and sum of
MUFA in serum phospholipid were significantly
increased, while 22:4ω6 level was significantly
reduced (P < 0.05).
Sprout Juice
Wheat sprout juice n = 14 healthy women, aged 100 mL of wheat sprout juice daily for 2 weeks. Reduction of urinary bisphenol A during the [37]
20–28 supplementation of wheat sprout juice was observed
(P = 0.02).
Fresh broccoli sprout n = 291 healthy subjects (62 143 participants consumed a placebo beverage and Statistically significant (P ≤ 0.01) increases in the [38]
juice men), aged 21–65 148 g broccoli sprout beverage for 12 weeks. levels of excretion of the glutathione-derived
conjugates of benzene, acrolein, but not
crotonaldehyde, were found in those receiving
broccoli sprout beverage compared with placebo.
Excretion of the benzene-derived mercapturic acid
was higher in participants who were GSTT1-positive
than in the null genotype, regardless of beverage type.
Broccoli sprout n = 81 patients with type 2 Subjects were randomly assigned into three groups for BSP in dose of 10 g/day, significantly decreased serum [6]
powder diabetes, aged 18–60 4 weeks TAG and oxidation LDL/LDL ratio (P < 0.05). HDL
G1: 10 g/day (BSP), n = 27. concentration was significantly higher (P < 0.01) in
G2: 5 g/day (BSP), n = 29. dose of 10 g/day, compared with group 5 g/day and
G3: 5 g/day corn starch (placebo), n = 25. placebo.
Broccoli sprout n = 81 patients with type 2 Subjects were randomly assigned into three groups for In the BSP treatment groups, there was a significant [39]
powder diabetes 4 weeks. decrease in serum hs-CRP and also nonsignificant
G1: 10 g/day (BSP), n = 27 decreases in serum IL-6 and TNFα concentration.
G2: 5 g/day (BSP), n = 29
G3: 5 g/day corn starch (placebo), n = 25
Abbreviations: Apo A, apolipoprotein A; Apo B, apolipoprotein B; BSP, broccoli sprout powder; GST, glutathione S-transferase;, GSTM1, glutathione S-transferase Mu 1; GSTT1, glutathi-
one S-transferase theta 1; HLD, high-density lipoprotein; hs-CRP, high sensitivity C-reactive protein; IL-1α, interleukin-1α; LDL, low-density lipoprotein; MDA, malondial-
dehyde; MUFA, monounsaturated fatty acids; NDMA, n-nitrosodimethylamine; TAG, triacylglycerols; TC, total cholesterol; TNFα, tumor necrosis factors-α.
49.4.1 Carrot Juice

According to human intervention studies, carrot juice can be used for the prevention of cardiovascu-
lar disease (CVD) [13,24], cancer [25,26], breast cancer recurrence [27], as well as the modulation of
immune functions [28].
Potter et al. [13] performed a study to evaluate if carrot juice has a positive effect on the antioxi-
dant status and cardiovascular risk markers (total cholesterol [TC], triacylglycerols [TAG], apolipopro-
tein A, apolipoprotein B, LDL, high-density lipoprotein [HDL], body fat percentage, insulin, leptin,
interleukin-1α [IL-1α], and high-sensitivity C-reactive protein [hs-CRP]) of humans with elevated plasma
TC and TAG levels. The researchers provided 16 oz (480 mL) of fresh carrot juice daily for 3 months to
eight males and nine females. Although no effect of drinking carrot juice was observed on the cardiovas-
cular risk markers evaluated, the antioxidant status was increased and lipid peroxidation (evaluated by
total malondialdehyde [MDA] production) was decreased. Therefore, Potter et al. [13] concluded that in
addition to drinking carrot juice, changes in dietary and lifestyle would be required to observe positive
impact on lipid profiles in subject with elevated TC and TAG levels.
Similarly, Abbey et al. [24] evaluated the effect of daily supplementation of carrot juice for 3 weeks on
plasma levels of β-carotene and oxidation of LDL in 15 normolipidemic male cigarette smokers. During
the study, the subjects were provided with a diet rich in polyunsaturated fatty acids (PUFA) and for the
washout period (3 weeks) consumed vitamin-free drinks. Carrot juice consumption for 3 weeks raised
plasma carotenoid levels and induced lower MDA levels in copper-oxidized LDL, suggesting that carrot
juice consumption partly protects LDL from oxidation in habitual cigarette smokers with a diet rich in
PUFA [24].
Plasma concentrations of total carotenoids have also been associated with protection against breast
cancer recurrence. Therefore, Butalla et al. [27] evaluated the effects of a carrot juice intervention on
plasma carotenoids, oxidative stress, and inflammation of 69 overweight breast cancer survivors. The
subjects consumed 8 oz (240 mL) of fresh carrot juice daily for 3 weeks. Two different carrot juices were
evaluated, one obtained from BetaSweet (anthocyanin-rich carrot) and the other from Balero orange
carrot juice. The hypothesis was that both juices would increase plasma total carotenoid concentra-
tions to levels previously shown to reduce breast cancer recurrence. The authors also hypothesized that
regular carrot juice intake would be associated with reductions in oxidative stress (8 isoform prosta-
glandin F2 alpha [8-iso-PGF2α]) and inflammation (thromboxane B2, prostaglandin E2 metabolites,
and hs-CRP). Consumption of carrot juice increased total plasma carotenoids, inducing higher levels
in the BetaSweet (1.65 µmol/L) than the Balero carrot (1.38 µmol/L) juice. Increases in total plasma
carotenoids were inversely correlated with 8-iso-PGFα levels and no differences existed between car-
rot varieties. However, no effect of carrot juice consumption was observed on inflammatory markers.
Therefore, results indicated that increasing plasma total carotenoids by consuming fresh carrot juice
could be used as an approach to reduce oxidative stress, but not inflammatory markers, in overweight
breast cancer survivors.
Likewise, β-carotene enhanced immune function in humans [29]. Based on this evidence, Watzi et al.
[28] determined if the supplementation of a low-carotenoid diet with carrot juice could modulate the
immune function in healthy men. To test their hypothesis, the authors provided 300 mL/day of carrot
juice to male subjects on a low-carotene diet with a 2-week depletion period after juice intervention. The
immune status was assessed by measuring the lytic activity of natural killer (NK) cells, secretion of
cytokines (interleukin 2 [IL-2], interleukin 4 [IL-4], tumor necrosis factors-α [TNFα]), and proliferation
by activated peripheral blood mononuclear cells. Juice consumption resulted in fast responses in plasma
carotenoid concentration, which were not accompanied by changes in immune function. The maximum
responses for IL-2, NK cell cytotoxicity, and lymphocyte proliferation were observed during depletion
periods. The highest production rate of TNFα was detected at the end of the first intervention period,
and carrot juice intervention did not modulate IL-4 secretion. Therefore, Watzi et al. [28] concluded that
increased plasma carotenoid concentrations after vegetable juice consumption are followed by a time-
delayed modulation of immune functions in healthy men consuming a low-carotenoid diet.
In order to determine if the consumption of vegetables containing different carotenoids could pro-
tect against cancer by preventing DNA damage, Pool-Zobel et al. [25] performed a human intervention
study with vegetable products including carrot juice. After 2 weeks of depletion period (weeks 1 and 2),
23 healthy, nonsmoking males (27–40 years old) were provided with 330 mL of tomato juice (with 40 mg
of lycopene) during 2 weeks (weeks 3 and 4). Thereafter, for the following 2 weeks (weeks 5 and 6),
the subjects consumed carrot juice containing 22.3 and 15.7 mg of β-carotene and α-carotene, respec-
tively. Finally, for the last 2 weeks (weeks 7 and 8), the subjects consumed 10 g of dried spinach powder
(in water or milk) with 11.3 mg lutein. Results indicated that supplementation with vegetable juices rich
in carotenoids induced significant decreases in endogenous levels of strand breaks in lymphocyte DNA.
Likewise, carrot juice intervention reduced oxidative damage to DNA bases. Results from this investiga-
tion provide evidence that plant foods with high levels of carotenoids could protect against cancer by
decreasing oxidative or other damages to DNA in humans [25].
Similarly, Lee et al. [26] evaluated the protective effect of carrot juice on lymphocyte DNA damage,
as well as on erythrocyte antioxidant enzymes and plasma lipid profiles in 48 Korean smokers. Subjects
were provided, during 8 weeks, with carrot juice (n = 18), purified β-carotene (n = 16) or placebo (n = 14),
after 14 days depletion. Carrot juice provided to the subjects (300 mL/day) contained approximately
20.49 mg of β-carotene/day and 1.2 mg of vitamin C/day. Likewise, carotene supplementation was per-
formed with capsules (1/day) containing 20.49 mg of β-carotene. The placebo group did not show any
protective effect, whereas carrot juice containing β-carotene or purified β-carotene itself showed great
antioxidative potential in preventing damage to lymphocyte DNA in smokers.
In addition to the clinical studies that demonstrate potential uses of carrot juice for the prevention
of CVD, breast cancer recurrence, enhanced immune function, and cancer prevention/recurrence,
other in vitro studies have demonstrated that carrot juice may be used for the treatment of leukemia
[30] and also induces an enhanced production of coenzyme Q10 [31], which has been used therapeu-
tically for CVD, as a supportive therapy in statin treatment and in mitochondrial respiratory-chain
diseases.
Additional in vivo and in vitro studies demonstrating the potential use of carrot juice on the prevention
of chronic diseases are summarized in Table 49.3. Khayatnouri et al. [32] compared the effect of carrot
juice and gliclazide (an oral hypoglycemic drug) on blood glucose level in diabetic mice. After 1 month,
glucose concentration was significantly decreased (P < 0.05) with the individual administration of carrot
juice and gliclazide, but there was a stronger antidiabetic effect when carrot juice and gliclazide were
coadministered. Poudyal et al. [11] evaluated a group of rats with a high-carbohydrate and high-fat diet
supplementing half of the study group with purple carrot juice or β-carotene. Rats without administra-
tion of carrot juice developed CVD as hypertension, cardiac fibrosis, endothelial dysfunction, increased
abdominal fat deposition, and plasma lipid profile.
49.4.2 Kale Juice

Kale juice has great potential for the prevention of CVD [33–35]. The effect of kale juice on the pre-
vention of coronary heart disease (CHD) in male subject with hyperlipidemia was evaluated [33].
During 3 months, 32 men with hypercholesterolemia consumed 150 mL of kale juice per day for a
12-week intervention period. HDL cholesterol increased by 27% and HDL to LDL cholesterol ratio was
increased by 52%. Likewise, LDL cholesterol concentration was reduced by 10%. Results also indicated
that kale juice could increase glutathione peroxidase activity and the serum selenium level. Therefore,
the authors concluded that meals’ supplementation with kale juice has a positive impact on serum
lipid profiles and antioxidant systems, in male subjects with hyperlipidemia, contributing to reduce the
risk of CHD.
In a similar human intervention study, Chung et al. [34] evaluated the effect of kale juice supplemen-
tation on serum lipid levels and phospholipid fatty acid compositions in 32 hypercholesterolemic men
who consumed 150 mL of kale juice per day for 3-month intervention period. The authors observed
that the levels of 12:0, 14:0, 18:1ω9, 18:3ω6, and sum of monounsaturated fatty acids (MUFA) in
serum phospholipid were significantly increased, whereas 22:4ω6 level was significantly reduced.
Therefore, the authors concluded that kale juice contributes to changes of serum phospholipid fatty
acid compositions, improving serum lipid profiles, and thus lowering the risks of CHD in men with
hypercholesterolemia.
TABLE 49.3
Potential Positive Health Effect of Vegetable-Containing Juices Evaluated in Cell Lines and Animal Studies
Vegetable- Experimental
Containing Juices Model Study Details Experimental Findings References
Carrot Juice
Carrot juice n = 50 diabetic adult Animals were randomly assigned into five groups (n = 10) Carrot juice consumption resulted in significant decreased [32]
Wistar male mice for 4 weeks, daily. (P < 0.05) in serum glucose in groups 3 and 4.
G1: Control (normal saline 10 mL/kg).
G2: Streptozocin (40 mg/kg).
G3: Streptozocin + carrot juice (10 mL/kg).
G4: Streptozocin + gliclazide (1 mg/kg).
G5: Streptozocin + carrot juice + gliclazide.
Carrot juice and Pseudomonas Various concentrations of juices were added in the culture Carrot juice and tomato juice acted as natural precursors [31]
tomato juice diminuta NCIM medium. and enhanced the yield of CoQ10, which has been
2865 successfully used therapeutically for CVD and in
mitochondrial respiratory-chain diseases.
Purple carrot juice n = 84 Wistar male Rats were randomly assigned into six groups (n = 12) and High-carbohydrate and high-fat diet-fed rats developed [11]
rats were fed for 8 weeks with hypertension, cardiac fibrosis, endothelial dysfunction,
G1: Maize starch. increased abdominal fat deposition, and plasma lipid
Vegetable-Containing Juices (Carrot, Kale, and Sprout)
G2: Maize starch + β-carotene. profile. Purple carrot juice diet reduced percentage in
G3: Maize starch + purple carrot juice. body weight, abdominal fat and circumference, enhanced
G4: High-carbohydrate and high-fat. glucose tolerance, and reduced plasma lipid profile.
G5: High-carbohydrate and high-fat + β-carotene.
G6: High-carbohydrate and high-fat + purple carrot juice.
Kale Juice
Kale juice Female BALB/c 14 days (2 mg/kg/day). An immunostimulating effect was observed on IL-5. The [36]
mice IL-5 improves the IgA secretion by B cells, IgM, and IgG.
Sprout Juice
Brussels sprouts, n = 32 male BALB/c 0.2 mL/10 g of juice for 14 days. Broccoli juice administered produced a significant (P < [19]
cauliflower, cabbage, mice Animals were randomly assigned into four groups (n = 8). 0.01) reduction in the number of micronuclei (clastogenic
and broccoli On day 14, they were given effect) in group 1 compared to group 3. The numbers of
G1: 20 mg/kg of IQ mutagen. micronuclei showed no statistically significant difference
G2: Control. between group 2 and group 4.
G3: 50 mg/kg of MNU mutagen.
G4: 7% DMSO (excipient).
(Continued)
619
620

Potential Positive Health Effect of Vegetable-Containing Juices Evaluated in Cell Lines and Animal Studies
Vegetable- Experimental
Containing Juices Model Study Details Experimental Findings References
Brussels sprout juice n = 40 Wistar male Animals were randomly assigned into two groups (n = 20). Treatment with DMH significantly (P < 0.01) increased [42]
rats G1: DMH (30 mg/kg body weight) injections of levels of mitosis and apoptosis. The elevated level of
colon-specific carcinogen. mitosis decreased if animals received raw sprout juice
G2: Injections of saline. after carcinogen treatment, but no effect was seen if
Thereafter, 10 DMH and 10 saline mice were fed with animals received raw juice in the absence of DMH.
2 mL freshly prepared sprout juice.
Broccoli sprout juice Human Cells were grown in DMEM/F-12 medium containing 10% Treatment of SH-SY5Y cells with both juices protected [43]
neuroblastoma fetal bovine serum and treated with 25 μM aggregated against Aβ-induced cytotoxicity and apoptotic cell death
SH-SY5Y cells Aβ25–35 in the presence or in the absence of 10 μL of as evidenced by cell viability, nuclear chromatin
juice. condensation, and apoptotic body formation
measurements.
Brussels sprout juice HT29 colorectal 1 × 106 cells—3 mL of warm DMEM medium HT29 cells treated with Brussels sprout juice resulted in [41]
cancer cells supplemented with10 μL juice/mL medium. the detaching from the surface after 24 h. There was a
decrease in the number of cells in the G1 phase of the cell
cycle, accompanied by a significant (P < 0.05) increase in
the number of cells in G2/M.
Abbreviations: CVD, cardiovascular disease; DMEM/F12, modified eagle medium: nutrient mixture F12; DMH, 1,2-dimethylhydrazine; DMSO, dimethyl sulfoxide; IgA, immunoglobulin
A; IgG, immunoglobulin G; IgM, immunoglobulin M; IL-5, interleukin-5; IQ, 2-amino-3-methyl-3H-imidazo-[4,5-f]quinoline; MNU, 2-nitroso-2-methylurea.
It is well known that glucosinolate breakdown products produced in kale after wounding have the
ability to activate glutathione S-transferase (GST) enzyme. This activation is higher in humans with the
glutathione S-transferase Mu 1 (GSTM1) than glutathione S-transferase theta 1 (GSTT1) polymorphism.
Based on that, Han et al. [35] evaluated if daily supplementation of kale juice can modulate blood pres-
sure, lipid profiles, and blood glucose level, and whether this modulation is influenced by GSTM1 and
GSTT1 polymorphisms. The study was performed with 84 subclinical hypertensive patients from South
Korea, who received 300 mL/day of kale juice for 6 weeks. Blood pressure was decreased in all patients.
However, blood glucose levels were decreased only in GSTM1 genotypes. Regarding the lipid profile,
GSTT1 patients showed increases in HDL cholesterol and decreases in LDL cholesterol. Results sug-
gest that the positive effect of kale juice on lipid profiles and blood glucose is dependent on GST genetic
polymorphisms.
Another in vivo study (Table 49.3) demonstrated that kale juice may be used for its immunostimulatory
effect [23]. Nishi et al. [36] evaluated kale juice consumption for 14 days in rats. The results showed an
immunoglobulin (Ig) production-stimulating effect in lymphocytes and Peyer’s patches.
49.4.3 Sprout Juices (Wheat and Broccoli)

According to human intervention studies (Table 49.2), the consumption of sprouts juices (wheat and
broccoli) can exert protection against endometriosis in young women [37] and prevent lung cancer [38].
Likewise, broccoli sprout juice can be used for the treatment of type II diabetes [39] and are effective on
the eradication of Helicobacter pylori [40].
Yi et al. [37] provided wheat sprout juice (100 mL/day) for 2 weeks to fertile young women (n = 14
and age = 24.4) and evaluated its effect on bisphenol A (BPA). BPA is an endocrine-disrupting chemical
that induces reactive oxygen species, which are involved on endometriosis, which precludes fertility. The
authors concluded that wheat sprout can exert protection against endometriosis in young women.
Exposure to air pollution is a risk factor for lung cancer. In order to evaluate if a broccoli sprout-
derived beverage, containing 600 μmol of glucoraphanin and 40 μmol of sulforaphane, could protect
against pollutants, Egner et al. [38] provided daily doses of the beverage to 291 subjects during 12 weeks.
The authors evaluated before and during the intervention the urinary excretion of mercapturic acids of
the pollutants, benzene, acrolein, and crotonaldehyde. The broccoli sprout‒derived beverage induced
significant increases in the levels of glutathione-derived conjugates of the pollutants (benzene and acro-
lein). This excretion was higher in GSTT1-positive subjects than in null genotypes. Likewise, pharma-
cokinetic studies indicated that the bioavailability of sulforaphane metabolites did not decline over the
12-week intervention period. The authors concluded that consumption of broccoli sprout-derived bever-
ages could enhance detoxification of airborne pollutants and, thus, protect against lung cancer.
Anti-inflammatory approaches have been used for the treatment of type II diabetes. Therefore,
Mirmiran et al. [39] provided broccoli sprouts powder (BSP) high in sulforaphane and evaluated
inflammatory markers during 4 weeks. Sixty three patients with type II diabetes were included in
the analysis and were divided into three groups. Group 1 consumed 10 g/day BSP (n = 21), group 2
consumed 5 g/day (n = 22), and group 3 was the placebo (n = 20). BSP treatment decreased serum
hs-CRP concentration (−20.5% and −16.4%) in group A and B, respectively. Likewise, serum hs-CRP
and IL-6 were lower in group A as compared to the placebo after intervention. Results indicated that
high-sulforaphane BSP had favorable effects on inflammatory markers in type II diabetic patients.
In addition, this high-sulforaphane BSP has proven effective for the eradication of H. pylori [40] and
also exerts favorable effects on lipid profiles and ox-LDL/LDL ratio (as risk factors for CVD) in type II
diabetic patients [6].
In addition to the aforementioned human intervention studies, the protective effect of sprout juices
has been evaluated in in vitro and in vivo animal models (Table 49.3). For instance, Smith et al. [41,42]
evaluated the effect of Brussels sprout juice in an animal model of colorectal neoplasia and found that
glucosinolate breakdown products derived from Brassica vegetables can exert a profound effect on the
balance of colorectal cell proliferation and death. Similarly, Masci et al. [43] determined that broccoli
sprout juice could also exert neuroprotective effects in vitro.
49.5 Vegetable-Containing Juices: Relationship between Chronic Disease,

Food, and Medicine
We have summarized, in Figures 49.4 and 49.5, the relationship between chronic diseases, food, and
medicine. It is well known that a healthy diet such as the “Mediterranean diet” can prevent a predisease
state, allowing in returning to a normal healthy state. However, it is unknown if a disease state can be
reversed by eating well as an intervention or therapeutic treatment; thus traditionally, a disease state is
treated with conventional medicinal drugs.
In this chapter, we presented clinical studies with carrot juice and juices of wheat and broccoli sprouts
that have a preventive effect on human health, while kale juice and broccoli sprout juice may exert a
therapeutic effect for different diseases. Similarly, we presented in vitro and in vivo studies with carrot
juice, kale juice, and broccoli sprout juice that have the potential to exert preventive effects, while carrot
juice and Brussels sprout juice have the potential as therapeutics for different diseases, which eventually
have to be confirmed with clinical studies.

In recent years, the fruit and vegetable juice industry has entered a period of rapid evolution, due to the
change which has occurred in the consumption patterns of the population in the world, especially in
developed countries, as currently consumers not only care about the taste but also the health benefits.
One of the major challenges in the vegetable juice industry is the development of processed juices that
preserves the health benefits of fresh juices after processing and during storage. To overcome this major
issue, research on the application of emerging technologies that avoid temperature to process vegetable
*Cardiovascular [23]
Reversal by eating well? *Cancer [25,26]
Reversal by eating well ***Cardiovascular [33–35]
**Endometriosis [37]
**Lung cancer [38]
**Type II diabetes [39]
*High cholesterol and TAG [23]
**CVD[6]
*Inflammation [27]
Food leading cause *DNA damage [25,26] **Ulcer [40]
high-fat [24] ***Hypercholesterolemia [33,34]
***Hypertensive [35]
**Inflammation [39]
state state state
*Carrot juice [23–28] ***Kale juice [33–35]
**Broccoli sprouts [6,39,40]
**Wheat [37] and
broccoli [38] sprouts
Conventional
*Less oxLDL [24] ***High HDL, high HDL/LDL, and low LDL [33]
***Increased glutathione peroxidase activity [33] medicinal drug
*Less oxidative stress [27]
*Delayed immune modulation [28] ***Improved serum lipid profile [34]
*Increased AOX status [23] ***Decreased blood pressure [35]
*Less lipid peroxidation [23] ***Less glucose levels–GSTM1 patients [35]
*Less DNA damage [25,26] ***High HDL and less LDL–GSTT1 patients [35]
**Less bisphenol (BPA) [37] **Decreased serum hs-CRP [39]
**Increased glutathione derived conjugates of **Decreased IL-6 [39]
pollutants-GSTT1 patients [38] **Favorable lipid profiles [6]
**Favorable oxLDL/LDL ratio [6]
**Eradicate H. pylori [40]
Prevention Intervention/Treatment/Therapeutics
FIGURE 49.4 Relationship between chronic diseases, food, and medicine in human health (based on clinical studies).
Carrot juice, wheat, and broccoli sprouts exert preventive effects in human health, while kale juice and broccoli sprouts
exert therapeutic effects in different chronic diseases. Preventive or therapeutic effects associated to *carrot juice, **wheat
and broccoli sprout juices, and ***kale juice.
*Leukemia [30]
*Cardiovascular [11]
Hypertension
Cardiac fibrosis
Endothelial dysfunction
Fat deposition
Reversal by eating well Reversal by eating well? Increased plasma lipid
*Diabetes [32]
**Colorectal neoplasia [41,42]
Food leading cause
High fat [11]
High carb [11]
state state state
*Carrot juice [11,31] *Carrot juice [30]
**Broccoli sprouts [43] *Carrot juice + gliclazide [32]
***Kale juice [36] **Brussels sprouts [41,42]
*High production of coenzyme Q10 [31] *Less glucose [32]

*Reversal effects [11] *Leukemia [30]
***Immunoglobulin production [36] **Reduced proliferation and
***Immune stimulation [36] induced cell death [41,42]
**Neuroprotective [43] Conventional
medicinal drug
FIGURE 49.5 Relationship between chronic diseases, food, and medicine in human health (based on in vitro and in vivo
studies). Carrot juice, kale juice, and broccoli sprouts exert preventive effects in human health, while carrot juice and
Brussels sprouts exert therapeutic effects in different chronic diseases. Preventive or therapeutic effects associated to
*carrot juice, **broccoli sprouts or Brussels sprouts, and ***kale juice.
juices is being performed. Examples of these technologies are high hydrostatic pressure, electric pulses,
and ultrasounds.
An additional trend in the vegetable juices is the development of convenience fresh vegetable juices
that allows their rapid preparation. For instance, there are individualized portions of “green juice” in
supermarkets, where the portion includes frozen vegetables and orange juice, which after defrosting can
be homogenized and is ready for consumption.
Another important trend is the production of dietary supplements with high concentrations of bio-
active compounds present in vegetables. As an example, glucosinolates may be found in the dietary
supplement market in several forms, such as extracts of broccoli and other cruciferous vegetables or
as broccoli sprouts rich in glucoraphanin. However, broccoli extracts may not be as effective because
glucosinolates have to be hydrolyzed to release isothiocyanates in order to be completely absorbed in the
gastrointestinal tract. Therefore, dietary supplements such as EnduraCell have preserved the activity of
myrosinase in order to yield isothiocyanates in an effective manner.
49.7 Conclusion
We presented clinical studies as well as in vitro and in vivo studies describing the relationship between
chronic diseases and their prevention or treatment with the consumption of vegetable-containing juices.
The preventive and/or therapeutic properties of vegetable-containing juices are mainly attributed to the
bioactive compounds present in the juices (e.g., carotenoids, phenolics, and glucosinolates). However,
it is important to point out that most of the studies suggest that consumption of vegetable-containing
juices should be accompanied with a balanced diet in order to observe a higher positive effect on human
health.
REFERENCES
1. U.S. Department of Agriculture (USDA), Dietary Guidelines for Americans, Center for Nutrition Policy
and Promotion, Washington, DC, 2010.
27, 2014. Published online at: http://ndb.nal.usda.gov/ndb/search (accessed April 28, 2015).
3. Chen, B.H., Peng, H.Y., and Chen, H.E., Changes of carotenoids, color, and vitamin A contents during
processing of carrot juice. J. Agric. Food Chem., 43, 1912–1918, 1995.
4. Ogawaa, K., Nakadab, K., Yamashitab, S., Hasegawaa, T., and Moriguchib, S., Beneficial effects of the
vegetable juice Aojiru on cellular immunity in Japanese young women. Nutr. Res., 24, 613–620, 2004.
5. Becerra-Moreno, A., Alanís-Garza, P.A., Mora-Nieves, J.L., Mora-Mora, J.P., and Jacobo-Velázquez,
D.A., Kale: An excellent source of vitamin C, pro-vitamin A, lutein and glucosinolates. CyTA J. Food,
12, 298–303, 2014.
6. Bahadoran, Z., Mirmiran, P., Hosseinpanah, F., Rajab, A., Asghari, G., and Azizi, F., Broccoli sprouts
powder could improve serum triglyceride and oxidized LDL/LDL-cholesterol ratio in type 2 diabetic
patients: A randomized double-blind placebo-controlled clinical trial. Diab. Res. Clin. Prac., 96,
348–354, 2012.
7. López, J., Tirado, L.G., Sánchez, D., Campas, O., Cantú, E., and Núñez, J., Biochemical composition
of broccoli seeds and sprouts at different stages of seedling development. Int. J. Food Sci. Technol., 48,
2267–2275, 2013.
8. Aires, A., Rosa, E., Carvalho, R., Haneklaus, S., and Schnug, E., Influence of nitrogen and sulfur fertil-
ization on the mineral composition of broccoli sprouts. J. Plant Nutr., 30, 1035–1046, 2007.
9. Alasalvar, C., Grigor, J.M., Zhang, D., Quantick, P.C., and Shahidi, F., Comparison of volatiles, phe-
nolics, sugars, antioxidant vitamins, and sensory quality of different colored carrot varieties. J. Agric.
Food Chem., 49, 1410–1416, 2001.
10. Vishwanath, K., Shweta, W., Meenakshi, S., and Charanjit, K., Black carrot (Daucus carota ssp. sativus)
juice: Processing effects on antioxidant composition and color. Food Bioprod. Process., 89, 482–486,
2011.
11. Poudyal, H., Panchal, S., and Brown, L., Comparison of purple carrot juice and β-carotene in a high-
carbohydrate, high-fat diet-fed rat model of the metabolic syndrome. Br. J. Nutr., 104, 1322–1332, 2010.
12. Turkyilmaz, M., Yemis, O., and Ozkan, M., Clarification and pasteurisation effects on monomeric
anthocyanins and percent polymeric colour of black carrot (Daucus carota L.) juice. Food Chem., 134,
1052–1058, 2012.
13. Potter, A.S., Foroudi, S., Stamatikos, A., Patil, B.S., and Deyhim, F., Drinking carrot juice increases
total antioxidant status and decreases lipid peroxidation in adults. Nutr. J., 10, 96, 2011.
14. Davinder, P.S., Joel, B., Jennifer, K.M., and Li, D., Impact of boron, calcium and genetic factors on vita-
min C, carotenoids, phenolic acids, anthocyanins and antioxidant capacity of carrots (Daucus carota).
Food Chem., 132, 1161–1170, 2012.
15. Mares-Perlman, J.A., Millen, A.E., Ficek, T.L., and Hankinson, S.E., The body of evidence to support a
protective role for lutein and zeaxanthin in delaying chronic disease: Overview. J. Nutr., 132, 518–524,
2002.
16. Weng, J.R., Tsai, C.H., Kulp, S.K., and Chen, C.S., Indole-3-carbinol as a chemopreventive and anti-
cancer agent. Cancer Lett., 262, 153–163, 2008.
17. Wang, X., He, H., Lu, Y., Ren, W., Teng, K., Chiang, C., Yang, Z. et al., Indole-3-carbinol inhibits
tumorigenicity of hepatocellular carcinoma cells via suppression of microRNA-21 and upregulation of
phosphatase and tensin homolog. Biochim. Biophys. Acta, 1853, 244–253, 2015.
18. Safe, S., Papineni, S., and Chintharlapalli, S., Cancer chemotherapy with indole-3-carbinol, bis(3′-
indolyl)methane and synthetic analogs. Cancer Lett., 269, 326–338, 2008.
19. Totušek, J., Lefnerová, D., Strohalm, J., Vrchotová, N., Zendulka, O., Průchová, J., Chaloupková, J.,
Novotná, P., and Houška, M., Contents of sulforaphane and total isothiocyanates, antimutagenic activ-
ity, and inhibition of clastogenicity in pulp juices from cruciferous plants. Czech. J. Food Sci., 29,
548–556, 2011.
20. Zhang, Y. and Callaway, E.C., High cellular accumulation of sulphoraphane, a dietary anticarcinogen,
is followed by rapid transporter-mediated export as a glutathione conjugate. Biochem. J., 364, 301–307,
2002.
21. Tarozzi, A., Angeloni, C., Malaguti, M., Morroni, F., Hrelia, S., and Hrelia, P., Sulforaphane as a poten-
tial protective phytochemical against neurodegenerative diseases. Ox. Med. Cell. Long., 2013, 415078,
2013.
22. Yang, F., Basu, T.K., and Ooraikul, B., Studies on germination conditions and antioxidant contents of
wheat grain. Int. J. Food Sci. Nutr., 52, 319–330, 2001.
23. Leoncini, E., Prata, C., Malaguti, M., Marotti, I., Segura-Carretero, A., Catizone, P., Dinelli, G., and
Hrelia, S., Phytochemical profile and nutraceutical value of old and modern common wheat cultivars.
PLoS One, 7, e45997, 2012.
24. Abbey, M., Noakes, M., and Nestel, P.J., Dietary supplementation with orange and carrot juice in ciga-
rette smokers lower oxidation products in copper-oxidized low-density lipoproteins. J. Am. Diet. Assoc.,
95, 671–675, 1995.
25. Pool-Zobel, B.L., Bud, A., Müller, H., Wollowski, I., and Rechkemmer, G., Consumption of vegetables
reduces genetic damage in humans: First results of a human intervention trial with carotenoid-rich
foods. Carcinogenesis, 19, 1847–1850, 1997.
26. Lee, H.J., Park, Y.K., and Kang, M.H., The effect of carrot juice, β-carotene supplementation on lym-
phocyte DNA damage, erythrocyte antioxidant enzymes and plasma lipid profiles in Korean smokers.
Nutr. Res. Pract., 5, 540–547, 2011.
27. Butalla, A.C., Crane, T.E., Patil, B., Wertheim, B.C., Thompson, P., and Thompson, C.A., Effects of a
carrot juice intervention on plasma carotenoids, oxidative stress, and inflammation in overweight breast
cancer survivors. Nutr. Cancer, 64, 331–341, 2012.
28. Watzi, B., Bub, A., Brivida, K., and Rechkemmer, G., Supplementation of a low-carotenoid diet with
tomato or carrot juice modulates immune function in healthy men. Ann. Nutr. Metab., 47, 255–261,
2003.
29. Hughes, D.A., Effects of carotenoids on human immune function. Proc. Nutr. Soc., 58, 713–718, 1999.
30. Zaini, R., Clench, M.R., and Le Maitre, C.L., Bioactive chemicals from carrot (Daucus carota) juice
extracts for the treatment of leukemia. J. Med. Food, 14, 1303–1312, 2011.
31. Bule, M. and Singhal, R.S., Use of carrot juice and tomato juice as natural precursors for enhanced
production of ubiquinone-10 by Pseudomonas diminuta NCIM 2865. Food Chem., 116, 302–305, 2009.
32. Khayatnouri, M., Nikmanesh, M., and Safarmashaei, S., Study of the effect of gliclazide and carrot juice
on blood sugar level in STZ–induced diabetic male mice. Adv. Environ. Biol., 5, 1742–1745, 2011.
33. Kim, S.Y., Yoon, S., Kwon, S.M., Park, K.S., and Lee-Kim, Y.C., Kale juice improves coronary artery
disease risk factors in hypercholesterolemic men. Biomed. Environ. Sci., 21, 91–97, 2008.
34. Chung, E.J., Shim, E., and Kim, S.Y., Effect of kale juice on serum lipid levels and phospholipid fatty
acid composition in hypercholesterolemic men. J. Life Sci., 22, 1538–1544, 2012.
35. Han, J.H., Lee, H.J., Kim, T.S., and Kang, M.H., The effect of glutathione S-transferase M1 and T1
polymorphisms on blood pressure, blood glucose, and lipid profiles following the supplementation of
kale (Brassica oleracea acephala) juice in South Korean subclinical hypertensive patients. Nutr. Res.
Pract., 9, 49–56, 2015.
36. Nishi, K., Kondo, A., Okamato, T., Nakano, H., Daifuku, M., Nishimoto, S., Ochi, K., Takaoka, T.,
and Sugahara, T., Immunoestimulatory in vitro and in vivo effects of a water-soluble extract from kale.
37. Yi, B., Kasai, H., Lee, H.S., Kang, Y., Park, J.Y., and Yang, M., Inhibition by wheat sprout (Triticum
aestivum) juice of bisphenol A-induced oxidative stress in young women. Mutat. Res., 724, 64–68, 2011.
38. Egner, P.A., Chen, J.G., Zarth, A.T., Ng, D.K., Wang, J.B., Kensler, K.H., Jacobson, L.P. et al., Rapid and
sustainable detoxification of airborne pollutants by broccoli sprout beverage: Results of a randomized
clinical trial in China. Cancer Prev. Res., 7, 813–823, 2014.
39. Mirmiran, P., Bahadoran, Z., Hosseinpanah, F., Keyzad, A., and Azizi, F., Effects of broccoli sprout
with high sulforaphane concentration on inflammatory markers in type 2 diabetic patients: A random-
ized double-blind placebo-controlled clinical trial. J. Funct. Foods, 4, 837–841, 2012.
40. Bahadoran, Z., Mirmiran, P., Yeganeh, M.Z., Hosseinpanah, F., Zojaji, H., and Azizi, F., Complementary
and alternative medicinal effects of broccoli sprouts powder on Helicobacter pylori eradication rate in
type 2 diabetic patients: A randomized clinical trial. J. Funct. Foods, 7, 390–397, 2014.
41. Smith, T.K., Lund, E.K., Clerke, R.G., Bennett, R.N., and Johnson, I.T., Effects of Brussels sprout juice
on the cell cycle and adhesion of human colorectal carcinoma cells (HT29) in vitro. J. Agric. Food
Chem., 53, 3895–3901, 2005.
42. Smith, T.K., Mithen, R., and Johnson, I.T., Effects of Brassica vegetable juice on the induction of apop-
tosis and aberrant crypt foci in rat colonic mucosal crypts in vivo. Carcinogenesis, 24, 491–495, 2003.
43. Masci, A., Mattioli, R., Costantino, P., Baima, S., Morelli, G., Punzi, P., Giordana, C., Pinto, A., Donini,
L.M., d’Erme, M., and Mosca, L., Neuroprotective effect of Brassica oleraceae sprouts crude juice in a
cellular model of Alzheimer’s disease. Ox. Med. Cell. Long., 781938, 2015.
44. Schnäbele, K., Brivida, K., Bub, A., Roser, S., and Pool-Zobel, B.L., Effects of carrot and tomato juice
consumption on faecal markers relevant to colon carcinogenesis in humans. Br. J. Nutr., 99, 606–613,
2008.
45. Chung, M.J., Hee, S.H., and Sung, N.J., Inhibitory effect of whole strawberries, garlic juice or kale juice
on endogenous formation of N-nitrosodimethylamine in humans. Cancer Lett., 182, 1–10, 2002.
Section IV
Caffeinated Beverages: Tea,

Coffee, and Cocoa/Chocolate
50
Teas (Green, Oolong, and Black)
Rui Jiao, Jingnan Chen, Yu Huang, and Zhen-Yu Chen
CONTENTS
50.1 Introduction................................................................................................................................... 629
50.3.1 Polyphenols in Tea.............................................................................................................631
50.3.2 Alkaloids in Tea................................................................................................................ 633
50.4 Health Effects................................................................................................................................ 633
50.4.1 Bioactivity......................................................................................................................... 633
50.4.1.1 Vasorelaxant Activity........................................................................................ 633
50.4.1.2 Hypotensive Activity........................................................................................ 634
50.4.1.3 Hypolipidemic Activity.................................................................................... 635
50.4.1.4 Antibacterial and Antiviral Activity................................................................. 635
50.4.1.5 Anti-Inflammatory Activity.............................................................................. 636
50.4.2 Chronic Diseases.............................................................................................................. 636
50.4.2.1 Cardiovascular Disease..................................................................................... 636
50.4.2.2 Cancer............................................................................................................... 636
50.4.2.3 Neurodegenerative Diseases............................................................................. 637
50.4.2.4 Other Health Benefits....................................................................................... 637
50.5 Tea Products/Formulations and Future Trends............................................................................. 637
50.6 Conclusion..................................................................................................................................... 639
References............................................................................................................................................... 639
50.1 Introduction
Tea is popularly consumed worldwide, second to water, and more than coffee and carbonated soft
drinks. As a most popular beverage worldwide, tea is mainly divided into three types based on the
degree of fermentation during manufacturing, namely, unfermented green tea, totally fermented black
tea, and partially fermented oolong tea. Green tea is consumed primarily in China, Japan, and India,
whereas black tea is consumed primarily in Western countries. Oolong tea production and consump-
tion are primarily confined to southeastern mainland China and Taiwan. White tea, yellow tea, and
postfermented tea such as pu-erh tea also become popular in recent years. Each type of tea has its own
processing method, which makes them different by quality characteristics, such as appearance, color,
aroma, and flavor.
Chemistry of tea is very complex. Tea contains over 4000 phytochemicals, some of which are bio-
active. The most important bioactive ingredients present in tea are a group of polyphenols, which are
believed to exert a diversity of beneficial effects, including the preventions of hypertension, atheroscle-
rosis, diabetes, hypercholesterolemia, and obesity. These phenolic compounds possess the antioxidative,
antithrombotic, anti-inflammatory, hypotensive, and hypocholesterolemic properties [1]. Among these
629
polyphenols, catechins are present in higher quantities in green tea than in black or oolong tea. This
chapter focuses on nutritional characteristics, bioactivities and antioxidant efficacy, health effects, novel
products/formulations, and future trends of green, oolong, and black teas.

Nutritional properties and chemical composition of tea have been well documented. Tea is a naturally
refreshing drink with almost no calories and no lipids but a good source of potassium and vitamin B
(Table 50.1) [2]. A serving of 8 oz of brewed tea (black, prepared) can provide above 80 mg potassium,
and 8 oz of powder tea (prepared) or commercial ice tea can provide about 40 mg potassium. According
to the current U.S. guidelines of 4700 mg Recommended Dietary Allowances (RDA) for an adult, 80 mg
would be about 1.7% of the RDA for potassium. Tea also provides some amounts of other minerals such
as calcium, manganese, fluoride, and phosphorus. B-complex vitamins are abundant in tea, such as
TABLE 50.1
Compositional and Nutritional Characteristics of Different Teas (per 100 g)
Brewed Tea Commercial Nestle
Nutrient Unit Tea Powder Prepared (Black) Powder Tea Prepared Ice Tea
Water g 5.09 99.70 99.62 91.18
Protein g 20.21 0.00 0.06 0.00
Lipid (fat) g 0.00 0.00 0.00 0.00
Carbohydrate g 58.66 0.30 0.17 9.09
Total sugars g 5.53 0.00 0.02 9.09
Minerals
Calcium mg 118 0 3 3
Iron mg 2.26 0.02 0.01 0.00
Phosphorus mg 239 1 1 36
Potassium mg 6040 37 18 19
Sodium mg 72 3 4 21
Zinc mg 1.69 0.02 0.01 0.06
Vitamins
Folate (DFE) µg 103 5 0 nr
Niacin mg 10.800 0.000 0.032 nr
Riboflavin mg 0.985 0.014 0.003 nr
Thiamin mg 0.000 0.000 0.000 nr
Vitamin A (REA) μg 0 0 0 nr
Vitamin B6 mg 0.356 0.000 0.001 nr
Vitamin C mg 0.0 0.0 0.0 nr
Vitamin E (ATE) mg 0.00 0.00 0.00 nr
Vitamin K μg 0.0 0.0 0.0 nr
Other
Caffeine mg 5714 20 11 3
Reference, Release 26, 2013, Published online at http://ndb.nal.usda.gov/ndb/foods (accessed January 24, 2014).
Abbreviations: DFE, dietary folate equivalents; RAE, retinol activity equivalents; ATE, alpha-tocopherol equivalents;
nr, not reported.
Teas (Green, Oolong, and Black) 631
riboflavin, niacin, B6, and folate. It has been estimated that six cups of natural tea may account for 10%
of the RDA for B-complex vitamins [3]. Vitamin C is also present in green tea but absent or little in black
tea and oolong tea due to degradation during fermentation [4].
Tea also contains many free amino acids. Amino acid degradation is involved in the biogenesis of the
tea aroma. Wang et al. [5] introduced solid-phase extraction combined with high-performance liquid
chromatography–diode array detector (HPLC-DAD) for the determination of free amino acids in different
Chinese teas, finding that green tea contained much higher amounts of free amino acids than fermented
ones. Theanine is a nonprotein amino acid found in tea, representing as much as 50% of the total free
amino acids in black tea and 1%–2% in green tea [5,6]. It not only plays an important role in the character-
istic flavor and delicate taste of tea but also has several biological functions, such as promoting relaxation,
inhibiting caffeine’s negative effects, and reducing blood pressure as well as enhancing antitumor activity
[7,8]. In addition, theanine has been reported to exhibit antiobesity and neuroprotective activities [9,10].

50.3.1 Polyphenols in Tea
Polyphenols in tea account for 25%–35% on a dry weight (Table 50.2) [11]. In addition to phenolic
acids, several groups of polyphenols have been identified in tea including flavanols, flavonols, antho-
cyanins, and flavones [12]. Among these flavanols, a group of catechins, namely, (–)-epicatechin (EC),
(–)-epicatechin gallate (ECG), (–)-epigallocatechin (EGC), and (–)-epigallocatechin gallate (EGCG)
exist in a large quantity [13]. In addition, flavonols, mainly quercetin, kaempferol, myricetin, and
their glycosides are also present in tea; however, their quantities are much smaller compared with
catechins [14]. Green tea is an excellent source of dietary antioxidants as it contains four epicate-
chin derivatives: EGCG, EGC, ECG, and EC (Figure 50.1). It is believed that the most active and
abundant catechin in green tea is EGCG [15]. Black tea is also rich in antioxidants such as EGCG,
EGC, ECG, and EC. However, it contains much lower concentrations of these catechins compared
to green tea [4]. The major polyphenols in black tea are thearubigins and theaflavins, two types of
complex polyphenols. Theaflavins in black tea consist of four dimeric catechins including theaflavin-1
(TF1), theaflavin-3-gallate (TF2A), theaflavin-3′-gallate (TF2B), and theaflavin-3,3′-digallate (TF3) [16]
(Figure 50.2). In contrast, oolong tea contains a lower concentration of polyphenols than green tea, but
it has a higher concentration of polyphenols than traditional black tea. Oolong tea also uniquely con-
tains theasinensins, which are a group of tea polyphenols different from green tea catechins and black
tea theaflavins (Figure 50.3) [17,18].
TABLE 50.2
Polyphenols in Green, Black, and Oolong Teas
Green Tea [15,18] Black Tea [16,18] Oolong Tea [17,18]
Gallic acid Gallic acid Gallic acid
(–)-Epigallocatechin (–)-Epigallocatechin (–)-Epigallocatechin
(–)-Epicatechin (–)-Epicatechin (–)-Epicatechin
(–)-Epigallocatechin gallate (–)-Epigallocatechin gallate (–)-Epigallocatechin gallate
(–)-Gallocatechin gallate (–)-Gallocatechin gallate (–)-Gallocatechin gallate
(–)-Epicatechin gallate (–)-Epicatechin gallate (–)-Epicatechin gallate
(–)-Gallocatechin Theaflavin-1 (–)-Gallocatechin
(–)-Catechin gallate Theaflavin-3-gallate Theasinensin A
(–)-Catechin Theaflavin-3′-gallate Theasinensin B
Theaflavin-3,3′-digallate Theasinensin C
Thearubigins Theasinensin D
Theasinensin E
OH
OH
HO O
R1
OR2
OH
FIGURE 50.1 Structures of green tea catechins: (–)-epicatechin: R1=R 2=H; (–)-epigallocatechin: R1=OH, R 2=H;
(–)-epicatechin gallate: R1=H, R 2=galloyl; and (–)-epigallocatechin gallate: R1=OH, R 2=galloyl.
OH
H
OR1
OH
HO O
H
O
H
HO O OH
OR2 OH
H
OH
FIGURE 50.2 Structures of black theaflavins: Theaflavin-1: R1=R2=H; theaflavin-3-gallate-A: R1=galloyl, R2=H;
theaflavin-3′-gallate-B: R1=H, R2=galloyl; and theaflavin-3,3′-digallate: R1=R2=galloyl.
OH OH
R 1O R3O
HO HO
O OH O
OH OH OH
OH OH
HO HO
OH OH
HO O HO O
OH OH
OR2 OR4
OH OH
R-configuration S-configuration
FIGURE 50.3 Structures of oolong tea theasinensins: Theasinensins A: R1=R2=galloyl; theasinensins B: R1=galloyl,
R2=H; theasinensins C: R1=R2=H; theasinensins D: R3=R4=galloyl; and theasinensins E: R3=R4=H.
50.3.2 Alkaloids in Tea

Teas contain three purine alkaloids: theobromine, theophylline, and caffeine. They are all of methylx-
anthine class of chemical compounds. Caffeine present in tea varies by the type of tea, being higher in
black tea, ranging from 25 to 50 mg/100 mL, followed by oolong tea, ranging 12–22 mg/100 mL [4].
In contrast, green tea has a lesser amount of caffeine, ranging from 10 to 16 mg/100 mL [19]. At moderate
doses, caffeine increases self-reported alertness and improves attention and psychomotor performance.
Tea also contains small amounts of theobromine and theophylline with the latter having the ability to
relaxing smooth muscles, so it is used as a treatment for asthma.

Oxidative modification of low-density lipoprotein (LDL) is believed to play a key role in the develop-
ment of atherosclerosis. α-Tocopherol functions as a major antioxidant in human LDL. Zhu et al. [20]
demonstrated that green tea catechins protected α-tocopherol, four pure epicatechin derivatives showed
varying protective activity against depletion of α-tocopherol in LDL with EC and ECG being more
effective than EGC and EGCG. Leung et al. [21] compared the relative antioxidant activities between
black tea theaflavins and green tea catechins and revealed that the antioxidant activity is in the following
order: TF3 > ECG > EGCG, TF2B, TF2A > TF1, EC > EG. This study has suggested that theaflavins
present in black tea have similar antioxidant potency as green tea catechins, and that the conversion
of catechins into theaflavins during fermentation is unlikely to reduce their free radical scavenging
activity. Xu et al. [22] showed that catechin gallate and its precursor EGC share similar antioxidant
activity, while gallocatechin is less potent than its precursor EGC as an antioxidant. While, EGCG and
gallocatechin gallate (GCG) exhibit comparable antioxidant potency as reflected by LDL oxidation and
1,1-diphenyl-2-picrylhydrazyl (DPPH) radical assays. However, GCG is less effective than EGCG in the
ferric-reducing antioxidant power (FRAP) assay. For EC and catechin, the latter has less free radical
scavenging activity in DPPH assay, but in LDL oxidation and FRAP assay, their antioxidant activity is
comparable. A few studies are emphatic that tea consumption reduces the oxidative damages to DNA in
the human cells. In view, those smokers could have high oxidative stress, but green tea consumption for
7 days could appreciably reduce DNA damage among smokers [23].
50.4 Health Effects

50.4.1 Bioactivity
50.4.1.1 Vasorelaxant Activity
One of the various biological functions associated with tea is its vasorelaxant activity (Figure 50.4). Many
pathophysiological conditions in cardiovascular system are duo to endothelial dysfunction characterized
by attenuated production of protective vasoactive substances. One major vasoprotective molecule produced
by endothelial cells is nitric oxide (NO). We have previously studied the vasorelaxative effect of green tea
epicatechins in isolated rat mesenteric arteries, finding that four purified epicatechin derivatives relax rat
arteries precontracted by phenylephrine and endothelin 1 [24]. At higher concentrations, these four epi-
catechins reduce the sustained tension developed by membrane depolarization in the presence of elevated
extracellular KCl, but they seem not to alter the caffeine-induced transient contractile response, indicating
that these polyphenols may inhibit Ca2+ influx without interfering with internal Ca2+ mobilization. Further
studies have elucidated a significant contributory role of endothelium-derived NO in relaxations induced
by purified green tea EC in rat mesenteric arteries based on the following observations: (1) EC-induced
relaxation is much dependent on the presence of a functional endothelium; (2) this endothelium-dependent
relaxation is largely inhibited by NG-nitro-l-arginine methyl ester (L-NAME), a competitive inhibitor of
nitric oxide synthase (NOS), and this inhibition is partially reversed by pre-treatment with l-arginine,
the NOS substrate; and (3) the endothelium-dependent relaxation is also attenuated by methylene blue,
a tentative inhibitor of NO-dependent guanylate cyclase [25,26]. The endothelium-dependent effect of
SREBP2
HIV
Tea polyphenols HBV Dental caries
Ch LDLR IFV Tea extracts -Streptococcus mutans
sy oles and S. sobrinus
nt te
PI3K PDK Akt he ro
sis l
Cholesterol
Enteric Probiotics and prebiotics
Fatty acids pathogens -Bifidobacteria

FAS
Antibacterial Cancer
Hypolipidemic and antiviral
Tumors NOS
Cardiovascular Diseases
Tea Consumption Cytokines NF-KB

Vasorelaxant Anti-inflammatory
Tea polyphenols
Vascular smooth muscle cell
Hypotensive Antioxidant Certain
[ K+] chemicals UVB
GC cGMP PKG
+
K channels α-tocopherol LDL
Inflammation
NO +
2+
eNOS [ Ca ] Tea polyphenols O2
L-arginine
Vasodilation
Endothelial cell Free radicals
ACE Neurodegenerative
Tea polyphenols DNA damage Diseases
α1-ARs
smokers
FIGURE 50.4 Bioactivity and functions of tea.
epicatechin appears to be causally associated with its stimulatory action on the intracellular Ca2+ levels
in the endothelium, which is required for the activation of endothelial NOS. In addition, EC-induced
endothelium-dependent relaxation is partially mediated through NO-dependent activation of iberiotoxin-
sensitive K+ channels. In this regard, Hodgson et al. [27] reported that regular ingestion of black tea in
human results in a significant and consistent increase in endothelium-dependent dilatation, suggesting
that tea may reduce cardiovascular risk by improving vasodilator function.
50.4.1.2 Hypotensive Activity

The blood pressure–lowering activity of tea has been extensively investigated (Figure 50.4). Tea cat-
echins have been shown to lower blood pressure in most studies using animal models. When stroke-
prone spontaneously hypertensive rats were given water containing green tea polyphenols and black tea
polyphenols, systolic and diastolic blood pressure were significantly lowered compared with the control
group [28]. Oral administration of tea saponin could decrease mean blood pressure very effectively at
a dose of 100 mg/kg of body weight in SHR rats [29]. In type 2 diabetic Goto–Kakizaki rats, dietary
catechins could regulate blood glucose and systolic blood pressure at lower levels compared with the con-
trol diet [30]. Results from cell culture and animal experiments suggest that tea catechins reduce blood
pressure probably by the following mechanisms. First, tea catechins inhibit the angiotensin-converting
enzyme (ACE). When cultured endothelial cells from human umbilical veins were incubated with green
tea, black tea, and four individual catechins (EC, EGC, ECG, and EGCG) for 10 min, a significant and
dose-dependent inhibition of ACE activity was seen [31]. Second, the antihypertensive activity of tea cat-
echins is mediated by its action on endothelial NO production. When the effects of green and black tea on
NO production and vasodilation were compared in bovine aortic endothelial cells, both green and black
teas equally stimulated endothelial NOS (eNOS) activity and phosphorylation as well as vasorelaxation
[32]. Third, it has been shown that tea catechin-induced endothelium-dependent relaxation is primarily
mediated by NO and partially through NO-dependent activation of iberiotoxin-sensitive K+ channels [33].
Fourth, green tea has been shown to cause a dose-dependent depressor action in anesthetized rats at least
partly through the blockade of α1-adrenergic receptors [34]. Finally, tea contains γ-glutamylmethylamide
and theanine, which are known to have some antihypertensive activity [35,36].
Data from tea studies on blood pressure in humans are not consistent. In a study conducted in Taiwan,
Yang et al. [37] examined the effect of tea drinking on the risk of newly diagnosed hypertension in 1507
subjects with no previous hypertensive history and found that compared with nonhabitual tea drinkers,
the risk of developing hypertension decreased by 46% for those who drank 120–599 mL/day and was
further reduced by 65% for those who drank 600 mL/day or more. Another study investigated the
relationship of tea with systolic blood pressure and mortality from coronary heart disease (CHD), dem-
onstrating that systolic blood pressure was inversely related with tea with drops of 2.1 mmHg in men and
3.5 mmHg in women [38]. However, some studies did not observe such favorable effect. For instance, the
results from a clinical trial did not demonstrate that drinking tea for 6 weeks was associated with any
favorable change in blood pressure compared with placebo [39]. Taubert et al. [40] summarized the find-
ings of five studies of tea consumption involving a total 343 subjects with a median duration of 4 weeks
and concluded that tea intake had no significant effects on blood pressure. Furthermore, tea may have
an acute blood pressure–raising effect as demonstrated from one study, which found that blood pressure
was significantly increased by tea alone in comparison to each of three other groups: water alone, meal
with water, and meal with tea beverage [41]. This acute effect is probably attributable to caffeine, which
is known to have a mild hypertensive effect for a few hours after use [42]. It, therefore, appears that tea
has both antihypertensive and hypertensive components. The former include tea catechins, theaflavins,
γ-glutamylmethylamide, and theanine, whereas the latter is caffeine. As the evidence for the blood
pressure–lowering activity of tea in humans is mixed, further additional clinical randomized, double-
blind, and crossover studies are needed to resolve the issue.
50.4.1.3 Hypolipidemic Activity

The synthesis of fatty acids is the key step for lipogenesis. Oolong, black, pu-erh, and green tea leaves
could decrease plasma triacylglycerols (TG), total cholesterol (TC), and LDL cholesterol in the experi-
mental animals [43]. Results showed that reduction in body weights by tea was in the order of oolong
tea > pu-erh tea > black tea > green tea. Pu-erh tea and oolong tea could lower plasma TG concentra-
tion more significantly than that of green and black teas, but pu-erh and green teas were more efficient
than oolong and black teas in lowering plasma TC. The molecular mechanism by which tea polyphe-
nols (EGCG, theaflavins) suppress the fatty acid synthase gene may be mediated by downregulation of
EGFR/PI3K/Akt/Sp-1 signal transduction pathways [44].
The cholesterol-lowering activity of tea catechins has been extensively investigated. Although the
mechanisms responsible for the cholesterol-lowering activity of tea catechins are not yet fully under-
stood, some evidence suggests that they reduce blood TC probably by the following mechanisms. First,
they upregulate the LDL receptor mediated by activation of sterol regulatory element-binding protein 2
[45]. In rats fed with a diet containing 2% tea catechins, LDL receptor-binding activity and its protein
mass were increased by 2.7- and 3.4-fold, respectively [46]. Second, tea catechins reduce the plasma
TC by increasing fecal bile acid and cholesterol excretion [47]. Third, tea catechins have been shown to
inhibit cholesterol synthesis in rabbits [46] but not in rats [47].
Data from studies on the cholesterol-lowering activity of tea catechins in humans are also supportive.
Epidemiological observations indicate that tea consumption is associated with reduced levels of plasma
TC and LDL cholesterols in Japanese [48] and Norwegian subjects [38]. One study demonstrated that
theaflavin-rich tea at a dose of 375 mg a day effectively reduced TC and LDL cholesterols in mild to
moderate hypercholesterolemia subjects [49].
50.4.1.4 Antibacterial and Antiviral Activity

Black, green, and pu-erh teas are able to inhibit the growth of both “Gram-positive” and “Gram-
negative” bacteria that are pathogenic to humans. Gram-positive bacteria are more sensitive than
Gram-negative bacteria to tea [50]. Tea inhibits enteric pathogens such as Staphylococcus aureus, S. epi-
dermidis, Plesiomonas shigelloides, Salmonella typhi, S. typhimurium, S. enteritidis, Shigella flexneri,
S. dysenteriae, Vibrio cholerae, V. parahaemolyticus, Helicobacter pylori, Campylobacter jejuni, and

C. coli [51–53], but they may not be effective in inhibiting Escherichia coli, Pseudomonas aeruginosa,
or Aeromonas hydrophila [50]. Relationship between consumption of tea with probiotics and prebiotics
has been the subject of some studies. In this regard, tea catechins may favor the growth of bifidobacteria
in large colon, possibly attributable to their low pH value. Association of tea consumption with low colon
cancer risk is probably due to the reduction in formation of bacteria-derived harmful compounds at low
pH conditions [54]. Drinking tea without sugar may inhibit the formation of dental caries caused by
mouth bacteria. It has been shown that tea catechins and tea volatile compounds can inhibit the growth
of bacteria causing tooth decay such as Streptococcus mutans and S. sobrinus [55,56]. Tea may help to
fight again flu due to its antiviral activity [57]. Previous research has demonstrated that tea, particularly
its active ingredient EGCG, has an inhibiting activity against hepatitis B virus probably due to suppres-
sion on viral replication [58].
50.4.1.5 Anti-Inflammatory Activity

Green tea polyphenols possess an anti-inflammatory activity both in vitro and in vivo. EGCG has been
shown to modulate the signal transduction pathway involved in inflammation and joint destruction
against arthritis [59]. Green tea polyphenols also have an activity against inflammation caused by cer-
tain chemicals or ultraviolet radiation B [60]. It has been shown that green tea polyphenols are associated
with low production of cytokines induced by tumors [61]. Research has demonstrated that TF3 from black
tea blocks the NO synthase by downregulating the activation of NF-kappaB in macrophages [62].
50.4.2 Chronic Diseases

50.4.2.1 Cardiovascular Disease
Effect of tea consumption on cardiovascular disease (CVD) has been the subject of many epidemiologic
studies without a definite conclusion. Most published studies have demonstrated that the green tea con-
sumption is associated with reduction in cardiovascular risk. The Ohsaki National Health Insurance
Cohort Study, which followed 40,530 Japanese adults for 11 years, demonstrated that green tea consump-
tion was inversely associated with mortality of CVD with stronger inverse association in women [63].
A case control study from China also suggested a favorable effect of green and oolong tea consumption
against ischemic stroke [64]. Recently, a very large prospective study involving 76,979 Japanese adults
examined the relationship between cardiovascular mortality and consumption of several different types
of tea, reporting a strong inverse relationship between cardiovascular mortality and consumption of more
than six cups of green tea per day [65]. The study also found that consumption of more than one cup of
oolong tea per day was also associated with a reduced risk in CVD.
Studies performed in Europe and the United States suggested a benefit from consumption of black tea.
In the determinants of Myocardial Infarction Onset Study, individuals consuming more than an average
of two cups of black tea per day had lower total and cardiovascular mortality during 3.8-year follow-
up compared to individuals consuming no or less tea [66]. A large cohort of 37,514 participants in the
Netherlands was prospectively followed for 13 years with end points of CVD morbidity and mortality [67].
This study demonstrated that consumption of tea (mainly black tea), 3–6 cups daily, was associated with
a reduced risk of CVD mortality. However, several epidemiological studies failed to show an association
between tea consumption and CVD. One meta-analysis of tea consumption in relation to stroke, myocardial
infarction, and all CHD was based on 10 cohort studies and seven case control studies, suggesting that
the geographic regions where the studies were conducted were a factor showing that tea consumption is
not always associated with positive beneficial effect [68].
50.4.2.2 Cancer
The role of tea as a cancer chemopreventive agent has been studied during the last 20 years. Epidemiological
and animal studies have shown an inverse association of tea consumption with the development of
certain cancer types. It has now been suggested that tea polyphenols can potently induce apoptotic cell
death and cell cycle arrest in tumor cells, but not in their normal cell counterparts by affecting several
biological pathways [69]. As supporting evidence, various animal studies have revealed that when tea is
given as only source of drinking solution, tumor incidence and multiplicity decreased in different organ
sites such as skin, lung, esophagus, stomach, liver, small intestine, pancreas, colon, bladder, prostate, and
mammary glands [70].
Tea consumption associated with reduction in cancers has been well documented in oriental coun-
tries, but not always with Western countries. A case control study carried out in Shanghai demonstrated
frequent tea drinkers had a lower incidence of esophageal cancer [71]. It has been shown that Japanese
women who daily consumed 10 cups of tea had lower risk for all cancers particularly low risk breast
cancer metastasis and recurrence [72]. In contrast, the Netherlands Cohort Study on Diet and Cancer
found that consumption of black tea had no clear effect on the risk for stomach, colorectal, lung, and
breast cancers [73]. An interesting observation is that most studies from Asian countries show beneficial
effect of green tea, whereas studies of black tea from European countries are inconclusive. One of the
explanations is that green tea is probably stronger than black tea against cancers. The second explanation
is that life style in different regions may interact differently with tea consumption, having the different
etiological cancer factors [70].
50.4.2.3 Neurodegenerative Diseases

Neurodegenerative disorders such as Alzheimer’s and Parkinson’s disease are age-related diseases that
are associated with protein aggregation, reactive oxygen species (ROS) production, oxidative damage,
mitochondrial dysfunction, and cell death. Human epidemiological and animal data suggest that drink-
ing green and black teas may help protect the aging brain and reduce the incidence of neurodegenerative
diseases. Alzheimer’s are characterized by intracellular aggregation of the Tau protein and extracellular
aggregation of amyloid beta peptides (Aβs). Rezai-Zadeh et al. [74] demonstrated that green tea reduced
Aβ production/aggregation in mice whose amyloid protein precursors (APP)/Aβ was overexpressed.
Similarly, Lee et al. [75] found that EGCG could reduce the Aβ aggregation and prevent neuron cell
death in Alzheimer’s disease. In addition to polyphenols, caffeine in tea has also some impact in brain
enhancing cognitive function in humans. There are some epidemiological and animal studies that suggest
that moderate intake of caffeine may delay or reduce the risk of Alzheimer’s disease [76]. In addition to
tea polyphenols and caffeine, theanine in tea also has decisive impact in brain function of humans [77].
Takeda et al. [78] demonstrated that theanine could counteract excitotoxicity and mitochondrial radical
formation, increase the dentate granule cell neurogenesis, and increase the recognition memory in the
developing hippocampal formation.
50.4.2.4 Other Health Benefits

Drinking tea may have some antiobesity activity. Oolong tea has been shown to prevent obesity and
fatty liver in diet-induced obese mice [79]. Green tea could induce additional brown adipose tissue ther-
mogenesis in addition to caffeine alone [80]. Among women with 65–76 years of age, tea consumption
was also associated with greater bone mineral density [81]. Tea was found to inhibit glucosyltransferase
activity of oral streptococci and the development of dental caries in rats [82]. As it contains fluoride, tea
may strengthen tooth enamel, improve dental health, and inhibit the development of dental caries [82].
50.5 Tea Products/Formulations and Future Trends

Tea is widely consumed all over the world. The main tea-producing countries according to FAOSTAT
2012 in descending order are China, India, Kenya, Sri Lanka, Turkey, Viet Nam, Iran, Indonesia,
Argentina, Japan, Thailand, Bangladesh, Malawi, Uganda, Tanzania, Myanmar, Rwanda, Mozambique,
Zimbabwe, and Nepal. Teas are mainly categorized into green, oolong, and black, regardless of various
brand names as described in Table 50.3.
TABLE 50.3
Selected Popular Brands of Teas in the World
Brand Names of Tea Tea Types Origin
Long Jing Green tea Zhejiang, China
Bi Luo Chun Green tea Jiangsu, China
Huangshan Maofeng Green tea Anhui, China
Gua Pian Green tea Anhui, China
Wuyuan Ming Mei Green tea Jiangxi, China
Shangrao Baimei Green tea Jiangxi, China
Jinggang Cuilv Green tea Jiangxi, China
Taiping Houkui Green tea Anhui, China
Twailay Green tea Anhui, China
Xinyang Maojian Green tea Henan, China
Enshi YuLu Green tea Hubei, China
Qi Hong (Keemun) Black tea Anhui, China
Zhengshan Xiaozhong (Lapsang Souchong) Black tea Fujian, China
Dian Hong Black tea Yunnan, China
Ying De Hong Black tea Guangdong, China
Tie Guan Yin Oolong tea Fujian, China
Da Hong Pao Oolong tea Fujian, China
Wuyi Shuixian Oolong tea Fujian, China
Fenghuang Dancong Oolong tea Guangdong, China
Dongding Oolong Oolong tea Taiwan, China
Pu erh Post fermented tea Yunnan, China
Liubao Post fermented tea Guangxi, China
Liu An Post fermented tea Anhui, China
Baihao Yinzhen (Silver Pekoe) White tea Fujian, China
Baimudan (White Peony) White tea Fujian, China
Shoumei (White Eyebrows) White tea Fujian, China
Junshan Yinzhen Yellow tea Hunan, China
Huoshan Huangya Yellow tea Anhui, China
Anhui Huangdacha Yellow tea Anhui, China
Himalaya Green Green tea India
Wagh Bakri Green Green tea India
Brooke Bond Taj Mahal Green tea India
Assam Black tea India
Darjeeling Black tea India
Munnar Black tea India
Kangra Green tea, black tea India
Nilgiri Black tea India
Matcha Green tea Japan
Gyokuro Green tea Japan
Sencha Green tea Japan
Matcha Green tea Japan
Ceylon Black tea Sri Lanka
Dilmah Black tea Sri Lanka
Betjeman and Barton Black tea France
Kusmi Green tea, black tea France
Mariage Frères Spiced tea France
Republic of Tea Black tea The United States
(Continued)

Selected Popular Brands of Teas in the World
Brand Names of Tea Tea Types Origin
Teavana Black tea, oolong tea The United States
Twinings Green tea, black tea The United Kingdom
Whittard of Chelsea Black tea The United Kingdom
Zealong Oolong tea, black tea New Zealand
Kenya Black tea Kenya
Thai Black tea Thailand
Rize (Çay) Black tea Turkey
Krasnodar Black tea Russia
Barry’s Green tea, black tea Ireland
Lipton Green tea, black tea Globally
Tetley Green tea, black tea Globally
Commercial tea beverages are usually bottled with addition of sugar or sweeteners and other addi-
tives including citric acid and synthetic antioxidant such as butylated hydroxytoluene (BHT). It should
be pointed out that the content of catechins or theaflavins, if black tea is starting material, is very low
in bottled commercial tea drinks, probably being only one-tenth of that homemade tea. A brewed cup
of natural tea without any formulation contains excellent source of polyphenol antioxidants and also a
moderate amount of caffeine, volatile oils, tannin, and several B-complex vitamins. Differing from for-
mulated tea beverages, each type of home-brewed teas has its unique flavor and aroma as the profile of
volatiles is somehow different from others. As the green tea undergoes lesser chemical changes during
the processing, its astringency and color are much lighter in proportion to its lesser tannin content. In
addition to its sliming activity, home-brewed tea gives a little energy with having less than 4 calories per
cup compared with coca cola and other soft drinks. Cautions must be given to the formulated tea bever-
ages when sugar and milk are added, a cup of tea may have 40 calories. Vitamin content and profile vary
with type of tea as these nutrients are susceptible to degradation [3]. It is obvious the naturally brewed
tea is preferred to the formulated commercial tea beverages.
50.6 Conclusion
Drinking tea is becoming popular worldwide. We begin to understand the molecular mechanisms by
which tea active ingredients such as catechins and theaflavins render such beneficial activity in humans.
Tea, regardless of being green or black, will continue to be a healthy beverage either homemade or
industry-made bottled drinks. In addition, catechins and theaflavins find more applications as health
supplements to treat and prevent chronic diseases including cardiovascular, hypertension, diabetes, obe-
sity, and neurodegenerative diseases.
REFERENCES
1. Yung, L.M., Leung, F.P., Wong, W.T., Tian, X.Y., Yung, L.H., Chen, Z.Y., Yao, X.Q., and Huang, Y., Tea
polyphenols benefit vascular function. Inflammopharmacology, 16, 230–234, 2008.
2. U.S. Department of Agriculture (USDA), USDA National Nutrient Standard Reference, Release 26,
2013. Published online at http://ndb.nal.usda.gov/ndb/foods (accessed January 24, 2014).
3. Hicks, A., Current status and future development of global tea. AU. J. Technol., 12, 251–264, 2009.
4. Cabrera, C., Artacho, R., and Gimenez, R., Beneficial effects of green tea—A review. J. Am. Coll. Nutr.,
25, 79–99, 2006.
5. Wang, L., Xu, R., Hu, B., Li, W., Sun, Y., Tu, Y., and Zeng, X., Analysis of free amino acids in Chinese
teas and flower of tea plant by high performance liquid chromatography combined with solid-phase
extraction. Food Chem., 123, 1259–1266, 2010.
6. Hara, Y., Luo, S.J., Wickremasinghe, R.L., and Yamanishi, T., Tea. Food Rev. Int., 11, 371–542, 1995.
7. Kimura, K., Ozeki, M., Juneja, L.R., and Ohira, H., l-Theanine reduces psychological and physiological
stress responses. Biol. Psychol., 74, 39–45, 2007.
8. Sugiyama, T. and Sadzuka, Y., Theanine and glutamate transporter inhibitors enhance the antitumor
efficacy of chemotherapeutic agents. Biochim. Biophys. Acta., 1653, 47–59, 2003.
9. Egashira, N., Hayakawa, K., Mishima, K., Kimura, H., Iwasaki, K., and Fujiwara, M., Neuroprotective
effect of gamma-glutamylethylamide (theanine) on cerebral infarction in mice. Neurosci. Lett., 363,
58–61, 2004.
10. Zheng, G., Bamba, K., Okubo, T., Juneja, L.R., Oguni, I., and Sayama, K., Effect of theanine, gamma-
glutamylethylamide, on bodyweight and fat accumulation in mice. Anim. Sci. J., 76, 153–157, 2005.
11. Balentine, D.A., Tea and health. Crit. Rev. Food Sci., 37, 691–692, 1997.
12. Mukhtar, H. and Ahmad, N., Tea polyphenols: Prevention of cancer and optimizing health. Am. J. Clin.
Nutr., 71(Suppl. 6), 1698S–1702S, 2000.
13. Hara, Y., Tea catechins and their applications as supplements and pharmaceutics. Pharmacol. Res., 64,
100–104, 2011.
14. Chaturvedula, V.S.P. and Prakash, I., The aroma, taste, color and bioactive constituents of tea. J. Med.
Plants Res., 5, 2110–2124, 2011.
15. Reto, M., Figueira, M.E., Filipe, H.M., and Almeida, C.M.M., Chemical composition of green tea
(Camellia sinensis) infusions commercialized in Portugal. Plant Foods Hum. Nutr., 62, 139–144, 2007.
16. Peng, C., Chan, H.Y.E., Li, Y.M., Huang, Y., and Chen, Z.Y., Black tea theaflavins extend the lifespan of
fruit flies. Exp. Gerontol., 44, 773–783, 2009.
17. Hou, D.X., Masuzaki, S., Tanigawa, S., Hashimoto, F., Chen, J., Sogo, T., and Fujii, M., Oolong tea
theasinensins attenuate cyclooxygenase-2 expression in lipopolysaccharide (LPS)-activated mouse
macrophages: Structure-activity relationship and molecular mechanisms. J. Agric. Food Chem., 58,
12735–12743, 2010.
18. Chen, Z.Y., Zhu, Q.Y., Tsang, D., and Huang, Y., Degradation of green tea catechins in tea drinks.
J. Agric. Food Chem., 49, 477–482, 2001.
19. Chin, J.M., Merves, M.L., Goldberger, B.A., Sampson-Cone, A., and Cone, E.J., Caffeine content of
brewed teas. J. Anal. Toxicol., 32, 702–704, 2008.
20. Zhu, Q.Y., Huang, Y., Tsang, D., and Chen, Z.Y., Regeneration of alpha-tocopherol in human low-density
lipoprotein by green tea catechin. J. Agric. Food Chem., 47, 2020–2025, 1999.
21. Leung, L.K., Su, Y.L., Chen, R.Y., Zhang, Z.H., Huang, Y., and Chen, Z.Y., Theaflavins in black tea and
catechins in green tea are equally effective antioxidants. J. Nutr., 131, 2248–2251, 2001.
22. Xu, J.Z., Yeung, S.Y.V., Chang, Q., Huang, Y., and Chen, Z.Y., Comparison of antioxidant activity and
bioavailability of tea epicatechins with their epimers. Br. J. Nutr., 91, 873–881, 2004.
23. Lee, I.P., Kim, Y.H., Kang, M.H., Roberts, C., Shim, J.S., and Roh, J.K., Chemopreventive effect of
green tea (Camellia sinensis) against cigarette smoke-induced mutations (SCE) in humans. J. Cell
Biochem., 27(Suppl. 27), 68S–75S, 1997.
24. Huang, Y., Zhang, A.Q., Lau, C.W., and Chen, Z.Y., Vasorelaxant effects of purified green tea epicat-
echin derivatives in rat mesenteric artery. Life Sci., 63, 275–283, 1998.
25. Huang, Y., Chan, N.W.K., Lau, C.W., Yao, X.Q., Chan, F.L., and Chen, Z.Y., Involvement of endothe-
lium/nitric oxide in vasorelaxation induced by purified green tea (−)-epicatechin. Biochim. Biophys.
Acta., 1427, 322–328, 1999.
26. Huang, Y., Yao, X.Q., Tsang, S.Y., Lau, C.W., and Chen, Z.Y., Role of endothelium/nitric oxide in
vascular response to flavonoids and epicatechin. Acta. Pharmacol. Sin., 21, 1119–1124, 2000.
27. Hodgson, J.M., Puddey, I.B., Burke, V., Watts, G.F., and Beilin, L.J., Regular ingestion of black tea
improves brachial artery vasodilator function. Clin. Sci., 102, 195–201, 2002.
28. Negishi, H., Xu, J.W., Ikeda, K., Njelekela, M., Nara, Y., and Yamori, Y., Black and green tea polyphe-
nols attenuate blood pressure increases in stroke-prone spontaneously hypertensive rats. J. Nutr., 134,
38–42, 2004.
29. Sagesaka Mitane, Y., Sugiura, T., Miwa, Y., Yamaguchi, K., and Kyuki, K., Effect of tea-leaf saponin on
blood pressure of spontaneously hypertensive rats. Yakugaku Zasshi, 116, 388–395, 1996.
30. Igarashi, K., Honma, K., Yoshinari, O., Nanjo, F., and Hara, Y., Effects of dietary catechins on glu-
cose tolerance, blood pressure and oxidative status in Goto-Kakizaki rats. J. Nutr. Sci. Vitaminol., 53,
496–500, 2007.
31. Persson, I.A.L., Josefsson, M., Persson, K., and Andersson, R.G.G., Tea flavanols inhibit angiotensin-
converting enzyme activity and increase nitric oxide production in human endothelial cells. J. Pharm.
Pharmacol., 58, 1139–1144, 2006.
32. Lorenz, M., Urban, J., Engelhardt, U., Baumann, G., Stangl, K., and Stangl, V., Green and black tea are
equally potent stimuli of NO production and vasodilation: New insights into tea ingredients involved.
Basic Res. Cardiol., 104, 100–110, 2009.
33. Naseri, M.K.G., Arabian, M., Badavi, M., and Ahangarpour, A., Vasorelaxant and hypotensive effects
of Allium cepa peel hydroalcoholic extract in rat. Pak. J. Biol. Sci., 11, 1569–1575, 2008.
34. Lim, D.Y., Lee, E.S., Park, H.G., Kim, B.C., Hong, S.P., and Lee, E.B., Comparison of green tea extract
and epigallocatechin gallate on blood pressure and contractile responses of vascular smooth muscle of
rats. Arch. Pharm. Res., 26, 214–223, 2003.
35. Yokogoshi, H. and Kobayashi, M., Hypotensive effect of gamma-glutamylmethylamide in spontane-
ously hypertensive rats. Life Sci., 62, 1065–1068, 1998.
36. Yokogoshi, H., Kato, Y., Sagesaka, Y.M., Takiharamatsuura, T., Kakuda, T., and Takeuchi, N., Reduction
effect of theanine on blood pressure and brain 5-hydroxyindoles in spontaneously hypertensive rats.
37. Yang, Y.C., Lu, F.H., Wu, J.S., Wu, C.H., and Chang, C.J., The protective effect of habitual tea consump-
tion on hypertension. Arch. Intern. Med., 164, 1534–1540, 2004.
38. Stensvold, I., Tverdal, A., Solvoll, K., and Foss, O.P., Tea consumption relationship to cholesterol, blood
pressure, and coronary and total mortality. Prev. Med., 21, 546–553, 1992.
39. Steptoe, A., Gibson, E.L., Vounonvirta, R., Williams, E.D., Hamer, M., Rycroft, J.A., Erusalimsky, J.D.,
and Wardle, J., The effects of tea on psychophysiological stress responsivity and post-stress recovery:
A randomised double-blind trial. Psychopharmacology, 190, 81–89, 2007.
40. Taubert, D., Roesen, R., and Schoemig, E., Effect of cocoa and tea intake on blood pressure—A meta
analysis. Arch. Intern. Med., 167, 626–634, 2007.
41. Hodgson, J.M., Burke, V., and Puddey, I.B., Acute effects of tea on fasting and postprandial vascular
function and blood pressure in humans. J. Hypertens., 23, 47–54, 2005.
42. Birkett, N.J. and Logan, A.G., Caffeine-containing beverages and the prevalence of hypertension.
J. Hypertens., 6(Suppl. 4), S620–S622, 1988.
43. Kuo, K.L., Weng, M.S., Chiang, C.T., Tsai, Y.J., Lin-Shiau, S.Y., and Lin, J.K., Comparative studies on
the hypolipidemic and growth suppressive effects of oolong, black, pu-erh, and green tea leaves in rats.
J. Agric. Food Chem., 53, 480–489, 2005.
44. Lin, J.K. and Lin-Shiau, S.Y., Mechanisms of hypolipidemic and anti-obesity effects of tea and tea
polyphenols. Mol. Nutr. Food Res., 50, 211–217, 2006.
45. Kuhn, D.J., Burns, A.C., Kazi, A., and Dou, Q.P., Direct inhibition of the ubiquitin-proteasome pathway
by ester bond-containing green tea polyphenols is associated with increased expression of sterol regula-
tory element-binding protein 2 and LDL receptor. Biochim. Biophys. Acta., 1682, 1–10, 2004.
46. Bursill, C.A. and Roach, P.D., Modulation of cholesterol metabolism by the green tea polyphenol
(−)-epigallocatechin gallate in cultured human liver (HepG2) cells. J. Agric. Food Chem., 54, 1621–1626,
2006.
47. Yang, T.T.C. and Koo, M.W.L., Chinese green tea lowers cholesterol level through an increase in fecal
lipid excretion. Life Sci., 66, 411–423, 2000.
48. Kono, S., Shinchi, K., Ikeda, N., Yanai, F., and Imanishi, K., Green tea consumption and serum lipid
profiles a cross-sectional study in Northern Kyushu, Japan. Prev. Med., 21, 526–531, 1992.
49. Maron, D.J., Lu, G.P., Cai, N.S., Wu, Z.G., Li, Y.H., Chen, H., Zhu, J.Q., Jin, X.J., Wouters, B.C.,
and Zhao, J., Cholesterol-lowering effect of a theaflavin-enriched green tea extract—A randomized
controlled trial. Arch. Intern. Med., 163, 1448–1453, 2003.
50. Toda, M., Okubo, S., Hiyoshi, R., and Shimamura, T., The bactericidal activity of tea and coffee. Lett.
Appl. Microbiol., 8, 123–125, 1989.
51. Toda, M., Okubo, S., Ikigai, H., Suzuki, T., Suzuki, Y., and Shimamura, T., The protective activity of tea
against infection by Vibrio cholerae-01. J. App. Bacteriol., 70, 109–112, 1991.
52. Diker, K.S. and Hascelik, G., The bactericidal activity of tea against Helicobacter pylori. Lett. Appl.
Microbiol., 19, 299–300, 1994.
53. Diker, K.S., Akan, M., Hascelik, G., and Yurdakok, M., The bactericidal activity of tea against
Campylobacter jejuni and Campylobacter coli. Lett. Appl. Microbiol., 12, 34–35, 1991.
54. Okubo, T., Ishihara, N., Oura, A., Serit, M., Kim, M., Yamamoto, T., and Mitsuoka, T., In vivo effects
of tea polyphenol intake on human intestinal microflora and metabolism. Biosci. Biotech. Biochem., 56,
588–591, 1992.
55. Sakanaka, S., Kim, M., Taniguchi, M., and Yamamoto, T., Antibacterial substances in Japanese green
tea extract against Streptococcus mutans, a cariogenic bacterium. Agric. Biol. Chem., 53, 2307–2311,
1989.
56. Sakanaka, S., Sato, T., Kim, M., and Yamamoto, T., Inhibitory effects of green tea polyphenols on
glucan synthesis and cellular adherence of cariogenic streptococci. Agric. Biol. Chem., 54, 2925–2929,
1990.
57. Nakayama, M., Toda, M., Okubo, S., and Shimamura, T., Inhibition of influenza-virus infection by tea.
Lett. Appl. Microbiol., 11, 38–40, 1990.
58. Xu, J., Wang, J., Deng, F., Hu, Z., and Wang, H., Green tea extract and its major component epigallocat-
echin gallate inhibits hepatitis B virus in vitro. Antiviral Res., 78, 242–249, 2008.
59. Singh, R., Akhtar, N., and Haqqi, T.M., Green tea polyphenol epigallocatechin-3-gallate: Inflammation
and arthritis. Life Sci., 87, 196–196, 2010.
60. Agarwal, R., Katiyar, S.K., Khan, S.G., and Mukhtar, H., Protection against ultraviolet-B radiation-
induced effects in the skin of SKH-1 hairless mice by a polyphenolic fraction isolated from green tea.
Photochem. Photobiol., 58, 695–700, 1993.
61. Katiyar, S.K., Rupp, C.O., Korman, N.J., Agarwal, R., and Mukhtar, H., Inhibition of
12-O-tetradecanoylphorbol-13-acetate and other skin tumor-promoter-caused induction of epidermal
interleukin1-alpha messenger-RNA and protein expression in sencar mice by green tea polyphenols.
J. Invest. Dermatol., 105, 394–398, 1995.
62. Lin, Y.L., Tsai, S.H., Lin-Shiau, S.Y., Ho, C.-.T., and Lin, J.K., Theaflavin-3,3′-digallate from black tea
blocks the nitric oxide synthase by down-regulating the activation of NF-kappaB in macrophages. Eur.
J. Pharmacol., 367, 379–388, 1999.
63. Kuriyama, S., Shimazu, T., Ohmori, K., Kikuchi, N., Nakaya, N., Nishino, Y., Tsubono, Y., and Tsuji, I.,
Green tea consumption and mortality due to cardiovascular disease, cancer, and all causes in Japan—
The Ohsaki Study. JAMA, 296, 1255–1265, 2006.
64. Liang, W., Lee, A.H., Binns, C.W., Huang, R., Hu, D., and Zhou, Q., Tea consumption and ischemic
stroke risk a case-control study in Southern China. Stroke, 40, 2480–2485, 2009.
65. Mineharu, Y., Koizumi, A., Wada, Y., Iso, H., Watanabe, Y., Date, C., Yamamoto, A. et al., Coffee,
green tea, black tea and oolong tea consumption and risk of mortality from cardiovascular disease in
Japanese men and women. J. Epidemiol. Community Health, 65, 230–240, 2011.
66. Mukamal, K.J., Maclure, M., Muller, J.E., Sherwood, J.B., and Mittleman, M.A., Tea consumption and
myocardial mortality after acute infarction. Circulation, 105, 2476–2481, 2002.
67. de Koning Gans, J.M., Uiterwaal, C.S.P.M., van der Schouw, Y.T., Boer, J.M.A., Grobbee, D.E.,
Verschuren, W.M.M., and Beulens, J.W.J., Tea and coffee consumption and cardiovascular morbidity
and mortality. Arterioscler. Thromb. Vasc. Biol., 30, 1665–1671, 2010.
68. Peters, U., Poole, C., and Arab, L., Does tea affect cardiovascular disease? A meta-analysis. Am. J.
Epidemiol., 154, 495–503, 2001.
69. Chen, D., Milacic, V., Chen, M.S., Wan, S.B., Lam, W.H., Huo, C., Landis-Piwowar, K.R., Cui, Q.C.,
Wali, A., Chan, T.H., and Dou, Q.P., Tea polyphenols, their biological effects and potential molecular
targets. Histol. Histopathol., 23, 487–496, 2008.
70. Yang, C.S., Chung, J.Y., Yang, G.Y., Chhabra, S.K., and Lee, M.J., Tea and tea polyphenols in cancer
prevention. J. Nutr., 130(Suppl. 2), 472S–478S, 2000.
71. Gao, Y.T., McLaughlin, J.K., Blot, W.J., Ji, B.T., Dai, Q., and Fraumeni, J.F., Reduced risk of esophageal
cancer-associated with green tea consumption. J. Natl. Cancer Inst., 86, 855–858, 1994.
72. Nakachi, K., Suemasu, K., Suga, K., Takeo, T., Imai, K., and Higashi, Y., Influence of drinking green
tea on breast cancer malignancy among Japanese patients. Jpn. J. Cancer Res., 89, 254–261, 1998.
73. Mukhtar, H., Consumption of black tea and cancer risk: A prospective cohort study. J. Natl. Cancer
Inst., 88, 768, 1996.
74. Rezai-Zadeh, K., Shytle, D., Sun, N., Mori, T., Hou, H., Jeanniton, D., Ehrhart, J. et al., Green tea
epigallocatechin-3-gallate (EGCG) modulates amyloid precursor protein cleavage and reduces cerebral
amyloidosis in Alzheimer transgenic mice. J. Neurosci., 25, 8807–8814, 2005.
75. Lee, J.W., Lee, Y.K., Ban, J.O., Ha, T.Y., Yun, Y.P., Han, S.B., Oh, K.W., and Hong, J.T., Green tea
(−)-epigallocatechin-3-gallate inhibits beta-amyloid-induced cognitive dysfunction through modifi-
cation of secretase activity via inhibition of ERK and NF-kappaB pathways in mice. J. Nutr., 139,
1987–1993, 2009.
76. Arendash, G.W., Schleif, W., Rezai-Zadeh, K., Jackson, E.K., Zacharia, L.C., Cracchiolo, J.R., Shippy,
D., and Tan, J., Caffeine protects Alzheimer’s mice against cognitive impairment and reduces brain
beta-amyloid production. Neuroscience, 142, 941–952, 2006.
77. Haskell, C.F., Kennedy, D.O., Milne, A.L., Wesnes, K.A., and Scholey, A.B., The effects of l-theanine,
caffeine and their combination on cognition and mood. Biol. Psychol., 77, 113–122, 2008.
78. Takeda, A., Sakamoto, K., Tamano, H., Fukura, K., Inui, N., Suh, S.W., Won, S.J., and Yokogoshi, H.,
Facilitated neurogenesis in the developing hippocampus after intake of theanine, an amino acid in tea
leaves, and object recognition memory. Cell. Mol. Neurobiol., 31, 1079–1088, 2011.
79. Han, L.K., Takaku, T., Li J., Kimura, Y., and Okuda, H., Anti-obesity action of oolong tea. Int. J. Obes.
Relat. Metab. Disord., 23, 98–105, 1999.
80. Dulloo, A.G., Seydoux, J., Girardier, L., Chantre, P., and Vandermander, J., Green tea and thermogen-
esis: Interactions between catechin-polyphenols, caffeine and sympathetic activity. Int. J. Obes. Relat.
Metab. Disord., 24, 252–258, 2000.
81. Hegarty, V.M., May, H.M., and Khaw, K.T., Tea drinking and bone mineral density in older women. Am.
J. Clin. Nutr., 71, 1003–1007, 2000.
82. Hamada, S., Ooshima, T., Fijiwara, T., Minami, T., and Kimura, S., Development of preventive mea-
sures based on the aetiology of dental caries: A review. Microb. Ecol. Health Dis., 9, 349–357, 1996.
51
Herbal Teas
Sha Li, Shu-Ke Li, Dong-Ping Xu, An-Na Li, and Hua-Bin Li
CONTENTS
51.1 Introduction................................................................................................................................... 645
51.3 Bioactivities and Antioxidant Efficacy......................................................................................... 647
51.3.2 Anti-Inflammatory Activity..............................................................................................651
51.3.3 Antiproliferative Activity................................................................................................. 652
51.3.4 Antimicrobial Activity..................................................................................................... 653
51.3.5 Immunological Efficacy................................................................................................... 653
51.3.6 Antiglycation Activity...................................................................................................... 654
51.3.7 Antimalarial Activity....................................................................................................... 654
51.3.8 Effects on Some Metabolic Enzymes............................................................................... 654
51.3.9 Other Benefits................................................................................................................... 655
51.4 Health Effects................................................................................................................................ 655
51.6 Conclusion..................................................................................................................................... 657
References............................................................................................................................................... 657
51.1 Introduction
Herbal teas, namely, herbal infusion or herbal tisane, which are commonly consumed beverages
brewed from the leaves, flowers, seeds, fruits, stems, or roots of plant species, other than Camellia
sinensis L., have been used for health promotion and disease prevention for thousands of years in
many countries, such as China, India, Japan, Thailand, Greece, and Turkey [1–4]. In China, espe-
cially, the history of using herbal teas may be as long as the usage of traditional Chinese medicines
(TCMs), and many TCMs are also consumed in the form of tea. A special kind of herbal infusion is
called cool tea (Liang cha in Chinese), which originated from South China and has been spread to
about 20 countries around the globe. It possesses the efficacies of clearing away heat, detoxification,
dewetting, moistening lungs, quenching thirst, relieving fever, alleviating pain, restoring strength,
modulating immunity, and reducing the risk of cardiovascular disease (CVD) and certain types of
cancer [5,6].
With increasing interest to achieve a healthy life, herbal teas have become popular as alternative to
caffeinated beverages during the past 2 decades [7]. The caffeine-free and comparatively low-tannin
status of herbal teas, combined with their potential health-promoting properties, most notably antioxi-
dant activity, contributes to their popularity [8]. This chapter focuses on the past and current research
about different kinds of herbal teas consumed widely in the world. The focus falls specifically on aspects
of nutritional characteristics, bioactivities and antioxidant efficacy, health effects, novel products/
formulations, and future trends.
645

These days, plant species commonly used as herbal teas may be derived from medicinal and food plants,
dietary supplement health food plants, and traditionally used non-Camellia plants [1]. The nutritional sub-
stances that have been reported in herbal teas are mainly micro- and macroelements, carbohydrate, vitamins,
minerals, fatty acids, and other bioactive components such as phenolics, essential oils, and terpenes.
Many research efforts have focused the micro- and macroelements, especially the minerals in herbal
teas [3,9–11]. The contents of micro- and macroelements of some herbal teas were studied [9]. The results
showed that the most abundant minerals present in all tea infusions were magnesium and phosphorus, in
the range of 981–3268 and 1292–2827 μg/g, respectively. The highest percentages of releases from the
teas to their infusions were noticed for magnesium, phosphorus, and copper (up to 60%), while the lowest
was devoted to iron. It indicated that the release of elements into tea infusions depends on whether they
are strongly bound to the organic matrix of tea or are more soluble in the solution. The daily intake of all
minerals from herbal tea infusions did not exceed the maximum permissible levels, hence not constituting
a health risk [9]. The contents of calcium, magnesium, iron, zinc, and copper in both dried herb samples
and prepared infusions of five herbs, namely, chamomile (flowers), mint (leaves), St. John’s worth (flow-
ers and leaves), sage (leaves), and nettle (leaves), were determined. The results showed that the contents
of individual elements in herbs and infusions depended on the type of raw material, as well as their origin.
Moreover, it was found that iron penetrated the herbal infusions to the lowest degree (4.4%–12.4%),
while copper did so to the highest (26.7%–50.7%) [10]. In another study, it was confirmed that most elements
in herbal tea powders were released into the infusions at different percentages depending on the types
of herbs [3]. Infusion of Gynostemma pentaphyllum contained minerals (such as magnesium, calcium,
and iron) at higher levels than those of C. sinensis and Morus alba. Furthermore, time parameter for the
minerals getting to both infusion and decoction indicated that higher concentrations were diffused into the
infusion at the 10th minute for copper, iron, and zinc, while at the 15th minute were copper, potassium,
and zinc, and at the 20th minute were copper, iron, and potassium. The results showed that 10th minute
was the optimum time for getting the minerals into the infusion, and it is apparent that herbal teas are good
sources of the minerals [11]. Minerals in some herbal teas are summarized in Table 51.1.
TABLE 51.1
Minerals in Certain Kinds of Herbal Teas
Organic
Chamomile St. John’s
Organic with Worth
Peppermint Lavender Chamomile Mint Flowers
(The United Peppermint (The United Flowers Leaves and Leaves
States) (Poland) States) (mg/100 (mg/100 (mg/100
Mineral (µg/g) (µg/g) (µg/g) mL) mL) mL) References
Aluminum 4.31 5.40 10.4 nr nr nr [9]
Barium 3.20 4.05 5.59 nr nr nr [9]
Calcium 3742 4457 3574 2.94 7.08 3.67 [9,10]
Copper 5.48 3.53 5.62 0.01 0.01 0.01 [9,10]
Iron 8.20 6.23 10.8 0.10 0.10 0.05 [9,10]
Magnesium 3268 2671 1908 3.20 2.97 1.28 [9,10]
Manganese 14.2 22.2 14.1 nr nr nr [9]
Nickel 1.16 2.69 2.37 nr nr nr [9]
Phosphorus 2827 2296 2114 nr nr nr [9]
Strontium 15.1 14.4 23.7 nr nr nr [9]
Titanium 0.07 0.03 <LOD nr nr nr [9]
Vanadium <LOD <LOD 0.15 nr nr nr [9]
Zinc 12.0 8.80 11.0 0.08 0.01 0.03 [9,10]
Abbreviations: nr, not reported; LOD, limit of detection.
Herbal Teas 647
Other nutrients in herbal teas such as carbohydrate, fatty acids, and vitamins have been investigated in
several studies. Yang et al. [12] characterized the water-soluble polysaccharides isolated from an herbal
tea, the leaves of Lycopus lucidus Turcz. High-performance liquid chromatography (HPLC) analysis
showed that L. lucidus polysaccharides were mainly composed of galactose (50.1%), followed by galact-
uronic acid (14.2%), accounting for 64.3% of all nine monosaccharides identified. In a study conducted
by Tulukcu [4], the fatty acid compositions of six medicinal plants used as herbal tea in Turkey were
investigated; 30 different fatty acids were determined. Moreover, the content of ascorbic acid (vitamin C)
in some common herbal teas was determined by Karasakal and Gurkan [13].
51.3 Bioactivities and Antioxidant Efficacy

Generally, herbal teas have been used for promoting human health and reducing the risk of chronic
diseases due to their multiple beneficial effects [1]. This section focuses on the antioxidant, anti-
inflammatory, antiproliferative, antimicrobial, immunological, antiglycation, and antimalarial activities
of selected herbal teas as well as their effects on some metabolic enzymes.
Additionally, it is usually considered that herbal teas can be used for therapeutic or nutritional
purposes, depending on their chemical constituents. Samali et al. [14] evaluated the phytochemical
constituents of herbal teas commonly consumed in Nigeria and showed the presence of tannins, steroids,
terpenoids, saponins, cardiac glycosides, flavonoids, alkaloids, and phlobatannins. The terpene fraction
in sage (Salvia officinalis L.) herbal tea was studied by Arceusz et al. [15]. They identified 20 com-
pounds, including α- and β-thujone, and several other oxygenated monoterpenes (1,8-cineole, linalool,
camphor, borneol, and bornyl acetate) and oxygenated sesquiterpenes. The terpene fraction of this plant
is responsible for many of its therapeutic and culinary properties. In addition, in classical phytotherapy,
the active principles of herbal teas are often attributed to their volatile constituents [16]. The volatile
compounds in some common herbal infusions were identified [17] and found that their composition
varied and depended on the raw material and the manufacturing process employed. The results showed
that herbal teas were characterized by the presence of a wide range of terpenes, such as guaiacol,
4-vinylguaiacol, eugenol, citral, phenol, carvone, menthol, 1,8-cineole, and citronellyl acetate.
Among various bioactive components in herbal teas, phenolic constituents have most widely been
investigated [1,2,5,6,8]. For example, Fabiana imbricata R. is a Chilean plant used as a tea in the Andean
regions of Chile and Argentina. The phenolic constituents identified in the tea were chlorogenic acid,
p-hydroxyacetophenone, scopoletin, and quercetin derivatives. The glycosides were mainly glucosides
from p-hydroxyacetophenone and scopoletin, and di- and triglycosides from quercetin were the main
flavonoids [18]. The qualitative and quantitative of terpenes and phenolics in some herbal tea or infusion
are summarized in Table 51.2. The chemical structures of common phenolic compounds found in herbal
teas are shown in Figure 51.1. Since the health-promoting properties reported for herbal tea can be
associated with the presence of several phenolics with known antioxidant, diuretic, and anti-inflammatory
activity, it would be described in detail for its constituent and contents in the next section of this chapter.

Herbal infusions often contain many different kinds of natural antioxidants. As an important category of
phytochemicals, phenolic compounds universally exist in plants. They have attracted increasing attention
as potential agents for preventing and treating many oxidative stress-related diseases.
Small centaury (Centaurium erythraea Rafin.) is an herbal species with a long use in traditional
medicine due to its digestive, stomachic, tonic, depurative, sedative, and antipyretic properties and con-
tains considerable amounts of polyphenolic compounds. The results demonstrated that small centaury
infusion exhibited interesting antioxidant properties, expressed both by its capacity to effectively scav-
enge hydroxyl radical and hypochlorous acid [20]. In another study, the total antioxidant capacity of
the aqueous extracts of some endemic herb infusions by steeping these herbs in hot water was assayed.
The highest antioxidant capacities of some herbal teas available in the Turkish market were for scarlet
TABLE 51.2
Content of Terpenes and Phenolics in Some Herbal Tea or Infusion
Concentration References
Salvia Officinalis L. (mg/kg)
Terpenes
1,8-Cineole 10.45 [15]
Linalool 7.92 [15]
α-Thujone 26.95 [15]
β-Thujone 10.75 [15]
Camphor 21.50 [15]
Pinocamphone 2.65 [15]
Isomenthone 1.35 [15]
Borneol 15.40 [15]
Terpinen-4-ol 7.20 [15]
α-Terpineol tr [15]
Carvone 2.00 [15]
Bornyl acetate 21.05 [15]
Thymol 1.10 [15]
Carvacrol 1.35 [15]
Caryophyllene oxide 4.75 [15]
Viridiflorol 11.55 [15]
Humulene epoxide I 5.25 [15]
Humulene epoxide II 10.53 [15]
Humulene epoxide III tr [15]
Unknown sesquiterpene alcohol 6.85 [15]
Aqueous infusion of Ficus deltoidea leaves (μmol/L)
Flavan-3-ols
Gallocatechin 44 [19]
Epigallocatechin 87 [19]
Catechin 98 [19]
(Epi)afzelechin-(epi)catechin 18 [19]
(Epi)afzelechin-(epi)afzelechin-(epi)catechin 5.7 [19]
(Epi)afzelechin-(epi)afzelechin 5.1 [19]
(Epi)catechin
Epicatechin 89 [19]
Flavones
Luteolin-6,8-C-diglucoside (lucenin-2) 13 [19]
Apigenin-6,8-C-diglucoside (vicenin-2) 15 [19]
Luteolin-6-C-hexosyl-8-C-pentoside 11 [19]
Luteolin-6-C-glucosyl-8-C-arabinoside 3.9 [19]
Apigenin-6-C-arabinosyl-8-C-glucoside 7.4 [19]
(isoschaftoside)
Luteolin-6-C-arabinosyl-8-C-glucoside 2.0 [19]
Apigenin-6-C-glucosyl-8-C-arabinoside (schaftoside) 33 [19]
Luteolin-8-C-glucoside (orientin) 10 [19]
Apigenin-6-C-pentosyl-8-C-glucoside 117 [19]
Apigenin-8-C-glucoside (vitexin) 7.8 [19]
Apigenin-6-C-glucosyl-8-C-pentoside 7 [19]
Apigenin-6,8-C-dipentoside isomer 2.2 [19]
(Continued)
Herbal Teas 649

Content of Terpenes and Phenolics in Some Herbal Tea or Infusion
Concentration References
Apigenin-6-C-glucoside (isovitexin) 15 [19]
Apigenin-6,8-C-dipentoside isomer 14 [19]
Hydroxycinnamates
4-p-Coumaroylquinic acid 139 [19]
Fabiana imbricata R. et. P. tea (mg/L)
Phenolics
Rutin 50–400 [18]
Scopoletin 8–24 [18]
p-Hydroxyacetophenone 12–50 [18]
Abbreviation: tr, trace.
OH
OH
OH O
OH
O
OH OH
O
OH
OH
Epigallocatechin gallate
OH
OH O
OHOH
OH
HO O
O O
H3C O
OH O
HO
HO OH
Rutin
O O OH
O
Scopoletin
FIGURE 51.1 Structures of common phenolic compounds found in herbal teas. (Continued)
OH
OH
HO O
OH
OH O
Quercetin
OH
OH
OH O
OH
OH
Catechin
FIGURE 51.1 (Continued) Structures of common phenolic compounds found in herbal teas.
pimpernel (Anagallis arvensis), sweet basil (Ocimum basilicums), and lemon balm (Melissa officina-
lis), in the order of 1.63, 1.18, and 0.99 mmol trolox equivalents (TE)/g, respectively [21]. Additionally,
the phenolic composition and antioxidant capacity of four oak leaf infusions from Quercus resinosa,
Q. sideroxyla, Q. eduardii, and Q. durifolia were investigated [22]. The herbal infusions were evaluated
for their total polyphenol content (TPC), total flavonoid content, trolox equivalent antioxidant capacity
(TEAC), and oxygen radical absorbance capacity (ORAC). Q. resinosa leaf infusions had the highest
TPC, TEAC, and ORAC values.
Lippia citriodora is a herbal species that contains several flavonoids and phenolic acids. The superoxide
radical, hydroxyl radical, and hypochlorous acid scavenging activities of L. citriodora infusion were
examined, displaying a potent superoxide radical scavenging activity and moderate scavenging activities of
hydroxyl radical and hypochlorous acid [23]. L. javanica and L. scaberrima are used as herbal remedies and
are commercially traded as health teas. The relationship between the presence of phenolic compounds and the
antioxidant activities of infusions prepared from four Lippia species (L. javanica, L. scaberrima, L. rehm-
annii, and L. wilmsii) indigenous to South Africa was evaluated. Of the four indigenous species, infusions of
L. javanica and L. wilmsii exhibited the highest antioxidant activities (EC50: 358 and 525 μg/mL, respec-
tively) and contained the most phenolic compounds (14.8 and 14.5 mg/mL, respectively) [24]. In another
study, 12 plants used traditionally in ethnomedicine from the South American Andes were investigated. All
herbal teas showed strong antioxidant capacities, and the results obtained with ferric-reducing antioxidant
power (FRAP), TEAC, 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity were very similar.
Moreover, upon correlating the polyphenolic contents with the antioxidant capacity a positive association
(TEAC-DPPH vs. gallic acid equivalents, r2 = 0.7437) for all plants was found [25].
Fifteen Thai herbal teas in comparison with tea of C. sinensis were investigated for their antioxi-
dant properties in relation with their total phenolics, flavonoids, and nonflavonoids contents. Significant
differences (P < 0.05) existed among the tea infusions. Stevia and sappan herbal teas had primary anti-
oxidant capacities comparable to that of C. sinensis. Principal component analysis showed that the total
phenolics, particularly flavonoids, highly correlated with primary antioxidant activities [26]. In another
study [5], total phenolic contents (TPC) of 28 kinds of Chinese commercial herbal teas were measured
by the Folin–Ciocalteu method, and their antioxidant capacities were evaluated. Twenty kinds of the
Herbal Teas 651
TABLE 51.3
Antioxidant Activities and Total Phenolic Content of 20 Commercial Herbal Teas
FRAP (mmol TEAC Total Phenolics
Name Fe[II]/L) (mmol TE/L) (g GAE/L)
Ping An Tang li yan cha 30.58 19.30 1.39
Ping An Tang shi gan cha 26.31 16.27 1.19
Qing Xin Tang jiang huo wang 25.45 6.47 1.03
Qing Xin Tang hou zheng tang 12.49 6.50 0.87
Qing Xin Tang er shi si wei 13.25 6.31 1.01
Qing Xin Tang zhi ke hua tan tang 11.70 6.70 0.91
Qing Xin Tang gan mao cha 10.38 6.19 0.84
Deng lao liang cha 8.34 3.44 0.44
Qing Xin Tang luo han guo wu hua cha 5.25 3.67 0.57
Ping An Tang shen ju cha 5.45 3.53 0.20
Ping An Tang xue li ju hua cha 4.69 2.99 0.41
Ben cao mi liang cha 4.28 2.35 0.16
Qing Xin Tang mao geng zhu zhe shui 3.16 2.48 0.35
Qing Xin Tang suan mei tang 3.11 2.18 0.39
Wang lao ji (ting zhuang) 3.51 2.08 0.15
Wang lao ji (he zhuang) 3.76 2.35 0.15
Qing Xin Tang ju hua xue li cha 2.72 2.04 0.25
Ping An Tang mao geng zhu zhe shui 7.23 0.50 0.05
Qing liang cha (he zhuang) 1.55 0.64 0.07
Bai Yun Shan liang cha 1.51 0.45 0.11
Source: Adapted from Fu, L. et al., Int. J. Mol. Sci., 12, 2112, 2011. With permission.
Abbreviations: FRAP, ferric-reducing antioxidant power; TEAC, trolox equivalent antioxidant
capacity; TE, trolox equivalents; GAE, gallic acid equivalents.
tested herbal teas in the study that possess the strongest antioxidant activity are summarized in Table
51.3. Generally, these herbal teas had high antioxidant capacities and TPC and could serve as important
dietary sources of antioxidants for prevention of diseases caused by oxidative stress.
It is often difficult to summarize and compare antioxidant capacities of herbal infusions because
different extraction and evaluation methods are used in the literature [27]. The antioxidant capacities
and TPC of 223 kinds of herbal infusions were evaluated using the same methodology [28]. Twenty
kinds of herbal infusions that possessed the strongest antioxidant activity are summarized in Table 51.4.
Furthermore, a very weak correlation (r 2 = 0.1563) existed between the TEAC and the FRAP values, sug-
gesting that the components capable of scavenging free radicals could be different from those reducing
oxidants in these herbal infusions. A significant correlation (r 2 = 0.7549) between TEAC value and TPC
as well as a very weak correlation (r 2 = 0.1966) between FRAP value and TPC implied that phenolic
compounds in these herbal infusions could be the main components contributing to free radical scaveng-
ing activities, but not be responsible for reducing oxidant abilities.

Wu et al. [29] investigated the water extract of Inulae flos, a herbal tea commonly consumed by Asians
for its inhibitory effects on the production of lipoxygenase-induced inflammatory mediators and its anti-
tyrosinase activity. The findings suggested that I. flos might serve as an alternative antioxidant source and
a noteworthy inhibitor of production of proinflammatory cytokines in a dose-dependent manner. Chen
et al. [30] reported that wogonin, a bioactive flavonoid in herbal tea, could inhibit inflammatory cyclo-
oxygenase-2 (COX-2) gene expression in human lung epithelial cancer cells. The results obtained sug-
gested that wogonin inhibits phorbol 12-myristate 13-acetate (PMA)-induced COX-2 gene expression
by inhibiting c-Jun expression and activator protein-1 (AP-1) activation in A549 cells. In another study,
TABLE 51.4
Antioxidant Activities and Total Phenolic Content of 20 Herbal Infusions
FRAP (µmol TEAC Total Phenolics
Scientific Name Fe[II]/g) (μmol TE/g) (mg GAE/g)
Salvia miltiorrhiza Bge. 601 1544 101
Cimicifuga foetida L. 707 349 25
Rhodiola sacra Fu. 541 472 51
Sargentodoxa cuneata Rehd. et Wils. 521 459 65
Fraxinus rhynchophylla Hance. 548 401 52
Sargentodoxa cuneata Rehd. et Wils. 521 459 65
Tussilago farfara L. 529 226 35
Paeonia lactiflora Pall. (red) 481 366 31
Dioscorea bulbifera L. 469 380 25
Acanthopanax gracilistylus W.W. Smith 455 306 30
Rosa chinensis Jacq. 1433 73 6
Sanguisorba officinalis L. 1845 9 1
Artemisia capillaris Thunb. 378 269 16
Lonicera japonica Thunb. (flower) 345 132 30
Magnolia officinalis Rehd. et Wils. 237 341 31
Morus alba L. (fruit). 310 234 19
Artemisia argyi Levl. et Vant. 215 239 24
Prunus persica (Linn) Batsch. 5 793 55
Picrorhiza scrophulariiflora Pennell 276 265 48
Paeonia suffruticosa Andr. 298 243 22
Source: Adapted from Li, S. et al., Ind. Crops Prod., 51, 289, 2013. With permission.
Abbreviations: FRAP, ferric-reducing antioxidant power; TEAC, trolox equivalent antioxidant
capacity; TE, trolox equivalents; GAE, gallic acid equivalents.
anti-inflammatory property of 16 water plant extracts used in the Limousin countryside as herbal teas
was studied by evaluating the inhibition of lipoxygenase activity. The results showed that Filipendula
ulmaria, Alchemilla vulgaris, and Rosmarinus officinalis belonged to the group with the highest a ctivity
(IC50 ~ 0.5 mg/mL); Equisetum arvense, Betula pendula, Hieracium pilosella, Achillea millefolium,
Lithospermum officinale, Cynara scolymus, Lamium album, Vaccinium myrtillus, Chamomilla recutita,
and Humulus lupulus had the next highest anti-inflammatory activity (IC50 ~ 1.5 mg/mL); while Melilotus
officinalis, Urtica dioı̈ca, and Lotus corniculatus were the least active extracts (IC50 > 2 mg/mL) [31].
51.3.3 Antiproliferative Activity

Li et al. [6] evaluated the in vitro antiproliferative activities of 32 kinds of herbal infusions on four cancer
cell lines by 3-(4,5-dimethylthiahiazole-2-y1)-2,5-diphenyl tetrazolium bromide (MTT) assay. The results
showed that some infusions strongly inhibited the proliferation of A549 (human lung cancer cells), MCF-7
(human breast cancer cells), HepG2 (human hepatoma cells), and HT-29 (human colon cancer cells), as
well as decreased the viability of these cancer cell lines in a dose-dependent manner. It is suggested that
some tea and herbal infusions may serve as potential dietary supplement for the prevention and treatment
of cancer. The chemoprotective properties of unfermented and fermented rooibos (Aspalathus linearis)
and honey bush (Cyclopia intermedia) herbal teas were investigated against fumonisin B-1 (FB-1) pro-
motion in rat liver utilizing diethylnitrosamine as cancer initiator. The results indicated that fermented
herbal teas and unfermented honey bush significantly (P < 0.05) decreased FB-1-induced lipid peroxida-
tion in the liver. It is suggested that differences in the major polyphenolic components and certain FB-1/
polyphenolic/tissue interactions may explain the varying effects of different herbal teas on the oxidative
parameters, hepatotoxic effects, and cancer promotion in rat liver [32]. Marnewick et al. [33] also studied
the inhibition of tumor promotion in mouse skin by the extracts of rooibos (A. linearis) and honey bush
Herbal Teas 653
(C. intermedia), which are unique South African herbal teas. The results showed that topical applica-
tion of the tea fractions prior to the tumor promoter on Institute of Cancer Research (ICR) mouse skin
initiated with 7,12-dimethylbenz[a]anthracene suppressed skin tumorigenesis significantly (P < 0.001)
with unprocessed honey bush by 90%, processed honey bush by 84.2%, processed rooibos by 75%, and
unprocessed rooibos by 60%. It is also indicated that differences in the flavanol/proanthocyanidin and
flavonol/flavone composition and/or nonpolyphenolic constituents are likely to be important determinants
in the inhibition of tumor promotion by the herbal tea ethanol/acetone fractions in mouse skin. Nseyo
et al. [34] investigated the use of the extract, specifically of the polar methanolic fraction of Hypericum
perforatum L, a kind of Greek herbal tea, in photodynamic therapy in the treatment of human bladder
cancer cells. The results showed that the polar methanolic fraction may be an effective agent for in vitro
photodynamic therapy. Confirmation of these results in preclinical studies may lead to clinical trials.
In another study, the effects of 16 water extracts of plants used in the Limousin countryside as herbal teas
on the proliferation of melanoma B16 cells were examined [31]. For low concentrations (<0.25 mg/mL),
there was no effect except for F. ulmaria, B. pendula, U. dioı c̈ a, A. millefolium, L. officinal, R. offi-
cinalis, H. pilosella, and C. scolymus, which showed increased proliferation. For high concentrations
(>0.5 mg/mL), six extracts (F. ulmaria, A. vulgaris, E. arvense, H. lupulus, A. millefolium, and L. album)
showed a significant antiproliferative effect.
Vinas and Smith [35] characterized the potential cytotoxicity of Prodigiosa herbal tea on HepG2
cells in vitro. The results indicated that Prodigiosa altered glucose transporter protein and cDNA levels
in HepG2 cells. Additionally, Marnewick et al. [36] found antimutagenic properties of South African
herbal teas. They suggested that two mechanisms might be involved in the antimutagenicity of herbal tea
extracts toward carcinogens that require metabolic activation (see succeeding text).
• The herbal tea components may interfere with cytochrome P350–mediated metabolism of
these mutagens.
• The direct interaction between the herbal tea constituents, presumably the polyphenolic com-
pounds, with the promutagens and/or the active mutagenic metabolites.
51.3.4 Antimicrobial Activity

Oh et al. [37] evaluated the antimicrobial activities of various leafy herbal tea (LHT) extracts. The
minimum inhibitory concentrations and minimum lethal concentrations were determined to verify the
antimicrobial activities of the LHT extracts against two oral pathogens (Streptococcus mutans and
S. sobrinus) and three food-borne pathogens (Listeria monocytogenes, Shigella flexneri, and Salmonella
enterica). Among the tested LHTs, the mate tea water extract was most effective against Gram-positive
bacteria. The results suggested their potential application as a health-promoting functional ingredient
or natural preservative in foods. L. javanica and L. scaberrima are used as herbal remedies and are
commercially traded as health teas in Southern Africa. Shikanga et al. [24] studied their antibacterial
activities against four human pathogens (Staphylococcus aureus, Enterococcus faecalis, Escherichia
coli, and Pseudomonas aeruginosa). The results showed that the extract of L. javanica was the most
active against all the pathogens tested. The study offers credence to the use of infusions of these Lippia
species for their general health benefits. In addition, the antimicrobial activities of herbal infusions from
four species of white oaks (Q. resinosa, Q. beta, Q. grisea, and Q. obtusata) were investigated and their
polar extracts showed different degrees of antimicrobial activity [38].
51.3.5 Immunological Efficacy

Systemic immunological efficacy of an herbal tea L. lucidus Turcz in mice was evaluated by Yang
et al. [12]. The results showed that L. lucidus polysaccharides could significantly enhance the plaque-
forming cells, serum hemolysin level, and delayed-type hypersensitivity in response to sheep red blood
cells in a dose-dependent manner (P < 0.01). In L. lucidus polysaccharide-treated mice, phagocytosis
capacity and concanavalin A–induced splenocyte proliferation were remarkably increased (P < 0.05).
It suggests that the polysaccharides derived from the leaves of L. lucidus improve the immune system
and might be regarded as a biological response modifier. Additionally, Guangdong herbal tea (GHT) has
been consumed as a traditional remedy for Shanghuo, a traditional Chinese concept, which is a physi-
ological process of uncoordinated response to stress with physical and mental fatigue syndromes, for a
long time in Southern China. He et al. [39] investigated the main components and antistress effects of
GHT in restrain-stressed mice. The results showed that oral administration of GHT could prevent stress-
induced immunocompromise. GHT also significantly improved the proportion of T-helper (Th)-type
lymphocyte, the production of Th cell-dependent cytokines, and the antioxidative activity of splenocytes.
Furthermore, these results indicated that GHT relieved stress by increasing the numbers and activities of
immunocytes in restraint mice, by exerting antioxidative activity directly or indirectly in immunocytes.
51.3.6 Antiglycation Activity

Ho et al. [40] evaluated the antiglycation activities of different herbal infusions. It was found that many
kinds of the tested herbal teas had potent antiglycation abilities that exceeded or equaled that of green
tea, which is a well-documented antiglycation beverage. They found that the amounts of phenolics and
flavonoid in the herbal infusions were highly correlated with their antiglycation activity. It revealed
that the antiglycation activity of herbal infusions was primarily attributable to phenolics, particularly
flavonoids. Furthermore, Ho et al. [41] also evaluated the antiglycation capacities of numerous flower
herbal infusions, based on their capacity to inhibit the formation of fluorescent advanced glycation end
products, which is involved in the pathogenesis of diabetic complications. The antiglycation capacities
of sweet osmanthus and rose infusions were better than that of chrysanthemum infusion, which is a
well-documented antiglycation herb. Lavender and chamomile infusions have an equivalent antiglyca-
tion capacity to that of chrysanthemum, with a moderate effect. Furthermore, it was also found that the
TPC of a particular flower herbal infusion was significantly correlated with its antiglycation capacity,
which is consistent with the results of their previous study [40]. In addition, Deetae et al. [26] studied
the antiglycation properties of 15 Thai herbal teas and found that stevia and sappan herbal teas had the
antiglycation capacities comparable to C. sinensis.
51.3.7 Antimalarial Activity

De Donno et al. [42] compared the antimalarial activity of Artemisia annua herbal tea and artemisinin.
The evaluation of the effects of A. annua tea on Plasmodium falciparum cultures in vitro was performed.
The concentration of artemisinin in the herbal tea preparation was also determined. The results of the
in vitro tests showed that it had potent antimalarial activity. However, in this study, they found that the
concentration of artemisinin in A. annua tea was far too low to be responsible for the antimalarial activ-
ity. The artemisinin present in the tea is probably cosolubilized with other ingredients, some of which
also have antimalarial activity and act synergistically with it. In fact, A. annua tea has proven to be a very
effective treatment for malaria in various clinical trials. Houel et al. [43] discovered that antimalarial
Quassia amara young leaf tea contained several quassinoids, namely, simalikalactone D, picrasin B,
picrasin H, neoquassin, quassin, picrasin I, and picrasin J.
51.3.8 Effects on Some Metabolic Enzymes

Dalar and Konczak [44] investigated the inhibitory activities against key metabolic syndrome relevant
enzymes of herbal teas from eastern Anatolia. The results showed that all herbal teas tested success-
fully suppressed the activities of key enzymes, namely, α-amylase, α-glucosidase, pancreatic lipase,
and angiotensin-converting enzyme, involved in metabolic syndrome. This study suggested potential
antidiabetic and antiobesity activity of Plantago lanceolata herbal infusion and supported its traditional
use as antidiabetic tea.
A study aimed to determine the function of Plectranthus barbatus (Lamiaceae) herbal tea as inhibi-
tor of the brain acetylcholinesterase (AChE) activity was carried out by Fale et al. [45]. They found that
when P. barbatus extract was administered intraperitoneally, all its compounds were found in plasma, and
Herbal Teas 655
rosmarinic acid was found in the brain. After intraperitoneal administration, an inhibition of 29.0% and 24.9%
in brain acetylcholinesterase activity was observed at 30 and 60 min, respectively. These results proved that
the administration of P. barbatus aqueous extract can reach the brain and act as AChE inhibitor. Furthermore,
they reported that rosmarinic acid, scutellarein 4′-methyl ether 7-O-glucuronide, and (16S)-coleon E were
the main compounds responsible for the anti-AChE activity in herbal tea of P. barbatus [46].
Li et al. [47] evaluated the effects of 41 kinds of commercial herbal teas on activities of alcohol
dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH), which are principal enzymes responsible
for the metabolism of ethanol. The results indicated that the effects of these herbal teas on ADH and
ALDH activities were highly different, and several herbal teas could markedly increase/reduce ADH
and ALDH activities, thus suggesting that some herbal teas should not be drunk after excessive alcohol
consumption. Furthermore, several herbal teas may serve as potential dietary supplements for the pre-
vention and treatment of harm from excessive alcohol consumption.
51.3.9 Other Benefits

In addition to the bioactive effects described earlier, herbal teas may render other beneficial effects, such
as antiaging, cardioprotective, chemopreventive, hepatoprotective, neuroprotective, anti-topoisomerase,
gastroprotective, and antiobesity activities [1,38]. For example, Sun and Yu [48] investigated the effect
of a special herbal tea on obesity and anovulation in androgen-sterilized rats (ASR) and their results
indicated that the herbal tea could reduce body weight and induce ovulation in ASR.
51.4 Health Effects

Most bioactive effects of herbal teas are just studied in vitro. However, their health effects need to be fur-
ther demonstrated in human clinical trials. In the literature, some health effects of herbal teas have been
reported. Thus, recent studies in Turkey have shown that spearmint tea has antiandrogenic properties
in females with hirsutism. Hirsutism in polycystic ovarian syndrome (PCOS) is consequent to elevated
androgen levels that lead to significant cosmetic and psychological problems. In a study conducted by
Grant [49], 42 volunteers were randomized to take spearmint tea twice a day for 1 month period and com-
pared with a placebo herbal tea. Free and total testosterone levels were significantly reduced over 30 days
in the spearmint tea group (P < 0.05). There was a clear and significant alteration in the relevant hormone
levels. It was confirmed that spearmint had antiandrogen properties. A much longer future study was also
proposed as the preliminary findings were encouraging due to the potential use of spearmint tea as a help-
ful and natural treatment for hirsutism in PCOS. In another study, to test an herbal supplement containing
black tea and caffeine for stimulation of thermogenesis, a double-blind, placebo-controlled, and crossover
study was conducted on 16 healthy, weight-stable, and nonsmoking subjects. The results indicated that the
herbal supplement could increase metabolic rate without changing substrate oxidation [50].
Cantonese herbal tea (CHT) has been consumed in South China to alleviate discomfort due to the
heat and humidity in the body according to the theory of traditional Chinese medicine. You et al. [51]
attempted to understand the mechanisms of action of CHT; therefore, a hydrogen-nuclear magnetic
resonance (1H-NMR)-based metabonomic approach was used to investigate the global biological char-
acterization of rat serum following the intake of CHT. A 20-day group study showed elevation of
gluconeogenesis and a shift in energy metabolism from carbohydrate metabolism to lipid metabolism.
Additionally, a notable decrease in pyruvate content with a consistent increase in lactate content and
significant decrease in both lipoprotein and glucose contents were observed for a 30-day group, indicat-
ing potential metabolic dysfunction.
Passiflora incarnata is a traditional herbal sedative, anxiolytic, and a popular sleep aid used for the
treatment of sleep disturbance. A double-blind and placebo-controlled investigation of the effects of
P. incarnata (passionflower) herbal tea on human sleep quality was carried out by Ngan and Conduit [52].
The results suggested that the consumption of a low dose of P. incarnata, in the form of tea, yielded
short-term subjective sleep benefits for healthy adults with mild fluctuations in sleep quality. In addi-
tion, a randomized, double-blind, and placebo-controlled study that aimed to determine the efficacy
of an echinacea herbal tea preparation given at early onset of cold or flu symptoms was carried out by
Lindenmuth and Lindenmuth [53]. The results showed that treatment with the echinacea herbal tea
preparation at early onset of cold or flu symptoms was effective for relieving these symptoms in a shorter
period of time than a placebo.
A randomized-controlled study involving 175 infants in Turkey was conducted to evaluate the effec-
tiveness of herbal tea for the treatment of infantile colic. In all intervention groups, a significant reduction
in crying hours per day was observed. It demonstrated that intervention of herbal tea was effective in the
treatment of colic, which can be used by nurses in neonatal and primary health-care settings as an aid
to families for the treatment of infantile colic [54]. Furthermore, another study evaluated the effects of
consumption of maternal herbal tea containing fenugreek on breast milk production and infants’ weight
gain pattern in the early postnatal period [55]. In this study, 66 mother–infant pairs were randomly and
evenly assigned to three groups. Group 1 received herbal tea containing fenugreek every day; Groups 2
and 3 were assigned as placebo and controls, respectively. The maximum weight loss was significantly
lower in infants in Group 1 (P < 0.05) who also regained their birth weight earlier than those in the control
and placebo groups (P < 0.05). The mean measured breast milk volume of the mothers in Group 1 was
significantly higher than the placebo and control groups (P < 0.05). The results indicated that maternal
galactagogue herbal tea supplementation was useful for enhancing breast milk production and facilitating
infant birth weight regain in early postnatal days.
In conclusion, from human trails, it has been proven that some kinds of herbal teas possess antiandro-
genic properties in females with hirsutism; some could increase metabolic rate, alleviate feelings of dis-
comfort due to the heat and humidity in the body, and relieve cold or flu symptoms; some could be used
for the treatment of sleep disturbance and infantile colic as well as for enhancing breast milk production
and facilitating infant birth weight regain.

Herbal teas are made from one or several kinds of herbs. Traditionally, herbal tea is infused only before
drinking. Today, a variety of herbal teas are produced and sold commercially. For example, a special kind of
herbal tea is called “cool tea” (Liang cha in Chinese), which originated in South China, and its vendition has
been from China to about 20 countries around the globe, such as Australia, Canada, France, Germany, the
United Kingdom, and the United States [5]. Furthermore, different formulations of cool teas are produced,
such as cool tea in a bottle, cool tea in a tin, cool tea granules, and cool tea bags. In addition, new cool teas
are being developed and produced continuously, which contain different kinds of herbs with various health
effects [5,6,47].
In the future, research and development of herbal teas should be carried out from several aspects, as
given in the succeeding text.
a.
Bioactive assessment of the plant: Different parts of the plant (such as the leaf, flower, fruit,
seed, stem, and root) should be assessed for different bioactivities, for example, antioxidant,
antistress, immunomodulator, antiaging, anti-inflammatory, antidepressant, anticancer, antidi-
abetic, antiobesity, antiviral, antihypertensive, memory enhancer, and antiflu symptom activi-
ties to support cardiovascular health and sleep quality as well as treatment for malaria. This
may lead to new findings that some common, but rarely used, herbs could be a good choice for
new herbal teas [28].
b.
Formulation of a new cool tea: A new cool tea could be formulated based on the bioactivity of
different herbs as well as the theory of traditional medicine (such as the theory of traditional
Chinese medicine). The new cool teas that contain different kinds of herbs with various health
efficiencies will be developed and produced continually.
c.
Effect of processing: Industrial processing affects the content of beneficial health compounds
(such as phenolic compounds and minerals) and bioactivities (such as antioxidant activity) as
well as the microbiological status and the concentrations of pollutants (heavy metals, nitrate,
Herbal Teas 657
nitrite, pesticide residues, and mycotoxins) of herbal teas. Therefore, it is important to study the
effects of the processing parameters (such as brewing water, brewing temperature, and brewing
duration) on the quality and safety of herbal teas.
d.
Formulation: Different formulations of cool teas could be developed, such as cool tea in a
bottle, cool tea in a tin, cool tea granules, and cool tea bags.
e.
New functions of herbal teas: For herbal teas that have been produced commercially, their
new bioactivities could be evaluated, which may lead to the discovery of some new functions
[5,6,47].
f.
Authentication of herbal tea ingredients: With increasing consumption of herbal teas, a number
of public health issues, including efficacy, safety, and quality assurance, have attracted wide
interest. Molecular biological techniques could be applied to the rapid authentication of plants,
which are used for the production of herbal tea, because some plants are frequently adulterated
by using related plant species. In addition, phytochemical analysis, as a key step to investigate
the chemical composition of herbal tea and ensure the quality, is very important [1]. Chemical
fingerprinting methodology and a metabolomic approach could be used for quality assessment
and the control of herbal teas [51,56].
51.6 Conclusion
Herbal teas contain various nutritional and bioactive substances such as minerals, polysaccharides, fatty
acids, and vitamins, among others (such as phenolics, essential oils, and terpenes). The presence of
diverse bioactive substances, especially phenolic constituents, makes herbal teas rendering different
effects such as antioxidant, anti-inflammatory, antiproliferative, antimicrobial, immunological, antigly-
cation, and antimalarial activities. In future, new cool teas should be developed and produced, which
contain different kinds of herbs with various health effects.
REFERENCES
1. Zhao, J., Deng, J.W., Chen, Y.W., and Li, S.P., Advanced phytochemical analysis of herbal tea in China.
J. Chromatogr. A, 1313, 2S–23S, 2013.
2. Kaliora, A.C., Kogiannou, D.A.A., Kefalas, P., Papassideri, I.S., and Kalogeropoulos, N., Phenolic
profiles and antioxidant and anticarcinogenic activities of Greek herbal infusions: Balancing delight
and chemoprevention. Food Chem., 142, 233–241, 2014.
3. Nookabkaew, S., Rangkadilok, N., and Satayavivad, J., Determination of trace elements in herbal tea
products and their infusions consumed in Thailand. J. Agric. Food Chem., 54, 6939–6944, 2006.
4. Tulukcu, E., Herbal tea fatty acid contents of some medicinal plants grown in Konya, Turkey. Asian J.
Chem., 23, 1369–1372, 2011.
5. Fu, L., Xu, B.T., Gan, R.Y., Zhang, Y., Xu, X.R., Xia, E.Q., and Li, H.B., Total phenolic contents and
antioxidant capacities of herbal and tea infusions. Int. J. Mol. Sci., 12, 2112–2124, 2011.
6. Li, F., Li, S., Li, H.B., Deng, G.F., Ling, W.H., and Xu, X.R., Antiproliferative activities of tea and herbal
infusions. Food Funct., 4, 530–538, 2013.
7. Kohno, H., Kouda, K., Tokunaga, R., and Sonoda, Y., Detection of estrogenic activity in herbal teas by
in vitro reporter assays. Eur. Food Res. Technol., 225, 913–920, 2007.
8. Joubert, E. and de Beer, D., Rooibos (Aspalathus linearis) beyond the farm gate: From herbal tea to
potential phytopharmaceutical. S. Afr. J. Bot., 77(Suppl. 4), 869S–886S, 2011.
9. Szymczycha-Madeja, A., Welna, M., and Zyrnicki, W., Multi-element analysis, bioavailability and
fractionation of herbal tea products. J. Braz. Chem. Soc., 24, 777–787, 2013.
10. Suliburska, J. and Kaczmarek, K., Herbal infusions as a source of calcium, magnesium, iron, zinc and
copper in human nutrition. Int. J. Food Sci. Nutr., 63, 194–198, 2012.
11. Ozcan, M.M., Unver, A., Ucar, T., and Arslan, D., Mineral content of some herbs and herbal teas by
infusion and decoction. Food Chem., 106, 1120–1127, 2008.
12. Yang, X.B., Lv, Y., Tian, L.M., and Zhao, Y., Composition and systemic immune activity of the polysac-
charides from an herbal tea (Lycopus lucidus Turcz). J. Agric. Food Chem., 58, 6075–6080, 2010.
13. Karasakal, A. and Gurkan, Y.Y., Determination of ascorbic acid in common fruits, herbal tea by using
cuprac and cerac methods. Asian J. Chem., 24, 2609–2612, 2012.
14. Samali, A., Kirim, R.A., and Mustapha, K.B., Qualitative and quantitative evaluation of some herbal
teas commonly consumed in Nigeria. Afr. J. Pharm. Pharmacol., 6, 384–388, 2012.
15. Arceusz, A., Occhipinti, A., Capuzzo, A., and Maffei, M.E., Comparison of different extraction
methods for the determination of alpha- and beta-thujone in sage (Salvia officinalis L.) herbal tea.
J. Sep. Sci., 36, 3130–3134, 2013.
16. Tschiggerl, C. and Bucar, F., The volatile fraction of herbal teas. Phytochem. Rev., 11(Suppl. 2–3),
245S–254S, 2012.
17. Lasekan, O. and Lasekan, A., Flavour chemistry of mate and some common herbal teas. Trends Food
Sci. Technol., 27, 37–46, 2012.
18. Quispe, C., Viveros-Valdez, E., and Schmeda-Hirschmann, G., Phenolic constituents of the Chilean
herbal tea Fabiana imbricata R. et P. Plant Foods Hum. Nutr., 67, 242–246, 2012.
19. Omar, M.H., Mullen, W., and Crozier, A., Identification of proanthocyanidin dimers and trimers, flavone
C-glycosides, and antioxidants in Ficus deltoidea, a Malaysian herbal tea. J. Agric. Food Chem., 59,
1363–1369, 2011.
20. Valentao, P., Fernandes, E., Carvalho, F., Andrade, P.B., Seabra, R.M., and Bastos, M.L., Hydroxyl
radical and hypochlorous acid scavenging activity of small centaury (Centaurium erythraea) infusion:
A comparative study with green tea (Camellia sinensis). Phytomedicine, 10, 517–522, 2003.
21. Apak, R., Guclu, K., Ozyurek, M., Karademir, S.E., and Ercag, E., The cupric ion reducing antioxidant
capacity and polyphenolic content of some herbal teas. Int. J. Food Sci. Nutr., 57, 292–304, 2006.
22. Rocha-Guzman, N.E., Medina-Medrano, J.R., Gallegos-Infante, J.A., Gonzalez-Laredo, R.F., Ramos-
Gomez, M., Reynoso-Camacho, R., Guzman-Maldonado, H., and Gonzalez-Herrera, S.M., Chemical
evaluation, antioxidant capacity, and consumer acceptance of several oak infusions. J. Food Sci., 77,
C162–C166, 2012.
23. Valentao, P., Fernandes, E., Carvalho, F., Andrade, P.B., Seabra, R.M., and Bastos, M.D., Studies on the
antioxidant activity of Lippia citriodora infusion: Scavenging effect on superoxide radical, hydroxyl
radical and hypochlorous acid. Biol. Pharm. Bull., 25, 1324–1327, 2002.
24. Shikanga, E.A., Combrinck, S., and Regnier, T., South African Lippia herbal infusions: Total phenolic
content, antioxidant and antibacterial activities. S. Afr. J. Bot., 76, 567–571, 2010.
25. Rojo, L.E., Benites, J., Lopez, J., Rojas, M., Diaz, P., Ordonez, J., and Pastene, E., Antioxidant
capacity and polyphenolic content of twelve traditionally used herbal medicinal infusions from the
South American Andes. Bol. Latin. Carib. Plant. Med. Arom., 8, 498–508, 2009.
26. Deetae, P., Parichanon, P., Trakunleewatthana, P., Chanseetis, C., and Lertsiri, S., Antioxidant and
anti-glycation properties of Thai herbal teas in comparison with conventional teas. Food Chem., 133,
953–959, 2012.
27. Li, S., Li, S.K., Li, H.B., Xu, X.R., Deng, G.F., and Xu, D.P., Antioxidant capacities of herbal infusions,
in Processing and Impact on Antioxidants in Beverages, Preedy, V.R., Ed., Elsevier, Amsterdam, the
Netherlands, 2014, pp. 41–50.
28. Li, S., Li, S.K., Gan, R.Y., Song, F.L., Kuang, L., and Li, H.B., Antioxidant capacities and total phenolic
contents of infusions from 223 medicinal plants. Ind. Crops Prod., 51, 289–298, 2013.
29. Wu, L.C., Jou, A.F.J., Chen, S.H., Tien, C.Y., Cheng, C.F., Fan, N.C., and Ho, J.A.A., Antioxidant, anti-
inflammatory and anti-browning activities of hot water extracts of oriental herbal teas. Food Funct.,
1, 200–208, 2010.
30. Chen, L.G., Hung, L.Y., Tsai, K.W., Pan, Y.S., Tsai, Y.D., Li, Y.Z., and Liu, Y.W., Wogonin, a bioactive
flavonoid in herbal tea, inhibits inflammatory cyclooxygenase-2 gene expression in human lung epithe-
lial cancer cells. Mol. Nutr. Food Res., 52, 1349–1357, 2008.
31. Trouillas, P., Calliste, C.A., Allais, D.P., Simon, A., Marfak, A., Delage, C., and Duroux, J.L.,
Antioxidant, anti-inflammatory and antiproliferative properties of sixteen water plant extracts used in
the Limousin countryside as herbal teas. Food Chem., 80, 399–407, 2003.
32. Marnewick, J.L., van der Westhuizen, F.H., Joubert, E., Swanevelder, S., Swart, P., and Gelderblom,
W.C.A., Chemoprotective properties of rooibos (Aspalathus linearis), honeybush (Cyclopia intermedia)
herbal and green and black (Camellia sinensis) teas against cancer promotion induced by fumonisin B-1
in rat liver. Food Chem. Toxicol., 47, 220–229, 2009.
Herbal Teas 659
33. Marnewick, J.L., Joubert, E., Joseph, S., Swanevelder, S., Swart, P., and Gelderblom, W.C.A., Inhibition
of tumour promotion in mouse skin by extracts of rooibos (Aspalathus linearis) and honeybush (Cyclopia
intermedia), unique South African herbal teas. Cancer Lett., 224, 193–202, 2005.
34. Nseyo, U., Kim, A., Stavropoulos, N.E., Skalkos, D., Nseyo, U.U., and Chung, T.D., Herbal tea extract
combined with light induced significant in vitro cytotoxicity of human bladder cancer cells, in Photonic
Therapeutics and Diagnostics, Vol. 5686, Bartels, K.E., Bass, L.A., DeRiese, W.T.W., Gregory,
K.W., Hirschberg, H., Katzir, A., Kollias, N. et al., Eds., Proceedings of the Society of Photo-optical
Instrumentation Engineers (SPIE) Series, Washington, DC, 2005, pp. 199–206.
35. Vinas, R. and Smith, E.E., Preliminary evaluation of Prodigiosa herbal tea: Cytotoxicity and GLUT2
expression in HepG2 cells. Toxicol. Environ. Chem., 95, 669–678, 2013.
36. Marnewick, J.L., Gelderblom, W.C.A., and Joubert, E., An investigation on the antimutagenic properties
of South African herbal teas. Mutat. Res., 471, 157–166, 2000.
37. Oh, J., Jo, H., Cho, A.R., Kim, S.J., and Han, J., Antioxidant and antimicrobial activities of various leafy
herbal teas. Food Control, 31, 403–409, 2013.
38. Sanchez-Burgos, J.A., Ramirez-Mares, M.V., Larrosa, M.M., Gallegos-Infante, J.A., Gonzalez-Laredo,
R.F., Medina-Torres, L., and Rocha-Guzman, N.E., Antioxidant, antimicrobial, antitopoisomerase and
gastroprotective effect of herbal infusions from four Quercus species. Ind. Crops Prod., 42, 57–62, 2013.
39. He, R.R., Tsoi, B., Li, Y.F., Yao, X.S., and Kurihara, H., The anti-stress effects of Guangdong herbal tea
on immunocompromise in mice loaded with restraint stress. J. Health Sci., 57, 255–263, 2011.
40. Ho, S.C., Wu, S.P., Lin, S.M., and Tang, Y.L., Comparison of anti-glycation capacities of several herbal
infusions with that of green tea. Food Chem., 122, 768–774, 2010.
41. Ho, S.C., Chang, P.W., Tong, H.T., and Yu, P.Y., Inhibition of fluorescent advanced glycation end-
products and N-carboxymethyllysine formation by several floral herbal infusion. Int. J. Food Proper.,
17, 617–628, 2014.
42. De Donno, A., Grassi, T., Idolo, A., Guido, M., Papadia, P., Caccioppola, A., Villanova, L., Merendino,
A., Bagordo, F., and Fanizzi, F.P., First-time comparison of the in vitro antimalarial activity of Artemisia
annua herbal tea and artemisinin. Trans. R. Soc. Trop. Med. Hyg., 106, 696–700, 2012.
43. Houel, E., Bertani, S., Bourdy, G., Deharo, E., Jullian, V., Valentin, A., Chevalley, S., and Stien, D.,
Quassinoid constituents of Quassia amara L. leaf herbal tea: Impact on its antimalarial activity and
cytotoxicity. J. Ethnopharmacol., 126, 114–118, 2009.
44. Dalar, A. and Konczak, I., Phenolic contents, antioxidant capacities and inhibitory activities against
key metabolic syndrome relevant enzymes of herbal teas from Eastern Anatolia. Ind. Crops Prod., 44,
383–390, 2013.
45. Fale, P.L.V., Madeira, P.J.A., Florencio, M.H., Ascensao, L., and Serralheiro, M.L.M., Function of
Plectranthus barbatus herbal tea as neuronal acetylcholinesterase inhibitor. Food Funct., 2, 130–136,
2011.
46. Fale, P.L.V., Borges, C., Madeira, P.J.A., Ascensao, L., Araujo, M.E.M., Florencio, M.H., and Serralheiro,
M.L.M., Rosmarinic acid, scutellarein 4′-methyl ether 7-O-glucuronide and (16S)-coleon E are the
main compounds responsible for the antiacetylcholinesterase and antioxidant activity in herbal tea of
Plectranthus barbatus (“falso boldo”). Food Chem., 114, 798–805, 2009.
47. Li, S., Gan, L.Q., Li, S.K., Zheng, J.C., Xu, D.P., and Li, H.B., Effects of herbal infusions, tea and
carbonated beverages on alcohol dehydrogenase and aldehyde dehydrogenase activity. Food Funct.,
5, 42–49, 2014.
48. Sun, F. and Yu, J., The effect of a special herbal tea on obesity and anovulation in androgen-sterilized
rats. Proc. Soc. Exp. Biol. Med., 223, 295–301, 2000.
49. Grant, P., Spearmint herbal tea has significant anti-androgen effects in polycystic ovarian syndrome.
A randomized controlled trial. Phytother. Res., 24, 186–188, 2010.
50. Roberts, A.T., de Jonge-Levitan, L., Parker, C.C., and Greenway, F.L., The effect of an herbal supple-
ment containing black tea and caffeine on metabolic parameters in humans. Altern. Med. Rev., 10,
321–325, 2005.
51. You, R., Xu, Z.B., Hu, S.Q., and Li, L., Characterization of temporary metabolic changes following
Cantonese herbal tea intervention. Phytother. Res., 26, 1097–1102, 2012.
52. Ngan, A. and Conduit, R., A double-blind, placebo-controlled investigation of the effects of Passiflora
incarnata (Passionflower) herbal tea on subjective sleep quality. Phytother. Res., 25, 1153–1159, 2011.
53. Lindenmuth, G.F. and Lindenmuth, E.B., The efficacy of echinacea compound herbal tea preparation on
the severity and duration of upper respiratory and flu symptoms: A randomized, double-blind placebo-
controlled study. J. Altern. Complement. Med., 6, 327–334, 2000.
54. Arikan, D., Alp, H., Gozum, S., Orbak, Z., and Cifci, E.K., Effectiveness of massage, sucrose solution,
herbal tea or hydrolysed formula in the treatment of infantile colic. J. Clin. Nurs., 17, 1754–1761, 2008.
55. Turkyilmaz, C., Onal, E., Hirfanoglu, I.M., Turan, O., Koc, E., Ergenekon, E., and Atalay, Y., The effect
of galactagogue herbal tea on breast milk production and short-term catch-up of birth weight in the first
week of life. J. Altern. Complement. Med., 17, 139–142, 2011.
56. Deng, J.W. and Yang, Y.Y., Chemical fingerprint analysis for quality assessment and control of Bansha
herbal tea using paper spray mass spectrometry. Anal. Chim. Acta, 785, 82–90, 2013.
52
Coffee
Iziar A. Ludwig, Michael N. Clifford, Michael E.J. Lean, and Alan Crozier
CONTENTS
52.1 Introduction....................................................................................................................................661
52.2 Composition and Nutritional Characteristics.................................................................................661
52.4 Health Effects................................................................................................................................ 665
52.4.1 Coffee and Antioxidant Status......................................................................................... 668
52.4.2 Coffee and Type 2 Diabetes............................................................................................. 669
52.4.3 Coffee and Cardiovascular Disease................................................................................. 669
52.4.4 Coffee and Cancer............................................................................................................ 670
52.4.5 Coffee and Parkinson’s Disease........................................................................................671
52.5 Conclusion......................................................................................................................................671
References............................................................................................................................................... 672
52.1 Introduction
Coffee is one of the most traded food products in the world, and coffee brew produced from the roasted
coffee beans is one of the most popular beverages in the world along with water and tea [1]. The generic
name Coffea covers approximately 70 species, of which only two are of economic importance, namely,
Coffea arabica, commonly known as Arabica coffee, which accounts for ~60% of world production, with
the remaining ~40% coming from C. canephora var. Robusta, which is used to produce Robusta coffee
[2]. Differences between these two species include ideal growing climate, physical aspects, chemical com-
position, and characteristics of the brew made with the ground-roasted beans. In general, Arabica coffee
brew is appreciated for its superior cup quality and aroma, whereas the Robusta beverage possesses a more
aggressive flavor and contains higher amounts of soluble solids, antioxidants, and caffeine [3]. The pleasant
aroma and taste of brewed coffee are a consequence of the roasting process, which transforms naturally
occurring substances in the green bean generating volatiles and a diversity of other compounds some of
which are derived from the Maillard reaction [4]. Although coffee is consumed primarily because of its
pleasant aroma, taste, and stimulating properties, more recent investigations indicate there may be potential
health benefits associated with the beverage including reduced incidences of several chronic and degenera-
tive diseases, such as cancer, cardiovascular disease (CVD), diabetes, and Parkinson’s disease [5,6]. This
chapter highlights the nutritional and phytochemical composition of coffee brews, along with potential
mechanisms by which the bioactive compounds of coffee may impact on human health. Finally, epidemio-
logical findings are discussed, which link coffee consumption with prevention of several chronic diseases.
52.2 Composition and Nutritional Characteristics

The composition of green coffee beans and the impact of roasting are well documented and will not be
discussed in this chapter [7–16]. The attractive aroma associated with coffee beverages develops during
roasting. Typical roasting conditions use air temperatures in the range of 180°C–250°C as a consequence
661
of which profound changes occur with carbohydrate caramelization, pyrolysis of organic compounds,
and the production of compounds derived from the Maillard reaction [4]. Upon roasting in excess of
700 volatile substances are produced, many being heterocycles rarely found elsewhere in the diet, albeit
only at trace levels. Nutritional characteristics of espresso, filter, and instant coffee are summarized in
Table 52.1. Several micronutrients are present in coffee brew, including magnesium, potassium, niacin,
and vitamin E. Depending on the coffeemaking procedure, one cup of coffee provides 7–24 mg of mag-
nesium and 34–116 mg of potassium [17]. During the roasting of coffee beans trigonelline is demeth-
ylated to form nicotinic acid (Figure 52.1) as well as being converted to several pyridine derivatives.
Coffee has been reported to provide 1–3 mg of nicotinic acid per cup contributing to 6%–18% of the
Recommended Dietary Allowances (RDA) of 16 mg/day for adult men [18].
TABLE 52.1
Compositional and Nutritional Characteristics of Different Coffees
Espresso Coffee Filter Coffee Instant Coffee
Nutrient Unit (1 fl oz: 30 g) (8 fl oz: 237 g) (6 fl oz: 179 g)
Water g 29 235 177
Energy kcal 3 2 4
Protein g 0.04 0.28 0.18
Lipid (fat) g 0.05 0.05 0.00
Total sugars g 0.00 0.00 0.00
Minerals
Calcium mg 1 5 7
Iron mg 0.04 0.02 0.07
Magnesium mg 24 7 7
Phosphorus mg 2 7 5
Sodium mg 4 5 7
Zinc mg 0.02 0.05 0.02
Vitamins
Folate (DFE) µg 0 5 0
Niacin mg 1.562 0.453 0.422
Riboflavin mg 0.053 0.180 0.002
Thiamin mg 0.000 0.033 0.000
Vitamin B6 mg 0.001 0.002 0.000
Vitamin C mg 0.1 0.0 0.0
Source: Adapted from the U.S. Department of Agriculture (USDA), USDA National
Nutrient Database for Standard Reference, Release 26, 2013, Published
online at: http://ndb.nal.usda.gov/ndb/search/list (accessed April 11, 2014).
CH3
N+ N
–OOC
HOOC
Trigonelline Nicotinic acid
FIGURE 52.1 Structures of trigonelline and nicotinic acid.

Coffee 663

The biologically active classes of compounds in coffee are usually considered to be chlorogenic acids
(CGAs) (Figure 52.2), caffeine, the diterpenes kahweol and cafestol (Figure 52.3), and melanoidins.
CGAs are the major phenolic compounds in coffee, and although losses occur during roasting, a cof-
fee beverage is a variable, but rich, probably the richest, dietary source of CGAs with a single serving of
espresso coffee supplying from 24 to 422 mg (Table 52.2) [19]. Nonespresso brews are also rich sources
and many regular coffee drinkers might easily consume 1 g or even 2 g of CGAs per day, greatly exceed-
ing the intake from fruits and vegetables. CGAs are frequently referred to as powerful antioxidants,
and this might be true in vitro. However, to have a biological effect, CGAs must reach organs or tissues
in a sufficient concentration that is maintained for an adequate time. After coffee ingestion by humans
unmetabolized CGAs achieve only transient and very low nM peak concentrations [20]. They, therefore,
OH
OH
OH OH
OH OH OH O
HOOC OH OH O
OH O O OH
HOOC HOOC
O OH OH O OH OH
3-O-Caffeoylquinic acid 4-O-Caffeoylquinic acid 5-O-Caffeoylquinic acid
OCH3
OH
OCH3 OCH3
OH OH OH O
OH OH O
HOOC
OH O OH
HOOC O HOOC
O OH OH O OH OH
3-O-Feruloylquinic acid 4-O-Feruloylquinic acid 5-O-Feruloylquinic acid
OH OH
OH
OH OH
HOOC OH
OH
OH O O
O O O
OH
HOOC OH OH OH
OH OH O OH
HOOC
OH
OH OH
O O O
O O O
3,4-O-Dicaffeoylquinic acid 3,5-O-Dicaffeoylquinic acid 4,5-O-Dicaffeoylquinic acid
OH
OH O
OH O
O HOOC OH
HOOC
OH OH O OH OH
4-O-p-Coumaroylquinic acid 5-O-p-Coumaroylquinic acid
FIGURE 52.2 Structures of the main chlorogenic acids in coffee beans.

O CH3
H3C N
N
O N N
CH3
Caffeine
CH2OH CH2OH
OH OH
O O
H H
Kahweol Cafestol
FIGURE 52.3 Structures of the purine alkaloid caffeine and the diterpenes kahweol and cafestol.
TABLE 52.2
Levels of CQA Isomers and Caffeine in Espresso Coffees Purchased from 20 Coffee Shops in Glasgow, UK
Volume of 4-CQA Total CQA per Caffeine per
Coffee Shop Coffee (mL) 3-CQA (mg) (mg) 5-CQA (mg) Serving (mg) Serving (mg)
Pattiserie Francoise 52 95 112 216 423 322
S’mug 32 62 78 160 300 173
Costa Coffee 25 48 61 118 227 157
Little Italy 23 37 59 121 217 129
Paperino’s 50 65 52 99 216 205
Peckhams 70 65 52 99 216 140
Chapter 1 26 45 58 112 215 140
University Cafe 49 40 54 93 187 230
Baguette Express 45 30 40 74 144 140
Kember & Jones 43 37 46 92 141 90
Heart Buchanan 24 22 37 67 126 156
Jellyhill 63 26 29 56 111 151
Coffee @ 491 49 23 31 55 109 98
Beanscene 48 19 25 49 93 77
Tinderbox 25 19 24 46 89 75
Café Cinnamon 59 17 23 41 81 242
Crepe á Croissant 34 17 23 41 81 95
Morton’s 35 13 16 27 56 73
Antipasti 36 8 15 21 44 72
Starbucks 27 5 7 12 24 51
Range 23–70 5–95 7–112 12–216 24–42 50–322
Median 43 36 37 67 126 140
Source: Adapted from Crozier, T.W.M. et al., Food Funct., 3, 30, 2012. With permission.
Abbreviation: CQA, caffeoylquinic acid.
cannot realistically compete with the much greater concentrations of the far more potent food-derived
antioxidants in the circulatory system such as the vitamins ascorbate and tocopherol [21]. That health
benefits of CGAs might arise from the over-simple direct-antioxidant theory has repeatedly been chal-
lenged [21–23], and alternative mode(s) of action must be sought. More recently, (poly)phenols have been
found to exert modulatory effects in cells through selective action on multiple cell-signaling pathways
Coffee 665
involved in pathogenesis of degenerative diseases, indicating that the health effects go well beyond sim-
ple antioxidant activity [24]. In this regard, it has been suggested that CGAs act as chemopreventive
agents by modulating the expression of genes encoding enzymes involved in phase II metabolism, which
form part of the endogenous antioxidant defenses, playing a critical role in the conversion of reactive
electrophiles and xenobiotics into less toxic products [25].
Coffee is also the main dietary source of caffeine with Robustas containing about twice as much as
Arabicas. Caffeine levels also decline during roasting, but to a much lesser extent than caffeoylquinic
acids, and typical amounts per cup of coffee range between 50 and 100 mg [3]. However, depending on the
roasted beans and barista procedures, significant variability between coffee brews has been reported with
values as high as 323 mg/cup (Table 52.2) [19]. In addition to its well-studied stimulatory effects, which
include enhanced perception, reduced fatigue, and an increased capacity to remain awake, caffeine has
been proposed as an antioxidant coffee component [26]. Lee [27] suggested as a source of caffeine’s anti-
oxidant capacity its degradation into the catabolites methylxanthine and methyluric acid, which at physi-
ological µM concentrations were highly effective antioxidants being able to prevent in vitro low-density
lipoprotein (LDL) oxidation. To what degree this reflects the situation in vivo remains to be determined.
The diterpenes kahweol and cafestol occur as fatty acyl esters in coffee and contribute to the bitter
taste of the beverage where they are found in quantities highly dependent on the brewing technique.
Filter and instant coffees contain less than 0.6 mg/cup, whereas French press and boiled coffees contain
6 to 12 mg [18]. While the presence of these diterpenes has been implicated in the cholesterol-raising
effect of coffee [28], they have also been shown to possess chemopreventive potential [29]. It has been
proposed that their mode of action includes induction of phase II detoxifying enzymes, and regulation of
nuclear erythroid 2-related factor 2/antioxidant response element signaling pathways, thereby enhancing
the endogenous defense systems against oxidative damage [30].
Melanoidins formed during roasting account for ~30% of the dry matter of coffee brew and are
generically defined as heterogeneous, brown-colored, nitrogen-containing, and high-molecular-
weight end products of the Maillard reaction [4]. The Maillard reaction involves the condensation of
the carbonyl group of reducing sugars, or other components, with the amino group of amino acids and
proteins. Through a complex network of chemical reactions that are modulated by pH, temperature,
and moisture, myriad products are formed, commonly known as Maillard reaction products (MRPs).
The polymeric, brown-colored final products of the Maillard reaction, the melanoidins, result from
cyclization, dehydration, retroaldolization, rearrangements, isomerization, and condensation of
MRPs. However, the exact composition of coffee melanoidins remains unresolved [31]. Different
potential biological activities have been ascribed to melanoidins including antioxidant and metal
chelating activity, antihypertensive and antiglycative activity, and the ability to modulate colonic
microflora. However, there is no evidence to suggest that even the low-mass melanoidins are absorbed
intact in the circulatory system. Thus, only effects exerted in the oro-gastrointestinal tract without
the need for absorption might be significant in vivo. Melanoidins could reach high concentra-
tions in the gastrointestinal tract where the data of Tagliazucchi et al. [32] suggest they might exert
beneficial effects by preventing the formation of oxidized lipids and decreasing the absorption of
these cytotoxic lipid peroxidation products. Furthermore, studies on melanoidins using model systems
have shown that they are able to evoke contractions of gastric smooth muscles by activating choliner-
gic receptors [33]. This finding implies that melanoidins might increase the colon motility, which is
linked with reduced colorectal cancer risk.
52.4 Health Effects

Earlier epidemiological studies reported that coffee consumption may increase the risk of coronary heart
disease (CHD) and several types of cancer, but more recent investigations have not confirmed these find-
ings. Table 52.3 summarizes the recent and most relevant epidemiological studies on the health effect
of coffee consumption. On balance, the currently available information from epidemiological research
TABLE 52.3
Epidemiological Studies on the Health Effects of Coffee Consumption
Model/Study Design Outcomes References
Antioxidant Status
Crossover intervention study/acute coffee FRAP and TRAP antioxidant activity in plasma [35]
intake of 200 mL instant coffee (4% w/v). increased 2.6% and 7.6%, respectively, after coffee
consumption.
Crossover intervention study/daily Coffee significantly increased the glutathione content [36]
consumption 1L of French press coffee in the colorectal mucosa by 8% and in plasma by
over 2 weeks. 15%.
Intervention study/5 cups of coffee daily Plasma glutathione increased by 16% on coffee [37]
over 1 week. consumption.
Intervention study/daily consumption 1 L of Weak induction of glutathione-S-transferase activity [38]
unfiltered boiled coffee over 5 days. in plasma.
Type 2 Diabetes Mellitus
Three-way randomized crossover Reduced glucose and insulin concentrations after [45]
intervention study/acute intake of 400 mL decaffeinated coffee consumption. Decrease in
caffeinated or decaffeinated coffee with glucose-dependent insulinotropic polypeptide
25 g of glucose. secretion and increase in glucagon-like peptide 1
secretion after decaffeinated coffee.
Double-blind randomized intervention Less pronounced increase in glucose and insulin [46]
study/acute ingestion of caffeine capsules, concentrations after caffeinated coffee consumption
caffeinated filter coffee, decaffeinated filter compared to caffeine capsules. Decaffeinated coffee
coffee, or placebo. resulted in a 50% lower glucose response than
placebo.
Intervention study/acute ingestion of Coffee did not affect the mean glycemic index value [47]
250 mL filter coffee together with white of foods compared to water.
bread, fruit leather, or cheese puffs.
Intervention study/daily consumption of four No changes were seen for markers of glucose [48]
cups (150 mL/cup) of filter coffee over 1 metabolism in an oral-glucose-tolerance test.
month and eight cups over the next month.
Prospective cohort study/17,111 Dutch men 50% lower risk for people consuming ≥7 cups of [41]
and women, aged 30–60 years. coffee per day compared to ≤2 cups.
Prospective cohort study/1361 Swedish 45% lower risk for people consuming 3–4 cups of [42]
women, aged 39–65 years, follow-up: coffee per day compared to ≤2 cups.
18 years.
Prospective cohort study (The Nurses’ 29% lower risk for people consuming ≥6 cups of [43]
Health Study)/84,276 women, aged coffee per day compared non–coffee drinkers.
43–59 years, follow-up: 18 years.
Prospective cohort study (Health 54% lower risk for people consuming ≥6 cups of [43]
Professionals’ Follow-up Study)/41,934 coffee per day compared to non–coffee drinkers.
men, aged 40–75 years, follow-up: 12 years.
Meta-analysis/nine prospective cohort 35% risk reduction for people consuming ≥6 cups of [39]
(193,473 participants) and six cross- coffee per day compared to ≤2 cups. Higher coffee
sectional studies (20,656 participants). consumption was consistently associated with a
lower prevalence of newly detected hyperglycemia.
Meta-analysis/18 prospective cohort studies Inverse log-linear relationship between coffee [44]
(457,922 participants). consumption and risk of diabetes. 7% relative risk
reduction with every additional cup of coffee
consumed in a day.
Cardiovascular Disease
Double-blind randomized crossover Caffeinated coffee increased brachial artery [53]
intervention study/acute ingestion of 1 cup flow-mediated dilation, systolic and diastolic blood
of caffeinated and 1 cup of decaffeinated pressure. This effect was not observed after
Italian espresso coffee. decaffeinated coffee ingestion.
(Continued)
Coffee 667

Case control study/848 cases with a first 30% lower risk for moderate coffee consumption [52]
symptom of coronary heart disease and (300 mL/day) and three times higher risk for heavy
1078 controls free of cardiovascular coffee consumption (>600 mL/day) compared to
disease. non–coffee drinkers.
Prospective cohort study (Honolulu Heart Risk of thromboembolic stroke was more than [50]
Study)/499 hypertensive men, aged doubled for men who consumed three cups of coffee
55–68 years, follow-up: 25 years. per day as compared to nondrinkers.
Prospective cohort study (The Nurses’ Long-term coffee consumption was not associated [51]
Health Study)/83,076 women, aged with an increased risk of stroke in women.
43–59 years, follow-up: 24 years. Decaffeinated coffee was associated with a trend
toward lower risk of stroke after adjustment for
caffeinated coffee consumption.
Meta-analysis/21 prospective cohort studies 18% lower risk in women consuming 1–3 cups of [49]
(407,806 participants). coffee per day compared to non–coffee drinkers.
Cancer
Endometrial Cancer
Case control study/2122 breast, 229 36% lower risk of endometrial cancer in women [55]
endometrial and 166 ovarian cancer cases, drinking 1–2 cups of coffee per day and 59% lower
and 12,425 women, free of cancer as risk in women drinking ≥3 cups compared to 0 cups.
control. No clear association for breast and ovarian
cancer risk.
Case control study/107 endometrial 40% lower risk of developing endometrioid [56]
endometrioid adenocarcinoma cases and adenocarcinoma in women drinking 5–6 cups/
214 women free of cancer as control. week-1 cup/day of coffee per day and 60% lower
risk in women drinking ≥2 cups/day compared to
≤4 cups/week.
Case control study/454 endometrial cancer 50% lower risk of endometrial cancer in women [57]
cases and 908 women free of cancer as drinking ≥4 cups/day compared to 1 cup.
control.
Prospective cohort study/53,724 Japanese 62% lower risk of endometrial cancer in women [58]
women, aged 40–69 years, follow-up: drinking ≥3 cups/day compared to ≤2 cup/week.
15 years.
Hepatocellular Cancer
Case control study/250 hepatocellular cases 20%, 60%, and 70% lower risk of developing [61]
and 500 controls hospitalized for any hepatocellular cancer for people consuming
reasons other than neoplasms, and liver and respectively, 1–2 cups, 3–4 cups, and >5 cups a day.
alcohol-related diseases.
Prospective cohort study/90,452 middle- Habitual coffee drinking (1–2 cups/day) was [59]
aged and elderly Japanese subjects, associated with 52% lower risk of hepatocellular
follow-up: 10 years. cancer.
Prospective cohort study (Japan Collaborative 50% lower risk of death due to hepatocellular cancer [60]
Cohort Study for Evaluation of Cancer for people consuming ≥1 cups/day compared to
Risk)/110,688 Japanese men and women non–coffee drinkers.
aged 40–79 years, follow-up: 10 years.
Meta-analysis of six case control (1,551 30% lower risk for moderate coffee drinkers and 55% [62]
cases and 4,330 controls) and four for heavy coffee drinkers compared to non–coffee
prospective cohort studies (235,525 drinkers. 23% relative risk reduction with every
participants). additional cup of coffee consumed in a day.
Meta-analysis of five case control (1,154 An increase in consumption of two cups of coffee per [63]
cases and 4,330 controls) and four day was associated with a 43% reduced risk of liver
prospective cohort studies (235,525 cancer
participants).
(Continued)

Colorectal Cancer
Meta-analysis of 24 case control studies 30% lower risk for colorectal cancer and 25% for [65]
(14,846 cases). colon cancer for heavy coffee drinkers when
compared to non/low drinkers.
Meta-analysis of 25 case control (15,522 Combined results from case control studies showed a [66]
cases and 46,550 controls) and 16 cohort significant lower risk of 15% for colorectal cancer
studies (953,669 participants). and 21% for colon cancer for heavy coffee drinkers
when compared to non/low drinkers.
Dose–response meta-analysis of 25 Significant lower risk of developing colorectal [67]
case control (15,522 cases and 46,550 (8%–11%) and colon cancer (17%–25%) for daily
controls) and 17 cohort studies (1,443,375 coffee intake of ≥4 cups. Potentially nonlinear
participants). dose–response relationship.
Breast Cancer
Case control study/1932 breast cancer cases 38% lower risk in premenopausal women consuming [68]
and 1895 controls hospitalized for any ≥4 cups of coffee per day.
reasons other than neoplasms.
Case control study/1690 women with a 70% lower risk of developing breast cancer in women [70]
BRCA1 or BRCA2, 845 women with a drinking ≥6 cups of coffee per day compared to
history of invasive breast cancer and 845 non–coffee drinkers. The effect was limited to
controls. caffeinated coffee.
Parkinson’s Disease
Prospective cohort study (The Nurses’ Among women using postmenopausal hormones [72]
Health Study)/77,713 postmenopausal heavy coffee drinkers (≥6 cups/day) had a four-fold
women, follow-up: 18 years. higher risk of Parkinson’s disease compared to
non–coffee drinkers.
Prospective cohort study (Cancer Prevention 34% lower risk of Parkinson’s disease mortality in [73]
Study II)/301,164 men and 238,058 men with regular coffee consumption. 53% lower
women, aged 30 or more years, follow-up: risk of Parkinson’s disease death in women who
10 years. never had used postmenopausal hormones and 31%
higher risk among hormone users.
Meta-analysis of eight case control and five 31% lower risk of developing Parkinson’s disease for [71]
cohort studies. coffee drinkers compared to non–coffee drinkers.
Strong linear dose–response relation in cohort
studies that included only men and null linear
relation in the study that included only women.
Abbreviations: FRAP, ferric-reducing antioxidant power; TRAP, total radical-trapping antioxidant parameter.
suggests that coffee consumption may help prevent several chronic diseases including type 2 diabetes,
CVD, and cancer, as well as neurodegenerative conditions such as Parkinson’s disease [18,34].
52.4.1 Coffee and Antioxidant Status

Several intervention studies have analyzed the impact of coffee consumption on the antioxidant activity
of plasma with increases being reported by Moura-Nunes et al. [35]. A change in the antioxidant status
in vivo is unlikely to be due to a direct antioxidant activity of coffee constituents and is more likely
to be a consequence of coffee compounds being able to increment endogenous antioxidant defenses
such as an increase in glutathione-S-transferase (GST) activity and glutathione concentration. After
daily consumption of coffee over 2 weeks, significant increases in glutathione concentration (15%) in
plasma and in colorectal mucosa (8%) have been observed [36]. Similar findings for plasma glutathione
concentration were reported after the ingestion of five cups of coffee over 7 days [37]. In a later study,
Coffee 669
consumption of unfiltered coffee showed a weak but significant increase in GST activity [38]. Thus, cof-
fee might prevent oxidative damage to cell components, DNA, proteins, and lipids, which contribute
to the pathogenesis of degenerative conditions, such as cardiovascular and neurodegenerative diseases
and cancers.
52.4.2 Coffee and Type 2 Diabetes

Based on meta-analysis of epidemiological studies, there is a strong relationship between regular coffee
intake and a reduced risk of type 2 diabetes mellitus (DM) [39,40]. A prospective cohort study in the
Netherlands, involving 17,000 men and women, reported a reduction in the risk of developing type 2 DM
of 50% for those who consumed at least seven cups of coffee per day compared to those who drank two
cups or less [41]. Similar results were found in a smaller 18-year cohort study of Swedish women with
a daily coffee intake of three cups compared with the consumption of two cups or less [42]. The two
largest prospective cohort studies to examine the relationship between coffee consumption and type 2
DM were the Health Professionals Follow-up Study (41,934 men) and the Nurses’ Health Study (84,276
women) in the United States. Men who drank at least six cups of coffee daily had a 54% lower risk of
developing type 2 DM than men who did not drink coffee, and women who drank ≧6 cups daily had a
29% lower risk than women who did not drink coffee. The inverse relation between coffee consump-
tion and type 2 DM was observed for caffeinated as well as decaffeinated coffee [43]. A more recent
meta-analysis of 18 prospective studies on coffee intake and type 2 DM found an inverse log–linear
relationship between coffee consumption and subsequent risk of diabetes. Every additional cup of coffee
consumed in a day was associated with a 7% reduction in the relative risk of diabetes [44]. However, not
all prospective cohort studies have observed a significant inverse relationship between coffee intake and
risk of developing type 2 DM. The inverse relationship found in epidemiological studies is supported by
results obtained in intervention studies, which found a positive effect of coffee on diabetic markers such
as serum glucose or insulin levels [45,46]. However, other investigators did not detect an effect of acute
coffee intake on these glucose metabolism markers [47,48].
Results of the effect of coffee on diabetes are not always clear cut, and until the relationship between
long-term coffee consumption and type 2 DM is better understood and the mechanism involved iden-
tified, it is premature, and possibly dangerous, to recommended coffee as a mean of preventing type
2 DM [18].
52.4.3 Coffee and Cardiovascular Disease

Several early investigations linked coffee consumption with an increased risk of developing CVD.
However, these findings are controversial and more recent studies have shown that the risk of CVD
seems to be related to the ingestion of the diterpenes cafestol and kahweol (Figure 52.3), which have
been shown to raise serum total and LDL cholesterols reversibly and can be found at high amounts in
boiled and unfiltered coffee. The consumption of boiled coffee has decreased as of late and a recent
meta-analysis including 21 cohort studies reported a lower risk of CVD in women who were moderate
coffee drinkers [49]. Nevertheless, most prospective cohort studies have not found a significant associa-
tion between coffee consumption and CVD risk [19], while a few have reported negative effects.
Results from the Honolulu Heart Study [50] indicated a doubling of the risk of thrombotic stroke in
hypertensive men who consumed at least 700 mL of coffee per day. Besides cafestol and kahweol, caf-
feine might also exert a negative effect on CVD health. Results obtained from the Nurses’ Health Study
[51] showed that long-term coffee consumption was not associated with stroke. However, decaffeinated
coffee was found to decrease the risk of stroke after adjustment for caffeinated coffee consumption.
Heavy coffee consumers (>600 mL daily) are reported to have a three times higher risk of develop-
ing acute coronary syndrome than those who did not drink coffee, while moderate coffee consumers
(<300 mL) had a 30% lower risk [52]. Results from intervention studies on coffee and CVD risk bio-
markers confirm negative effects for caffeinated coffee, whereas beneficial effects have been linked with
decaffeinated coffee consumption [34,53].
The results of epidemiological and intervention studies on the effects of coffee on CVD risk vary and
coffee seems to have a mix of beneficial and harmful effects. Nevertheless, long-term studies suggest that
harmful effects are unlikely for moderate coffee consumption and it is possible that other compounds in
coffee counteract the negative effects of caffeine and diterpenes on CVD health.
52.4.4 Coffee and Cancer

There are numerous epidemiological studies relating coffee to the risk of cancer and the available evi-
dence suggests that coffee might prevent some cancers. Strong and consistent protective association has
been found between coffee consumption and reduced risk of endometrial and hepatocellular cancer.
In the case of breast and colorectal cancer, a modest or borderline negative association with coffee
consumption was reported, whereas no association was found for pancreatic, ovarian, prostate, or gastric
cancer [5,34,54].
Several case control studies on coffee and the risk of endometrial cancer conducted in Italy and Japan
revealed risk reductions of 50%–60% for coffee consumption of three or more cups per day and a statisti-
cally significant linear association between the amount of coffee consumed and the relative risk [55–57].
Similar results were obtained in a Japanese population-based cohort study involving 53,724 women with
15 years follow-up, which showed a risk reduction of 60% for those consuming at least three cups of cof-
fee per day compared to those who drank less than two cups [58].
In the case of hepatocellular cancer (HCC), a significant risk reduction was observed for habitual
coffee consumers in several cohort and case control studies. Two Japanese cohort studies, one involv-
ing more than 90,000 men and women during 10 years of follow up and the other with more than
100,000 men and women over a 10–12 year period, found a significant inverse association between
coffee drinking and HCC risk [59,60]. Results from these studies suggest that the consumption of 1 or
more cups of coffee on a daily basis reduces the possibility of developing HCC, or death due to HCC,
by 50%. Both studies found a clear dose-dependent decrease between the risk of HCC and the amount
of coffee consumed. Results of case control studies corroborate these findings [54]. Among these, one
hospital-based control studies conducted in Italy concluded that compared to nondrinking subjects, the
risk of HCC decreased by 20% for 1–2 cups/day, by 60% for 3–4 cups a day, and by 70% for >5 cups
a day [61]. Most of these data were summarized in two meta-analyses that concluded that an increase
in coffee consumption of one cup per day was associated with a 22%–23% reduced risk of liver cancer
[62,63].
Although following ingestion a significant amount of potentially anticarcinogenic coffee compounds,
including CGAs, reaches the colon, controversial results have been reported on coffee consumption and
reduced risk of colorectal cancer. A review including 15 case control and three cohort studies concluded
that a significant risk reduction linked to coffee consumption was found for some case control studies,
but the results were inconsistent in the cohort studies [64]. No consistent dose–response was observed
among the studies and no relationship emerged for rectal cancer. More recently, two meta-analyses, one
including 24 case control studies and the other one including 25 case control, and 16 cohort studies,
showed a moderate inverse relation between colorectal and colon cancer for case control studies, but no
association was found with rectal cancer [65,66]. Another recent study on the dose–response effect of
coffee consumption and colorectal cancer confirmed these findings and reported a significant associa-
tion between colorectal and colon cancer for daily coffee intake of at least four cups [67]. Although not
all studies were consistent, accumulated evidence suggests at least a modest inverse association between
coffee and the risk of colorectal and colon cancer.
Several studies have analyzed the effect of coffee consumption on breast cancer but no association was
found for case control or cohort studies among postmenopausal women [5,54]. However, coffee seems
to reduce the risk of breast cancer in premenopausal women [68]. In addition, a strong negative asso-
ciation was observed between coffee consumption and breast cancer among high-risk women carrying
BRCA gene mutations. Women with BRCA mutations have an estimated risk of developing breast cancer
as high as 80% by the age of 70 [69]. However, women with BRCA mutations who habitually drank six
or more cups of coffee per day showed a 70% risk reduction compared to non–coffee drinkers [70].
Coffee 671
No association was observed for decaffeinated coffee, suggesting that caffeine might be responsible for
the protective effect.
Overall, the available evidence indicates a potential protective effect of coffee consumption in endo-
metrial and hepatocellular cancer. A modestly favorable effect was observed in the risk reduction of
colorectal and colon cancer, whereas protective effects in breast cancer have only been reported among
premenopausal women and women carrying BRCA mutations.
52.4.5 Coffee and Parkinson’s Disease

Results from epidemiological studies reveal an inverse association between coffee consumption and the
risk of Parkinson’s disease. A meta-analysis of eight case control and five cohort studies conducted in
four countries between 1968 and 2001 concluded that coffee drinkers had a 31% lower risk of Parkinson’s
disease than non–coffee drinkers [71]. Interestingly, the two cohort studies that included only men, the
“Honolulu Heart Study” and the “Health Professionals Follow-up Study,” found a strong inverse linear
relation between the number of cups of coffee consumed and risk of Parkinson’s disease with 49% lower
risk per three additional cups per day, whereas the cohort study that included only women (Nurses’
Health Study) found no such “three additional cups per day” effect. The sex difference observed could be
due to hormonal effects and, indeed, further analysis of the “Nurses’ Health Study” revealed that coffee
reduced the risk of Parkinson’s disease among women who did not use postmenopausal hormones, but
increased the risk among hormone users [72]. These findings were corroborated by the results obtained
from the “Cancer Prevention Study II” cohort, which showed a significant inverse association between
coffee consumption and Parkinson’s disease mortality in men. In women, however, this association was
dependent on postmenopausal estrogen use, with a risk reduction of 53% for women drinking four or
more cups of coffee (≥600 mL) per day compared with nondrinkers and a risk increment of 31% among
estrogen users [73]. Although the cause of this adverse effect of estrogen is not yet understood, it might
be linked with the fact that estrogen replacement therapy has been found to inhibit cytochrome P4501-
mediated caffeine metabolism [74].
The positive effect of coffee on Parkinson’s disease has been ascribed to its caffeine content. Parkinson’s
disease is a neuropathological disorder involving the degeneration of dopaminergic neurons in the sub-
stantia nigra, with the subsequent loss of their terminals in the striatum. Caffeine and other A2 adenosine
receptor antagonists have proven to protect against dopaminergic neurotoxicity in animal models [75,76].
A possible explanation is that caffeine has been shown to aid in improving the performance of dopaminer-
gic system by blocking the A2 adenosine receptors and so stimulating the dopamine release [77].
Results obtained to date in epidemiological studies indicate a strong association between coffee con-
sumption and a reduced risk of Parkinson’s disease. However, the mechanisms involved are not fully
understood, and it is premature to recommend increasing coffee consumption to prevent Parkinson’s
disease, especially in women taking postmenopausal hormones.
52.5 Conclusion
Coffee is one of the most popular beverages in the world and, consequently, its impact on human health
is of great interest. Among the vast array of compounds present in coffee brew, the biologically active
classes are usually considered to be CGAs, melanoidins, caffeine, and the diterpenes cafestol and kah-
weol. Epidemiological studies have associated coffee consumption with potential beneficial effects
including reduced instances of type 2 DM, hepatocellular, endometrial, colorectal, and premenopausal
breast cancer. Results on the effect on CVD are conflicting, but negative effects of caffeine, kahweol,
and cafestol seem to be reduced or counteracted by CGAs. A reduced risk of Parkinson’s disease was
found in men but only in women who never used postmenopausal estrogen. Before these epidemiological
observations can be confirmed, and used as a sound basis for dietary advice, further research is needed
on the bioavailability and pharmacokinetics of coffee components in order to elucidate the compounds
responsible for those effects and mechanisms involved.
REFERENCES
1. Bond, T.J., The origins of tea, coffee and cocoa beverages, in Teas, Cocoa and Coffee: Plant Secondary
Metabolites and Health, Crozier, A., Ashihara, H., and Tomás-Barberán, F., Eds., Wiley-Blackwell,
Oxford, UK, 2012, pp. 1–24.
2. International Coffee Organisation (ICO), Statistics. Total Production by all Exporting Countries
2011–2014, July 2015. Published online at: http://www.ico.org/prices/po-production.pdf (accessed
October 15, 2015).
3. Farah, A., Coffee constituents, in Coffee: Emerging Health Effects and Disease Prevention, Chu, Y.F.,
Ed., Wiley-Blackwell, Oxford, UK, 2012, pp. 21–58.
4. Belitz, H.D., Grosch, M., and Schieberle, P., Food Chemistry, 4th ed., Springer-Verlag, Berlin, Germany,
2009.
5. Nkondjock, A., Coffee consumption and the risk of cancer: An overview. Cancer Lett., 277, 121–125,
2009.
6. Bøhn, S.K., Ward, N.C., Hodgson, J.M., and Croft, K.D., Effects of tea and coffee on cardiovascular
disease risk. Food Funct., 3, 575–591, 2012.
7. Clarke, R.J. and Macrae, R., Coffee 1, Chemistry, Elsevier Applied Science, London, UK, 1985.
8. Clifford, M.N. and Willson, K.C., Coffee: Botany, Biochemistry and Production of Beans and Beverage,
Chapman & Hall, London, UK, 1985.
9. Clarke, R.J. and Vitzthum, O.G., Coffee: Recent Developments, Blackwell Science, Oxford, UK, 2001.
10. Clifford, M.N. and Knight, S., The cinnamoyl-amino acid conjugates of green Robusta coffee beans.
Food Chem., 87, 457–463, 2004.
11. Clifford, M.N., Knight, S., Surucu, B., and Kuhnert, N., Characterization by LC-MSn of four new classes
of chlorogenic acids in green coffee beans: Dimethoxycinnamoylquinic acids, diferuloylquinic acids,
caffeoyl-dimethoxycinnamoylquinic acids, and feruloyl-dimethoxycinnamoylquinic acids. J. Agric.
Food Chem., 54, 1957–1969, 2006.
12. Clifford, M.N., Marks, S., Knight, S., and Kuhnert, N., Characterization by LC–MSn of four novel
classes of p-coumaric acid-containing diacyl chlorogenic acids in green coffee beans. J. Agric. Food
Chem., 54, 4095–4101, 2006.
13. Jaiswal, R. and Kuhnert, N., Hierarchical scheme for liquid chromatography/multi-stage spectrometric
identification of 3,4,5-triacyl chlorogenic acids in green Robusta coffee beans. Rapid Commun. Mass
Spectrom., 24, 2283–2294, 2010.
14. Jaiswal, R., Matei, M.F., Golon, A., Witt, M., and Kuhnert, N., Understanding the fate of chlorogenic
acids in coffee roasting using mass spectrometry based targeted and non-targeted analytical strategies.
Food Funct., 3, 976–984, 2012.
15. Stalmach, A., Clifford, M.N., Williamson, G., and Crozier, A., Phytochemicals in coffee and the
bioavailability of chlorogenic acids, in Teas, Cocoa and Coffee: Plant Secondary Metabolites and
Health, Crozier, A., Ashihara, H., and Tomás-Barberán, F., Eds., Wiley-Blackwell, Oxford, UK, 2012,
pp. 143–168.
16. Jaiswal, R., Matei, M.F., Subedi, P., and Kuhnert, N., Does roasted coffee contain chlorogenic acid lac-
tones or/and cinnamoylshikimate esters? Food Res. Int., 61, 214–227, 2014.
Release 26, 2013. Published online at: http://ndb.nal.usda.gov/ndb/search/list (accessed April 11, 2014).
18. Higdon, J.V. and Frei, B., Coffee and health: A review of recent human research. Crit. Rev. Food Sci.
Nutr., 46, 101–123, 2006.
19. Crozier, T.W.M., Stalmach, A., Lean, M.E.J., and Crozier, A., Espresso coffees, caffeine and chloro-
genic acid intake: Potential health implications. Food Funct., 3, 30–33, 2012.
20. Stalmach, A., Mullen, W., Barron, D., Uchida, K., Yokota, T., Cavin, C., Steiling, H., Williamson, G.,
and Crozier, A., Metabolite profiling of hydroxycinnamate derivatives in plasma and urine after inges-
tion of coffee by humans: Identification of biomarkers of coffee consumption. Drug Metab. Disp., 37,
1749–1758, 2009.
21. Clifford, M.N., Diet-derived phenols in plasma and tissues and their implications for health. Planta
Medica, 12, 1103–1114, 2004.
22. Halliwell, B., Rafter, J., and Jenner, A., Health promotion by flavonoids, tocopherols, tocotrienols, and
other phenols: Direct or indirect effects? Antioxidant or not? Am. J. Clin. Nutr., 81, 268S–276S, 2005.
Coffee 673
23. Scalbert, A., Johnson, I.T., and Saltmarsh, M., Polyphenols: Antioxidants and beyond. Am. J. Clin.
Nutr., 81, 215S–217S, 2005.
24. Del Rio, D., Rodrigues-Mateos, A., Spencer, J.P.E., Tognolini, M., Borges, G., and Crozier, A., Dietary
(poly)phenolics in human health and disease: Structures, bioavailability, evidence of protective effects
and potential mechanisms. Antioxid. Redox Signal., 18, 1818–1892, 2013.
25. Feng, R., Lu, Y., Bowman, L.L., Qian, Y., Castranova, V., and Ding, M., Inhibition of activator Protein-1,
NF-κB, and MAPKs and induction of phase 2 detoxifing enzyme activity by chlorogenic acid. J. Biol.
Chem., 280, 27888–27895, 2005.
26. Devasagayam, T., Kamat, J., Mohan, H., and Kesavan, P.C., Caffeine as an antioxidant: Inhibition of
lipid peroxidation induced by reactive oxygen species. Biochim. Biophys. Acta, 1282, 63–70, 1996.
27. Lee, C., Antioxidant ability of caffeine and its metabolites based on the study of oxygen radical absorb-
ing capacity and inhibition of LDL peroxidation. Clin. Chim. Acta., 295, 141–154, 2000.
28. Urgert, R. and Katan, M.B., The cholesterol-raising factor from coffee beans. Ann. Rev. Nutr., 17,
305–324, 1997.
29. Cavin, C., Holzhaeuser, D., Scharf, G., Constable, A., Huber, W.W., and Schilter, B., Cafestol and kah-
weol, two coffee specific diterpenes with anticarcinogenic activity. Food Chem. Toxicol., 40, 1155–1163,
2002.
30. Bøhn, S.K., Blomhoff, R., and Paur, I., Coffee and cancer risk, epidemiological evidence, and molecular
mechanisms. Mol. Nutr. Food Res., 2013, 58, 915–930, 2014.
31. Moreira, A.S.P., Nunes, F.M., Domingues, M.R., and Coimbra, M.A., Coffee melanoidins: Structure,
mechanisms of formation and potential health impacts. Food Funct., 3, 903–915, 2012.
32. Tagliazucchi, D., Verzelloni, E., and Conte, A., Effect of dietary melanoidins on lipid peroxidation dur-
ing simulated gastric digestion: Their possible role in the prevention of oxidative damage. J. Agric. Food
Chem., 58, 2513–2519, 2010.
33. Argirova, M.D., Stevanova, I.D., Krustev, A.D., and Turiiski, V.I., Testing biological activity of model
Maillard reaction products: Studies on gastric smooth muscle tissues. Amino Acids, 38, 797–803,
2010.
34. Williamson, G., Coffee and health, in Teas, Cocoa and Coffee: Plant Secondary Metabolites and
Health, Crozier, A., Ashihara, H., and Tomás-Barberán, F., Eds., Wiley-Blackwell, Oxford, UK, 2012,
pp. 169–181.
35. Moura-Nunes, N., Perrone, D., Farah, A., and Donangelo, C.M., The increase in human plasma anti-
oxidant capacity after acute coffee intake is not associated with endogenous non-enzymatic antioxidant
components. Int. J. Food Sci. Nutr., 60, 173–181, 2009.
36. Grubben, M.J., van den Braak, C.C., Broekhuizen, R., de Jong, R., van Rijt, L., de Ruijter, E., Peters,
W.H., Katan, M.B., and Nagengast, F.M., The effect of unfiltered coffee on potential biomarkers for
colonic cancer risk in healthy volunteers: A randomized trial. Aliment. Pharmacol. Ther., 14, 1181–1190,
2000.
37. Esposito, F., Morisco, F., Verde, V., Ritieni, A., Caporaso, N., and Fogliano, V., Moderate coffee con-
sumption increases plasma glutathione but not homocysteine in healthy subjects. Aliment. Pharmacol.
Therap., 17, 595–601, 2003.
38. Steinkellner, H., Hoelzl, H., Uhl, M., Cavin, C., Haidinger, G., Gsur, A., Schnid, R., Kundi, M., Bichler,
J., and Knasmüller, S., Coffee consumption induces GSTP in plasma and protects lymphocytes against
(+/−)-anti-benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide induced DNA-damage: Results of controlled
human intervention trials. Mutat. Res. Fund. Mol. Mech. Mutagen., 591, 264–275, 2005.
39. van Dam, R.M. and Hu, F.B., Coffee consumption and risk of type 2 diabetes—A systematic review.
JAMA, 294, 97–104, 2005.
40. van Dam, R.M., Coffee and type 2 diabetes: From beans to ß-cells. Nutr. Metab. Cardiovasc. Dis., 16,
69–77, 2006.
41. van Dam, R.M. and Feskens, E.J., Coffee consumption and risk of type 2 diabetes mellitus. Lancet, 360,
1477–1478, 2002.
42. Rosengren, A., Dotevall, A., Wilhelmsen, L., Thelle, D., and Johansson, S., Coffee and incidence of
diabetes in Swedish women: A prospective 18-year follow-up study. J. Int. Med., 255, 89–95, 2004.
43. Salazar-Martinez, E., Willett, W.C., Ascherio, A., Mansonm, J.E., Leitzmann, M.F., Stampfer, M.J., and
Hu, F.B., Coffee consumption and risk for type 2 diabetes mellitus. Ann. Intern. Med., 140, 1–8, 2004.
44. Huxley, R., Lee, C.M.Y., Barzi, F., Timmermeister, L., Czernichow, S., Perkovic, V., Grobbee, D.E.,
Batty, D., and Woodward, M., Coffee, decaffeinated coffee, and tea consumption in relation to incident
type 2 diabetes mellitus A systematic review with meta-analysis. Arch. Int. Med., 169, 2053–2063. 2009.
45. Johnston, K.L., Clifford, M.N., and Morgan, L.M., Coffee acutely modifies gastrointestinal hormone
secretion and glucose tolerance in humans: Glycemic effects of chlorogenic acid and caffeine. Am. J.
Clin. Nutr., 78, 728–733, 2003.
46. Battram, D.S., Arthur, R., Weekes, A., and Graham, T.E., The glucose intolerance induced by caf-
feinated coffee ingestion is less pronounced than that due to alkaloid caffeine in men. J. Nutr., 136,
1276–1280, 2006.
47. Aldughpassi, A. and Wolever, T.M.S., Effect of coffee and tea on the glycaemic index of foods: No effect
on mean but reduced variability. Br. J. Nutr., 101, 1282–1285, 2009.
48. Kempf, K., Herder, C., Erlund, I., Kolb, H., Martin, S., Carstensen, M., Koenig, W. et al., Effects of
coffee consumption on subclinical inflammation and other risk factors for type 2 diabetes: A clinical
trial. Am. J. Clin. Nutr., 91, 950–957, 2010.
49. Wu, J.N., Ho, S.C., Zhou, C., Ling, W.H., Chen, W.Q., Wang, C.L., and Chen, Y.M., Coffee consumption
and risk of coronary heart diseases: A meta-analysis of 21 prospective cohort studies. Int. J. Cardiol.,
137, 216–225, 2009.
50. Hakim, A.A., Ross, G.W., and Curb, J.D., Coffee consumption in hypertensive men in older middle-age
and the risk of stroke. The Honolulu Heart Program. J. Clin. Epidemiol., 51, 487–494, 1998.
51. Lopez-Garcia, E., Rodriguez-Artalejo, F., and Rexrode, K.M., Coffee consumption and risk of stroke in
women. Circulation, 119, 1116–1123, 2009.
52. Panagiotakos, D.B., Pitsavos, C., Chrysohoou, C., Kokkinos, P., Toutouzas, P., and Stefanadis, C.,
The J-shaped effect of coffee consumption on the risk of developing acute coronary syndromes: The
CARDIO2000 case–control study. J. Nutr., 133, 3228–3232, 2003.
53. Buscemi, S., Verga, S., Batsis, J.A., Donatelli, M., Tranchina, M.R., Belmonte, S., Mattina, A., Re, A.,
and Cerasola, G., Acute effects of coffee on endothelial function in healthy subjects. Eur. J. Clin. Nutr.,
64, 483–489, 2010.
54. Arab, L., Epidemiologic evidence on coffee and cancer. Nutr. Cancer, 62, 271–283, 2010.
55. Hirose, K., Niwa, Y., Wakai, K., Matsuko, K., Nakanishi, T., and Tajima, K., Coffee consumption and
the risk of endometrial cancer: Evidence from a case–control study of female hormone-related cancers
in Japan. Cancer Sci., 98, 411–415, 2007.
56. Koizumi, T., Nakaya, N., Okamura, C., Sato, Y., Shimazu, T., Nagase, S., Nijikura, H. et al., Case–control
study of coffee consumption and the risk of endometrial endometrioid adenocarcinoma. Eur. J. Cancer
Prev., 17, 358–363, 2008.
57. Bravi, F., Scotti, L., Bosetti, C., Zucchetto, A., Talamini, R., Montella, M., Greggi, S. et al., Food groups
and endometrial cancer risk: A case–control study from Italy. Am. J. Obst. Gynecol., 200, 293.e1–293.
e7, 2009.
58. Shimazu, T., Inoue, M., Sasazuki, S., Iwasaki, M., Kurahashi, N., and Yamaji, S., Coffee consumption
and risk of endometrial cancer: A prospective study in Japan. Int. J. Cancer, 123, 2406–2410, 2008.
59. Inoue, M., Yoshimi, I., Sobue, T., and Tsugane, S., Influence of coffee drinking on subsequent risk of
hepatocellular carcinoma: A prospective study in Japan. J. Nat. Cancer Inst., 97, 293–300, 2005.
60. Kurozawa, Y., Ogimoto, I., Shibata, A., Nose, T., Yoshimura, T., Suzuki, H., Sakata, R. et al., Coffee and
risk of death from hepatocellular carcinoma in a large cohort study in Japan. Br. J. Cancer, 93, 607–610,
2005.
61. Gelatti, U., Covolo, L., Franceschini, M., Pirali, F., Tagger, A., Ribero, M.L., Tevisi, P., Martelli, C.,
Nardi, G., and Donato, F., Coffee consumption reduces the risk of hepatocellular carcinoma indepen-
dently of its aetiology: A case–control study. J. Hepatol., 4, 528–534, 2005.
62. Bravi, F., Boscetti, C., Tavani, A., Bagnardi, V., Gallus, S., Negri, E., Franceschi, S., and La Vecchia, C.,
Coffee drinking and hepatocellular carcinoma risk: A meta-analysis. Hepatology, 46, 430–435, 2007.
63. Larsson, S.C. and Wolk, A., Coffee consumption and risk of liver cancer: A meta-analysis.
Gastroenterology, 132, 1740–1745, 2007.
64. Tavani, A. and La Vecchia, C., Coffee, decaffeinated coffee, tea and cancer of the colon and rectum:
A review of epidemiological studies, 1990–2003. Cancer Causes Control, 15, 743–757, 2004.
65. Galeone, C., Turati, F., La Vecchia, C., and Tavani, A., Coffee consumption and risk of colorectal
cancer: A meta-analysis of case–control studies. Cancer Causes Control, 21, 1949–1959, 2010.
Coffee 675
66. Li, G.W., Ma, D.F., Zhang, Y.M., Zheng, W., and Wang, P., Coffee consumption and risk of colorectal
cancer: A meta-analysis of observational studies. Public Health, 16, 346–357, 2013.
67. Tian, C., Wang, W., Hong, Z., and Zhang, X., Coffee consumption and risk of colorectal cancer: A dose-
response analysis of observational studies. Cancer Causes Control, 24, 1265–1268, 2013.
68. Baker, J.A., Beehler, G.P., Sawant, A.C., Jayaprakash, V., McCann, S.E., and Moysich, K.B., Consumption
of coffee, but not black tea, is associated with decreased risk of premenopausal breast cancer. J. Nutr.,
136, 166–171, 2006.
69. Easton, D.F., Ford, D., and Bishop, D.T., Breast and ovarian cancer incidence in BRCA1-mutation carri-
ers. Breast Cancer Linkage Consortium. Am. J. Hum. Genet., 56, 265–271, 1995.
70. Nkondjock, A., Ghadirian, P., Kotsopoulos, J., Lubinski, J., Lynch, H., Kim-Sing, C., Horsman, D. et al.,
Coffee consumption and breast cancer risk among BRCA1 and BRCA2 mutation carriers. Int. J. Cancer,
118, 103–107, 2006.
71. Hernán, M.A., Takkouche, B., Caamaño-Isorna, F., and Gestal-Otero, J.J., A meta-analysis of coffee
drinking, cigarette smoking, and the risk of Parkinson’s disease. Ann. Neurol., 52, 276–284, 2002.
72. Ascherio, A., Chen, H., Schwarzschild, M.A., Zhang, S.M., Colditz, G.A., and Speizer, F.E., Caffeine,
postmenopausal estrogen, and risk of Parkinson’s disease. Neurology, 60, 790–795, 2003.
73. Ascherio, A., Weisskopf, M.G., O’Reilly, E.J., McCullough, M.L., Calle, E.E., Rodriguez, C., and Thun,
M.J., Coffee consumption, gender, and Parkinson’s disease mortality in the cancer prevention study II
cohort: The modifying effects of estrogen. Am. J. Epidemiol., 160, 977–984, 2004.
74. Pollock, B.G., Wylie, M., Stack, J.A., Sorisio, D.A., Thompson, D.S., Kirshner, M.A., Folan, M.M., and
Condifer, K.A., Inhibition of caffeine metabolism by estrogen replacement therapy in postmenopausal
women. J. Clin. Pharmacol., 39, 936–940, 1999.
75. Chen, J.F., Xu, K., Petzer, J.P., Staal, R., Xu, Y.H., Beilstein, M., Sonsalla, P.K., Castagnoli, Jr, N., and
Schwarzschild, M.A., Neuroprotection by caffeine and A2A adenosine receptor inactivation in a model
of Parkinson’s disease. J. Neurosci., 21, 1–6, 2001.
76. Ikeda, K., Kurokawa, M., Aoyama, S., and Kuwana, Y., Neuroprotection by adenosine A2A receptor
blockade in experimental models of Parkinson’s disease. J. Neurochem., 80, 262–270, 2002.
77. Trevitt, J., Kawa, K., Jalali, A., and Larsen, C., Differential effects of adenosine antagonists in two
models of Parkinsonian tremor. Pharmacol. Biochem. Behav., 94, 24–29, 2009.
53
Beverages from Green Coffee Beans
Yuanyuan Ma and Ronald B. Pegg
CONTENTS
53.1 Introduction................................................................................................................................... 677
53.4 Health Effects................................................................................................................................ 680
53.4.1 Diabetes Mellitus.............................................................................................................. 680
53.4.2 Weight Loss...................................................................................................................... 680
53.4.3 Antihypertension.............................................................................................................. 680
53.4.4 Anticarcinogenesis........................................................................................................... 681
53.4.5 Neuroprotective Potential................................................................................................. 681
53.5.1 Potential Health Risks of Green Coffee Consumption.................................................... 682
53.6 Conclusion..................................................................................................................................... 682
References............................................................................................................................................... 683
53.1 Introduction
Coffee beans are the seeds of red or yellow cherries of evergreen shrubs belonging to the family
Rubiaceae, subfamily Cinchonoideae tribe, Coffeae (Figure 53.1). Coffea arabica (Arabica) and C.
canephora (Robusta) are two species of worldwide commercial importance, with C. liberica (Liberian)
being only a small player on the world stage. To process coffee, the handpicked coffee cherries are first
pulped (e.g., removal of the pericarp and mesocarp); the beans then fermented to remove their mucilage
layer, dried to reduce their moisture content to <8%, and stored for at least 3 months to allow for proper
flavor development; parchment layer (e.g., endocarp) was removed; and the resultant green beans then
roasted before brewing and consumption. During the roasting process, the phenolic constituents, namely,
the chlorogenic acids (CGAs), are progressively destroyed with roughly an 8%–10% loss per 1% loss of
dry matter [1].
Countries in which coffee is harvested (involving roughly 60 tropical and subtropical countries) ship
green beans to various regions of the world where eventually they will be roasted and then sold. Green
coffee beans (GCBs), therefore, refer to “raw” coffee beans that have not yet been roasted. Once the
parchment layer is removed from the bean, the GCBs still possess a silver skin layer on them. This
silver skin comes off if GCBs are subjected to a polishing step or when the GCBs are roasted. The raw
or green coffee beans possess higher levels of CGAs compared to their roasted counterparts. Although
new compounds with antioxidant activity are formed during roasting via the Maillard reaction, such as
melanoidins, the high loss of CGAs cannot be compensated [2,3]. Therefore, GCB extracts typically
possess higher levels of CGAs.
In recent years, there has been interest in the purported health benefits of extracts or decoctions pre-
pared by steeping green, rather than roasted, coffee beans in water. Such preparations have demonstrated
higher anti-inflammatory efficacy in animal models compared to roasted coffee extract counterparts [4].
Industry has taken note and is using extracts from GCBs (NB, not the GCB itself) in a number of energy
677
FIGURE 53.1 Ripe red and yellow Catuai and Caturra coffee cherries of the Coffea arabica variety.
drink formulations, supplements, and so-called functional food products. One of the challenges facing
the industry is the poor sensory characteristics of GCB extracts compared to their roasted counterparts.
The alleged health benefits of GCB extracts are believed to stem from a number of bioactives that are
present in greater concentrations in raw coffee beans than their roasted counterparts. The main com-
ponents of interest are the CGAs. This chapter reports on some of the bioactives present and the health
benefits associated with GCB extracts.

The resultant brews, extracts, or decoctions prepared from GCBs are generally formulated into specialty
beverages for health promotion, such as weight loss, the details of which are discussed later. When GCB
extracts are included to various formulations, the quantity added may be standardized to a known level
of CGA equivalents. Depending on the type of GCBs, brewing time, and strength of the resultant extract,
the level of nutrients will vary. Yet it is important to note that the function of adding GCB extract to a
specialty beverage is to provide known types and quantities of bioactives, not nutrients. To that end, there
appears to be no nutritional properties of GCB extracts reported in the literature.

Confusion exists as to what a CGA actually is. It is a natural product found in foods, which is the ester
of caffeic acid and l-quinic acid. The term CGAs also refers to a related family of esters of trans-
hydroxycinnamic acids with quinic acid. In other words, in this chapter, we are not referring to CGA
as a single compound; rather, we are considering it as part of a family of natural compounds with com-
monalities relating to their chemistry. The main groups of chlorogenic isomers present in GCBs include
caffeoylquinic acids (CQAs) with three isomers (3-CQA, 4-CQA, and 5-CQA), dicaffeoylquinic acids
(diCQAs) with three isomers (3,4-diCQA; 3,5-diCQA; 4,5-diCQA), feruloylquinic acids (FQAs) with
three isomers (3-FQA, 4-FQA, and 5-FQA), p-coumaroylquinic acids (pCoQA) with three isomers
(3-pCoQA, 4-pCoQA, and 5-pCoQA), and six mixed diesters of caffeoyl-feruloylquinic acids (CFAQ)
[5–7]. Moreover, four new classes of CGAs were identified in GCBs by a liquid chromatography–mass
spectrometry (LC-MS) study and include the compounds of dimethoxycinnamoylquinic acids, diferu-
loylquinic acids, caffeoyl-dimethoxycinnamoylquinic acids, and feruloyl-dimethoxycinnamoylquinic
acids [8]. The content of CGAs in GCBs is roughly 3.4–7.9 g/100 g dry matter in C. arabica and 6.1–14.4
g/100 g dry matter in C. canephora (Table 53.1) [5,7]. CGAs, a family of esters formed between l-quinic
Beverages from Green Coffee Beans 679
TABLE 53.1
Content of Chlorogenic Acids in Green Coffee Beans (g/100 g Dry Weight)
Green Coffee Beans CQA FQA diCQA Total CGA References
Wild C. arabica (average) 3.26 0.19 0.60 4.10 [5]
Wild C. arabica (max) 3.97 0.27 0.88 3.40 [5]
Wild C. arabica (min) 2.61 0.12 0.43 4.80 [5]
C. arabica (Angola) 5.67 0.79 1.39 7.85 [7]
Wild C. canephora (average) 7.66 1.43 2.31 11.3 [5]
Wild C. canephora (max) 9.50 2.23 3.03 14.4 [5]
Wild C. canephora (min) 5.12 0.77 1.57 7.88 [5]
C. canephora cv Robusta (Angola) 3.43 0.54 1.20 6.08 [7]
Abbreviations: CQA, caffeoylquinic acids (including 3-, 4-, and 5-isomers); FQA, feruloylquinic
acids (including 3-, 4-, and 5-isomers); diCQA, dicaffeoylquinic acids (including
3,4-, 3,5-, and 4,5-isomers); CGA, chlorogenic acids.
R
O
O 5
6 OH
OH
4
HO 1 3 OH
2
OH
FIGURE 53.2 Chlorogenic acid isomers: R=H, 5-p-coumaroylquinic acid (5-pCoQA); R=OH, 5-caffeoylquninic acid
(5-CQA); and R=OCH3, 5-feruloylquinic acid (5-FQAs).
acid and trans-hydroxycinnamic acids (Figure 53.2), are the dominant phenolic compounds present in
coffee [1]. They have attracted increasing attention for their preventative effects against diabetes [9,10],
obesity [11,12], and hypertension [13–15]. These topics are discussed later in detail.
In vitro studies have shown CGAs and caffeic acid from coffee beans to be antioxidative in nature
[16,17], but the antioxidant activity they possess, if at all in vivo, remains unclear because of extensive
metabolization. Unfortunately, the metabolites are often much weaker antioxidants compared to the
parent compounds [18]. A study performed on healthy ileostomy subjects revealed that the intestinal
absorption for CGAs was only ~33%, while for caffeic acid it was ~95%. Two-thirds of the ingested
CGAs reached the colon, where they may act as an antioxidant agent and be further metabolized by
colonic microflora [18,19].
The colonic metabolites can potentially be reabsorbed into the circulatory system and then be
excreted in urine as hippuric acid. Farah et al. [20] evaluated the pharmacokinetic profile and appar-
ent bioavailability of CGAs in both the plasma and urine of 10 healthy adults after administration of a
decaffeinated GCB extract containing 170 mg of CGA. The results indicated that CGAs of green coffee
are highly absorbed and metabolized in humans. Three CQAs, three diCQAs, as well as caffeic, ferulic,
isoferulic, and p-coumaric acids were identified in plasma, whereas 4-CQA, 5-CQA, as well as sinapic,
p-hydroxybenzoic, gallic, vanillic, dihydrocaffeic, caffeic, ferulic, iso-ferulic, and p-coumaric acids
were identified in urine after treatment. Over 30% of the ingested trans-cinnamic acid moieties were
recovered in the plasma, but only 5.5% were recovered in the urine.
According to Svilaas et al. [21], among the various foods consisting of total antioxidant intake in
humans measured by an in vitro ferric reducing ability of plasma assay, coffee contributed ~65% of the
total antioxidant intake and is the single greatest contributor. Another study reported that after supple-
mentation of filter-brewed coffee, an increase in plasma antioxidant capacity resulted from the direct
absorption of coffee polyphenols and their metabolites, such as uric acid [22]. CGAs are the most abun-
dant phenolics present in coffee and they play a major role with regard to antioxidant activity.
Coffee consumption, therefore, might help prevent the development of chronic disease states by its anti-
oxidant benefits. For instance, GCB hydroxycinnamic acids have been shown to bestow protective effects
in human HepG2 cells against oxidative stress [23].
53.4 Health Effects

53.4.1 Diabetes Mellitus
The CGAs present in coffee help to balance hepatic glucose production by inhibiting the enzyme known
as glucose-6-phosphatase [24]. They can also restrict intestinal glucose absorption by inhibiting Na+-
dependent glucose transport across the brush-border membranes and by altering intestinal glucose
transport hormone secretion [10]. This basically implies that long-term consumption of coffee and coffee-
containing energy drinks is inversely associated with the risk of one developing type 2 diabetes.
The likelihood to develop type 2 diabetes mellitus is reduced by half when people drank at least seven
cups of coffee instead of two cups or less a day [25]. A long-term study on coffee consumption including
regular coffee, decaffeinated coffee, and other caffeinated beverages revealed a significant lower risk of type
2 diabetes with coffee intake for both men and women [26]. To illustrate this, coffee consumption in Finland
is among the highest in the world: a study for an average of 12 years followed greater than 14,000 men and
women in Finland. The results indicated that 55% of men and 79% of women had a lower risk of developing
type 2 diabetes when they drank at least 10 cups of coffee per day than those who drank two cups or less [27].
53.4.2 Weight Loss

GCB extract has become a “hot” weight loss trend on the market and has been promoted on television
shows in the United States. One tends to lose weight when taking GCB extract on account of the high
content of CGAs, which have a pronounced effect on both glucose and fat metabolism. A 22-week cross-
over study of 16 overweight individuals with no significant changes to daily diet or exercise patterns
indicated significant reductions in body weight, body mass index, and percent body fat, particularly
abdominal fat reduction, after administration of a commercial GCB extract rich in CGAs [12]. The
results are further consistent with a meta-analysis on GCB extract that also concluded a significant
reduction in body weight with overweight individuals compared to a placebo [11].
The potential mechanisms for the effect of GCB on weight loss are multifaceted and are as follows: (1) they
inhibit Na+-dependent glucose transport across the brush-border membranes in the small intestine, (2) they
reduce glucose absorption by altering patterns of intestinal hormone secretion (e.g., decrease the secretion
of insulinotropic polypeptide and increase the concentration of glucagon-like peptide-1), and (3) they inhibit
the activity of glucose-6-phosphatase, which thereby limits the plasma glucose level and restricts fat deposit
in adipose tissue due to a decreased glucose availability as an energy source as well as the decreased insulin
activity [28,29]. A more recent study conducted on a commercial GCB extract showed CGAs to be respon-
sible for long-term lipolytic activity in human adipocytes instead of caffeine and therefore associated with
long-term health benefits [30]. Current findings suggest that GCB extracts and functional beverages contain-
ing this elixir could be an economic and effective way for weight loss. While adult obesity continues to climb
in the United States, GCB extracts might truly be helpful in a weight management program.
53.4.3 Antihypertension
Another frontier for GCB extracts is their preventative effect against hypertension. It is established that
caffeine consumption acutely raises blood pressure, but GCB extracts seem to act in an opposite manner,
possibly due to their marked content of CGAs. Several studies have reported that GCB extract intake
was effective in decreasing blood pressure in both spontaneously hypertensive rats [14] and humans
[13,15]. Even a small reduction such as 5 mmHg in one’s systolic blood pressure reduces the risk of stroke
and coronary heart disease (CHD) mortality by 14% and 9%, respectively, whereas a reduction of only
2 mmHg of diastolic blood pressure is associated with 14% fewer strokes and 8% less CHD.
According to Suzuki et al. [14], nitric oxide (NO)-mediated vasodilation might be involved in the
hypotensive effect of CGAs, in which CGAs improve NO bioavailability through their superoxide anion
radical scavenging activity of ferulic acid, a metabolite of 5-CQA. A study conducted on healthy male
subjects demonstrated an improvement in vasoreactivity after ingestion of a drink containing GCB
extract, which is induced by direct reaction between GCB extract and blood vessels [31]. This study
further suggested that the preventative effect of GCB extract was not only on hypertension but also on
arteriosclerosis.
Kozuma et al. [32] conducted a multicenter, randomized, double-blind, and placebo-controlled parallel
group study to evaluate the dose–response relationship between GCB extract intakes in 117 male volun-
teers suffering from mild hypertension. The subjects were randomized into four groups: a placebo and
three drug groups that received 46, 93, and 185 mg of GCB extract once a day. After 4 weeks, systolic
and diastolic blood pressures were significantly reduced in a dose-dependent manner by GCB extracts
(P < 0.001), and no adverse effects were caused by the consumption of GCB extracts. The findings suggest
that daily use of GCB extract has a blood pressure‒lowering effect in patients with mild hypertension.
53.4.4 Anticarcinogenesis
The activations of phase II detoxifying and antioxidant enzymes such as glutathione S-transferase (GST),
NAD(P)H quinone oxidoreductase 1 (NQO1), γ-glutamylcysteine ligase, or heme oxygenase 1 are one
of the primary ways to protect cells and tissues from carcinogens and carcinogenic metabolites [33].
Recently, Boettler et al. [34] identified two potent activators of the Nrf2/ARE (antioxidant response
element) pathway using HT29, a human colon carcinoma cell line. They were Nrf2 nuclear translocation
and ARE-dependent gene expression of selected antioxidative phase II enzymes from coffee constituents,
namely, 5-O-CQA and pyridinium derivatives such as the N-methylpyridinium ion. 5-O-CQA is a CGA
found in GCBs and N-methylpyridinium is a typical roasting product. According to Volz et al. [35], a cof-
fee blend combining GCB constituents demonstrated chemopreventive effects by promoting the nuclear
Nrf2 translocation and resulted in increased transcription levels of the Nrf2-dependent phase II enzymes
GST1 and NQO1 as well as increased GST enzyme activity in vitro. In a subsequent human intervention
trial, significantly increased mean Nrf2 gene transcripts and a decrease in oxidative DNA damage in
peripheral blood lymphocytes were observed in test subjects after 4 weeks of coffee consumption [35].
53.4.5 Neuroprotective Potential

Caffeine in coffee has been shown to possess a protective effect against Alzheimer’s disease [36]
and Parkinson’s disease [10]. Apart from caffeine, coffee is a rich source of antioxidant constituents,
especially CGAs, which contribute to many of the health benefits associated with coffee consumption.
CGAs, or their metabolites, provide antioxidant capacity as well as regulation of enzymes and thereby
ameliorate oxidative stress, inflammation, and carcinogenesis. According to Daglia et al. [37], the neu-
roprotective effects established by epidemiological studies for coffee consumption are attributed to the
antioxidants present in coffee and GCB preparations such as 5-CQA, which demonstrated significant
antihydroxyl radical activity in vitro and ex vivo. Furthermore, decaffeinated GCB coffee was found
to be able to improve brain mitochondrial energy metabolism by modulating a number of genes in
the brain, which are implicated in cellular energy metabolism. This could benefit the progressively
impaired glucose energy metabolism in the brain caused by cognitive dysfunction in aging as well as
the neurodegenerative disorders [38].

GCB extracts, possessing high levels of CGAs, are prepared by steeping ground GCBs in hot water.
The purported health benefits of the brew are why companies have begun coming out with supplements
(e.g., All-Nutra™ GCB extract powder from AuNutra, Chino, CA) or beverages made with an elixir of
GCBs. Unfortunately, the resulting brew does not have a particularly pleasant taste, so this offers many
challenges to product developers. It is very important in the preparation of brews to try and extract as many
of the CGAs as possible. One study compared microwave-assisted extraction (MAE) and conventional
heat reflux methods. The authors reported that the yields of CGAs by MAE under optimum conditions
(5 min, 800 W, and 50°C, using water as the solvent for extraction) were higher than conventional reflux
methods [39]. Therefore, the MAE process has a potential in industrial applications and could be an alter-
native to conventional solvent extraction methods for the isolation of phenolic constituents from plants.
A number of products made with GCB extracts exist on the market today. These include Skinny
Coffee™ from Genesis Today, which has been standardized to contain at least 50% CGAs (3-CQA,
4-CQA, and 5-CQA) and claims support for energy and healthy weight; Starbucks Refreshers, a sparkling
ready-to-drink beverage made with real fruit juice and GCB extract in strawberry lemonade, raspberry
pomegranate, and orange melon varieties; PURE Café from Genesis Pure, which is a stevia-sweetened
beverage providing a robust mocha flavor including a blend of roasted coffee beans and 900 mg of GCB
extract; and finally, DaVinci Gourmet® Invigorators™, a fruit-flavored beverage concentrate with a hint
of GCB extract. If claims persist that consumption of GCB beverages aid in the reduction of one’s body
weight, then likely there will be an exponential growth in GCB-containing nutraceutical beverages, by
either the diversification of existing beverage lines or the creation of new brands.
53.5.1 Potential Health Risks of Green Coffee Consumption

GCBs and their extracts contain caffeine, which can cause numerous potential health risks when consumed
in large amounts. Even though significant associations between coffee consumption and the development
of hypertension have not been established, it is well known that blood pressure can increase by acute con-
sumption of caffeine at dietary levels [10]. About 200–250 mg of caffeine are contained in 2–3 cups of
coffee, and in normotensive individuals, this can raise one’s systolic blood pressure by 3–14 mmHg and
one’s diastolic blood pressure by 4–13 mmHg [40]. Based on the available data, women who are having dif-
ficulty conceiving or are pregnant and lactating should limit caffeine consumption to less than 300 mg/day
as well as eliminate tobacco use and decrease alcohol consumption [10]. Furthermore, older adults are
more vulnerable to the side effects of caffeine, and they should ensure adequate calcium and vitamin D
intake and limit coffee consumption to 3 cups/day (300 mg/day of c affeine). This may help lower the risk of
osteoporosis and osteoporotic fractures [10]. Children are another special risk group, who are susceptible
to the effects of caffeine. A high dose of caffeine (e.g., >3.0 mg/kg) has been reported to result in some
behavioral effects, such as increased nervousness or anxiety and sleep disturbances [10].
On the other hand, CGAs have a laxative effect and may cause diarrhea if consumed in too large of
a quantity [41]. Coffee polyphenols can also inhibit the intestinal absorption of nonheme iron [42]. Iron
absorption was reported to have decreased by 39% after drinking a cup of coffee with a hamburger
meal [43]. Brewed coffee contains a brownish zinc-chelating polymer, formed by the Maillard reaction
involving the polymerization of phenolics or their degradation products during roasting and designated as
ApV [44]. The content of ApV is elevated owing to the roasting process. ApV can inhibit the bioavailability
of zinc in vitro by 21%–32% [45]. Further research is needed to examine the effect of GCBs on zinc absorp-
tion. Consuming unfiltered coffee (e.g., prepared by the French press method) can increase both serum total
and low-density lipoprotein (LDL) cholesterol concentrations because of the presence of diterpenes such as
cafestol and kahweol [46]. Moreover, the CGAs and caffeine of coffee can result in elevated plasma total
homocysteine concentrations, which are considered as another cardiovascular disease risk factor [47,48].
Although extremely rare, GCB can also initiate an allergic reaction in coffee workers by a class III chitin-
ase of C. arabica, which was identified by Manavski et al. [49] to be the first known coffee allergen Cof a 1.
53.6 Conclusion
Although many people believe significant health risks exist with excessive consumption of caffein-
ated coffee, the science does not support this hypothesis. Processing techniques such as decaffeination
via the Swiss water treatment and paper filtration of coffee are at hand to remove, or at least reduce,
bioactives such as caffeine and diterpenes, which may present health issues at too high concentrations
to a small segment of the population. GCB extracts are rich in CGAs and have attracted interest for
their preventative effects against diabetes mellitus, obesity, hypertension, cancer, and Parkinson’s and
Alzheimer’s disease. A number of health benefits have been associated with an intake of 3–4 cups of
coffee daily [10]. Consumption of lesser amounts, such as 2–3 cups of coffee per day, can improve
cognitive function, the sense of sensation, and digestion and also reduce the risk of developing chronic
diseases [50]. In fact, the Pan American Health Organization recommends coffee consumption in the
amount of 3–4 cups per day for overall health improvement and disease prevention. Nevertheless,
special risk groups to the effects of caffeine such as pregnant and lactating women, children, and older
adults perhaps should be more careful with regard to their consumption of coffee or nutraceutical
beverages containing GCB extracts. More research on determining the health effects of GCB extracts,
specifically, is highly desirable to draw a conclusive result and to eliminate the ambiguities.
REFERENCES
1. Clifford, M.N., Chlorogenic acids and other cinnamates—Nature, occurrence and dietary burden.
J. Sci. Food Agric., 79, 362–372, 1999.
2. Del Castillo, M.D., Ames, J.M., and Gordon, M.H., Effect of roasting on the antioxidant activity of
coffee brews. J. Agric. Food Chem., 50, 3698–3703, 2002.
3. Del Pino-García, R., Gónzalez-SanJosé, M.L., Rivero-Pérez, M.D., and Muñiz, P., Influence of the
degree of roasting on the antioxidant capacity and genoprotective effect of instant coffee: Contribution
of the melanoidin fraction. J. Agric. Food Chem., 60, 10530–10539, 2012.
4. de Castro Moreira, M.E., Pereira, R.G.F.A., Dias, D.F., Gontijo, V.S., Vilela, F.C., de Oliveira Isac de
Moraes, G., Giusti-Paiva, A., and dos Santos, M.H., Anti-inflammatory effect of aqueous extracts of
roasted and green Coffea arabica L. J. Funct. Foods, 5, 466–474, 2013.
5. Ky, C.-L, Louarn, J., Dussert, S., Guyot, B., Hamon, S., and Noirot, M., Caffeine, trigonelline,
chlorogenic acids and sucrose diversity in wild Coffea arabica L. and C. canephora P. accessions.
Food Chem., 75, 223–230, 2001.
6. Clifford, M.N., Johnston, K.L., Knight, S., and Kuhnert, N., Hierarchical scheme for LC-MSn identifica-
tion of chlorogenic acids. J. Agric. Food Chem., 51, 2900–2911, 2003.
7. Farah, A. and Donangelo, C.M., Phenolic compounds in coffee. Braz. J. Plant Physiol., 18, 23–36, 2006.
8. Clifford, M.N., Knight, S., Sururu, B., and Kuhnert, N., Characterization by LC-MS n of four new classes
of chlorogenic acids in green coffee beans: Dimethoxycinnamoylquinic acids, diferuloylquinic acids,
caffeoyl-dimethoxycinnamoylquinic acids, and feruloyl-dimethoxycinnamoylquinic acids. J. Agric.
Food Chem., 54, 1957–1969, 2006.
9. Ranheim, T. and Halvorsen, B., Coffee consumption and human health—Beneficial or detrimental?
Mechanisms for effects of coffee consumption on different risk factors for cardiovascular disease and
type 2 diabetes mellitus. Mol. Nutr. Food Res., 49, 274–284, 2005.
10. Higdon, J.V. and Frei, B., Coffee and health: A review of recent human research. Crit. Rev. Food Sci.
Nutr., 461, 101–123, 2006.
11. Onakpova, I., Terry, R., and Ernst, E., The use of green coffee extract as a weight loss supplement:
A systematic review and meta-analysis of randomised clinical trials. Gastroenterol. Res. Pract., Article
ID 382852, 6 p., 2011.
12. Vinson, J.A., Burnham, B.R., and Nagendran, M.V., Randomized, double-blind, placebo-controlled,
linear dose, crossover study to evaluate the efficacy and safety of a green coffee bean extract in overweight
subjects. Diabetes Metab. Syndr. Obes., 5, 21–27, 2012.
13. Saito, I., Tsuchida, T., Watanabe, T., Arai, Y., Mitsui, Y., Okawa, W., and Kajihara, Y., Effect of coffee
bean extract in essential hypertension. Jpn. J. Med. Pharm. Sci., 47, 67–74, 2002.
14. Suzuki, A., Kagawa, D., Ochiai, R., Tokimitsu, I., and Saito, I., Green coffee bean extract and its metabolites
have a hypotensive effect in spontaneously hypertensive rats. Hypertens. Res., 25, 99–107, 2002.
15. Watanabe, T., Arai, Y., Mitsui, Y., Kusaura, T., Okawa, W., Kajihara, Y., and Saito, I., The blood pressure-
lowering effect and safety of chlorogenic acid from green coffee bean extract in essential hypertension.
Clin. Exp. Hypertens., 28, 439–449, 2006.
16. Rice-Evans, C.A., Miller, N.J., and Paganga, G., Structure–antioxidant activity relationships of fl avonoids
and phenolic acids. Free Radic. Biol. Med., 20, 933–956, 1996.
17. Iwai, K., Kishimoto, N., Kakino, Y., Mochida, K., and Fujita, T., In vitro antioxidative effects and
tyrosinase inhibitory activities of seven hydroxycinnamoyl derivatives in green coffee beans. J. Agric.
Food Chem., 52, 4893–4898, 2004.
18. Olthof, M.R., Hollman, P.C.H., Buijsman, M.N.C.P., van Amelsvoort, J.M.M., and Katan, M.B.,
Chlorogenic acid, quercetin-3-rutinoside and black tea phenols are extensively metabolized in humans.
J. Nutr., 133, 1806–1814, 2003.
19. Olthof, M.R., Hollman, P.C.H., and Katan, M.B., Chlorogenic acid and caffeic acid are absorbed in
humans. J. Nutr., 131, 66–71, 2001.
20. Farah, A., Monteiro, M., Donangelo, C.M., and Lafay, S., Chlorogenic acids from green coffee extract
are highly bioavailable in humans. J. Nutr., 138, 2309–2315, 2008.
21. Svilaas, A., Sakhi, A.K., Andersen, L.F., Svilaas, T., Ström, E.C., Jacobs Jr, D.R., Ose, L., and Blomhoff, R.,
Intakes of antioxidants in coffee, wine, and vegetables are correlated with plasma carotenoids in humans.
J. Nutr., 134, 562–567, 2004.
22. Natella, F., Nardini, M., Giannetti, I., Dattilo, C., and Scaccini, C., Coffee drinking influences plasma
antioxidant capacity in humans. J. Agric. Food Chem., 50, 6211–6216, 2002.
23. Baeza, G., Amigo-Benavent, M., Sarriá, B., Goya, L., Mateos, R., and Bravo, L., Green coffee hydroxy-
cinnamic acids but not caffeine protect human HepG2 cells against oxidative stress. Food Res. Int., 62,
1038–1046, 2014.
24. Arion, W.J., Canfield, W.K., Ramos, F.C., Schindler, P.W., Burger, H.-J., Hemmerle, H., Schubert,
G., Below, P., and Herling, A.W., Chlorogenic acid and hydroxynitrobenzaldehyde: New inhibitors of
hepatic glucose 6-phosphatase. Arch. Biochem. Biophys., 339, 315–322, 1997.
25. van Dam, R.M. and Feskens, E.J.M., Coffee consumption and risk of type 2 diabetes mellitus. Lancet,
360, 1477–1478, 2002.
26. Salazar-Martinez, E., Willett, W.C., Ascherio, A., Manson, J.E., Leitzmann, M.F., Stampfer, M.J., and
Hu, F.B., Coffee consumption and risk for type 2 diabetes mellitus. Ann. Intern. Med., 140, 1–8, 2004.
27. Tuomilehto, J., Hu, G., Bidel, S., Lindström, J., and Jousilahti, P., Coffee consumption and risk of type
2 diabetes mellitus among middle-aged Finnish men and women. JAMA, 291, 1213–1219, 2004.
28. Thom, E., The effect of chlorogenic acid enriched coffee on glucose absorption in healthy volunteers
and its effect on body mass when used long-term in overweight and obese people. J. Inter. Med. Res., 35,
900–908, 2007.
29. Tunnicliffe, J.M. and Shearer, J., Coffee, glucose homeostasis, and insulin resistance: Physiological
mechanisms and mediators. Appl. Physiol. Nutr. Metab., 33, 1290–1300, 2008.
30. Flanagan, J., Bily, A., Rolland, Y., and Roller, M., Lipolytic activity of Svetol®, a decaffeinated green
coffee bean extract. Phytother. Res., 28, 946–948, 2014.
31. Ochiai, R., Jokura, H., Suzuki, A., Tokimitsu, I., Ohishi, M., Komai, N., Rakugi, H., and Ogihara, T.,
Green coffee bean extract improves human vasoreactivity. Hypertens. Res., 27, 731–737, 2004.
32. Kozuma, K., Tsuchiya, S., Kohori, J., Hase, T., and Tokimitsu, I., Antihypertensive effect of green coffee
bean extract on mildly hypertensive subjects. Hypertens. Res., 28, 711–718, 2005.
33. Talalay, P., Chemoprotection against cancer by induction of phase 2 enzymes. BioFactors, 12, 5–11, 2000.
34. Boettler, U., Sommerfeld, K., Volz, N., Pahlke, G., Teller, N., Somoza, V., Lang, R., Hofmann, T.,
and Marko, D., Coffee constituents as modulators of Nrf2 nuclear translocation and ARE (EpRE)-
dependent gene expression. J. Nutr. Biochem., 22, 426–440, 2011.
35. Volz, N., Boettler, U., Winkler, S., Teller, N., Schwarz, C., Bakuradze, T., Eisenbrand, G. et al., Effect
of coffee combining green coffee bean constituents with typical roasting products on the Nrf2/ARE
pathway in vitro and in vivo. J. Agric. Food Chem., 60, 9631–9641, 2012.
36. Arendash, G.W. and Cao, C., Caffeine and coffee as therapeutics against Alzheimer’s disease.
J. Alzheimers Dis., 20(Suppl.), S117–S126, 2010.
37. Daglia, M., Racchi, M., Papetti, A., Lanni, C., Govoni, S., and Gazzani, G., In vitro and ex vivo antihy-
droxyl radical activity of green and roasted coffee. J. Agric. Food Chem., 52, 1700–1704, 2004.
38. Ho, L., Varghese, M., Wang, J., Zhao, W., Chen, F., Knable, L.A., Ferruzzi, M., and Pasinetti, G.M.,
Dietary supplementation with decaffeinated green coffee improves diet-induced insulin resistance and
brain energy metabolism in mice. Nutr. Neurosci., 15, 37–45, 2012.
39. Upadhyay, R., Ramalakshmi, K., and Rao, L.J.M., Microwave-assisted extraction of chlorogenic acids
from green coffee beans. Food Chem., 130, 184–188, 2012.
40. Nurminen, M.-L., Niittynen, L., Korpela, R., and Vapaatalo, H., Coffee, caffeine and blood pressure:
A critical review. Eur. J. Clin. Nutr., 53, 831–839, 1999.
41. Stacewicz-Sapuntzakis, M., Bowen, P.E., Hussain, E.A., Damayanti-Wood, B.I., and Farnsworth, N.R.,
Chemical composition and potential health effects of prunes: A functional food? Crit. Rev. Food Sci.
Nutr., 41, 251–286, 2001.
42. Fairweather-Tait, S.J., Iron nutrition in the UK: Getting the balance right. Proc. Nutr. Soc., 63, 519–528,
2004.
43. Morck, T.A., Lynch, S.R., and Cook, J.D., Inhibition of food iron absorption by coffee. Am. J. Clin.
Nutr., 37, 416–420, 1983.
44. Wen, X., Enokizo, A., Hattori, H., Kobayashi, S., Murata, M., and Homma, S., Effect of roasting on
properties of the zinc-chelating substance in coffee brews. J. Agric. Food Chem., 53, 2684–2689, 2005.
45. van Dyck, K., Tas, S., Robberecht, H., and Deelstra, H., The influence of different food components
on the in vitro availability of iron, zinc and calcium from a composed meal. Int. J. Food Sci. Nutr., 47,
499–506, 1996.
46. Urgert, R. and Katan, M.B., The cholesterol-raising factor from coffee beans. Ann. Rev. Nutr., 17,
305–324, 1997.
47. Verhoef, P., Pasman, W.J., van Vliet, T., Urgert, R., and Katan, M.B., Contribution of caffeine to the
homocysteine-raising effect of coffee: A randomized controlled trial in humans. Am. J. Clin. Nutr., 76,
1244–1248, 2002.
48. Olthof, M.R., Hollman, P.C., Zock, P.L., and Katan, M.B., Consumption of high doses of chlorogenic
acid, present in coffee, or of black tea increases plasma total homocysteine concentrations in humans.
Am. J. Clin. Nutr., 73, 532–538, 2001.
49. Manavski, N., Peters, U., Brettschneider, R., Oldenburg, M., Baur, X., and Bittner, C., Cof a 1:
Identification, expression and immunoreactivity of the first coffee allergen. Int. Arch. Allergy Immunol.,
159, 235–242, 2012.
50. Butt, M.S. and Sultan, M.T., Coffee and its consumption: Benefits and risks. Crit. Rev. Food Sci. Nutr.,
51, 363–373, 2011.
54
Cocoa and Hot Chocolate
Beatriz Sarriá, Raquel Mateos, and Laura Bravo
CONTENTS
54.1 Introduction................................................................................................................................... 687
54.4 Health Effects................................................................................................................................ 692
54.6 Conclusion..................................................................................................................................... 699
References............................................................................................................................................... 700
54.1 Introduction
The worldwide production of raw cocoa beans in 1895 was about 75,000 metric tons (MT). One hundred
years later, raw cocoa production has increased almost 40-fold, up to 2,760,000 MT. The majority of today’s
world cocoa supply comes from West Africa, followed by Central and South America. For commercial
purposes, the varieties developed through breeding and cultivation are more important than the species
of Theobroma. These varieties are often classified into three or four broad categories: Criollo, Forastero,
Trinitario, and Nacional, based on the sharp distinction in the kind of flavor and color they exhibit in the
manufacturing process [1].
Cocoa is the dried and fermented fatty seed of the Theobroma cacao L. tree from which chocolate is made.
In pre–Columbian times, the fruity pulp surrounding cacao seeds was used in South America as “a refresh-
ing source of liquid or was fermented to produce chicha (an alcoholic beverage often made from maize or
manioc/cassava)” [2]. Before initial European–Mexican contact in 1519, ancient Central Americans prepared
cocoa only as a beverage, which was a very valuable and prestigious food reserved for nobility. Chocolate,
prepared as a beverage, was introduced to the Spanish court in 1544 [3]. Within a century, the culinary and
medical uses of chocolate had spread to France, England, and elsewhere in Western Europe. In 1648, choco-
late was commonly drunk throughout the West Indies, as well as in Flanders, Italy, and Spain [4].
The first important technical development in the chocolate manufacturing process occurred when water-
powered mills superseded the use of manual labor to grind cocoa beans. This led to the establishment of many
chocolate factories from 1804 to 1840. Early production consisted entirely of a type of chocolate beverage that
was somewhat indigestible since cocoa butter was not removed during processing. In 1828, the cocoa press was
invented, which facilitated the production of cocoa powder by partial removal of the cocoa butter from beans,
resulting in a beverage more palatable and easier to prepare. In the nineteenth century, the production of cocoa
using this new technology increased and its consumption raised dramatically [1]. In the first half of the twen-
tieth century, cocoa beverages were regarded as a bedtime drink, an idea that has remained until recent years,
although the high fat and, in the case of drinking chocolate, high sugar content has limited its consumption.
Today, cocoa is considered as to be a dry, powdered, and nonfat component product manufactured
from cocoa tree seeds. Cocoa liquor contains cocoa as well as cocoa butter (∼55%); it is used in choco-
late confectionery manufacturing along with other ingredients such as sugar, emulsifiers, and milk
protein, depending on the desired product [5]. Novel cocoa beverages have been developed consisting
of reduced caloric products made from defatted cocoa powder with added artificial sweeteners and
687
products enriched with bioactive components that are perceived as higher quality and healthier drinks.
This chapter highlights the nutritional characteristics and bioactive content of cocoa and hot chocolate,
as well as the health effects related to acute or regular consumption, showing that moderate amounts of
cocoa beverages may contribute to the prevention of chronic diseases in the context of a healthy diet.

Unroasted cocoa beans present high energy and nutritional density. Although the chemical composition
varies depending on geographical origin, in general fat is the major nutrient (>40%), followed
by carbohydrate, mainly sugars (>32%), and protein (12%–13%). Table 54.1 shows the compositional
TABLE 54.1
Compositional and Nutritional Characteristics of Cocoa Mix Powder and Hot
Chocolate (per 100 g)
Nutrient Unit Cocoa Mix Powder Hot Chocolate
Water g 1.50 86.34
Energy kcal 398 55
Protein g 6.67 0.92
Lipid (fat) g 4.00 0.55
Minerals
Calcium mg 133 21
Copper mg 0.29 0.048
Iron mg 1.19 0.17
Magnesium mg 83 12
Manganese mg 0.27 0.037
Phosphorus mg 315 43
Potassium mg 712 99
Selenium µg 5.0 0.7
Sodium mg 504 73
Zinc mg 1.46 0.21
Vitamins
Choline mg 33.1 4.6
Niacin mg 0.586 0.081
Pantothenic acid mg 0.893 0.123
Vitamin A IU 4 1
Vitamin B12 µg 0.35 0.05
Vitamin C mg 0.2 0
Database for Standard Reference, Release 27, USDA, Springfield, VA, 2014.
Abbreviations: DFE, dietary folate equivalents; RAE, retinol activity equivalents; ATE,
Cocoa and Hot Chocolate 689
and nutritional characteristics of cocoa mix powder and cocoa mix powder prepared with water
(hot chocolate) [6]. However, carbohydrate is the main nutrient in hot chocolate (12%) due to added
sugar (9%), followed by protein (0.9%) and lipid (0.6%). Regarding the micronutrient composition, cocoa
powder is a rich source of minerals so that cocoa and chocolate can provide significant amounts of the
Recommended Dietary Allowances (RDA) of iron, magnesium, and zinc [7]. Cocoa powder is also a rich
source of vitamins, particularly choline and folate.

Cocoa presents high contents of flavonoids, theobromine, and dietary fiber that show extraordinary
bioactivity (Tables 54.1 and 54.2) [6]. In cocoa beans, the main flavonoids are flavan-3-ols, present as
monomeric (−)-epicatechin and (+)-catechin, together with type-B proanthocyanidins, formed from
monomeric flavanols by oxidative coupling between the C4 position of the heterocyclic ring and the C6
or C8 positions of the adjacent unit to create oligomers and polymers [5]. Procyanidins in cocoa include
the B1 and B2 types [8] (Figure 54.1), as well as B5 dimers and the C1 trimer [5], together with high levels
of longer-chain polymers with four or more monomeric units. Other flavonoids and phenolic compounds
are also present in minor proportions [8]. Raw cocoa contains significantly more (−)-epicatechin than
(+)-catechin, and the procyanidins are, therefore, believed to contain a high density of epicatechin sub-
units. Interestingly, epicatechin and its related dimeric procyanidins are unstable in some thermal and
extreme pH environments, such as that in food processing practices associated with the manufacture of
cocoa and chocolate products, and significant conversion of epicatechin to catechin can occur, and the
flavanols and procyanidins originally present in the cocoa bean can be substantially reduced or elimi-
nated. When the lipophilic and hydrophilic antioxidant capacities of several types of cocoa and chocolate
products were determined, using both hydrophilic and lipophilic oxygen radical absorbance capacity
(ORAC) assays, hydrophilic fractions contributed >90% to the total antioxidant capacity in all cocoa
products, and the procyanidin content highly correlated with the antioxidant capacity (r 2 = 0.92), thus
suggesting these compounds were the dominant antioxidants [9]. On the way from fresh beans to finished
cocoa products, the concentration of flavan-3-ols and procyanidins can be affected by biological and
processing conditions, such as fermentation, treatment with alkali (“dutching”), and roasting (usually up
to 120°C), as well as the addition of sugar, milk, vanilla, and emulsifiers; therefore, both the origin of the
cocoa bean and the processing determine the antioxidant properties of cocoa products [10].
TABLE 54.2
Carotenoids, Alkaloids, and Flavonoids in Cocoa Mix Powder and Hot Chocolate (per 100 g)
Unit Cocoa Mix Powder Hot Chocolate
Carotenoids
Lutein + zeaxanthin µg 5 1
Alkaloids
Caffeine mg 18 2
Theobromine mg 323 44
Flavonoids
(+)-Catechin mg 21.5 0.7
(−)-Epicatechin mg 31.2 0.6
Quercetin mg 2.0 na
Epigallocatechin mg na 0.0
Epigallocatechin 3-gallate mg na 0.0
Epicatechin 3-gallate mg na 0.0
Gallocatechin mg na 0.0
Source: Adapted from the U.S. Department of Agriculture (USDA), USDA National Nutrient Database
for Standard Reference, Release 27, USDA, Springfield, VA, 2014.
FLAVANOLS
Monomers
OH OH
OH OH
O HO O
HO
OH OH
OH OH
(+)-Catechin (–)-Epicatechin
Polymers
OH
OH
OH HO O
OH OH
OH
HO O O OH
OH HO OH
OH
OH HO O
OH OH OH
OH OH OH
OH OH
HO O HO O OH
OH OH
HO O
OH OH
OH OH OH
OH
Procyanidin B1 Procyanidin B2 Procyanidin C1
METHYLXANTHINE
O CH3
HN N
O N N
CH3
Theobromine
FIGURE 54.1 Chemical structures of major phenolic compounds and methylxanthine in cocoa.
Comparing cocoa to other phenolic-rich beverages, although the predominant phenolic compounds
are different, cocoa contains much higher levels of total phenolics (611 mg of gallic acid equivalents
[GAE]) and flavonoids (564 mg of epicatechin equivalents [ECE]) per serving (7.3 g of Ghanaian cocoa
beans in 200 mL of distilled water) than black tea (124 mg of GAE and 34 mg of ECE, respectively),
green tea (165 mg of GAE and 47 mg of ECE; the serving of both teas consisted 2 g of tea [tea bag]
in 200 mL of distilled water), and red wine (340 mg of GAE and 163 mg of ECE; the serving of wine
was 140 mL). Accordingly, cocoa exerted the highest antioxidant activity among the beverages studied.
The 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl
(DPPH) radical scavenging assays indicated values of 1128 and 836 mg of ascorbic acid equivalents
(AAE) per serving, respectively. The total antioxidant capacities from ABTS and DPPH assays highly
correlated with phenolic content (r 2) of 0.981 and 0.967, respectively, and flavonoid content (r 2) of 0.949
and 0.915, respectively [11]. Cocoa products were among the food groups richest in polyphenols accord-
ing to the values collected in the Phenol-Explorer database [12]. In Table 54.3, the polyphenol content
and antioxidant activity of cocoa products, including a chocolate beverage with milk, alcoholic bever-
ages, and nonalcoholic beverages, which have been included among the 100 richest dietary sources of
polyphenols, are shown. In the original list, the 100 richest foods were ranked according to their poly-
phenol contents and antioxidant activities [12]. Cocoa powder and dark chocolate appeared at the 4th
and 8th positions (according to polyphenol content) and 24th and 13th positions (according to antioxidant
activity), respectively. When the same 100 foods were ranked according to the amount of polyphenols in
TABLE 54.3
Polyphenols and Antioxidant Activity in the Cocoa Products and Beverages Included in the 100 Richest
Dietary Sources of Polyphenols (mg/100 g or mg/100 mL)
Food Food Group Polyphenolsa Polyphenols AE Antioxidantsb
Cocoa powder Cocoa products 3448 3294 1104
Dark chocolate Cocoa products 1664 1618 1860
Milk chocolate Cocoa products 236 236 854
Coffee (filter) Nonalcoholic beverages 214 110 267
Black tea Nonalcoholic beverages 102 90 104
Red wine Alcoholic beverages 101 91 215
Green tea Nonalcoholic beverages 89 82 62
Pure apple juice Nonalcoholic beverages 68 61 34
Pure pomegranate juice Nonalcoholic beverages 66 37 204
Pure blood orange juice Nonalcoholic beverages 56 28 72
Pure grapefruit juice Nonalcoholic beverages 53 23 54
Pure blond orange juice Nonalcoholic beverages 46 20 —
Pure lemon juice Nonalcoholic beverages 42 20 —
Chocolate beverage with milk Nonalcoholic beverages 21 21 —
Soy milk Nonalcoholic beverages 18 11 —
Pure pomelo juice Nonalcoholic beverages 18 7.9 —
White wine Alcoholic beverages 10 8.6 32
Rosé wine Alcoholic beverages 10 7.8 82
Source: Adapted from Pérez-Jiménez, J. et al., Eur. J. Clin. Nutr., 64(Suppl.), S112, 2010. With permission.
a Sum of the content of individual polyphenols as determined by chromatography and of proanthocyanidin oligomers as
determined by direct-phase high-performance liquid chromatography.
b Determined by the Folin assay. Some foods with a high antioxidant content as determined by the Folin assay are not
included in the table due the absence of documented data on their polyphenol content as obtained by chromatography.
Abbreviation: AE, aglycone equivalents.
a serving (according to the Food Standards Agency, UK), a second list was produced in which cocoa
powder and dark chocolate appeared at the 24th and 14th positions, close to black tea, green tea, and
red wine (16th, 17th, and 22nd positions, respectively). In contrast, when the serving amount of the
chocolate beverage with milk was considered, the beverage changed from position 87 in the former list,
to position 39 in the latter, proceeded by coffee (filter), black and green tea, and pure apple, pomegranate,
grapefruit, and blood orange juices, as well as red wine, and ahead of soy milk, pure pomelo juice, and
beer, in addition to white and rosé wine, dark beer, and pure lemon juice.
The antioxidant properties of flavonoids are based, in part, on their structural characteristics, including
the degree of hydroxylation of the B-ring (catechol structure), the oligomer chain length, and the stereo-
chemical features of the molecule. The structural characteristics of flavonols represent the molecular
bases for both their hydrogen-donating (radical scavenging) and metal-chelating antioxidant properties.
For example, cocoa and purified cocoa flavonoids and procyanidins have been reported to attenuate the
copper-mediated and endothelial cell–mediated oxidation of low-density lipoprotein (LDL), to reduce the
production of reactive oxygen species by activated leukocytes, to protect against erythrocyte hemolysis,
and to inhibit ultraviolet-C (UV-C)-induced DNA oxidation [13].
Although cocoa has a potent antioxidant activity in vitro, the critical question is whether the same
effects can be observed in vivo. Cocoa’s antioxidant action could not be confirmed in 19 controlled
intervention studies in healthy participants and specific patient groups in which markers of plasma anti-
oxidant capacity and oxidative stress were analyzed after bolus and/or regular cocoa consumption [14].
In contrast, other studies show that acute dark chocolate consumption may affect the antioxidant status
and oxidative stress responses to prolonged exercise [15], the antioxidant capacity enhancement being
greatest 1–2 h after cocoa administration and gradually decreasing to reach baseline levels at about 6 h
postingestion, probably due to the short plasma half-life of flavonoids. Similarly, cocoa modulated lipid
peroxidation inducing an acute decrease in isoprostane levels 2–4 h after intake, disappearing after 6 h,
although this effect could not be confirmed in another acute cocoa intake study. In chronic studies, the
lipid peroxidation biomarker level, oxidized LDL, in hypercholesterolemic volunteers, was decreased
after consuming cocoa powders with three polyphenolic levels (13, 19.5, and 26 g/day) for 4 weeks [16],
although no effects were observed in healthy participants or smokers [17]. The possible mechanisms
that contribute to the antioxidant protection of cocoa phenolic compounds are due to the presence of a
catechol group on the B ring in flavanols, epicatechin, and catechin, which can trap free radicals and
chelate redox-active metals. In addition, these small molecules are able to diffuse across the cell mem-
brane, in contrast to longer-chain oligomers, and affect enzymes and signaling cascades, augmenting the
antioxidant defense system. Regarding cocoa procyanidins, due to the higher number of catechol groups,
the antioxidant activity of these phenolics could increase with the oligomer chain length; however, there
are studies that do not support this relation, and certainly procyanidins have different interactions with
biological cell membranes than flavanols do [13].
The cocoa bean is the major natural source of theobromine in the diet. Although methylxanthine con-
centrations in cocoa beans are broadly variety dependent, theobromine represents about 2.5% of cocoa
bean dry weight, whereas caffeine is approximately 10-fold lower and theophylline is present at low and
negligible concentrations [5]. Theobromine is not degraded during cocoa processing and can be used as
a marker of cocoa content and a biomarker of cocoa intake [18]. Cocoa methylxanthines are bioavailable,
partially metabolized, and rapidly eliminated, presenting sustained urinary excretion for as long as 24 h
after intake [19]. Some authors report that theobromine and caffeine are neither pro-oxidant nor antioxi-
dant [20], although others indicate that these compounds display low antioxidant capacity compared with
other cocoa fractions, and their presence could reduce the antioxidant capacity of flavonoids in cocoa [21].
The cocoa bean has a seed coat or bran that is usually removed, constituting a considerable by-product,
which is particularly rich in insoluble dietary fiber (IDF) [22]. As part of the bran remains in cocoa
powder, it is a good source of dietary fiber, in contrast to chocolate [23]. Moreover, cocoa powder may be
supplemented with dietary fiber without negatively affecting its organoleptic properties. In view of cocoa
husks’ health potential, a fiber-rich product has been produced from cocoa husks as a potential functional
food ingredient, containing 10% (dry matter) of soluble dietary fiber, 50% of IDF, and 2.36% of soluble
polyphenolic compounds, which provide the cocoa fiber product with intrinsic antioxidant activity and,
if absorbed in the gastrointestinal tract, could contribute to the antioxidant status in vivo [22]. More
research is required to better understand how chronic daily cocoa consumption affects the biomarkers of
oxidative stress mainly in different population groups.
54.4 Health Effects

For many years, because of cocoa’s high saturated fatty acid content, it was considered to be a hyper-
cholesterolemic foodstuff. However, in 1994 [24], stearic acid (C18:0), the main saturated fatty acid in
cocoa, was shown to have neutral effects on serum total cholesterol and LDL cholesterol. Today, cocoa’s
beneficial cardiovascular effects are widely accepted and have been principally attributed to its contents
in flavonoids, in addition to theobromine and dietary fiber (Table 54.4). Nevertheless, the reported effects
of cocoa on blood lipid levels are not consistent; in 10 intervention studies (durations ranging from 2 to
12 weeks), cocoa decreased total and LDL cholesterol levels, whereas no major effects were observed
on high-density lipoprotein (HDL) cholesterol. Interestingly, the positive effects were stronger when the
studies using cocoa beverages were excluded, pointing to a discrepancy in the effective dose of flavanols
when delivered in a chocolate or beverage matrix [25]. In contrast, other 4-week-long studies showed
positive effects on HDL cholesterol levels after consuming soluble cocoa products in milk [23–27], and,
in addition, a decrease of LDL susceptibility to oxidation was also noted [16,17]. Flavonoids, particularly
flavanols, were the main bioactive compounds in all these studies responsible for the beneficial effects
on lipid profile. Flavanols may lower LDL cholesterol by inhibiting its biosynthesis, suppressing hepatic
secretion of apolipoprotein B (Apo B), and increasing hepatic expression of LDL cholesterol receptors,
whereas the pathways through which flavanols elevate HDL cholesterol involve the inhibition of vascu-
larw endothelial activation via apolipoprotein A1 (Apo A1) [28]. However, the health effects of cocoa
TABLE 54.4
Summary of the Most Relevant Human Clinical Trials on the Effects of Cocoa Beverages on Health
Bioactive Compounds Provided/
Intervention Study Design Dose(s) of Cocoa Consumed Outcomes References
LF, MF, or HF cocoa beverage Double blind and placebo controlled; Cocoa powder containing low-polyphenolic compounds ↓ Apo B concentration in middle [16]
(twice per day for 4 weeks) 160 normo- and mildly (placebo–cocoa group) or three levels of polyphenolic and high cocoa groups
hypercholesterolemic subjects. cocoa powder (13, 19.5, and 26 g/day for LF, MF, and ↓ Oxidized LDL-C in all groups
HF) and 7.8 g of total dietary fiber in all products.
Cocoa beverage consumed Randomized and controlled; 25 12 g sugar/day + 26 g of cocoa/day that provided ↓ LDL-C susceptibility to [17]
(twice per day for 12 weeks) healthy males. 26.9 g fiber, 7.7 g minerals, 377 mg epicatechin, oxidation
135 mg catechin, 158 mg procyanidin B2, 96.1 mg ↑ HDL-C
procyanidin C1, 2192 mg TB, and 470 mg caffeine
powder or 12 g sugar/day (control).
Soluble, fiber-rich cocoa Randomized, controlled, crossover, 30 g of cocoa/day that provided 10.17 g total dietary ↑ HDL-C [23]
powder in milk consumed and free-living study; healthy (n = 24) fiber, 43.8 mg flavanols, and 168.6 mg ↓ Glucose
(twice per day for 4 weeks) and moderately hypercholesterolemic methylxanthines or 500 mL of semiskimmed ↓ IL-1β
(>200 mg/dL, n = 20) subjects. milk/day (control). ↓ IL-10
Soluble, flavanol-rich cocoa Randomized, controlled, and The product provided daily 3.74 g, 45.3 mg, and ↑ HDL-C [27]
powder in milk free-living study; healthy (n = 24) and 109.8 mg of total dietary fiber, flavanols, and ↓ IL-10
moderately hypercholesterolemic methylxanthines, respectively.
(>200 mg/dL, n = 20).
Soluble, fiber-rich cocoa Free-living, noncontrolled, The cocoa product provided 12 g of dietary fiber and ↓ Glucose [33]
powder in milk (twice per nonrandomized, and open 283 mg of soluble polyphenols/day. ↓ SBP
day for 8 weeks) intervention trial; 21 moderately ↓ DBP (P = 0.001)
hypercholesterolemic volunteers. ↓ Lipid peroxidation
Acute ingestion of a beverage Double blind and randomized; 23 TB: 0, 27, 66, 138, or 279 mg. Dose-dependent greater change in [37]
containing either 0 (placebo) healthy subjects. Total polyphenols: 330, 420, 420, 840, or 1470 mg. FMD (5, 13, and 26 g)
or 2, 5,13, or 26 g of cocoa Flavan-3-ols: 0.0, 9.3, 25.8, 66.6, or 146.0 mg. ↑ SBP (2 and 26 g)
Total procyanidins: 0.0, 69, 180, 465, or 1095 mg. ↑ DBP (2, 13, and 26 g)
↑ MAP (2, 13, and 26 g)
Glucose (0, 2, and 5 g)
Acute ingestion for up to 6 h 11 healthy male subjects with Acutely, single doses of a cocoa drink with 28–918 mg Acutely, dose-dependent increases [38]
after single-dose intake; smoking-related endothelial of flavanols. in FMD and nitrite, maximal
chronic consumption of a dysfunction. Chronically, flavanol-rich cocoa drink (3 × 306 mg FMD at 2 h after consumption
flavanol-rich cocoa drink flavanols/day). ↑ FMD and circulating nitrite
(three times per day for
7 days)
(Continued)
693
694
HF and LF cocoa beverage Double blind, randomized, and Dietary HF intervention (375 mg) and a macronutrient- ↑ FMD (both conditions) [39]
(twice per day for 30 days) crossover; 16 coronary artery and micronutrient-matched LF intervention (9 mg). FMD postintervention higher than
disease patients. postcontrol
↑ % of circulating angiogenic cells
↓ Plasma nitrite
↑ SBP
SF cocoa beverage or an SS Double blind, randomized, and Catechin: 21, 21, and 0 mg. ↑ FMD (both conditions) [40]
cocoa beverage (twice per crossover; 39 overweight, Epicatechin: 48, 48, and 0 mg.
day for 6 weeks vs. a placebo healthy subjects. Total procyanidins (total flavanols): 805, 805, and 9 mg.
noncocoa beverage) TB: 436, 436, and 0 mg for SF, SS, and placebo,
respectively.
Cocoa beverage (twice per Double blind, randomized, and Flavanol-rich cocoa drink (~900 mg flavanols/day) or ↑ Insulin-stimulated brachial artery [41]
day for 2 weeks) crossover; 20 hypertensive subjects. flavanol-poor placebo (~28 mg flavanols/day). diameter
Flavanol-containing beverage Double blind and randomized; 41 type Flavanol-rich cocoa (321 mg flavanols per dose) or a ↑ FMD [42]
(three times per day for 2 diabetic subjects. nutrient-matched control (25 mg flavanols per dose).
30 days)
DC and cocoa beverage (once Five type 2 diabetic patients with stage DC and the cocoa beverage contained ~100 mg of ↑ HDL-C [43]
per day for 3 months) II and III heart failure. epicatechin/day. Enhanced expression of
mitochondrial structure markers
in skeletal muscle
LF or HF beverage (twice per Double blind and randomized; 49 HF cocoa (902 mg flavanols), HF, and exercise, LF ↑ FMD (combined exercise and [46]
day with or without 45 min overweight and obese subjects. cocoa (36 mg flavanols) or LF and exercise. nonexercise results)
of physical activity 3 days ↓ Insulin resistance, DBP, and
for 12 weeks) MAP (flavanol treatment nested
in time)
DC and cocoa beverage Double blind, placebo controlled, 37 g dark chocolate bar (~11 g natural cocoa, 397 mg ↑ Pulse rate at midpoint and end of [49]
(for 6 weeks) parallel group, and randomized; 101 total proanthocyanidins/g, and 237 mL of an artificially treatment
healthy subjects. sweetened cocoa beverage also ~11 g of cocoa and
357 mg proanthocyanidins/g) or similar placebo
products containing 0.2 and 40.9 mg
proanthocyanidins.
(Continued)
Cocoa beverage containing Double blind and randomized; 52 33, 372, 712, or 1052 mg total flavanols/day ↓ 24 h ambulatory MAP, SBP, and [50]
LF, MF, or HF contents mildly hypertensive subjects (20 for 6 weeks. DBP (1052 mg)
females and 32 males, 42–74 years). ↓ Overnight ambulatory SBP, DBP,
and HR
SC cocoa beverage or SF Single blind, randomized, and SC cocoa (containing 22 g cocoa powder), sugared ↑ FMD [51]
cocoa beverage (two crossover; 45 healthy subjects. cocoa (containing 22 g cocoa powder), or a placebo ↓ SBP and DBP
cups per day) (containing 0 g cocoa powder). Greatest FMD improvement
following SF cocoa beverage
LF or HF cocoa beverage, Double blind, randomized, and HF (701 mg) or a LF (22 mg) cocoa beverage. ↑ FMD [52]
followed by 10 min cycling crossover; 21 healthy overweight/ ↓ AUC for DBP and MAP in
obese subjects. response to exercise
FRC or cocoa beverage (for Randomized, double-blind, and Subjects consumed a FRC bar and cocoa beverage daily Unaltered vascular function [56]
6 weeks) placebo-controlled study; 40 (total flavanols, 444 mg/day) or matching isocaloric
volunteers with coronary artery placebos (total flavanols, 19.6 mg/day).
disease.
Cocoa beverage: 20 g cocoa Randomized and crossover; 42 40 g of cocoa/day: ↑ HDL-C [26,59]
powder in 250 mL skimmed high-risk CVD subjects. (+)-catechin: 10.41 mg. ↓ Expression of adhesion
milk (twice per day for (−)-epicatechin: 46.08 mg. molecules on the surface of
4 weeks) Procyanidin B2: 36.54 mg. monocytes and concentration of
Total proanthocyanidins: 425.7 mg. circulating soluble adhesion
Total polyphenols: 495 mg. molecules
HF or LF cocoa beverage Randomized, double-blind, and HF cocoa beverage (446 mg of total flavanols) or ↑ Brachial artery hyperemic blood [60]
(for 6 weeks) parallel (n = 16/group) study; LF cocoa beverage (43 mg of total flavanols). flow
32 postmenopausal
hypercholesterolemic women.
Acute consumption of a cocoa Double blind and crossover; 30 healthy 520 or 994 mg cocoa flavanols and a matched control. Both beverages improved serial [64]
beverage subjects. threes performance. The 994 mg
cocoa flavanol beverage speeded
RVIP
Increases in “mental fatigue” were
attenuated by the 520 mg CF
beverage
695
(Continued)
696

FRC beverage (for 1 week) Double blind and randomized; 21 FRC provided 900 mg flavanols/day versus a flavanol ↑ Cerebral blood flow in response [65]
healthy subjects. poor cocoa (36 mg flavanol/day). Both cocoas to acute dose of cocoa beverage
presented similar TB and magnesium contents.
MF or HF chocolate beverage Double blind and randomized; 63 250 mg (MF) or 500 mg (HF) cocoa flavanol drink ↑ Posterior parietal activity, [66]
(per day for 30 days) subjects. versus a 0 mg flavanol cocoa drink (placebo). synaptic excitation, and neural
information processing speed
Dairy-based flavanol-rich Double blind, randomized, and Flavanol-rich cocoa with natural dose of TB: 106 mg of ↑ 24 h DBP (flavanol-rich cocoa [55]
cocoa beverage containing crossover; 42 pre-/stage 1 TB or flavanol-rich cocoa with natural dose of TB with natural dose of TB)
either natural dose of TB or hypertensive and healthy subjects. enriched: 979 mg of TB. ↑ 24 h SBP, daytime DBP, 24 h,
TB enriched once per day day- and nighttime HR
for 3 weeks (TB-enriched flavanol-rich cocoa)
↓ Central. SBP, HR, and stroke
volume (TB-enriched flavanol-
rich cocoa)
Abbreviations: Apo B, apolipoprotein B; AUC, area under the curve; CVD, cardiovascular disease; DBP, diastolic blood pressure; DC, dark chocolate; FMD, flow-mediated dilatation; FRC,
flavanol-rich chocolate; HDL-C, high-density lipoprotein cholesterol; HF, high flavanol; HR, heart rate; IL, interleukin; LDL-C, low-density lipoprotein cholesterol; LF, low
flavanol; MAP, mean arterial pressure; MF, moderate flavanol; RVIP, rapid visual information processing; SBP, systolic blood pressure; SC, sugar-containing; SF, sugar-free;
SS, sugar-sweetened; TB, theobromine.
polyphenols depend on the amount consumed and their bioavailability (absorption, distribution, metabo-
lism, and elimination), which is influenced by the food matrix among other factors. Common ways to
prepare a cocoa beverage imply dissolving cocoa powder in milk or in hot water. Some studies indicate
that milk does not affect the bioavailability of cocoa flavonoids analyzed in plasma [29], whereas others
suggest that interaction between milk proteins and chocolate flavonoids negatively affects the absorption
of flavonoids [30].
Cocoa’s content of dietary fiber may also contribute to this effect, possibly through upregulation
of HDL cholesterol [31] or hindering digestion and absorption of dietary fats through diluting gas-
trointestinal contents, in the case of the insoluble fiber fraction [22]. In hyperlipidemic rats, a cocoa
product rich in IDF induced hypocholesterolemic and hypotriacylglycerolemic effects [32], but con-
versely when the same cocoa product was regularly consumed as a beverage mixed with milk by
moderately hypercholesterolemic subjects, blood lipid levels were unaffected and only a slight increase
in HDL cholesterol was observed [33]. In addition, theobromine in a cocoa beverage was shown to
possess HDL-cholesterol-raising properties by increasing the concentration of Apo A1 [34]. However,
when the health effects of consuming a fiber-rich cocoa product [23] versus flavanol-rich cocoa prod-
uct [27] were comparatively studied (both interventions were carried out in the same volunteers), the
higher insoluble fiber and theobromine contents of the former product did not seem to have any major
effect on the HDL cholesterol metabolism [35]. Conversely, a positive effect of the cocoa beverage rich
in fiber on glucose homeostasis was observed, as fasting serum glucose levels decreased [23], in agree-
ment with a previous study in hypercholesterolemic subjects using another fiber-rich cocoa product
[33] (in neither of these studies insulin changes were controlled). These results contrast with the meta-
analysis performed by Hooper et al. [36] in which only reductions in fasting serum insulin concentra-
tions were described. Thus, the effects of flavonoids on nitric oxide–dependent vascular function seem
to be linked to that on insulin sensitivity; in fact, a dose-dependent change relation existed between
cocoa and flow-mediated dilation (FMD) [37–39]. Accordingly, in overweight and healthy subjects, an
increase in FMD was described after consuming either a sugar-sweetened or sugar-free cocoa beverage
for 6 weeks [40]. However, daily consumption of flavanol-rich cocoa for 2 weeks was insufficient to
reduce blood pressure or improve insulin resistance in subjects with essential hypertension [41]. Other
benefits of cocoa consumption lie in its ability to reduce diabetes-related metabolic complications.
In type 2 diabetic patients, cocoa consumption reversed vascular dysfunction [42] and enhanced the
expression of mitochondrial structure markers in the skeletal muscle [43].
There is consistent evidence of the beneficial effects of cocoa and its related products on endothe-
lium function via the modulation of nitric oxide bioavailability, as shown in studies in healthy [44],
cardiovascular-risk [45], insulin-resistant [42], or obese subjects [46]. In fact, there was enough scientific
evidence to approve a health claim on the relationship between cocoa flavanols and maintenance of
endothelium-dependent vasodilation, which contributes to normal blood flow [47]. Nitric oxide pro-
duction has also been pointed out as the potential mechanism responsible for the reduction in blood
pressure associated with cocoa’s hypotensive effects that have been observed in healthy adults [48] and
patients with essential hypertension [42], although not in normotensive, healthy adults after consuming
dark chocolate and a cocoa beverage for 6 weeks [49]. In order to establish the minimum dose required
to reduce blood pressure, reconstituted cocoa beverages containing 33, 372, 712, or 1052 mg/day of
cocoa flavanols were assessed in untreated mild hypertensive subjects for 6 weeks, observing that only
the highest lowered blood pressure [50], although this positive effect could be attenuated by the sugar
content in the cocoa [51]. Flavanols in cocoa, by facilitating vasodilation, were also able to attenuate
exercise-induced increases in blood pressure [52]. Another mechanism that may contribute to cocoa’s
antihypertensive effect is the angiotensin-converting enzyme inhibition by cocoa flavanols [53], stearic
acid, or theobromine [54]. Recently, a theobromine-enriched cocoa significantly increased 24 h ambula-
tory systolic blood pressure (SBP), while lowering central SBP [55]. However, in contrast to the previous
studies, subjects with coronary heart disease (CHD) showed unaltered vascular function after consuming
a flavanol-rich chocolate bar and cocoa beverage daily [56].
Cocoa flavanols can also modulate the transcription and secretion of inflammatory cytokines in
human peripheral blood mononuclear cells, macrophages, and lymphoid cell lines, in addition to having
anti-inflammatory effects via the lipoxygenase pathway. Monomeric (−)-epicatechin and (+)-catechin
and dimeric flavanols may reduce nuclear factor kappa Β activation (NF-κB), thus resulting in reduced
pro-inflammatory cytokine production and oxidative burst [57]. In particular, polymeric fractions from
cocoa appear to have immunomodulatory effects on the production of cytokines interleukin (IL)-1β,
IL-2, and IL-4. Larger oligomeric fractions (hexamer through decamer) have also been reported to inhibit
IL-5 release, which may promote immunoglobulin A [58]. Dihydroxylated phenolic acids derived from
cocoa colonic microbial metabolism could act as an anti-inflammatory agents [18]. However, the anti-
inflammatory effects derived from regular consumption of cocoa in cardiovascular-risk subjects were
very modest compared with those observed for other polyphenol-rich foods [59], and even neutral or
contradictory inflammatory effects have been described [23,27,60]. On the other hand, cocoa favorably
affects intermediary factors in cancer progression [61], probably by counteracting chronic inflamma-
tion and oxidative stress that contribute to carcinogenesis. Numerous in vitro and animal studies sup-
port the anticancer effect of cocoa; however, in some human studies, an inverse relationship between
the incidences of cancer and flavonoid or epicatechin consumption has been described, whereas in
others, it fails [62].
Cocoa flavanols’ positive effects on insulin sensitivity [48], endothelial function [38,39], reduced
platelet aggregation [63], and blood pressure [45] can lead to a range of cognition-relevant benefits,
such as acute improvements in mood and cognitive performance during sustained mental effort [64],
being the mechanisms underlying these effects related to changes in the pool of bioavailable nitric oxide
[42] as well as the subchronic and acute increases in cerebral blood flow following cocoa ingestion
[42,46,65]. Increased neural efficiency in spatial working memory function has also been associated
with chronic cocoa beverage consumption [66]. Other positive health effects related to vasodilation
induced by consuming cocoa flavanols [5] are improvement in dermal blood circulation, leading to
higher skin density and hydration as well as increased resistance against UV-induced erythema. In addi-
tion, beneficial effects on oral health have been described, attributed to cocoa polyphenols’ capacity to
delay acid production by Streptococcus and cocoa tannin’s antibacterial and antienzymatic properties.
As indicated in the first section, unroasted cocoa beans present high energy density, which can increase
if sugar is added to the cocoa powder mix. However, cocoa polyphenols have been shown to prevent
diet-induced obesity by modulating lipid metabolism, especially by decreasing fatty acid synthesis and
transport systems, and enhancing thermogenesis in hepatic and white adipose cells [67]. In addition,
certain constituents in cocoa also have the potential to modulate glucocorticoid metabolism, which could
be of relevance to obesity-related complications [68]. Conversely, in a study carried out in obese subjects,
regular intake of high-flavanol cocoa did not directly affect body composition nor augment the effect of
exercise on body composition [46]. In agreement, the consumption of a sugar-sweetened and sugar-free
cocoa did not lead to differences in body weight or body mass index in overweight subjects or a decrease
in waist circumference with consumption of sugar-free cocoa. Therefore, it was concluded that healthy,
overweight adults can make a sugar-free or a sugar-sweetened cocoa beverage part of their diet without
adverse effects on body weight regulation [40]. Accordingly, no anthropometric changes were observed
in normoweight subjects who consumed a sugar-sweetened fiber-rich cocoa product [23] or a sugar-low
flavanol-rich product [27].

Several compounds contribute to the flavor of processed cocoa products. During fermentation, enzy-
matic hydrolysis liberates free amino acids (and reducing sugars), which, during roasting, participate
in Maillard reactions to produce 3-methylbutanal, phenylacetaldehyde, 2-methyl-3-(methylthio)furan,
2-ethyl-3,5-dimethylpyrazine, and 2,3-diethyl-5-methylpyrazine. Cocoa’s astringency is due to flavanols
and procyanidins, which also contribute to cocoa’s bitterness, although major bitter flavors in cocoa come
from theobromine, caffeine, and protein thermal degradation products. Astringency is attenuated during
the latter stages of fermentation, when flavonoids become susceptible to oxidation followed by condensa-
tion reactions with amino acids [7]. Fermentation, drying, and roasting develop the precursors of choco-
late flavor, but also many undesirable chemical compounds that give rise to acidic and astringent tastes in
the mouth. In addition, the grinding process creates many new surfaces, particularly of sugar, which are
not yet covered with fat, preventing chocolate from flowing properly. The conching process coats these
new surfaces with fat, developing the desired flow properties as well as removing undesirable flavors [69].
In recent years, the cocoa and chocolate industry is introducing changes in response to consumers’ increas-
ing demands for healthy products, especially low-fat, low-sugar, or sugar-free cocoa products, as well as dark
and high flavonoid content products. In order to reduce the cocoa fat content, techniques for processing
cocoa to produce low-fat cocoa powder have been designed leading to numerous patents, which in general
terms consist in extracting the cocoa fat to produce lower-fat cocoa butter and low-fat cocoa powder or low-
fat cocoa powder with high polyphenol content. In cocoa fat reduction, normally gaseous solvent extraction
is used to overcome the limits of hydraulic pressing, yielding a cocoa of superior functional properties (con-
centrated color and flavor, improved fat compatibility, ease of handling, and fine grinding). Alternatively,
different types of dietary fibers are used as fat replacers, such as oat β-glucan-rich hydrocolloid (C-trim30),
or soluble cocoa fiber, resulting in products with lower fat content and higher nutritional value.
Sucrose-free products are still limited and focused mainly to consumers with health concerns, such
as overweight and diabetes. Although the sweetness of sugar-free chocolate is equal to the conventional
products containing sucrose, these chocolates often exhibit poorer sensory properties, which limit their
wider use for all population [70]. As a replacement for sucrose, bulk sweeteners that are commonly used
in foods and pharmaceuticals can be used, providing body, texture, and sweet flavor to products with
reduced caloric value. However, replacement of sucrose with sugar alcohols such as maltitol, xylitol,
lactitol, sorbitol, and mannitol affects rheological properties and consequently the quality of chocolates.
In order to compensate the lower relative sweetness of sugar alcohols, intense sweeteners, such as aspar-
tame, acesulfame K, sucralose, and steviosides derived from Stevia rebaudiana Bertoni, have also been
added in the production of sucrose-free chocolates. Although high-intensity sweeteners are calorie free,
some of these sweeteners impart undesirable flavor and aftertaste, especially bitterness, which can limit
their application in foods and beverages. Many other natural alternatives to sugar are available, though
not widely used, despite the fact that natural nonrefined sugar alternatives potentially contain other
beneficial bioactive compounds, such as polyphenolic compounds [70].
In addition to low-fat and low-sugar cocoa products, dark chocolate and cocoa products enriched
with polyphenols are gaining popularity. Consumer perception of cocoa as healthy received a major
boost when the EU Commission approved a health claim on cocoa flavanols [47]. However, catechin and
procyanidin enrichment over a certain level may lead to an undesired bitter and astringent taste. This
limitation may be overcome by encapsulating the cocoa polyphenol extract. This technology allows the
addition of bioactive compounds to food, guaranteeing their protection during processing and permitting
their release over time and/or at particular sites also masking unwanted flavors [71]. Recently, encapsula-
tion of cocoa polyphenols with high-amylose maize starch effectively masked bitter taste and allowed
delivering of flavanol monomers into the gut; thus, encapsulated cocoa polyphenols were considered as a
functional prebiotic ingredient [71]. Increasing flavonoid intake without increasing overall energy intake
results in nutritional advantage.
New soluble cocoa products enriched with other dietary components, such as dietary fiber [25], methylx-
anthines [29], and other micronutrients such as calcium, iron, and vitamins (A, C, E, B6, and B12) are being
introduced in the food market in order to enhance cocoa’s beneficial health properties. Regular consump-
tion of a cocoa product rich in dietary fiber produced cardiometabolic benefits [23] in addition to improving
bowel habits [72]. In future, in order to assess the health effects of novel-enriched cocoa products, long-term
studies that investigate nutritional doses are required. However, controlling the diet of humans in the long
term remains a challenge, and it is necessary to design an adequate placebo, which is not easy. In these
studies, it is relevant to consider the additional calories provided by the novel cocoa products [5].
54.6 Conclusion
Cocoa beverages have evolved from a prestigious drink to one consumed by less affluent groups, partly
because for some time it was considered to be an unhealthy foodstuff associated to increased cardiovascular
disease (CVD) risk. However, today, cocoa is being considered as a beverage with cardioprotective proper-
ties, among other beneficial health effects. Cocoa presents a high energy and nutritional density, providing
mainly carbohydrates and fats, but it is also rich in proteins and micronutrients, especially minerals. More
importantly, cocoa provides exceptional value from a health viewpoint, particularly due to its high content of
flavonoids, which show extraordinary bioactivity, as well as theobromine, in addition to being a good source
of dietary fiber. Beyond cocoa’s antioxidant activity in vitro, which needs to be further studied in vivo, the
consumption of cocoa beverages has shown to improve lipid profiles, increase FMD and insulin sensitivity,
and reduce blood pressure and inflammation that leads to cardiovascular, cognition, and dermal improve-
ments in addition to reducing diabetes and obesity-related metabolic complications. Novel low-fat, low-sugar
cocoa products, as well as those supplemented with bioactive components, are being formulated to address
the present demands for healthy foods and beverages, meeting the needs of different consumers groups.
REFERENCES
1. Apgar, J.L. and Tarka, S.M., Methylxanthine composition and consumption patterns of cocoa and chocolate
products, in Caffeine, Spiller, G.E., Ed., CRC Press/Taylor & Francis Group, Boca Raton, FL, 1998.
2. Henderson, J.S., Joyce, R.A., Hall, G.R., Hurst, W.J., and McGovern, P.E., Chemical and archaeological
evidence for the earliest cacao beverages. Proc. Natl. Acad. Sci. U.S.A., 104, 18937–18940, 2007.
3. Coe, S.D. and Coe, M.D., The True History of Chocolate, Thames & Hudson, London, UK, 1996.
4. Dillinger, T.L., Barriga, P., Escárcega, S., Jimenez, M., Salazar, L.D., and Grivetti, L.E., Food of the
gods: Cure for humanity? A cultural history of the medicinal and ritual use of chocolate. J. Nutr.,
130(Suppl.), 2057S–2072S, 2000.
5. Ellam, S. and Williamson, G., Cocoa and human health. Annu. Rev. Nutr., 33, 105–128, 2013.
Release 27. USDA, Springfield, VA, 2014.
7. Schmitz, H.H., Kelm, M.A., and Hammerstone, J.F., Effect of cocoa and chocolate beverage consump-
tion on human cardiovascular health, in Beverages in Nutrition and Health, Wilson, T. and Temple,
N.J., Eds., Humana Press, Inc., Totowa, NJ, 2004, pp. 157–169.
8. Tomas-Barberán, F.A., Cienfuegos-Jovellanos, E., Marín, A., Muguerza, B., Gil-Izquierdo, A., Cerda,
B., Zafrilla, P. et al., A new process to develop a cocoa powder with higher flavonoid monomer content
and enhanced bioavailability in healthy humans. J. Agric. Food Chem., 55, 3926–3935, 2007.
9. Gu, L., House, S.E., Wu, X., Ou, B., and Prior, R.L., Procyanidin and catechin contents and antioxidant
capacity of cocoa and chocolate products. J. Agric. Food Chem., 54, 4057–4061, 2006.
10. Jalil, A.M. and Ismail, A., Polyphenols in cocoa and cocoa products: Is there a link between antioxidant
properties and health? Molecules, 13, 2190–2219, 2008.
11. Lee, K.W., Kim, Y.J., Lee, H.J., and Lee, C.Y., Cocoa has more phenolic phytochemicals and a higher
antioxidant capacity than teas and red wine. J. Agric. Food Chem., 51, 7292–7295, 2003.
12. Pérez-Jiménez, J., Neveu, V., Vos, F., and Scalbert, A., Identification of the 100 richest dietary
sources of polyphenols: An application of the Phenol-Explorer database. Eur. J. Clin. Nutr., 64(Suppl.),
S112–S120, 2010.
13. Keen, C.L., Holt, R.R., Oteiza, P.I., Fraga, C.G., and Schmitz, H.H., Cocoa antioxidants and cardiovas-
cular health. Am. J. Clin. Nutr., 81(Suppl.), 298S–303S, 2005.
14. Scheid, L., Reusch, A., Stehle, P., and Ellinger, S., Antioxidant effects of cocoa and cocoa products
ex vivo and in vivo: Is there evidence from controlled intervention studies? Curr. Opin. Clin. Nutr.
Metab. Care, 6, 737–742, 2010.
15. Davison, G., Callister, R., Williamson, G., Cooper, K.A., and Gleeson, M., The effect of acute pre-exercise
dark chocolate consumption on plasma antioxidant status, oxidative stress and immunoendocrine
responses to prolonged exercise. Eur. J. Nutr., 51, 69–79, 2012.
16. Baba, S., Natsume, M., Yasuda, A., Nakamura, Y., Tamura, T., Osakabe, N., Kanegae, M., and Kondo, K.,
Plasma LDL and HDL cholesterol and oxidized LDL concentrations are altered in normo- and hypercho-
lesterolemic humans after intake of different levels of cocoa powder. J. Nutr., 137, 1436–1441, 2007.
17. Baba, S., Osakabe, N., Kato, Y., Natsume, M., Yasuda, A., Kido, T., Fukuda, K., Muto, Y., and Kondo, K.,
Continuous intake of polyphenolic compounds containing cocoa powder reduces LDL oxidative
susceptibility and has beneficial effects on plasma HDL-cholesterol concentrations in humans.

Am. J. Clin. Nutr., 85, 709–717, 2007.
18. Khan, N., Khymenets, O., Urpí-Sardà, M., Tulipani, S., Garcia-Aloy, M., Monagas, M., Mora-Cubillos, X.,
Llorach, R., and Andres-Lacueva, C., Cocoa polyphenols and inflammatory markers of cardiovascular
disease. Nutrients, 6, 844–880, 2014.
19. Martínez-López, S., Sarriá, B., Gomez-Juaristi, M., Goya, L., Mateos, R., and Bravo, L.,
Theobromine, caffeine, and theophylline metabolites in human plasma and urine after consump-
tion of soluble cocoa products with different methylxanthine contents. Food Res. Inter., 63(Special
issue), 446–455, 2014.
20. Vinson, J.A., Proch, J., and Zubik, L., Phenol antioxidant quantity and quality in foods: Cocoa, dark
chocolate, and milk chocolate. J. Agric. Food Chem., 47, 4821–4824, 1999.
21. Badrie, N., Bekele, F., Sikora, E., and Sikora, M., Cocoa agronomy, quality, nutritional, and health
aspects. Crit. Rev. Food Sci. Nutr., 55, 620–659, 2015.
22. Lecumberri, E., Mateos, R., Izquierdo-Pulido, M., Rupérez, P., Goya, L., and Bravo, L., Dietary fibre
composition, antioxidant capacity and physico-chemical properties of a fibre-rich product from cocoa
(Theobroma cacao L.). Food Chem., 104, 948–954, 2007.
23. Sarriá, B., Martínez-López, S., Sierra-Cinos, J.L., García-Diz, L., Mateos, R., and Bravo, L., Regular
consumption of a cocoa product improves the cardio-metabolic profile in healthy and moderately
hypercholesterolemic adults. Br. J. Nutr., 111, 122–134, 2014.
24. Kris-Etherton, P.M. and Mustad, V.A., Chocolate feeding studies: A novel approach for evaluating the
plasma lipid effects of stearic acid. Am. J. Clin. Nutr., 60(Suppl. 6), 1029S–1036S, 1994.
25. Tokede, O.A., Gaziano, J.M., and Djoussé, L., Effects of cocoa products/dark chocolate on serum lipids:
A meta-analysis. Eur. J. Clin. Nutr., 65, 879–886, 2011.
26. Khan, N., Monagas, M., Andres-Lacueva, C., Casas, R., Urpí-Sardà, M., Lamuela-Raventós, R.M., and
Estruch, R., Regular consumption of cocoa powder with milk increases HDL cholesterol and reduces
oxidized LDL levels in subjects at high-risk of cardiovascular disease. Nutr. Metab. Cardiovasc. Dis., 22,
1046–1053, 2012.
27. Martínez-López, S., Sarriá, B., Sierra-Cinos, J.L., Goya, L., Mateos, R., and Bravo, L., Realistic intake
of a flavanol-rich soluble cocoa product increases HDL-cholesterol without inducing anthropometric
changes in healthy and moderately hypercholesterolemic subjects. Food Funct., 2, 364–374, 2014.
28. Latham, L.S., Hensen, Z.K., and Minor, D.S. Chocolate—Guilty pleasure or healthy supplement?
J. Clin. Hypertens., 2, 101–106, 2014.
29. Roura, C., Andrés-Lacueva, R., Estruch, M.L., Mata-Bilbao, M., Izquierdo-Pulido, A.L., Waterhouse,
A.L., and Lamuela-Ráventos, R.M., Milk does not affect the bioavailability of cocoa powder flavonoid
in healthy human. Ann. Nutr. Metab., 51, 493–498, 2007.
30. Keogh, J.B., McInerney, J., and Clifton, P.M., The effect of milk protein on the bioavailability of cocoa
polyphenols. J. Food Sci., 72, S230–S233, 2007.
31. Wu, H., Dwyer, K.M., and Fan, Z., Dietary fiber and progression of atherosclerosis: The Los Angeles
atherosclerosis study. Am. J. Clin. Nutr., 78, 1085–1091, 2003.
32. Lecumberri, E., Goya, L., and Mateos, R., A diet rich in dietary fiber from cocoa improves lipid profile
and reduces malondialdehyde in hypercholesterolemic rats. Nutrition, 23, 332–341, 2007.
33. Sarriá, B., Mateos, R., Sierra-Cinos, J.L., Goya, L., García-Diz, L., and Bravo, L., Hypotensive, hypoglycae-
mic and antioxidant effects of consuming a cocoa product in moderately hypercholesterolemic humans.
Food Funct., 3, 867–874, 2012.
34. Neufingerl, N., Zebregs, Y.E., Schuring, E.A., and Trautwein, E.A., Effect of cocoa and theobromine
consumption on serum HDL-cholesterol concentrations: A randomized controlled trial. Am. J. Clin. Nutr.,
97, 1201–1209, 2013.
35. Sarriá, B., Martínez-López, S., Sierra-Cinos, J.L., Garcia-Diz, L., Goya, L., Mateos, R., and Bravo, L.,
Effects of bioactive constituents in functional cocoa products on cardiovascular health in humans.
Food Chem., 174, 214–218, 2015.
36. Hooper, L., Kroon, P.A., Rimm, E.B., Cohn, J.S., Harvey, I., Le Cornu, K.A., Ryder, J.J., Hall, W.L., and
Cassidy, A., Flavonoids, flavonoid-rich foods, and cardiovascular risk: A meta-analysis of randomized
controlled trials. Am. J. Clin. Nutr., 88,38–50, 2008.
37. Monahan, K.D., Feehan, R.P., Kunselman, A.R., Preston, A.G., Miller, D.L., and Lott, M.E., Dose-
dependent increases in flow-mediated dilation following acute cocoa ingestion in healthy older adults.
J. Appl. Physiol., 111, 1568–1574, 2011.
38. Heiss, C., Finis, D., Kleinbongard, P., Hoffmann, A., Rassaf, T., Kelm, M., and Sies, H., Sustained increase
in flow-mediated dilation after daily intake of high-flavanol cocoa drink over 1 week. J. Cardiovasc.
Pharmacol., 49, 74–80, 2007.
39. Heiss, C., Jahn, S., Taylor, M., Real, W.M., Angeli, F.S., Wong, M.L., Amabile, N. et al., Improvement
of endothelial function with dietary flavanols is associated with mobilization of circulating angiogenic
cells in patients with coronary artery disease. J. Am. Coll. Cardiol., 56, 218–24, 2010.
40. Njike, V.Y., Faridi, Z., Shuval, K., Dutta, S., and Lay, C.D., Effects of sugar-sweetened and sugar-free
cocoa on endothelial function in overweight adults. Int. J. Cardiol., 149, 83–88, 2011.
41. Muniyappa, R., Hall, G., Kolodziej, T.L., Karne, R.J., Crandon, S.K., and Quon, M.J., Cocoa consump-
tion for 2 wk enhances insulin-mediated vasodilatation without improving blood pressure or insulin
resistance in essential hypertension. Am. J. Clin. Nutr., 88, 1685–1696, 2008.
42. Balzer, J., Rassaf, T., Heiss, C., Kleinbongard, P., Lauer, T., Merx, M., Heussen, N. et al., Sustained
benefits in vascular function through flavanol-containing cocoa in medicated diabetic patients a dou-
ble-masked, randomized, controlled trial., J. Am. Coll. Cardiol., 51, 2141–2149, 2008.
43. Taub, P.R., Ramirez-Sanchez, I., Ciaraldi, T.P., Perkins, G., Murphy, A.N., Naviaux, R., Hogan,
M. et al., Alterations in skeletal muscle indicators of mitochondrial structure and biogenesis in
patients with type 2 diabetes and heart failure: Effects of epicatechin rich cocoa. Clin. Transl. Sci.,
5, 43–47, 2012.
44. Schroeter, H., Heiss, C., Balzer, J., Kleinbongard, P., Keen, C.L., Hollenberg, N.K., Sies, H., Kwik-
Uribe, C., Schmitz, H.H., and Kelm, M., (−)-Epicatechin mediates beneficial effects of flavanol-rich
cocoa on vascular function in humans. Proc. Natl. Acad. Sci. U.S.A., 103, 1024–1029, 2006.
45. Grassi, D., Necozione, S., Lippi, C., Croce, G., Valeri, L., Pasqualetti, P., Desideri, G., Blumberg,
J.B., and Ferri, C., Cocoa reduces blood pressure and insulin resistance and improves endothelium-
dependent vasodilation in hypertensives. Hypertension, 46, 398–405, 2005.
46. Davison, K., Coates, A.M., Buckley, J.D., and Howe, P.R.C., Effect of cocoa flavanols and exercise on
cardiometabolic risk factors in overweight and obese subjects. Int. J. Obes., 32, 1289–1296, 2008.
47. European Food Safety Agency (EFSA), Scientific opinion on the substantiation of a health claim related
to cocoa flavanols and maintenance of normal endothelium-dependent vasodilation pursuant to Article
13(5) of Regulation (EC) No 1924/20061. EFSA J., 10, 2809–2821, 2012.
48. Grassi, D., Lippi, C., Necozione, S., Desideri, G., and Ferri, C., Short-term administration of dark
chocolate is followed by a significant increase in insulin sensitivity and a decrease in blood pressure in
healthy persons. Am. J. Clin. Nutr., 81, 611–614, 2005.
49. Crews, W.D., Harrison, D.W., and Wright, J.W., A double-blind, placebo-controlled, randomized trial of
the effects of dark chocolate and cocoa on variables associated with neuropsychological functioning and
cardiovascular health: Clinical findings from a sample of healthy, cognitively intact older adults. Am.
J. Clin Nutr., 87, 872–880, 2008.
50. Davison, K., Berry, N.M., Misan, G., Coates, A.M., Buckley, J.D., and Howe, P.R., Dose-related effects
of flavanol-rich cocoa on blood pressure. J. Hum. Hypertens., 24, 568–576, 2010.
51. Faridi, Z., Njike, V.Y., Dutta, S., Ali, A., and Katz, D.L., Acute dark chocolate and cocoa ingestion and
endothelial function: A randomized controlled crossover trial. Am. J. Clin. Nutr., 88, 58–63, 2008.
52. Berry, N.M., Davison, K., Coates, A.M., Buckley, J.D., and Howe, P.R., Impact of cocoa flavanol
consumption on blood pressure responsiveness to exercise. Br. J. Nutr., 103, 1480–1484, 2010.
53. Actis-Goretta, L., Ottaviani, J.I., and Fraga, C.G., Inhibition of angiotensin converting enzyme activity
by flavanol-rich foods. J. Agric. Food Chem., 54, 229–234, 2006.
54. Kelly, C.J., Effects of theobromine should be considered in future studies. Am. J. Clin. Nutr., 82,
486–487, 2005.
55. van den Bogaard, B., Draijer, R., Westerhof, B.E., van den Meiracker, A.H., van Montfrans, G.A., and
van den Born, B.J., Effects on peripheral and central blood pressure of cocoa with natural or high-dose
theobromine: A randomized, double-blind crossover trial. Hypertension, 56, 839–846, 2010.
56. Farouque, H.M., Leung, M., Hope, S.A., and Baldi, M., Acute and chronic effects of flavanol-rich cocoa
on vascular function in subjects with coronary artery disease: A randomized double-blind placebo-
controlled study. Clin. Sci. (London), 111, 71–80, 2006.
57. Selmi, C., Cocchi, C.A., Lanfredini, M., Keen, C.L., and Gershwin, M.E., Chocolate at heart: The
anti-inflammatory impact of cocoa flavanols. Mol. Nutr. Food Res., 52, 1340–1348, 2008.
58. Mao, T.K., Van de Water, J., Keen, C.L., Schmitz, H.H., and Gershwin, M.E., Effect of cocoa flavanols
and their related oligomers on the secretion of interleukin-5 in peripheral blood mononuclear cells.
J. Med. Food., 5, 17–22, 2002.
59. Monagas, M., Khan, N., Andres-Lacueva, C., Casas, R., Urpí-Sardà, M., Llorach, R., Lamuela-Raventós,
R.M., and Estruch, R., Effect of cocoa powder on the modulation of inflammatory biomarkers in patients
at high risk of cardiovascular disease. Am. J. Clin. Nutr., 90, 1144–1150, 2009.
60. Wang-Polagruto, J.F., Villablanca, A.C., Polagruto, J.A., and Lee, L., Chronic consumption of flavanol-rich
cocoa improves endothelial function and decreases vascular cell adhesion molecule in hypercholesterol-
emic postmenopausal women. J. Cardiovasc. Pharmacol., 47(Suppl. 2), S177–S186, 2006.
61. Maskarinec, G., Cancer protective properties of cocoa: A review of the epidemiologic evidence. Nutr.
Cancer, 61, 573–579, 2009.
62. Martin, M.A., Goya, L., and Ramos, S., Potential for preventive effects of cocoa and cocoa polyphenols
in cancer. Food Chem. Toxicol., 56, 336–351, 2013.
63. Holt, R., Schramm, D., Keen, C., Lazarus, S., and Schmitz, H., Chocolate consumption and platelet
function. J. Am. Med. Assoc., 287, 2212–2213, 2002.
64. Scholey, A.B., French, S.J., Morris, P.J., Kennedy, D.O., Milne, A.L., and Haskell, C.F., Consumption
of cocoa flavanols results in acute improvements in mood and cognitive performance during sustained
mental effort. J. Psychopharmacol., 24, 1505–1514, 2010.
65. Sorond, F.A., Lipsitz, L.A., Hollenberg, N.K., and Fisher, N.D., Cerebral blood flow response to flava-
nol-rich cocoa in healthy elderly humans. Neuropsychiatr. Dis. Treat., 4, 433–440, 2008.
66. Camfield, D.A., Scholey, A., Pipingas, A., Silberstein, R., Kras, M., Nolidin, K., Wesnes, K., Pase, M.,
and Stough, C., Steady state visually evoked potential (SSVEP) topography changes associated with
cocoa flavanol consumption. Physiol. Behav., 105, 948–957, 2012.
67. Matsui, N., Ito, R., Nishimura, E., Yoshikawa, M., Kato, M., Kamei, M., Shibata, H., Matsumoto, I.,
Abe, K., and Hashizume, S., Ingested cocoa can prevent high-fat diet-induced obesity by regulating the
expression of genes for fatty acid metabolism. Nutrition, 21, 594–601, 2005.
68. Almoosawi, S., Fyfe, L., Ho, C., and Al-Dujaili, E., The effect of polyphenol-rich dark chocolate on
fasting capillary whole blood glucose, total cholesterol, blood pressure and glucocorticoids in healthy
overweight and obese subjects. Br. J. Nutr., 103, 842–850, 2010.
69. Beckett, S.T., Industrial Chocolate Manufacture and Use, 4th edn., John Wiley & Sons Ltd., Chichester,
UK, 2009.
70. Belščak-Cvitanović, A., Komes, D., Dujmović, M., Karlović, S., Biškić, M., Brnčić, M., and Ježek, D.,
Physical, bioactive and sensory quality parameters of reduced sugar chocolates formulated with natural
sweeteners as sucrose alternatives. Food Chem., 167, 61–70, 2015.
71. Vitaglione, P., Barone Lumaga, R., Ferracane, R., Sellitto, S., Morello, J.R., Reguant, M.J., Shimoni, E.,
and Fogliano, V., Human bioavailability of flavanols and phenolic acids from cocoa-nut creams enriched
with free or microencapsulated cocoa polyphenols. Br. J. Nutr., 109, 1832–1843, 2013.
72. Sarriá, B., Martínez-López, S., Fernández-Espinosa, A., Gómez-Juaristi, M., Goya, L., Mateos, R., and
Bravo, L., Effects of regularly consuming dietary fibre rich soluble cocoa products on bowel habits
in healthy subjects: A free-living, two-stage, randomized, crossover, single-blind intervention. Nutr.
Metab. (London), 9, 33, 2012.
Section V
Dairy and Soy Beverages

55
Dairy Beverages
Ranjan Sharma
CONTENTS
55.1 Introduction................................................................................................................................... 707
55.2 Functionality, Applications, and Health Effects of Bioactive Components................................. 708
55.2.1 Colostrum........................................................................................................................ 709
55.2.2 Glycomacropeptide..........................................................................................................711
55.2.3 Lactoferrin.......................................................................................................................712
55.2.4 Lactoperoxidase...............................................................................................................713
55.2.5 Casein and Whey Protein Hydrolysates..........................................................................714
55.2.6 Milk Minerals..................................................................................................................716
55.2.7 Micellar Casein................................................................................................................716
55.2.8 α-Lactalbumin.................................................................................................................717
55.2.9 Osteopontin......................................................................................................................717
55.2.10 Caseinophosphopeptide...................................................................................................717
55.3 Functional Dairy Foods.................................................................................................................717
55.3.1 Nutrient Content Claims..................................................................................................718
55.3.2 Probiotics.........................................................................................................................718
55.3.3 Prebiotic Fiber..................................................................................................................719
55.3.4 Omega-3 Fatty Acids.......................................................................................................719
55.3.5 Phytosterols..................................................................................................................... 720
55.3.6 Antioxidants and Plant Extracts..................................................................................... 720
55.3.7 Dietary Fiber................................................................................................................... 720
55.4 Technical Challenges in Developing Functional Dairy Beverages.............................................. 720
55.5 Conclusion......................................................................................................................................721
References................................................................................................................................................721
55.1 Introduction
Global health and wellness market was estimated to be worth US$734 billion in 2013 [1]. This market is
made up of natural health products (US$284 billion), functional/fortified foods (US$246 billion), better-
for-you products (US$166 billion), organic foods (US$30 billion), and food intolerance market (US$8
billion). In this market, functional/fortified foods are shown to be leading the growth with an annual
rate of over 4%. Due to their wider availability, superior nutrition, flavor, and convenience, dairy foods
and beverages represent nearly 40% of the global functional beverages market and up to 70% in some
European countries.
Milk and dairy beverages contain several components with physiological functions, such as proteins,
peptides, fats, oligosaccharides, vitamins, and minerals, among others. These different milk components
could be used as functional and nutraceutical ingredients to promote good health and reduce disease risk.
Dairy is not only considered as a source of bioactive ingredients but is also a preferred vehicle for delivery
of nondairy functional/health ingredients due to their superior flavor, nutritional, and bioactive profiles.
707
A significant number of reviews have recently been published highlighting dairy components as func-
tional foods [2–5]. This chapter highlights dairy as a source of bioactive components and as a delivery
vehicle for nondairy bioactive components.
55.2 Functionality, Applications, and Health Effects of Bioactive Components

Milk contains several physiological functional and bioactive components that are buried inside the com-
plex structures of dairy components and largely remain untapped commercially. In order to isolate,
fractionate, and concentrate bioactive components from milk, the first step is the separation of fresh
milk into cream and skim milk. The cream part contains almost all of the fat globules and the associ-
ated fat globule membrane that is a rich source of several bioactive components (Figure 55.1). Among
these are ingredients enriched in whole proteins (e.g., β-lactoglobulin, α-lactalbumin, lactoferrin, and
Bioactive ingredients from milk
Milk Early lactation milk

Colostrum powder
Separation Colostrum
Fractionation
Cream separation
Cream Immunoglobulins
Anhydrous Lactoferrin
milkfat (AMF) Lactoperoxidase
Insulin-like growth factors
Fat globule membrane (IGF-1)
material Skim milk
Transforming growth
factor beta-2 (TFG-β2)
Growth hormones
Milk fat globule Acid Lysozyme
membrane (MFGM) Rennet
Phospholipids
Cheese/rennet casein
Enzymes Acid casein
Membrane proteins Acid
whey Sweet
Mucins whey
Glycoproteins
Hydrolysis Glyco-
macropeptide
(GMP)
Hydrolysis
Casein Casein Fractionation
hydrolysates peptides
β-Lactoglobulin
Lactose α-Lactalbumin
Bovine serum albumin (BSA)
Whey protein Whey
hydrolysates Whey growth factors (WGF)
peptides
Lactoferrin
Mucins
Milk minerals
Immunoglobulins
Oligosaccharides Milk calcium
FIGURE 55.1 Bioactive components in milk.

Dairy Beverages 709
TABLE 55.1
Compositional and Nutritional Characteristics of Cow’s Milk and Commercial Functional Dairy Products
Cow’s Colostrum Milk Minerals
Component Unit Milk (22% IgG) GMP Lactoferrin Lactoperoxidase (24% Calcium)
Moisture % 87.4 5 5 5 5 10
Fat % 3.5 2 <1 <1 <1 1
Protein % 3.4 75 80 92 91 5
Ash % 0.7 6 6 1 2 78
Lactose % 5.0 10 <1 <1 <1 5
Calcium % 1.1 1.5 <0.1 <0.1 <0.1 24
Iron mg/100 g <0.1 <1 <1 12,000 <1 11
IgG % <0.1 22 0 0.0 0.0 0.0
GMP % of protein 0.0 0.0 90 0.0 0.0 0.0
Lactoferrin % of protein <0.1 0.5 <0.1 83 <0.1 0.0
Sialic acid % <0.1 <0.1 4 <0.1 <0.1 0.0
Lactoperoxidase U/mg <0.1 0.0 0.0 0.0 270 0.0
activity
Source: Based on commercial specification sheets.
Abbreviations: IgG, immunoglobulin G; GMP, glycomacropeptide.
lactoperoxidase), hydrolyzed forms of proteins (e.g., whey protein hydrolysates with varying degrees of
hydrolysis), and milk minerals [5,6].
Compositional and nutritional characteristics of key bioactive components from cow’s milk and com-
mercial functional dairy products are shown in Table 55.1. Functional and bioactive properties of these
components as well as their potential applications are shown in Table 55.2.
55.2.1 Colostrum
Colostrum is the first milk produced by a cow after the birth of a calf. Colostrum provides life-
supporting immune and growth factors that ensure the health and vitality of the newborn. It is a rich
source of several bioactive components including immunoglobulins, lactoferrin, lactoperoxidase, lyso-
zyme, and several growth factors [7,8]. Colostrum obtained from the first few milking is generally
pooled and either frozen or transported chilled to the dairy processing facility. After separation of
colostrum cream (which can be fed to the calf), the skim colostrum can be concentrated and dried into
colostrum powder. In order to remove casein, skim colostrum is usually treated with rennet or acid, and
the casein-free whey is used for the development of colostrum whey powder or colostrum whey protein
concentrate. Colostrum whey protein concentrate is particularly valuable as it is rich in bioactive protein
components and contains low levels of lactose. Commercial drying of colostrum is either carried out
by freeze-drying or by spray-drying under mild temperatures. Bovine colostrum is marketed in several
forms. Commercial colostrum products are available with immunoglobulin contents ranging from 16%
to 50% (Unpublished data).
Colostrum contains over 10-fold the amount of immunoglobulins present in normal milk (Table 55.1).
Immunoglobulins are very heat-sensitive proteins, which makes the processing of colostrum into an
ingredient a difficult process. Nevertheless, many commercial products are available. Colostrum is also
a rich source of growth factors that are key regulators of a variety of cellular functions and are involved
in the control of tissue growth and repair [9,10]. Extensive research has identified a number of applica-
tions for their use in clinical medicine and biotechnology. The most important of these is likely to be a
therapeutic potential in wound healing [11].
Colostrum products can provide a number of physiologically functional properties. The benefits of
colostrum to the human body—from boosting the immune system to promoting cell repair—are continu-
ally being researched and discovered. Major functionality and applications of colostrum include source
TABLE 55.2
Bioactive Ingredients in Milk, Their Biological Functions, and Potential Applications
Bioactive Ingredient Potential Biological Function Potential Food Applications References
Colostrum Immune factors, growth factors, and Sports formulation, hospital, and [7,8,12,14]
antimicrobial. medical nutrition.
Glycomacropeptide or Satiety, low phenylalanine, and Dental care products such as [17,18,22]
caseinomacropeptide branched-chain amino acids. toothpaste and mouthwash for
prevention of dental caries and
remineralization.
Supplements and diets for
phenylketonuria sufferers.
Prebiotic for probiotic supplements
and foods.
Sports nutrition products as a source
of branched-chain amino acids.
High-protein diets for weight control.
Lactoferrin Iron-binding ability responsible for Health supplements, supplements for [24,25]
many functions such as the elderly or immune-compromised
bacteriostatic effect, cell growth patients, functional foods and drinks,
promotion, antioxidation and iron infant formulas, cosmetics and oral
delivery, and absorption. care products, and supplements for
recovery from gastrointestinal
infections.
Lactoperoxidase Preservation effect: bacteriostatic Food preservation in general, meat [29,30,32,33]
effect against Gram-positive products.
bacteria and bactericidal effect
against Gram-negative bacteria,
e.g., Pseudomonas, Coliforms,
Salmonella, and Listeria.
Casein and whey Reduced allergenicity, increased Infant and enteral formulation, [35–37]
protein hydrolysate protein absorption, increased geriatric products, sports beverages,
peptide bioactivity, and lowering and weight control diets.
blood pressure.
Milk minerals and Prevention of osteoporosis and Dairy products such as recombined [41–44]
milk calcium growth of healthy bones and teeth. milk, flavored milk, yogurt, and
Blood pressure and CVD control. cheese.
Lower effect on hypertension. Nutritional and functional foods such
Prevention of colon cancer. as sports and adult nutritional
Control of weight gain and obesity. beverages, weight loss products, and
sports bars.
Bakery products such as breads and
cakes.
Confectionery products.
Breakfast cereals.
Convenience foods such as soups,
sauces, and frozen desserts.
Food supplements such as capsules
and tablets.
α-Lactalbumin Sleep enhancement: ACE-I activity, Drinks, beverages, and infant formula. [45,46]
immunomodulation, antimicrobial,
opioid agonist, and anticancer.
Caseinophopeptide Remineralization and mineral High-mineral beverages, chewing [47,48]
absorption through the help of gum, and breakfast cereals.
mineral carrier, protection against
dental caries, and antibacterial.
Abbreviations: ACE-I, angiotensin-converting enzyme inhibitor; CVD, cardiovascular disease.
Dairy Beverages 711
of growth factors, antimicrobial components, immune-enhancing properties and intestinal benefits, and
sports and performance components [7,12–15].
Source of growth factors: In addition to being a rich source of essential nutrients such as amino
acids, carbohydrate, lipids, and minerals, colostrum contains a range of growth factors that can
stimulate healthy development of cells and tissues. Growth factors from colostrum and whey
have commercially been available for some time. Growth factors present in colostrum include
insulin-like growth factors 1 and 2 (IGF-1 and IGF-2), transforming growth factors β1 and β2,
and epidermal growth factor. Some of these factors are also present in regular milk but in very
small amounts (100–1000 times less than colostrum). The IGFs stimulate the immune system,
promote cell repair and growth, and influence how the body uses fat, protein, and sugar. The
IGFs while acting as endocrine, autocrine, and paracrine hormones enhance cellular glucose
uptake stimulating synthesis of proteins, DNA, RNA, and lipids. The amino acid sequence of
bovine IGF-1 is identical to that of human IGF-1, and bovine IGF-2 differs from human IGF-2
by only three amino acid residues. Both IGF-1 and IGF-2 are heat-stable proteins that help in
growth and differentiation of cells [11].
Antimicrobial components: Colostrum is a rich source of antimicrobial compounds that can help
in protection against infections. Antimicrobial components present in colostrum include lac-
toferrin, lactoperoxidase, lysozyme, and immunoglobulins. Antibodies from colostrum in oral
immunotherapy have aided treatment of various human infections, including those caused by
antibiotic resistant bacteria [13].
Immune-enhancing properties and intestinal benefits: One of the main reasons for hospital admis-
sion of infants and young children is infectious diarrhea, usually caused by a rotavirus infec-
tion. Infants can also acquire rotavirus in hospital neonatal and pediatric wards. The infection
can also be transmitted to adult members of the family. Colostrum has been successfully tried
out in the protection against rotavirus and diarrhea [12]. It has been suggested that infant for-
mulas could be fortified with colostral immunoglobulins [14]. Colostrum may help protect the
gastrointestinal tract against stomach cancers and ulcers [15]. It has also been shown to prevent
nonsteroidal anti-inflammatory drug (NSAID)-induced gut damage. NSAIDs are given for the
treatment of pain and are a common cause of gastritis [16].
Sports and performance components: Colostrum appears to aid strength and speed in athletes,
plus increase insulin, which has anabolic effects. Studies have shown that supplementation with
bovine colostrum (20 g/day) in combination with exercise training for 8 weeks may increase
bone-free lean body mass in active men and women [8]. Colostrum has been shown to enhance
serum IGF-1, immunoglobulin G (IgG), hormone, and saliva immunoglobulin A during ath-
letes’ training. In clinical trials, IGF-1 is known to have strong anabolic effects on muscle tis-
sue, as it is able to mimic most of the actions of growth hormones. IGF-I is of benefit to athletes,
body builders, and people concerned about weight because it can help burn fat and encourage
lean muscle tissue.
55.2.2 Glycomacropeptide
Glycomacropeptide (GMP) is a hydrophilic peptide (amino acid residue 102–169) of κ-casein that pro-
vides stability to casein micelles in milk. When rennet acts on κ-casein during the manufacture of cheese,
GMP is released into the whey. It makes up about 15%–20% of the whey proteins. Recent advances in
fractionation have allowed separation of GMP from cheese whey into commercial GMP-enriched ingre-
dients. Due to the highly negative charge of GMP at low pH where whey proteins are positively charged,
an ion-exchange process can isolate GMP. When whey at pH 3 is contacted with a cation exchanger, the
GMP is not adsorbed by the cation exchanger and may be concentrated and desalted by ultrafiltration.
Alternatively, GMP from whey at pH less than 4 can be bound to an anion exchanger, while the rest of the
whey proteins are not bound. Pure GMP can then be eluted from the ion-exchange column [17].
GMP is unique among some of the whey proteins in that it is a glycoprotein and, thus, has an oligosac-
charide chain attached to it. It is also unique since it contains no phenylalanine, tryptophan, or tyrosine.
GMP has also high levels of the branched-chain amino acids (leucine, isoleucine, and valine), which
have been shown to benefit muscle recovery after strenuous exercise [18].
The composition of a commercial GMP is compared with milk in Table 55.1. The composition of
GMP gives it some unique characteristics that can be utilized in a variety of interesting applications. A
small population has phenylketonuria (PKU), meaning they are unable to digest phenylalanine. GMP is
one of the few amino acid sources PKU patients can tolerate because the pure GMP does not contain
phenylalanine [19].
GMP has been linked with many physiological functions, including promotion of bifidobacterial
growth, suppression of gastric secretions, inhibition of bacterial and viral adhesion, modulation of
immune system responses, and binding of cholera and Escherichia coli enterotoxins. In simpler terms,
GMP offers potential benefits to intestinal health, appetite control, reduced dental caries, enhanced
immunity, and protection against diarrhea. Some of the bioactive properties of GMP include anti-
inflammation, toxin binding, inhibition of bacterial and viral adhesion, immune modulation, protec-
tion against diarrhea, prebiotic effects, source of amino acids for population suffering from PKU, and
improved satiety [20–23].
55.2.3 Lactoferrin
Lactoferrin is an iron-binding glycoprotein present in colostrum, milk, and whey. It exists as a single
peptide chain with a molecular weight of 77,000 Da. It is folded into two globular units with each unit
able to bind 1.4 mg of iron per gram of protein. The iron-binding ability of lactoferrin is responsible for
many biological functions such as bacteriostatic effect, growth-promoting effect on certain cell lines,
and prevention of lipid peroxidation and promotion of iron absorption in the body. In addition, lactoferrin
is one of few proteins in whey that are positively charged at pH 7.0 (isoelectric point of approximately
pH 7.9), while most other proteins are negatively charged. This feature of lactoferrin has been exploited
in commercial isolation of lactoferrin. Using cation-based resins and selective salt solutions, lactoferrin
can be separated from other positively charged proteins attached to the resin. Further concentration of
lactoferrin is carried out using ultrafiltration and spray-drying. Table 55.1 shows the compositional and
nutritional characteristics of a commercial lactoferrin powder.
Lactoferrin can provide several physiological functional (bioactive) properties, which are mainly
derived from its ability to bind iron. Each molecule of lactoferrin can bind two atoms of iron. The
functional properties of lactoferrin include antibacterial and antiviral properties, antioxidant properties,
immune modulation and iron transport, and absorption [24,25].
Lactoferrin inhibits the growth of pathogenic bacteria and fungi, due to its ability to bind large quan-
tities of iron. It binds iron very strongly, thus rendering this essential nutrient unavailable to support
microbial growth. Lactoferrin also disrupts bacterial digestion of carbohydrates, further limiting their
growth. In addition, the action of pepsin in the stomach converts lactoferrin into lactoferricin, which has
a broad spectrum of activity against pathogenic bacteria and yeast. It has also the ability to bind to para-
sites and the outer membrane of Gram-negative bacteria, making the cell wall more permeable, and thus
improving the efficiency of antibiotics [24]. Additionally, segments of the lactoferrin molecule can exert
a direct bactericidal effect on certain strains of bacteria and are also thought to inhibit the attachment
of bacteria to the gut wall, therefore reducing the probability of infection. Antiviral effects of bovine
lactoferrin against several types of viruses have been reported. Lactoferrin appears to achieve this effect
by inhibiting virus absorption and its penetration into cells [24].
Lactoferrin can be used as a natural antioxidant and may reduce the susceptibility to aging pro-
cesses and disease. It provides protection against oxidative damage by scavenging excess iron, which
catalyzes the undesired formation of free radicals from hydrogen peroxide produced as a result of
microbial respiration, thus allowing the cell’s own peroxidase to harmlessly break down the hydro-
gen peroxide [25]. Lactoferrin contributes to the defense against pathogens by activation of cells
involved in the anti-inflammatory response during the course of microbial infection, thus enhancing
the self-immunity [26].
Dairy Beverages 713
The addition of lactoferrin to food products may be based on the amount of lactoferrin or the desired
level of iron and generally can be added in the range of 1–100 mg/100 g of product. The desired level of
iron could vary from as little as 1 mg/100 mL for infant formula to as high as 7 mg/100 mL for sports
formulation. Recently, the United Sates Drug Administration (USDA) has recently approved the use of
activated lactoferrin on fresh beef to enhance its food safety. Activated lactoferrin or lactoferricin is a
pepsin hydrolysate of lactoferrin. Lactoferricin has enhanced antimicrobial action in comparison to lac-
toferrin. This activated form has been shown to protect fresh beef against E. coli 0157:H7, Salmonella,
Campylobacter, and more than 30 types of other pathogenic bacteria. It is stable at low pH and can be
added to drinking yogurt after preheat treatment, after fermentation, or through mixing with the fruit
preparation. In fruit yogurt, lactoferrin can be added after the heat treatment of milk together with the
starter or with the fruit preparation (Unpublished data).
Like other whey proteins, lactoferrin is sensitive to high-temperature heat treatment. However, for-
mulation containing lactoferrin can be pasteurized at normal pasteurization temperature, for example,
72°C for 16 s with less than 5% loss of bioactivity. Although pasteurization causes minimal reduction
in the activity, excessive heat treatment during processing of food products can reduce the activity of
lactoferrin [27,28].
55.2.4 Lactoperoxidase
Lactoperoxidase [EC 1.11.1.7] is an enzyme present in colostrum and milk, with a molecular weight of
approximately 77.5 kDa. Bovine colostrum and milk contain about 11–45 and 13–30 mg/L lactoperoxi-
dase, respectively [29]. In whey, lactoperoxidase constitutes approximately 0.5% of whey proteins. The
biological significance of lactoperoxidase is its involvement in the natural host defense system against
invading microorganisms [30]. The separation of lactoperoxidase from whey is based on the same prin-
ciple as used for isolation of lactoferrin. Lactoperoxidase is positively charged at the normal pH of whey
(isoelectric point in the pH range 9.0–10.0) and can be bound to cation-exchange resins and fractionated
from the rest of the whey proteins.
Lactoperoxidase inactivates or kills a wide spectrum of microorganisms through an enzymatic action.
This reaction involves two cofactors, hydrogen peroxide and thiocyanate ions, which together with lac-
toperoxidase constitute the lactoperoxidase system. Activation of the enzyme results in the formation of
hypothiocyanite ions, which are responsible for the antimicrobial action. The reaction of lactoperoxidase
system relies on the production of short-lived intermediary oxidation products of the thiocyanate ion
that reacts with bacterial cytoplasmic membranes, as well as impairs the function of metabolic enzymes.
As the addition of H2O2 is not permitted in certain countries, in situ development of H2O2 is carried out by
the addition of glucose oxidase (a permitted additive). The source of thiocyanate ion can be either natu-
rally present (as in the case of animal tissues and plant) or added as sodium or potassium thiocyanate [24].
Commercially, lactoperoxidase is isolated from either skim milk or whey using an ion-exchange pro-
cess similar to that used for the isolation of lactoferrin. The basic principle underlying the process is the
fact that lactoperoxidase has an isoelectric pH in the alkaline range (9.0–9.5), which means that it is posi-
tively charged at the normal pH of cheese whey (6.0–6.6), while the rest of the proteins are negatively
charged. This difference in pH is used to adsorb lactoperoxidase to an anion-exchange column that is
subsequently separated from other proteins [24]. The compositional and nutritional characteristics of a
commercial lactoperoxidase are shown in Table 55.1.
Lactoperoxidase when used in the form of lactoperoxidase system has a broad spectrum of antibacte-
rial activity, having a bacteriostatic effect against Gram-positive bacteria and a bactericidal effect against
Gram-negative microorganisms (e.g., Pseudomonas, Coliforms, Salmonella, and Listeria) [31–34]. The
following are the examples of potential applications of lactoperoxidase:
• Use of lactoperoxidase system to improve the yield of aquaculture by bactericidal effect on fish
pathogens.
• Application of lactoperoxidase together with lactoperoxidase system–activating ingredi-
ents (thiocyanate and hydrogen peroxide) in toothpaste formulations to protect against oral
Streptococci.
• Activation of salivary peroxidase antimicrobial system in toothpaste and mouth rinse reduces
acid formation by oral microorganisms, and clinical studies have shown that plaque accumula-
tion, gingivitis, and early carious lesions and aphthous lesions may all be reduced by appropri-
ate applications of the applied enzyme preparations.
• Promising results have been obtained when activating components are included in calf feed
with the aim of activating the lactoperoxidase system in the intestinal tract.
• Using lactoperoxidase system to protect contamination of Campylobacter jejuni in poultry
during slaughter.
• Lactoperoxidase can be used improving the shelf life in meat products by creating conditions
that allow activation of the lactoperoxidase system.
• Lactoperoxidase can successfully be used for controlling lactose fermentation and acidity
development during storage of yogurt.
• Application of the lactoperoxidase system for preservation of cosmetics showed a broad-
spectrum antimicrobial activity against bacteria, yeasts, and molds.
55.2.5 Casein and Whey Protein Hydrolysates

The enzymatic hydrolysis process of milk proteins produces ingredients that have several benefits for
nutritional, dietetic, and medical foods.
For the manufacture of protein hydrolysates, milk proteins (casein, caseinate, milk protein concen-
trate, lactalbumin, whey protein concentrate, and whey protein isolate) are first dispersed and solubilized
in water, and then the pH and temperature are adjusted to the desired levels (generally to the optimum
temperature for the enzyme). An appropriate enzyme is then added to the protein solution (substrate) at
a certain enzyme: substrate ratio that optimizes the enzymatic reaction. Under controlled conditions, the
enzyme cleaves the peptide bonds and produces the desired level of protein hydrolysis. The hydrolyzed
protein is optionally processed through steps such as clarification, flavor reduction, and concentration
and subsequently is spray dried [35].
There are a number of variables that need to be considered and controlled during the manufacture
of high-quality protein hydrolysate. The first consideration is the selection of appropriate substrate,
for example, protein and most commonly either whey protein concentrate or whey protein isolate are
used, as these result in hydrolysates that are of high quality and have bland flavor. Selection of an
appropriate enzyme is the next consideration and is one of the most critical steps in the development
of high-quality protein hydrolysate. Considerable efforts go into selecting an approved, food-grade
enzyme and also in the desired amount (enzyme/substrate ratio) required for hydrolysis. Hydrolysis of
protein results in a decrease in the pH of the protein solution, and, therefore, regular pH adjustment
is needed during manufacture. If a low-sodium hydrolysate is desired, potassium hydroxide is used
for pH adjustment; otherwise, sodium hydroxide is used. Throughout the process, pH, time, and tem-
perature are monitored and controlled that helps in producing high-quality, nutritional, and functional
protein hydrolysates [36].
A common way of differentiating between protein hydrolysates is the assessment of the degree of
hydrolysis (DH), where the higher value reflects higher level of hydrolysis. Users of protein hydrolysates
need an understanding of the desired attributes, and selection should be based on flavor, DH, required
bioactivity, and nutritional composition [37].
The bitter taste of protein hydrolysates is a major barrier to their use in food and health-care products.
The intensity of the bitterness is proportional to the number of hydrophobic amino acids in the hydro-
lysate. The presence of a proline residue in the center of the peptide also contributes to the bitterness.
The peptidases that can cleave hydrophobic amino acids and proline are valuable in debittering protein
hydrolysates [35].
Aminopeptidases from lactic acid bacteria (LAB) are available under the trade name Debitrase.
Carboxypeptidase A has a high specificity for hydrophobic amino acids and hence has a great potential for
debittering. A careful combination of an endoprotease for the primary hydrolysis and an aminopeptidase
Dairy Beverages 715
TABLE 55.3
Composition and Potential Functionality and Applications of Milk Protein Hydrolysates
Whey Protein or Casein Hydrolysate
Degree of <5 6–10 11–20 >20
hydrolysis (%)
Protein (%) 80–92 80–92 80–92 80–92
Amino nitrogen (%) 1–2 1–3 1–4 3–10
pH, 5% solids 6.0–7.6 6.0–7.6 6.0–7.6 6.0–7.6
Fat (%) 0.1–3.5 0.1–3.5 0.1–1.0 0.1–1.0
Lactose (%) 0.1–3.0 0.1–3.0 01–1.0 0.1–1.0
Ash (%) 2.0–4.0 3.0–4.0 3.0–4.0 3.0–5.0
Major differences Improved High levels of High levels of short- to High levels of di- and
physical medium-chain medium-chain peptides, tripeptides, free
functionality peptides, high reduced protein allergy, amino acids, high
(solubility, solubility, and heat high heat stability, low heat stability, low
emulsification, stability. lactose, low fat, and lactose, low fat, and
and foaming). reduced allergenicity. reduced allergenicity.
Potential food Dry and liquid High-protein Hypoallergenic infant, Medical and clinical
applications beverages, beverage powders, sports, and enteral nutritional
infant formula, powdered diet formulations, formulations,
and sports supplements, and high-protein hypoallergenic infant
nutritional infant and sports formulations, and and sports nutritional
products. and enteral lactose-free formulations, and
nutritional products. formulations. lactose-free
formulations.
Source: Based on commercial specification sheets.
for the secondary hydrolysis is required for the production of a functional hydrolysate with reduced
bitterness. Commercial milk protein hydrolysates are available in a range of DH and molecular weight
profiles (Unpublished data). Table 55.3 shows the composition, potential functionality, and applications
of milk protein hydrolysates.
Milk protein hydrolysates can be used for their bioactive or physiologically functional properties such
as reduced allergenicity and antigenicity, increased protein absorption, and release of bioactive peptides
[35,37–39].
Reduced allergenicity and antigenicity: Due to the differences in the protein composition of human
and cow milk, feeding of cow milk to newborn babies can cause allergic reactions. Hydrolysis
of cow milk proteins into smaller peptides reduces the risk of allergenicity and allows the use
of hydrolysate as a substitute for human milk protein in infant formula. Milk protein hydroly-
sates are also suitable for replacement of intact proteins in adult nutritional formulations where
reduced allergenicity is needed. The antigenicity of a protein, for example, its ability to induce
an allergic reaction, is related to the size of protein, amino acid sequence, and the presence
of secondary and tertiary structures. The antigenicity of hydrolysates can be measured by an
enzyme-linked immunosorbent inhibition assay that measures the amount of immunologically
active protein. Intact casein shows a high value for immunologically active casein; increasing
the DH decreases this value. Benefits of reduced allergenicity have been demonstrated in food
products such as infant, adult nutritional, and enteral formulations making them suitable for
consumers allergic to milk proteins.
Increased protein absorption: Depending on the DH, type of enzyme and conditions during
hydrolysis, a range of peptides can be obtained during hydrolysis. The decrease in the peptide
size generally leads to an increase in the absorption of peptides. Milk protein hydrolysates con-
taining mostly di- and tripeptides are absorbed more rapidly than free-form amino acids and
much more rapidly than intact proteins. This is desirable for athletes who want to maximize the
amino acid delivery to muscles, and to the patients with impaired absorption system.
Release of bioactive peptides: Hydrolysis of milk proteins may produce biologically active pep-
tides that are usually buried inside the aggregated structures of protein molecules. Milk protein
hydrolysates, for example, contain several biologically active peptides such as antihypertensive
peptides. The antihypertensive effect of several peptides has been related to the inhibition of
the angiotensin-converting enzyme (ACE). ACE activity results in blood pressure increase via
conversion of angiotensin I to angiotensin II, which is a vasoconstrictive peptide, and via deg-
radation of bradykinin, which is a vasodilatative peptide. Inhibition of ACE (e.g., by peptides
in milk protein hydrolysates) results in a decrease in blood pressure.
55.2.6 Milk Minerals

Calcium and phosphorus are the major minerals required for the growth and development of the bones
and teeth. Calcium deficiency is far too common in diet, and awareness of this deficiency has led to
calcium fortification of a range of food products including breakfast cereals and fruit juices. Although
consumers consider milk and dairy products to be the richest sources of calcium, many have limited
their consumption to reduce fat in their diets or because of their intolerance to lactose. Milk minerals are
a rich source of calcium used for calcium fortification of food and beverage products [40]. Commercial
milk mineral complex is obtained from cheese whey after removal of proteins, which are converted into
protein concentrates, and lactose, which is dried into lactose powder (Unpublished data). A typical com-
position of a commercial milk mineral product (milk calcium) with 24% calcium is shown in Table 55.1.
Milk minerals are a natural milk calcium complex product manufactured from milk and whey.
Calcium in milk minerals is a highly bioavailable calcium phosphate that is a natural milk calcium
complex. Dietary calcium has been linked to osteoporosis and bodily functions such as regulation of
cell function, nerve conduction, muscle contraction, and blood coagulation. Milk minerals are rich in
calcium, and milk calcium has been suggested to have a number of bioactive functional properties such
as prevention of osteoporosis and growth of healthy bones and teeth, blood pressure and cardiovascu-
lar disease (CVD) control, lowering effect on hypertension, prevention of colon cancer, and control of
weight gain and obesity [41–44]. Suggested applications of milk minerals include:
• Dairy products such as recombined milk, flavored milk, yogurt, and cheese.
• Nutritional and functional foods such as sports and adult nutritional beverages, weight loss
products, and sports bars.
• Bakery products such as breads and cakes.
• Confectionery products.
• Breakfast cereals.
• Convenience foods such as soups, sauces, and frozen desserts.
• Food supplements such as capsules and tablets.
55.2.7 Micellar Casein

Keeping high concentrations of casein in micellar form can help create new opportunities for dairy
proteins that exploit colloidal behavior and the naturally high calcium content. Kerry Ingredients has
commercialized micellar casein under the brand name Micellnor™ that is specifically developed for use
in sports nutrition and weight loss applications. According to Kerry Ingredients, Micellnor™ generates
a positive nitrogen balance in the body and promotes satiety. Slow amino acid release promotes muscle
growth and recovery, thus preventing muscle breakdown. The slow release principle ensures that casein
coagulates and aggregates in the stomach. This results in slow and prolonged digestion of proteins. Kerry
Ingredients’ Micellnor™ is claimed as a native micellar casein product, produced from skim milk using
a proprietary combination of membrane filtration technologies.
Dairy Beverages 717
55.2.8 α-Lactalbumin
Alpha-lactalbumin in cow milk has considerable resemblance to that of human milk that makes it a valu-
able ingredient for construction of infant formulas with amino acid compositions that are very close to
that of human milk. A small number of dairy companies have isolated α-lactalbumin from cheese whey
using an ion-exchange technology and developed ingredients rich in α-lactalbumin. Davisco Foods in the
United States has been marketing tryptophan-rich α-lactalbumin based on its ability to improve sleep and
early-morning performance by increased alertness. A study carried out by Markus et al. [45] showed a
130% increase in plasma tryptophan after the evening intake of α-lactalbumin-enriched standard diet as
compared with a placebo. This was accompanied by reduced sleepiness and higher task-related brain
activity the following morning, which suggests improved alertness due to better sleep. The effects
of poor sleep on performance may be partly mediated by a biochemical imbalance of brain serotonin
(5-hydroxytryptamine [5-HT]). Brain 5-HT seems to be involved in the regulation of sleep and cognitive
processes, and sleep abnormalities and cognitive decline in clinical populations are partly attributable to
deficient brain 5-HT activity. Accordingly, reduced 5-HT concentrations resulting from the exhaustion
of its plasma precursor tryptophan has been found to provoke sleep abnormalities seen in depression,
whereas increases in available plasma tryptophan for uptake into the brain improved sleep in different
subjects. These sleep-promoting effects of tryptophan may be stronger in subjects with sleep complaints,
because they are vulnerable to the sleep-reducing effects of tryptophan depletion. Sleep-enhancing
attributes of α-lactalbumin are attributed to its enhanced levels of tryptophan [46].
55.2.9 Osteopontin
Osteopontin is a heavily phosphorylated and acidic glycoprotein with strong calcium-binding proper-
ties. Although its exact function is not yet known, the ubiquitous presence of osteopontin in the human
body has led many academic researchers to focus on investigating the glycoprotein in a variety of physi-
ological functions. Preliminary tests suggest that osteopontin plays a role in the development of infant
immune function. In infant nutrition products, it is thought to help nonbreastfed infants achieve immune
modulation comparable to that of breastfed infants. The whey protein osteopontin was identified in
human milk only a few years after being discovered in the human body. Small quantities have since
been identified in bovine milk and are now available from Arla Foods Ingredients in a 95% pure form.
Lacprodan OPN-10 is the first commercially available osteopontin product on the market. The target
application for osteopontin is infant formula.
55.2.10 Caseinophosphopeptide
Buried inside casein is a phosphopeptide (caseinophosphopeptide [CPP]) sequence -Pse-Pse-Pse-Glu-
Glu-, where Pse is a phosphoseryl residue which stabilizes calcium and phosphate ions in aqueous
solution and makes them bioavailable [47,48]. The CPP can be isolated from casein by tryptic hydro-
lysis, and ingredients so obtained have found several commercial applications. CPP has been shown to
enhance mineral absorption from food and beverage products, improving consumers’ overall uptake of
important minerals such as calcium. Studies have also suggested that CPP may improve dental health
by reducing the risk of dental caries. Currently, a number of commercial CPP-rich dairy powders are
available on the market, one of which is sold by Arla Foods Ingredients in Denmark under the brand
Lacprodan®. Commercial CPP-rich ingredients, Recaldent™, can be found in a range of toothpaste,
beverage, and chewing gum products.
55.3 Functional Dairy Foods

In past the few years, innovation and growth in the dairy industry has been driven by consumer health
concerns such as obesity, coronary heart disease (CHD), and diabetes. Apart from being a rich source
of bioactive ingredients, milk and dairy products, due to their wider availability, superior nutrition, and
TABLE 55.4
Key Trends in Functional Dairy Beverages
Major Modification and/or
Key Functional Ingredient Consumer Milk Product Proposed Benefits
Nutrient content claims—reduced Liquid milk, yogurt, and cheese General health and wellness and weight
fat, sodium or carbohydrate, and with nutritional claims. management.
low calories
Probiotics Yogurt, drinking yogurt, yogurt Digestive health, immune support (or “natural
smoothie, and cheese. defense”), and optimal health and wellness
maintenance.
Prebiotic fiber Yogurt and drinking yogurt. Act as soluble fiber in the colon for the
growth of probiotic bacteria.
Omega-3 fatty acids Milk, cheese, yogurt, drinking Heart health and brain function.
yogurt, and cheese.
Phytosterols Milk and yogurt. Cholesterol reduction.
Antioxidants Milk and drinking yogurt. Breast, colon, and prostate cancers.
Dietary fiber Milk and drinking yogurt. Able to enhance gut function and may act as
prebiotics.
Source: Based on own experience.
flavor, remain one of the biggest vehicles for the delivery of functional ingredients. This has led to the
growth of functional dairy products containing nondairy bioactive ingredients such as omega-3 (ω-3)
fatty acids and plant sterols. A summary of trends in consumer dairy products is shown in Table 55.4.
55.3.1 Nutrient Content Claims

Nutrient content claims in foods include “low fat,” “low saturated fat,” “low sodium,” “low carbohydrate,”
and “low calories.” In the few years, products with “low” claims have grown significantly, often taking the
market share of regular products. For the dairy industry, the main focus for “low” claims has been related
to the reduction of fat, including cholesterol. Some companies also market low or no-lactose milk prod-
ucts. Milk and other dairy products such as cheese and yogurt are routinely available in low-fat versions.
55.3.2 Probiotics
Probiotics are living microflora or beneficial bacteria, which are critical in maintaining good health
by supporting intestinal balance. They help to inhibit the growth of harmful bacteria. When harmful
bacteria increase and outnumber the good microflora or bacteria within the digestive system, the diges-
tive system becomes unbalanced, which can impede nutritional absorption, accelerate aging, promote
cancer (particularly in the colon), reduce immune response, and lead to other ailments such as infection,
diarrhea, constipation, colitis, and lactose intolerance [49]. Furthermore, beneficial bacteria is depleted
in any number of ways including antibiotic use and infections, as well as lifestyle factors such as stress
or following a “modern” diet filled with highly processed and low-fiber foods. Combined with the high
levels of pesticides, preservatives, and toxins in our food and environment, our digestive balance is
compromised, debilitating our digestive capacity, slowing decomposition, and further weakening our
immune systems. These facts are increasingly being recognized by consumers who are searching for
natural solutions.
Probiotics’ healthy benefits include [49,50]:
• Speeding the breakdown of organic waste fragments and cleaning the intestinal tract by
dislodging decayed matter and toxin buildup through metabolic processes.
• Increasing the production of important enzymes and the availability of vitamins and nutrients.
• Helping regulate digestion, relieving constipation and diarrhea conditions.
Dairy Beverages 719
• Helping to kill viruses and parasites.

• Strengthening the immune system to alleviate allergies, skin problems, chronic fatigue syn-
drome, and many other disease conditions.
Dairy industry has managed to have an upper hand in the delivery of probiotics through yogurt, although
challenges from other delivery vehicles such as supplements, juices, and drinks have been noticed recently.
Yogurt is considered a preferred vehicle for the delivery of probiotics because of the flavor and healthy
image of yogurt. The most common food-based delivery systems for probiotics are yogurt and fermented
milk drinks such as Yakult, Actimel, Müller Vitality and Healthy Balance, Petit Filous Plus, and Danone
Activia®. These probiotic foods can be eaten at any time of the day, and many people take them as part of
their breakfast routine. At present, there is only limited scientific evidence to suggest that healthy people
will benefit from a regular intake of probiotics; however, there is strong growth of probiotics in dairy
due to the preferred taste profile and convenient delivery format (such as single-serve bottles and drinks).
Europe is by far the leading market for the consumption of probiotic dairy products due to the long his-
tory of fermented milk in the region. However, the U.S. consumers are becoming increasingly aware of
the benefits of probiotics because of the influential marketing and consumer awareness efforts of com-
panies such as Danone (known as Dannon in the United States) and Yakult and General Mills (Yoplait).
Although yogurts and yogurt drinks are the preferred vehicle for delivery of probiotics because of the
flavor and healthy image, other dairy products such as cheese and milk have also been used. Recently,
Kraft Foods Canada launched probiotic cheddar and cottage cheese under the brand name “LiveActive.”
As probiotics are predominantly LAB and prefer lactose a as source of nutrition, milk and its derivatives
will remain at the forefront of probiotics delivery market.
55.3.3 Prebiotic Fiber

Prebiotics are carbohydrates that cannot be broken down by the human digestive system. The main types
are inulin and fructooligosaccharides. They occur naturally in some foods such as chicory, artichoke,
leeks, onions, and asparagus. Prebiotics help feed the good bacteria already in the digestive system,
which can improve the health of the digestive system and may help in strengthening the immune system.
There is a growing body of evidence on the health benefits of prebiotics; however, further research is
required to confirm the actual amounts that are needed for optimal health. The products including prebi-
otic fiber are yogurt drinks such as Früzinga, Flora pro.active, and Nestle Bliss.
55.3.4 Omega-3 Fatty Acids

In recent years, there has been a trend toward the addition of ω-3 fatty acids to dairy products. These
fatty acids are essential that the human body is unable to produce on its own and must be incorporated
into the diet. Fish is the usual source of ω-3, but it is well recognized that most consumers are not con-
suming sufficient quantities of ω-3, this important nutrient. The benefits of ω-3 fatty acids include the
reduction of triacylglycerols (TAG); prevention of certain illnesses, such as CVD; improved immune
reactions against allergies; and a reduction in the threat of blood clots [51].
Since 2004, the FDA has allowed a limited health claim for reduced risk of CHD on conventional foods
that contain eicosapentaenoic acids and docosahexaenoic acids. The role of ω-3 fatty acids in growth and
development as well as in health and disease, particularly CHD, is currently one of the fastest-growing
research areas in nutritional science. Commercially, fish oil, algae, and flaxseed oil are the most com-
monly available sources of ω-3 fatty acids. Leading companies marketing ω-3 fatty acids are Martek/
DSM (algal source), Ocean Nutrition Canada/DSM (fish oil), and Denomega (fish oil). Dairy products,
such as milk and yogurt, offer convenient delivery system for ω-3 fatty acids. However, the delivery of
ω-3 fatty acids in dairy products remains a challenge as these fatty acids have a strong odor and poor
oxidative stability and tend to deteriorate during homogenization and pasteurization. Consequently, fish
oil, algae, or flaxseed oil are normally preemulsified and admixed with antioxidant blends using a tech-
nology called microencapsulation that helps protect ω-3 fatty acids during milk processing. Milk proteins
have successfully been used as encapsulants for ω-3 fatty acids due to their ability to emulsify and pro-
tect fat from oxidation. Recent trends suggest that one of the fastest-growing markets for ω-3 fatty acids
is the infants’ and kids’ dairy market where manufacturers are promoting the brain health benefits of
ω-3. Dairy products, especially milk and yogurt drinks, may remain important delivery vehicles for ω-3
fatty acids, and more innovation in this area is anticipated [52].
55.3.5 Phytosterols
Phytosterols are plant materials chemically similar to cholesterol but not found in any significant abun-
dance in the human body. They are clinically proven to reduce the absorption of cholesterol from the gut,
resulting in a lowering of low-density lipoprotein (LDL) cholesterol. Lowering LDL cholesterol can help
maintain a healthy heart. Plant sterols are found naturally in very small amounts in fruits, vegetables,
vegetable oils, grains, nuts, and seeds. Benecol, Danone Danacol, Minicol, and Flora pro.activ are some
of the many dairy products available with plant sterols.
As rates of heart disease continue to rise worldwide, makers of plant sterols are optimistic about the
success of their products. The heart health benefits of phytosterols have been recognized by the FDA since
2003, and claims on a broad range of food products are allowed when formulated with 0.65 g of phytos-
terol esters or 0.4 g free phytosterols per serving. Companies active in marketing plant sterols to the dairy
industry are Forbes MediTech (Pharmatech), Raisio, Cognis, and Cargill. With a strong interest in heart
health and approval by regulatory authorities, more dairy companies are likely to introduce milk prod-
ucts with phytosterols. However, challenges still remain for dairy companies to convince consumers that
dairy products can be as effective as cholesterol-lowering margarines, which are leading the cholesterol-
lowering markets around the world.
55.3.6 Antioxidants and Plant Extracts

Dairy products especially yogurts have seen the addition of antioxidants in the form of so-called super
fruits or plant extracts. Antioxidants are promoted for their role in protecting human cells from free radical
damage that occurs during aging and infections. Free radicals are formed from cellular reactions in the
human body, and they are cited as an important cause of most chronic diseases. Antioxidants, by scaveng-
ing the free radicals, have been suggested to protect body cells. Examples of popular plant extracts that have
been added to yogurt drinks are green tea extract, berries, black currant extract, and grape seed extract.
55.3.7 Dietary Fiber

Dietary fiber, found mainly in fruits, vegetables, whole grains, and legumes, is recognized for health
benefits such as lowering the risk of diabetes and CHD. The American Dietetic Association recommends
an intake of 20–35 g for adults, 25 g daily for girls aged 9–18 years, and 31–38 g for boys aged 9–18. The
American Heart Association recommends 25–30 g daily. On average, consumers in Western countries
consume less than 50% of the dietary fiber levels required for good health. Recognizing the growing
scientific evidence for the physiological benefits of increased fiber intake, the FDA has given approval
to health claims for fiber. Although dairy products are not generally associated with fiber, some milk
products have recently been launched with added fiber. Opportunities exist for incorporating the soluble
fiber beta-glucan (from oats and barleys), as beta-glucan has cholesterol-lowering function in humans
and this is an approved health claim in the United States.
55.4 Technical Challenges in Developing Functional Dairy Beverages

Although milk and dairy products remain preferred vehicles for the delivery of bioactive ingredients,
potentially a number of challenges are faced by manufacturers when incorporating bioactive ingredients
in dairy beverages. Most of these related to the physical stability or sensory attribute that can make the
final product undesirable to consumers. Table 55.5 summarizes potential challenges for dairy beverages
containing bioactive ingredients.
Dairy Beverages 721
TABLE 55.5
Potential Challenges for Dairy Beverages Containing Bioactive Ingredients
Bioactive Potential Formulation Challenge Precautions Needed
Probiotics Exposure to heat, oxygen, low pH, moisture, Selection of appropriate species of bacteria,
and direct light can reduce activity. Ensuring providing sterile environment for growth, and
enough live probiotic bacteria reaches the gut. avoiding inactivation postprocessing.
Omega-3 fatty Undesirable flavor, sensitivity to heat, light, Selection of protected (encapsulated) omega-3
acids and air. fatty acids, avoiding excessive heating, and
homogenization during processing.
Phytosterols Insolubility and difficulty in incorporation into Dispersion of phytosterols in small amounts of fat,
low- or no-fat beverages. continuous shearing, and temperature control.
Bioactive Bitterness, stability toward processing, poor Avoiding excessive heating and high-pressure
peptides from emulsifying properties, and flavor. homogenization. Use of emulsifier to enhance
milk emulsification.
Milk calcium Insolubility, sedimentation and grittiness, and Using micronized (finely milled) milk calcium,
protein instability. use of stabilizer such as carboxymethyl cellulose
and microcrystalline cellulose.
Antioxidant and Bitterness, beany taste, and poor solubility in Avoiding extreme heat, pH, and shearing cycles
isoflavones water. during processing.
Dietary fiber Poor suspension and sedimentation. Use of finely milled powder and addition of
stabilizers such as starch, guar gum, and acacia
gum.
Source: Based on own experience.
55.5 Conclusion
Milk is a rich source of nutrients and contains numerous bioactive components both in the lipid and in
the skim part. By far, skim milk and whey are the major sources of bioactive components. Thus far,
many of the bioactive components from milk remain unexploited as commercial ingredients. However,
developments in new processing technologies and a desire to add value to dairy have stimulated sig-
nificant commercialization of healthy bioactive components from milk. Functional ingredients from
dairy including colostrum, GMP, lactoferrin, and protein hydrolysates have been successfully commer-
cialized and found applications in infant formula, sports nutrition, antibacterial, and immune enhance-
ment products. Although milk and dairy products remain preferred vehicles for the delivery of bioactive
ingredients, potentially a number of challenges are faced by manufacturers when incorporating bioac-
tive ingredients in dairy beverages. Most of these related to the physical stability or sensory attribute,
although not impossible to overcome, can make the final product undesirable to consumers.
REFERENCES
1. Euromonitor, Opportunities to Target the Ageing through Functional Food and Drink, Euromonitor,
London, UK, 2013.
2. Özer, B.H. and Kirmaci, H.A., Functional milks and dairy beverages. Int. J. Dairy Technol., 63, 1–15,
2010.
3. Bhat, Z.F. and Bhat, H., Milk and dairy products as functional foods: A review. Int. J. Dairy Sci., 6,
1–12, 2011.
4. Weaver, C.M., Role of dairy beverages in the diet. Phys. Behav., 100, 63–66, 2010.
5. Fosset, S. and Tome, D., Nutraceuticals from milk, in Encyclopedia of Dairy Sciences, Fuquay, J.W.,
Fox, P.F., and MacSweeny, P., Eds., Academic Press, London, UK, 2011, pp. 1062–1066.
6. Playne, M.J., Bennett, L.E., and Smithers, G.W., Functional dairy foods and ingredients. Aust. J. Dairy
Technol., 58, 242–264, 2003.
7. Antonio, J., Sanders, M., and Van Gammeren, D., The effects of bovine colostrum supplementation on
body composition and exercise performance in active men and women. Nutrition, 17, 243–247, 2001.
8. Stelwagen, K., Carpenter, E., Haigh, B., Hodgkinson, A., and Wheeler, T.T., Immune components of
bovine colostrum and milk. J. Anim. Sci., 87(Suppl. 13), S3–S9, 2009.
9. Elfstranda, L., Lindmark-Månssonb, H., Paulsson, M., Nybergc, L., and Åkessond, B., Immunoglobulins,
growth factors and growth hormone in bovine colostrum and the effects of processing. Int. Dairy J., 12,
879–887, 2002.
10. Blum, J.W. and Baumrucker, C.R., Insulin-like growth factors (IGFs), IGF binding proteins, and other
endocrine factors in milk: Role in the newborn. Adv. Exp. Med. Biol., 606, 397–422, 2008.
11. Uruakpa, F.O., Ismonda, M.A.H., and Akobundua, E.N.T., Colostrum and its benefits: A review. Nutr.
Res., 22, 755–767, 2002.
12. Davidson, G.P., Whyte, P.B., Daniels, E., Franklin, K., Nunan, H., McCloud, P.I., Moore, A.G., and
Moore, D.J., Passive immunisation of children with bovine colostrum containing antibodies to human
rotavirus. Lancet, 2, 709–712, 1989.
13. Stephan, W., Dichtelmuller, H., and Lissner, R., Antibodies from colostrum in oral immunotherapy.
J. Clin. Chem. Clin. Biochem., 28, 19–23, 1990.
14. Seung, C.B. and Jae, H.Y., Separation of immunoglobulin from Holstein colostrum and its immunologi-
cal response. Food Biotechnol., 4, 117–121, 1995.
15. Playford, R.J., Macdonald, C.E., and Johnson, W.S., Colostrum and milk-derived peptide growth factors
for the treatment of gastrointestinal disorders. Am. J. Clin. Nutr., 72, 5–14, 2000.
16. Lilius, E.-M. and Marnila, P., The role of colostral antibodies in prevention of microbial infections.
Curr. Opin. Infect. Dis., 14, 295–300, 2001.
17. Xua, Y., Sleigh, R., Hourigan, J., and Johnson, R., Separation of bovine immunoglobulin G and glyco-
macropeptide from dairy whey. Process Biochem., 36, 393–399, 2000.
18. Brody, E.P., Biological activities of bovine glycomacropeptide. Br. J. Nutr., 84(Suppl. 1), S39–S46,
2000.
19. Ney, D.M., Gleason, S.T., van Calcar, S.C., MacLeod, E.L., Nelson, K.L., Etzel, M.R., Rice, G.M., and
Wolff, J.A., Nutritional management of PKU with glycomacropeptide from cheese whey. J. Inherit.
Metab. Dis., 32, 32–39, 2009.
20. Neeser, J.R., Chambaz, A., Vedovo, S.D., Prigent, M.J., and Guggenheim, B., Specific and nonspecific
inhibition of adhesion of oral actinomyces and Streptococci to erythrocytes and polystyrene by caseino-
glycopeptide derivatives. Infect. Immun., 56, 3201–3208, 1988.
21. Kawasaki, Y., Isoda, H., Tanimoto, M., Dosako, S., Idota, T., and Ahiko, K., Inhibition by lactoferrin and
κ-casein glycomacropeptide of binding of cholera toxin to its receptor. Biosci. Biotechnol. Biochem., 56,
195–198, 1992.
22. Otani, H., Monnai, M., Kawasaki, Y., Kawakami, H., and Tanimoto, M., Inhibition of mitogen-induced
proliferative responses of lymphocytes by bovine κ-caseinoglycopeptides having different carbohydrate
chains. J. Dairy Res., 62, 349–357, 1995.
23. Daddaoua, A, Puerta, V., Zarzuelo, A., Suárez, M.D., Sánchez de Medina, F., and Martínez-Augustin,
O., Bovine glycomacropeptide is anti-inflammatory in rats with hapten-induced colitis. J. Nutr., 35,
1164–1170, 2005.
24. Farnaud, S. and Evans, R.W., Lactoferrin—A multifunctional protein with antimicrobial properties.
Mol. Immun., 40, 395–405, 2003.
25. Mulder, A.M., Connellan, P.A., Oliver, C.J., Morris, C.A., and Stevenson, L.M., Bovine lactoferrin sup-
plementation supports immune and antioxidant status in healthy human males. Nutr. Res., 28, 583–589,
2008.
26. Debbabi, H., Dubarry, M., Rautureau, M., and Tomé, D., Bovine lactoferrin induces both mucosal and
systemic immune response in mice. J. Dairy Res., 65, 283–293, 1998.
27. Paulsson, M.A., Thermal behavior of bovine lactoferrin in water and its relation to bacterial interaction
and antibacterial activity. J. Dairy Sci., 76, 3711–3720, 1993.
28. González-Chávez, S.A., Arévalo-Gallegos, S., and Rascón-Cruz, Q., Lactoferrin: Structure, function
and applications. Int. J. Antimicrob. Agents, 33, 301.e1–301.e8, 2009.
29. Korhonen, H., Antimicrobial factors in bovine colostrum. J. Sci. Agric. Soc. Fin., 49, 434–447, 1977.
30. de Wit, J.N. and van Hooydonk, A.C.M., Structure, functions and applications of lactoperoxidase in
natural antimicrobial systems. Neth. Milk Dairy J., 50, 227–244, 1996.
31. Reiter, B. and Harnulv, G., Lactoperoxidase antibacterial system: Natural occurrence, biological func-
tion and practical applications. J. Food Protect., 47, 724–732, 1984.
Dairy Beverages 723
32. Nakada, M., Dosako, S., Hirano, R.O., and Nakajima, I., Lactoperoxidase suppresses acid production in
yogurt during storage under refrigeration. Int. Dairy J., 6, 33–42, 1996.
33. Kussendrager, K.D. and van Hooijdonk, A.C.M., Lactoperoxidase: Physico-chemical properties, occur-
rence, mechanism of action and applications. Br. J. Nutr., 84(Suppl. 1), S19–S25, 2000.
34. Min, S., Harris, L.J., and Krochta, J.M., Antimicrobial effects of lactoferrin, lysozyme, and the lac-
toperoxidase system and edible whey protein films incorporating the lactoperoxidase system against
Salmonella enterica and Escherichia coli O157:H7. J. Food Sci., 70, m332–m338, 2005.
35. Sinha, R., Radha, C., Prakash, J., and Kaul, P., Whey protein hydrolysate: Functional properties, nutri-
tional quality and utilization in beverage formulation. Food Chem., 101, 1484–1491, 2007.
36. FitzGerald, R.J. and Meisel, H., Milk protein-derived peptide inhibitors of angiotensin-I-converting
enzyme. Br. J. Nutr., 84(Suppl. 1), S33–S37, 2000.
37. Mahmoud, M.I., Malone, W.T., and Cordle, C.T., Enzymatic hydrolysis of casein: Effect of degree of
hydrolysis on antigenicity and physical properties. J. Food Sci., 57, 1223, 1992.
38. Peña-Ramos, E.A. and Xiong, Y.L., Antioxidative activity of whey protein hydrolysates in a liposomal
system. J. Dairy Sci., 84, 2577–2583, 2001.
39. Otte, J, Shalaby, S.M., Zakora, M., Pripp, A.H., and El-Shabrawy, S.A., Angiotensin-converting enzyme
inhibitory activity of milk protein hydrolysates: Effect of substrate, enzyme and time of hydrolysis.
Int. Dairy J., 17, 488–503, 2007.
40. Gaucheron, F., The minerals of milk. Reprod. Nutr. Dev., 45, 473–483, 2005.
41. Murphy, S., Khaw, K.-T., May, H., and Compston, J.E., Milk consumption and bone mineral density in
middle aged and elderly women. Br. Med. J., 308, 939–941, 1994.
42. Cadogan, J., Eastell, R., Jones, N., and Barker, M.E., Milk intake and bone mineral acquisition in ado-
lescent girls: Randomised, controlled intervention trial. Br. Med. J., 315, 1255–1260, 1997.
43. Cashman, K.D., Milk minerals (including trace elements) and bone health. Int. Dairy J., 16, 1389–1398,
2006.
44. Scholz-Ahrens, K.E. and Schrezenmeir, J., Milk minerals and the metabolic syndrome. Int. Dairy J., 16,
1399–1407, 2006.
45. Markus, C.R., Jonkman, L.M., Lammers, J.H., Deutz, N.E., Messer, M.H., and Rigtering, N., Evening
intake of alpha-lactalbumin increases plasma tryptophan availability and improves morning alertness
and brain measures of attention. Am. J. Clin. Nutr., 81, 1026–1033, 2005.
46. Silbera, M.Y. and Schmitt, J.A.J., Effects of tryptophan loading on human cognition, mood, and sleep.
Neurosci. Biobehav. Rev., 34, 387–407, 2010.
47. Reynolds, E.C., Anticariogenic complexes of amorphous calcium phosphate stabilized by casein phos-
phopeptides: A review. Spec. Care Dentist., 18, 8–16, 1998.
48. Cross, K.J., Huq, N.L., Palamara, J.E., Perich, J.W., and Reynolds, E.C., Physicochemical character-
ization of casein phosphopeptide-amorphous calcium phosphate nanocomplexes. J. Biol. Chem., 280,
15362–15368, 2005.
49. Goldin, B.R., Health benefits of probiotics. Br. J. Nutr., 80(Suppl.), S203–S207, 1998.
50. Parvez, S., Malik, K.A., Kang, S., and Kim, K.-Y., Probiotics and their fermented food products are
beneficial for health. J. Appl. Microb., 100, 1171–1185, 2006.
51. Simopoulos, A.P., Omega-3 fatty acids in health and disease and in growth and development. Am. J.
Clin. Nutr., 54, 438–463, 1991.
52. Schmidt, E.B. and Dyerberg, J., Omega-3 fatty acids. Drugs, 47, 405–424, 1994.
56
Soybean Beverages
Tzou-Chi Huang and Chi-Tang Ho
CONTENTS
56.1 Introduction................................................................................................................................... 725
56.3.1 Isoflavones........................................................................................................................ 727
56.3.2 Anthocyanins.................................................................................................................... 729
56.3.3 Phenolic Acids.................................................................................................................. 729
56.3.4 Antioxidants and Bioactives Responsible for Antioxidant Activity of Soybean Milk.... 729
56.4 Health Effects................................................................................................................................ 730
56.4.1 Bioavailability of Bioactives from Soybean Milk............................................................ 730
56.4.2 Animal Studies with Soybean Milk................................................................................. 730
56.4.3 Human Intervention Studies with Soybean Milk..............................................................731
56.6 Conclusion..................................................................................................................................... 733
References............................................................................................................................................... 733
56.1 Introduction
Soybean (Glycine max (L.) [Merrill] germplasms) is a valuable crop and is the most commonly consumed
legume in the world. Especially in Asia, soybean is used in various foods, such as soybean sprouts, soy
paste, soy milk, tofu, and soybean oil. It is considered to be a perfect crop since it contains essential
amino acids, which are not synthesized in the human body [1].
Soybean milk is an important food in China where it is commonly used as a hot breakfast drink. It is
consumed extensively throughout China and the vast oversea Chinese community [2]. The beans are washed,
ground, extracted with water, then boiled for about 20 min. Pentanal, hexanal, and other aldehydes are respon-
sible for a beany flavor of soybean milk. Various techniques, such as high-pressure processing (800 MPa), use
of antioxidant [3], and thermal treatment [4], have been applied to produce beany flavor-free soybean milk.
The inactivation of lipoxygenase in soy milk and crude soybean extract leads to inhibition of generation of
volatile aldehydes via the lipoxygenase-assisted oxidation of polyunsaturated fatty acids (PUFA) present.
Setchell [2] reported a substantial difference between the intake of soy food items in the Japanese,
Chinese, and Indonesian populations compared with the U.S. and European populations. A substantial
difference between the intake of soy food items in the Oriental populations compared with the Western
populations is that in the Asian countries, people consume a traditional diet that is rich in fermented soy
product, whereas in the Western countries, mainly nonfermented soy products or soy supplements are
consumed. Oriental fermented soy products contain mainly isoflavone aglycones. Western nonfermented
soy foods have mainly total isoflavone as β‐glycoside conjugates and its concentration is similar to those
in the raw soybean. Asians, especially Japanese, have a much larger daily intake of these isoflavone
aglycones compared to Americans on a body weight basis. Coward et al. [1] attributes the lower inci-
dence and mortality from breast cancer in Asian women to the higher intake of dietary isoflavones. This
chapter highlights the nutritional characteristics, bioactives, and health benefits of soy beverages.
725

Soybean, which is rich in protein, carbohydrate, fat, and isoflavone, has attracted much attention because
of its potential health benefits. The composition of soy milk covers a relatively wide range, depending
on varieties, the extraction procedures, and the temperature of extraction. Data from various sources
on the compositional and nutritional characteristics (proximate compositions, minerals, vitamins, and
essential amino acids) of soybean milk and fermented soybean milk were compiled and are summarized
in Table 56.1 [5]. Fermentation of soybean leads to increases in essential amino acids, especially
phenylalanine, tyrosine, lysine, and threonine.
Soy milk is made by combining dried soybeans with water, then finely ground, and subsequently
filtered. The nutrient composition of soy milk is affected by numerous factors including varieties of
soybeans, the extraction procedure, ratio of the water or salt solution to the beans, and the temperature
of extraction. The major nutritional composition of soybean milk is similar to that of regular cow’s
TABLE 56.1
Compositional and Nutritional Characteristics of Soybean Milk and Fermented Soybean Milk (per 100 g)
Nutrient Unit Soybean Milk Fermented Soybean Milk
Water g 91.53 84.67
Energy kcal 41 66
Protein g 2.88 2.64
Lipid (fat) g 1.65 1.76
Minerals
Calcium mg 123 132
Iron mg 0.44 13
Magnesium mg 16 na
Zinc mg 0.25 na
Vitamins
Vitamin A IU 206 53
Vitamin B12 μg 1.23 1.3
Vitamin D μg 1.20 na
Essential Amino Acids
Cystine g 0.51 0.57
Isoleucine g 1.76 1.79
Leucine g 2.62 2.73
Lysine g 1.94 3.01
Methionine g 0.57 0.69
Phenylalanine g 1.98 2.31
Threonine g 0.88 1.42
Tyrosine g 1.26 1.51
Valine g 1.67 1.66
Soybean Beverages 727
milk with fewer calories (80 cal/oz vs. 103 cal/oz cup), saturated fat (1.5 g vs. 4.5 g), and cholesterol
(0 mg vs. 12 mg) [6]. The American Heart Association (AHA) recommends soy milk as a good low-fat
choice for vegetarian and vegan diets.
The principal soluble carbohydrates of mature soybeans are the disaccharide sucrose (range
2.5%–8.2%), trisaccharide raffinose (0.1%–1.0%), and tetrasaccharide stachyose (1.4%–4.1%) [7]. One
cup of plain soy milk contains almost 4 g of carbohydrate, only 1 g of sugar, and is free from lactose. Soy
milk is a good choice for those who are lactose intolerant or allergic to dairy.
One cup of unfortified soymilk contains at least 7 g of soybean. Soybean protein contains all the
essential amino acids and is regarded as a complete protein [8]. Nutritional quality of soybean has been
evaluated using the protein digestibility corrected amino acid score and found to be almost equivalent
to meat, eggs, and casein for human growth and health. Whole soybean has a biological value of 96,
soybean milk 91, and eggs 97 [9].
Soy protein plays a vital role in protecting the body from cardiovascular disease (CVD). In 1999,
the U.S. Food and Drug Administration (FDA) approved labeling for foods containing soy protein as
protective against coronary heart disease (CHD) [10]. The AHA contradicted Dr. Anderson’s 1995
[11] findings. They claimed that soy protein and isoflavones have little effect on cholesterol levels or
other lipids. However, in 2013, the FDA approved labeling for health claims. One of them is “Diets
low in saturated fat and cholesterol that include 25 g of soy protein a day may reduce the risk of heart
disease” [12].
The health-promoting capacities of soybeans are totally dependent on processing history [13]. For
human consumption, soybeans must be cooked to destroy antinutritional factors, such as Kunitz trypsin
inhibitor (KTI) and Bowman–Birk inhibitor [14]. Thermal treatment not only improves the aroma qual-
ity of soybean milk but also increases the bioavailability of nutrients. Kwok et al. [15] reported that the
heating time needed for destroying 90% of soymilk trypsin inhibitors was temperature dependent (100°C,
40 min; 125°C, 5 min; and 143°C, 60 s). The thermal treatment causes rapid KTI incorporation into pro-
tein aggregates leading to a 100% inactivation of KTI.

A substantial difference between the intake of soy food items in the Japanese, Chinese, and
Indonesian populations compared with the U.S. and European populations is that in the Asian
countries, people consume a traditional diet that is rich in fermented soy product, whereas in
the Western countries, mainly nonfermented soy products or soy supplements are consumed [2].
Fermented soy products, for instance, natto or tempeh, have aglycones as the major form, whereas
in nonfermented soy products, for example, soy milk or soy supplements, the glucoside form pre-
dominates [1].
56.3.1 Isoflavones
The most abundant isoflavones in soybean are malonyl-, acetyl-, and β-glucoside conjugates of genis-
tein and daidzein as shown in Figure 56.1 [5]. The isoflavone contents of the raw yellow soybean, raw
black soybean, yellow soybean milk, and black soybean milk are listed in Table 56.2 [16,17]. The
total isoflavones in the raw yellow soybean and black soybean are 1700–2200 and 1000–1200 μg/g
on dry weight (DW), respectively. The major isoflavones in raw yellow soybean exist as gluco-
sides. The highest content is 6-O-malonyl-β-glucosides, followed by 7-O-β-glucosides, whereas
6-O-acetyl-β-glucosides and aglycones occur in only very small amounts. Malonylglucosides in soy
milk are transformed into 7-O-β-glucosides and acetylglucosides after traditional thermal process-
ing [16,17]. Thermal processing methods cause significant decreases in aglycone level. This is con-
sistent with the findings of Kao et al. [18] that aglycones (daidzein and glycitein) decrease rapidly
during the early stage of heating. Traditional thermal processing significantly affects the content,
retention, and distribution of phenolic compounds and antioxidant activity of soy milk.
Skeleton Compounds R1 R2
OH O
Daidzein H H
R2 Glycitein H OCH3
R1 O Genistein OH H
OH
Skeleton Compounds R1 R2
OH
HO H
OH Daidzein
O O O 6”-O-acetyldaidzin H H
HO 6”-O-malonyldaidzin
R2
R1 O
OH
OH
HO H
OH
Glycitein
O O O 6”-O-acetylglycitin H OCH3
6”-O-malonylglycitin
O R2
O
R1 O
OH
OH
HO H
OH
Genistein
O O O
6”-O-acetylgenistin OH H
O R2 6”-O-malonylgenistin
O
R1 O
OH
O
HO
FIGURE 56.1 Chemical structure of soy isoflavones found in soy beverages.

TABLE 56.2
Isoflavones in Raw Soybeans and Soybean Milks (μg/g Dry Weight)
Yellow Soybean Yellow Soybean Black Soybean Black Soybean
Nutrient (Raw) Milk (Raw) Milk References
Total Isoflavones 1673 1782 2564 1144 [16]
7-O-β-Glucoside
Daidzin 121 357 92.6 134 [17]
Genistin 189 626 152 204 [17]
Glycitin 68.1 109 62.8 88.6 [17]
Malonylglucosides [17]
Malonyldaidzin 927 667 587 486 [17]
Malonylgenistin 1630 1187 921 769 [17]
Malonylglycitin 138 99.8 205 138 [17]
Acetylglucosides
Acetyldaidzin 4.69 27.0 7.10 2.01 [17]
Acetylgenistin 18.8 57.2 47.3 10.8 [17]
Acetylglycitin 34.3 45.0 24.9 16.5 [17]
Aglycones [17]
Daidzein 15.7 11.4 32.3 15.1 [17]
Genistein 36.7 22.5 65.4 31.4 [17]
Glycitein nd nd nd nd [17]
56.3.2 Anthocyanins
Three major anthocyanins, namely, delphinidin‐3‐glucoside, cyanidin‐3‐glucoside, and petunidin‐3‐
glucoside, as well as minor anthocyanins such as pelargonidin‐3‐O‐glucoside, pelargonidin‐3‐O‐
(6″‐malonylglucoside), and cyanidin, have been characterized in black soybean. The contents of
delphinidin‐3‐glucoside, cyanidin‐3‐glucoside, petunidin‐3‐glucoside, and total anthocyanins in black
soybeans are in the ranges of 0.0–3.71, 0.94–15.98, 0.0–1.41, and 1.58–20.18 mg/g, respectively [19].
Anthocyanins, such as cyanindin-3-glucoside and peonidin-3-glucoside, cannot be detected in processed
black soy milk.
56.3.3 Phenolic Acids

Two phenolic acids of the benzoic acid type (gallic acid and p-hydroxybenzoic acid) and three phenolic
acids of the cinnamic acid type (chlorogenic acid, p-coumaric acid, and trans-cinnamic acid) have been
detected in both raw and processed soy milk. Thermal processing causes significant increases in free
gallic, 2,3,4-trihydroxybenzoic, and p-hydroxybenzoic acids as well as protocatechualdehyde but causes
significant decreases in protocatechuic, caffeic, chlorogenic, and p-coumaric acids [16]. There are also
no significant differences between different processing methods in terms of (+)-catechin content in pro-
cessed soy milk. About 50–60 μg/g (+)-catechin has been detected in the raw and thermally processed
soy milk. These data indicate that flavan-3-ol is stable upon soy milk processing.
56.3.4 Antioxidants and Bioactives Responsible for Antioxidant Activity of

Soybean Milk
Isoflavones act as powerful antioxidant and anti-inflammatory agents. In addition to the estrogenic
effects, isoflavones and their oxidative metabolites are reported to have antioxidant and free radical
scavenging properties at physiological concentrations [20].
Interest in the possible health benefits of flavonoids has increased owing to their potent antioxidant
and free radical scavenging activities observed in vitro. Antioxidant activities of the raw and processed
soy milk, using 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferric-reducing antioxidant power (FRAP), and
oxygen radical absorbance capacity (ORAC) parameters, have been compared. Traditional processes
cause significant increases in DPPH and FRAP, whereas they significantly decrease ORAC values.
Traditional cooking methods (100°C for 20 min) cause significant decreases in ORAC in the
black soybean milk. Thermal treatment transforms malonylglucosides into 7-O-β-glucosides and
acetylglucosides and results in increased overall antioxidant activities (DPPH and FRAP) of pro-
cessed black soy milk products. These results indicate that daidzin, glycitin, and genistin may play
a more dominant role in the overall antioxidant activities of soy milk than that of phenolic acids and
isoflavones. The differences may be attributable, in part, to the differences of chemical composition
between black and yellow soybeans. The anthocyanins in black soybeans could easily be degraded
upon thermal processing [21].
Lactic acid bacteria (LAB), isolated from humans, have been tested for their ability to convert isofla-
vone glucosides to aglycones in soy milk [22]. Soy milk fermented with kefir, LAB, and bifidobacteria
improves the antioxidant properties dramatically [23]. Isoflavones act as powerful antioxidant and anti‐
inflammatory agents. Aglycone isoflavones have been reported as dietary modulators of cardiovascular
function by the regulation of vascular tone, oxidative stress, and antioxidant gene transcription [24] as
well as inflammatory processes [25].
The content of various isoflavones (aglycones, glucoside, and acetyl‐ and malonylglucosides) is signifi-
cantly decreased in fermented soymilk products. Regardless of starter organism employed, fermentation
causes a major reduction in the contents of various isoflavones along with a significant increase of aglycone
isoflavones contents. The level of change in the content of various isoflavones and β‐glucosidase activity
after fermentation varies with the starter organism.
56.4 Health Effects

Most of the flavonoids present in plants exist as the glycosides type rather than as aglycones. Human
feeding studies indicate that the absorption and bioavailability of specific flavonoids is much higher
than originally believed. However, epidemiologic studies exploring the role of flavonoids in human
health have been inconclusive. Pharmacokinetic studies show that bioavailability of pure β-glycosides
daidzin and genistin is significantly greater than that of the corresponding aglycones. Similar observa-
tions are well established for flavonoids, especially quercetin [26].
56.4.1 Bioavailability of Bioactives from Soybean Milk

Several human studies have been conducted to investigate the bioavailability of isoflavones in the aglycone
and glucoside forms; however, contradictory results were obtained. Some studies found that the isoflavone
aglycones of soymilk were absorbed faster and in greater amounts than their glucosides in healthy adults
[27], whereas Setchell et al. [28] reported a more efficient use of glucosides. Other studies showed that
absorption of aglycones and glucosides did not differ significantly [29]. The metabolism of isoflavones
might be affected by the type of soymilk consumed. This is possibly due to sample variation from pure
isoflavone extracts in a pharmaceutical formulation; others used soy food items such as soy milk [30].
Comprehensive overviews about the pharmacokinetics, including absorption mechanisms, bioavailability,
metabolism, and elimination, of the soy isoflavone daidzein and its bacterial and oxidative metabolites
after consumption of the pure, nonformulated aglycone and glucoside are available [31].
56.4.2 Animal Studies with Soybean Milk

A wide range of biological properties for genistein and daidzein has been reported [32]. Studies with
animal models provide valuable understanding of the pharmacokinetic data, especially information about
the absorption, metabolism, and distribution patterns of soybean isoflavones [33]. Biological activity of
phytoestrogens depend on multiple factors, for example, the chemical form of the phytoestrogen, the route
of administration, the metabolism, the intrinsic estrogenic status, and the time and the level of exposure.
TABLE 56.3
List of Animal Model Studies with Soybean Beverages and Their Key Results
Key Results References
Combination of Bifidobacteria and oligofructose reduces colon cancer risk in carcinogen-treated rats. [37]
Feeding of soy milk fermented with the Bifidobacterium breve reduced the sizes of mammary gland [38]
tumors in female Sprague–Dawley rats.
Soya yogurt containing Bifidobacteria increases feces bifidobacterial count and prolongs the lifespan [39]
of female Swiss albino mice.
The habitual consumption of soy isoflavones in combination with the probiotic Lactobacillus casei [40]
decreases the risk of breast cancer in Female Sprague–Dawley rats.
Fermented soy milk attenuates bone loss in ovariectomized mice and lower the risk of osteoporosis. [41]
Soymilk prevents bone loss in rats induced by ovarian hormone deficiency rats. [42]
Fermented soymilk modulates the cholesterol metabolism in rats fed a high cholesterol diet. [43]
Soy yogurt prevents hepatic lipid accumulation in Male Sprague–Dawley rats. [44]
On the market, there are several preparations containing varying concentrations of individual phytoestro-
gens of different origin, and their use as dietary supplements may result in high levels of phytoestrogens,
relative to those provided in whole foods. In addition, the concept that “natural is safe” may not prove
valid [34].
In vivo beneficial effects of phytoestrogens in animal models of postmenopausal estrogen deficiency
have been reported. Bone-conserving effects in animal models of the synthetic isoflavone was demon-
strated, which was later approved for clinical use for the prevention of osteoporosis [35]. Soybean food
has attracted much attention because of its potential health benefits. Soy milk contains bioactive com-
pounds that may have beneficial roles in cardiovascular health promotion. Its protein supplemented to a
high‐fat diet lowered concentrations of plasma lipids, total cholesterol, triacylglycerols (TAG), and free
fatty acids in C 57BL/6N mice [36]. Table 56.3 lists the animal model studies with soybean beverages
and their key results.
56.4.3 Human Intervention Studies with Soybean Milk

Significantly lower incidence of osteoporosis‐related fracture in Southern and Eastern Asian women
than that in Western women has been reported [45]. Epidemiologic evidence of reduced rates of hip
fractures in Asians consuming soy protein despite the lower calcium intakes has been reported [46].
Setchell [2] reported a substantial difference between the intake of soy food items in the Japanese,
Chinese, and Indonesian populations compared with the U.S. and European populations. Asians
consume traditional fermented soy products, have aglycones as the major form of isoflavones, and may
be responsible for the high potential for the bone loss prevention [1], menopausal symptoms, [47], and
osteoporosis [2,48,49].
Isoflavones are nonsteroidal phytoestrogenic and antioxidative polyphenolic molecules and have the
potential to protect against hormone‐dependent diseases. Barnes [50] reported lower incidence of hor-
monally dependent breast cancer in Asian populations compared to Americans and Europeans. Other
epidemiological and clinic investigations on health benefits of isoflavone-rich diets showed lower inci-
dences of colonic, prostate, and breast cancers in Asian populations as listed in Table 56.4. Isoflavones
are natural plant phytoestrogens with structural similarities to 17‐β‐estradiol and are able to bind to
estrogen receptors (ERs) [56].
A high‐phytoestrogen diet may result in various health effects; however, some of the results obtained
are still controversial. High levels of phytoestrogen promote breast and endometrial cancer in women,
whereas the loss of estrogen during menopause is correlated with osteoporosis, CHD, depression, and
neurodegeneration. Potential adverse effects and toxicity from early estrogenic exposure in infants and
an elevated breast cancer risk in woman have raised [57,58]. Many of the aforementioned benefits are
derived from the interaction between isoflavones and ERs. Interaction of isoflavones with ERs leads to
the activation of the so‐called estrogen response elements.
TABLE 56.4
List of Human Intervention studies with Soybean Beverages and Their Key Results
Key Results References
A daily soy beverage decreases trend or more than two times prolongation of prostatic specific [48]
antigen doubling time in 41% of prostate cancer patients.
Consumption of soy foods reduces risk of ovarian cancer in southern Chinese women. [49]
Frequent consumption of soy milk causes 70% reduction of the risk of prostate cancer in 225 [51]
incident cases of prostate cancer.
Powdered whole soy milk and its hydrolysate decreases plasma lipid concentrations and hepatic [52]
fatty acid synthase and glucose-6-phosphate dehydrogenase activities.
A declining trend or more than two times prolongation of PSA doubling time in 41% of subjects. [53]
Daily consumption of soymilk for 21 days decreases atherogenic plasma cholesterol [54]
concentration in 42 apparently healthy young to middle-aged subjects.
Low and moderate-fat plant sterol-enriched soy beverages reduce circulating lipid concentrations [55]
in normal to hypercholesterolemic individuals.

Novel products derived from soybean milk have been developed. Varieties of LAB isolated from
human’s fecal have been applied to improve the functionalities and bioavailability of soybean milk.
Lactobacillus paraplantarum, L. acidophilus, L. paracasei, Weissella confusa, Enterococcus durans,
Streptococcus salivarius, S. thermophilus, Bifidobacterium longum, and B. infantis have been used in
single or mixed cultures for soymilk fermentation. Other isolates have been found to possess the ability
to produce peptides and free isoflavones in soy milk. The antihypertensive, antioxidant, and anti-
inflammatory properties of the resulting fermented soy milks were observed [59].
Several studies have reported that isoflavone glucosides in soybean milk are converted to aglycones
during the fermentation process [22,60]. The content of various isoflavones (aglycones, glucoside, and
acetyl‐ and malonylglucosides) significantly decreased in fermented soymilks. Regardless of the starter
organism employed, fermentation causes a major reduction in the contents of various isoflavones along
with a significant increase of aglycone isoflavones content. The level of change in the content of various
isoflavones and β‐glucosidase activity after fermentation varies with the starter organism.
The rates of hydrolysis vary depending on the species of LAB. It corresponds well with a sharp
increase in β‐glucosidase activity during fermentation. Among all the starters tested, S. thermophilus
showed high β‐glucosidase activity, whereas, L. paraplantarum KM seems to be a promising starter
for conversion of soymilk glucosides to corresponding aglycones [61]. It has been shown that daid-
zein is converted by the gut microflora to dihydrodaidzein, which can be further metabolized to both
S‐equol and O‐desmethylangolensin. The level of various aglycone isoflavones plays an important
role in the absorption, bioavailability, antioxidative activity, and some functionalities of soybean
beverages [62].
The LAB proteolytic system, including endopeptidases, aminopeptidases, tripeptidases, and dipep-
tidases, is well characterized [63]. Several studies [64] have shown that starter cultures including
E. faecium, S. thermophilus, L. helveticus, and L. plantarum could be applied to produce fermented
soy milks with angiotensin-I-converting enzyme (ACE) inhibitory activity. ACE inhibitory peptides
are released from precursor inactive soybean proteins by the action of microbial proteases during
the fermentation of soy milk. Among them, L. helveticus is commercialized for the production of
fermented soybean products with potent antihypertensive functionalities. The amino acid sequences
of these inhibitors were determined as Ile-Phe-Leu (IC50, 44.8 µM), Trp-Leu (IC50, 29.9 µM), and
Val-Leu-Ile-Val-Pro (IC50, 1.69 µM); the sequences were found in the β-conglycinin α- and β-subunits,
glycinin B-, B1A-, and BX-subunits, and glycinin subunit G2, respectively, from soybean [65]. It is
possible that supplementation with inulin may influence plasma concentrations of soybean isoflavones,
possibly by increasing their absorption.
Inulin, fructo-oligosaccharides, and other oligosaccharides are included as fiber in food labels in the
United States. Among them, oligosaccharides are well characterized as prebiotics that change both
composition and/or activity of the gastrointestinal microflora that confers benefits upon the host’s well-
bring and health. Studies have provided evidence that inulin, oligofructose, lactulose, and resistant starch
may stimulate the growth of Bifidobacterium in the colon [66]. Lee et al. [67] reported that both lactic and
acetic acids were detected from the Bifidobacterium species, whereas, only lactic acid was found in LAB
strains. Zhao and Cheung [59] found that B. infantis produced significant amounts of total short‐chain
fatty acids that had a relatively higher proportion of propionic and butyric acids, but less acetic acid.
Bang et al. [68] demonstrated that intake of soy oligosaccharide (3 g/day) increases fecal Bifidobacteria
counts, short‐chain fatty acid concentrations, and fecal lipid output in young Korean women.
56.6 Conclusion
Soybean is regarded as a valuable crop and is the most commonly consumed legume all over the world,
especially in Asia. It is used in various food types, among which fermented soybean beverage deserves
special attention. Soybean beverage provides health benefits such as modulating cholesterol and lipid
metabolism, reducing CVD, osteoporosis, and reducing cancer risk. Soybean milk contains active com-
ponents such as isoflavones, anthocyanins, and oligosaccharides that promote health. Fermented soymilk
is a good source of bioactive anti-ACE peptides, antioxidative aglycone isoflavones, anticancer phytoes-
trogens, and immunomodulatory peptides. The chemistry, absorption, metabolism, and mechanisms of
action of plant isoflavones in soy beverage were reviewed. These various properties may explain the much
lower incidence of hormonally dependent diseases in Oriental populations compared to Americans and
Europeans.
REFERENCES
1. Coward, L., Barnes, N.C., Setchell, K.D.R., and Barnes, S., Genistein, daidzein, and their beta-glycoside
conjugates: Antitumor isoflavones in soybean foods from American and Asian diets. J. Agric. Food
Chem., 41, 1961–1967, 1993.
2. Setchell, K.D., Phytoestrogens: The biochemistry, physiology, and implications for human health of soy
isoflavones. Am. J. Clin. Nutr., 68(Suppl.), 1333S–1346S, 1998.
3. van der Ven, C., Matser, A.M., and van den Berg, R.W., Inactivation of soybean trypsin inhibitors and
lipoxygenase by high-pressure processing. J. Agric. Food Chem., 53, 1087–1092, 2005.
4. Yang, H.W., Hsu, C.K., and Yang, Y.F., Effect of thermal treatments on anti-nutritional factors and
antioxidant capabilities in yellow soybeans and green-cotyledon small black soybeans. J. Sci. Food
Agric., 94, 1794–1801, 2014.
6. Messina, M.J., Legumes and soybeans: Overview of their nutritional profiles and health effects. Am.
J. Clin. Nutr., 70(Suppl.), 439S–450S, 1999.
7. Obendorf, R.L. and Kosina, S.M., Soluble carbohydrates in soybean, in Soybean-Biochemistry,
Chemistry and Physiology, Ng, T.B., Ed., InTech, Rijeka, Croatia, 2011, pp. 201–228.
8. Yagasaki, K., Takagi, T., Sakai, M., and Kitamura, K., Biochemical characterization of soybean protein
consisting of different subunits of glycinin. J. Agric. Food Chem., 45, 656–660, 1997.
9. Food and Agriculture Organization (FAO), Protein quality evaluation: Report of the joint FAO/WHO
expert consultation, FAO Food and Nutrition Paper 51, FAO, Bethesda, MD, 1989.
10. Food and Drug Administration (FDA), Food labeling: Health claims; soy protein and coronary heart
disease. Food and Drug Administration, HHS. Final rule. Fed. Regist., 64, 57700–57733, 1999.
11. Anderson, J.W., Johnstone, B.M., and Cook-Newell, M.E., Meta-analysis of the effects of soy protein
intake on serum lipids. N. Engl. J. Med., 333, 276–282, 1995.
12. BeMiller, J.N. and Huber, K.C., Carbohydrates, in Fennema’s Food Chemistry, 4th edn, Damodarin, S.,
Parkin, K.L., and Fennema, O.R., Eds., CRC Press, Boca Raton, FL, 2007, pp. 83–154.
13. Zou, Y. and Chang, S.K., Effect of black soybean extract on the suppression of the proliferation
of human AGS gastric cancer cells via the induction of apoptosis. J. Agric. Food Chem., 59, 4597–
4605, 2011.
14. Chen, Y., Xu, Z., Zhang, C., Kong, X., and Hua, Y., Heat-induced inactivation mechanisms of Kunitz
trypsin inhibitor and Bowman–Birk inhibitor in soymilk processing. Food Chem., 154, 108–116, 2014.
15. Kwok, K.C., Liang, H.H., and Niranjan, K., Optimizing conditions for thermal processes of soy milk.
J. Agric. Food Chem., 50, 4834–4838, 2002.
16. Kim, E.H., Ro, H.M., Kim, S.L., Kim, H.S., and Chung, I.M., Analysis of isoflavone, phenolic, soyasa-
pogenol, and tocopherol compounds in soybean [Glycine max (L.) Merrill] germplasms of different seed
weights and origins. J. Agric. Food Chem., 60, 6045–6055, 2012.
17. Xu, B. and Chang, S.K., Isoflavones, flavan-3-ols, phenolic acids, total phenolic profiles, and antioxi-
dant capacities of soy milk as affected by ultrahigh-temperature and traditional processing methods.
J. Agric. Food Chem., 57, 4706–4717, 2009.
18. Kao, T.H., Lu, Y.F., Hsieh, H.C., and Chen, B.H., Stability of isoflavone glucosides during processing of
soymilk and tofu. Food Res. Int., 37, 891–900, 2004.
19. Koh, K., Youn, J.E., and Kim, H.S., Identification of anthocyanins in black soybean [Glycine max (L.)
Merr.] varieties. J. Food Sci. Technol., 51, 377–381, 2014.
20. Ng, L.T., Ko, H.H., and Lu, T.M., Potential antioxidants and tyrosinase inhibitors from synthetic
polyphenolic deoxybenzoins. Bioorg. Med. Chem., 17, 4360–4366, 2009.
21. Xu, B. and Chang, S.K., Total phenolics, phenolic acids, isoflavones, and anthocyanins and antioxidant
properties of yellow and black soybeans as affected by thermal processing. J. Agric. Food Chem., 56,
7165–7175, 2008.
22. Chun, J., Jeong, W.J., Kim, J.S., Lim, J., Park, C.S., Kwon, D.Y., Choi, I., and Kim, J.H., Hydrolysis
of isoflavone glucosides in soymilk fermented with single or mixed cultures of Lactobacillus para-
plantarum KM, Weissella sp. 33, and Enterococcus faecium 35 isolated from humans. J. Microbiol.
Biotechnol., 18, 573–578, 2008.
23. Wang, Y.C., Yu, R.C., and Chou, C.C., Antioxidative activities of soymilk fermented with lactic acid
bacteria and Bifidobacteria. Food Microbiol., 23, 128–135, 2006.
24. Siow, R.C., Li, F.Y., Rowlands, D.J., de Winter, P., and Mann, G.E., Cardiovascular targets for estrogens
and phytoestrogens: Transcriptional regulation of nitric oxide synthase and antioxidant defense genes.
Free Radic. Biol. Med., 42, 909–925, 2007.
25. Pan, M.H., Lai, C.S., Dushenkov, S., and Ho, C.-T., Modulation of inflammatory genes by natural dietary
bioactive compounds. J. Agric. Food Chem., 57, 4467–4477, 2009.
26. Hollman, P.C., vd Gaag, M., Mengelers, M.J., van Trijp, J.M., de Vries, J.H., and Katan, M.B., Absorption
and disposition kinetics of the dietary antioxidant quercetin in man. Free Radic. Biol. Med., 21, 703–
707, 1996.
27. Kano, M., Takayanagi, T., Harada, K., Sawada, S., and Ishikawa, F., Bioavailability of isoflavones after
ingestion of soy beverages in healthy adults. J. Nutr., 136, 2291–2296, 2006.
28. Setchell, K.D., Brown, N.M., Desai, P., Zimmer-Nechemias, L., Wolfe, B.E., Brashear, W.T., Kirschner,
A.S., Cassidy, A., and Heubi, J.E., Bioavailability of pure isoflavones in healthy humans and analysis of
commercial soy isoflavone supplements. J. Nutr., 131(Suppl.), 1362S–1375S, 2001.
29. Zubik, L. and Meydani, M., Bioavailability of soybean isoflavones from aglycone and glucoside forms
in American women. Am. J. Clin. Nutr., 77, 1459–1465, 2003.
30. Nielsen, I.L. and Williamson, G., Review of the factors affecting bioavailability of soy isoflavones in
humans. Nutr. Cancer, 57, 1–10, 2007.
31. Ross, J.A. and Kasum, C.M., Dietary flavonoids: Bioavailability, metabolic effects, and safety. Annu.
Rev. Nutr., 22, 19–34, 2002.
32. Barnes, S., Evolution of the health benefits of soy isoflavones. Proc. Soc. Exp. Biol. Med., 217,
386–392, 1998.
33. Ronis, M.J., Little, J.M., Barone, G.W., Chen, G., Radominska-Pandya, A., and Badger, T.M., Sulfation
of the isoflavones genistein and daidzein in human and rat liver and gastrointestinal tract. J. Med. Food,
9, 348–355, 2006.
34. Moutsatsou, P., The spectrum of phytoestrogens in nature: Our knowledge is expanding. Hormones
(Athens, Greece), 6, 173–193, 2007.
35. Agnusdei, D., Gennari, C., and Bufalino, L., Prevention of early postmenopausal bone loss using low
doses of conjugated estrogens and the non-hormonal, bone-active drug ipriflavone. Osteoporos. Int., 5,
462–466, 1995.
36. Scheiber, M.D., Liu, J.H., Subbiah, M.T., Rebar, R.W., and Setchell, K.D., Dietary inclusion of whole soy
foods results in significant reductions in clinical risk factors for osteoporosis and cardiovascular disease
in normal postmenopausal women. Menopause, 8, 384–392, 2001.
37. Gallaher, D.D. and Khil, J., The effect of synbiotics on colon carcinogenesis in rats. J. Nutr., 129(Suppl.),
1483S–1487S, 1999.
38. Ohta, T., Nakatsugi, S., Watanabe, K., Kawamori, T., Ishikawa, F., Morotomi, M., Sugie, S., Toda,
T., Sugimura, T., and Wakabayashi, K., Inhibitory effects of Bifidobacterium-fermented soy milk
on 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine-induced rat mammary carcinogenesis, with a
partial contribution of its component isoflavones. Carcinogenesis, 21, 937–941, 2000.
39. Abd el-Gawad, I.A., el-Sayed, E.M., Hafez, S.A., el-Zeini, H.M., and Saleh, F.A., Inhibitory effect of
yoghurt and soya yoghurt containing Bifidobacteria on the proliferation of Ehrlich ascites tumour cells
in vitro and in vivo in a mouse tumour model. Br. J. Nutr., 92, 81–86, 2004.
40. Kaga, C., Takagi, A., Kano, M., Kado, S., Kato, I., Sakai, M., Miyazaki, K. et al., Lactobacillus casei
Shirota enhances the preventive efficacy of soymilk in chemically induced breast cancer. Cancer Sci.,
104, 1508–1514, 2013.
41. Chiang, S.S. and Pan, T.M., Antiosteoporotic effects of Lactobacillus-fermented soy skim milk on bone
mineral density and the microstructure of femoral bone in ovariectomized mice. J. Agric. Food Chem.,
59, 7734–7742, 2011.
42. Taguchi, H., Chen, H., Yano, R., and Shoumura, S., Comparative effects of milk and soymilk on bone
loss in adult ovariectomized osteoporosis rat. Okajimas Folia Anat. Jpn., 83, 53–59, 2006.
43. Kobayashi, M., Hirahata, R., Egusa, S., and Fukuda, M., Hypocholesterolemic effects of lactic acid-
fermented soymilk on rats fed a high cholesterol diet. Nutrients, 4, 1304–1316, 2012.
44. Kitawaki, R., Nishimura, Y., Takagi, N., Iwasaki, M., Tsuzuki, K., and Fukuda, M., Effects of
Lactobacillus fermented soymilk and soy yogurt on hepatic lipid accumulation in rats fed a cholesterol-
free diet. Biosci. Biotechnol. Biochem., 73, 1484–1488, 2009.
45. Tham, D.M., Gardner, C.D., and Haskell, W.L., Clinical review 97: Potential health benefits of dietary
phytoestrogens: A review of the clinical, epidemiological, and mechanistic evidence. J. Clin. Endocrinol.
Metab., 83, 2223–2235, 1998.
46. Lau, E.M. and Cooper, C., The epidemiology of osteoporosis. The oriental perspective in a world
context. Clin. Orthop. Relat. Res., 323, 65–74, 1996.
47. Cassidy, A., Albertazzi, P., Lise Nielsen, I., Hall, W., Williamson, G., Tetens, I., Atkins, S. et al., Critical
review of health effects of soyabean phyto-oestrogens in post-menopausal women. Proc. Nutr. Soc., 65,
76–92, 2006.
48. Setchell, K.D. and Lydeking-Olsen, E., Dietary phytoestrogens and their effect on bone: Evidence from
in vitro and in vivo, human observational, and dietary intervention studies. Am. J. Clin. Nutr., 78(Suppl.),
593S–609S, 2003.
49. Potter, S.M., Baum, J.A., Teng, H., Stillman, R.J., Shay, N.F., and Erdman, J.W., Jr., Soy protein and
isoflavones: Their effects on blood lipids and bone density in postmenopausal women. Am. J. Clin.
Nutr., 68(Suppl.), 1375S–1379S, 1998.
50. Barnes, S., The biochemistry, chemistry and physiology of the isoflavones in soybeans and their food
products. Lymphat. Res. Biol., 8, 89–98, 2010.
51. Jacobsen, B.K., Knutsen, S.F., and Fraser, G.E., Does high soy milk intake reduce prostate cancer
incidence? The Adventist Health Study (United States). Cancer Causes Control, 9, 553–557, 1998.
52. Choi, J.Y., Jeon, J.E., Jang, S.Y., Jeong, Y.J., Jeon, S.M., Park, H.J., and Choi, M.S., Differential effects
of powdered whole soy milk and its hydrolysate on antiobesity and antihyperlipidemic response to high-fat
treatment in C57BL/6N mice. J. Agric. Food Chem., 59, 2584–2591, 2011.
53. Lee, A.H., Su, D., Pasalich, M., Tang, L., Binns, C.W., and Qiu, L., Soy and isoflavone intake associated
with reduced risk of ovarian cancer in southern Chinese women. Nutr. Res., 34, 302–307, 2014.
54. Onuegbu, A.J., Olisekodiaka, J.M., Onibon, M.O., Adesiyan, A.A., and Igbeneghu, C.A., Con
sumption of soymilk lowers atherogenic lipid fraction in healthy individuals. J. Med. Food, 14,
257–260, 2011.
55. Rideout, T.C., Chan, Y.M., Harding, S.V., and Jones, P.J., Low and moderate-fat plant sterol fortified
soymilk in modulation of plasma lipids and cholesterol kinetics in subjects with normal to high cholesterol
concentrations: Report on two randomized crossover studies. Lipids Health Dis., 8, 45, 2009.
56. Pilsakova, L., Riecansky, I., and Jagla, F., The physiological actions of isoflavone phytoestrogens.
Physiol. Res., 59, 651–664, 2010.
57. Sheehan, D.M., Herbal medicines, phytoestrogens and toxicity: Risk:benefit considerations. Proc. Soc.
Exp. Biol. Med., 217, 379–385, 1998.
58. Bouker, K.B. and Hilakivi-Clarke, L., Genistein: Does it prevent or promote breast cancer? Environ.
Health Perspect., 108, 701–708, 2000.
59. Zhao, J. and Cheung, P.C., Fermentation of beta-glucans derived from different sources by Bifidobacteria:
Evaluation of their bifidogenic effect. J. Agric. Food Chem., 59, 5986–5992, 2011.
60. Chien, H.L., Huang, H.Y., and Chou, C.C., Transformation of isoflavone phytoestrogens during the fer-
mentation of soymilk with lactic acid bacteria and Bifidobacteria. Food Microbiol., 23, 772–778, 2006.
61. Chang, Y.C. and Nair, M.G., Metabolism of daidzein and genistein by intestinal bacteria. J. Nat. Prod.,
58, 1892–1896, 1995.
62. Xu, X., Harris, K.S., Wang, H.J., Murphy, P.A., and Hendrich, S., Bioavailability of soybean isoflavones
depends upon gut microflora in women. J. Nutr., 125, 2307–2315, 1995.
63. Christensen, J.E., Dudley, E.G., Pederson, J.A., and Steele, J.L., Peptidases and amino acid catabolism
in lactic acid bacteria. Antonie Leeuwenhoek, 76, 217–246, 1999.
64. Martinez-Villaluenga, C., Torino, M.I., Martin, V., Arroyo, R., Garcia-Mora, P., Estrella Pedrola, I.,
Vidal-Valverde, C., Rodriguez, J.M., and Frias, J., Multifunctional properties of soy milk fer
mented by Enterococcus faecium strains isolated from raw soy milk. J. Agric. Food Chem.,
60, 10235–10244, 2012.
65. Kuba, M., Tanaka, K., Tawata, S., Takeda, Y., and Yasuda, M., Angiotensin I-converting enzyme
inhibitory peptides isolated from tofuyo fermented soybean food. Biosci. Biotechnol. Biochem., 67,
1278–1283, 2003.
66. Slavin, J., Fiber and prebiotics: Mechanisms and health benefits. Nutrients, 5, 1417–1435, 2013.
67. Lee, K.Y., So, J.S., and Heo, T.R., Thin layer chromatographic determination of organic acids for rapid
identification of bifidobacteria at genus level. J. Microbiol. Methods, 45, 1–6, 2001.
68. Bang, M.H., Chio, O.S., and Kim, W.K., Soyoligosaccharide increases fecal Bifidobacteria counts,
short-chain fatty acids, and fecal lipid concentrations in young Korean women. J. Med. Food, 10,
366–370, 2007.
Section VI
Alcoholic Beverages and Water

57
Wine
Andrew L. Waterhouse, Rosa M. Lamuela-Raventós, Paola Quifer-Rada, and Creina S. Stockley
CONTENTS
57.1 Introduction................................................................................................................................... 739
57.3.1 Antioxidant Activity of Wine Polyphenols.......................................................................742
57.3.2 Anti-Inflammatory Activity of Wine Polyphenols............................................................745
57.3.3 Polyphenols and Platelet Function....................................................................................745
57.3.4 Mechanistic Effects of Polyphenols on Blood Pressure................................................... 746
57.4 Health Effects................................................................................................................................ 746
57.4.1 Epidemiological Evidence................................................................................................ 746
57.4.2 Clinical Intervention Trials...............................................................................................747
57.4.2.1 Wine Polyphenols and Oxidative Status............................................................747
57.4.2.2 Wine Polyphenols and Inflammation................................................................747
57.4.2.3 Wine Polyphenols and Blood Pressure..............................................................747
57.4.2.4 Wine Polyphenols and Lipid Profile................................................................. 748
57.4.2.5 Wine Polyphenols and Diabetes Mellitus......................................................... 748
57.4.2.6 Wine Prebiotic Effects...................................................................................... 748
57.4.2.7 Wine Polyphenols and Cancer...........................................................................749
57.4.2.8 Red Wine and Dementia....................................................................................749
57.5 Current Government Policies Affecting Wine Consumption........................................................749
57.7 Conclusion......................................................................................................................................751
References................................................................................................................................................751
57.1 Introduction
Wine is certainly the oldest functional beverage. Archaeological evidence suggests the production as
far back as 7000 years ago in the middle east and over 9000 years ago in China [1], and that evidence
is limited by the contemporaneous production of wine containers that can survive in the soil. It is pos-
sible that animal hide containers were used to hold wine in earlier times, but the containers have all
decomposed.
Wine is a very complex liquid and the potential functional ingredients are many. Ethanol is of course
a major component, being present at 12%–15% in most wines, and that alone imparts some physiological
effects that are beneficial. There is also some literature on the potential benefit of glycerol, also a major
constituent with levels approaching 1% in most wines. However, today’s research focuses on the polyphe-
nolic substances. Unfortunately, too much attention is paid to the simple antioxidant effects, as they are
easy to measure, but provide little more than a suggestion that something interesting might be present.
Luckily, some labs are delving into the specific mechanisms of action available to phenolics, and very
739
importantly, their metabolites, which are many. The short residency time of phenolics is a two-edged
sword—daily consumption is necessary to retain active circulating levels.
Wine holds much social significance, with important historical legacies regarding the use of wine. For
instance, wine is typically used to toast newlyweds, honor military victories, seal business deals, and
denote many other significant events. It also has religious connotations in Western countries. So, it is
clear that wine is not just one of many alternative beverages, but one with deep cultural importance. In
part for that reason, data on the consumption of wine are reliable because most people can recall their
consumption patterns. By comparing that consumption versus health outcomes, epidemiologists now
have good data on the health effects of wine.
Since alcohol is present at significant levels, wine falls under social, regulatory, and taxation scru-
tiny in virtually all jurisdictions. This alters the marketing and availability of wine compared to other
functional beverages. Despite sound science showing that moderate wine drinkers have lower rates of
many chronic diseases and lower total mortality, many government agencies continue to treat wine as
just another alcoholic beverage. This is because a minority of individuals drink alcohol abusively with
both negative health and social consequences that are perceived by the agencies as negating any positive
health benefits.
Abusive alcohol consumption should be discouraged, not all wine consumption. Government officials
might consider the insight of Thomas Jefferson: “I think it is a great error to consider a heavy tax on
wines as a tax on luxury. On the contrary, it is a tax on the health of our citizens’’. This chapter highlights
the documented health effects of wine consumption, studies that have shown possible mechanisms for
those effects, and the regulatory framework for producing and marketing wine.

Wine is a fermented beverage that has functional properties due to its complex composition, mainly for
its minor compounds. Considering the nutritional characteristics of red and white wines, both wines
are fat-free drinks, without cholesterol, that may contain low levels of carbohydrate, depending on
the winemaking process, according to National Nutrient Database for Standard Reference Release 27
(Table 57.1). Per serving (147 g) of red and white wines have around 3.8 g of carbohydrate.
One serving, containing 13% (v/v) of ethanol, provides ~121–125 kcal, coming mainly from the
alcohol. This caloric contribution would increase if a sweet wine is consumed, as it may contain
upto 19 g of total carbohydrate per drink (additional 76 kcal). Thus, to better inform the consumer,
alcohol calories should be included in our dietary calculations. However, epidemiological data and
short-term intervention clinical trials demonstrate that light-to-moderate wine intake seems to protect
against weight gain [2]. However, further research is needed to understand how this dietary pattern
affects weight gain. This is surprising because it is reported that calories coming from ethanol do
not provide satiety and as alcoholic beverages contain few other nutrients, they are also considered
“empty” calories. However, a better understanding of wine is that it is a functional food since it
contains many minor bioactive compounds, as it will be noted later in this chapter, and these have
interesting nutritional impacts. Water is the major component of wine, representing ~86%, followed
by ethanol (~13%) and then glycerol (~0.5%–1.0%), a secondary fermentation product that gives
density to the wine.
Wine is an acidic drink with a pH around 3.5. The main acid present in wine is tartaric acid, a very
good biomarker of wine intake [3,4], as it is almost exclusively naturally found in grapes and wines.
In addition to ethanol, acidity is key to the preservation of the wine; and probably for this reason, wine
is considered a digestive aid [5]. In addition, these substances and others impart to wine antimicrobial,
antioxidant, and prebiotic properties [6].
Red and white wines contain some essential nutrients such as minerals and vitamins, mostly niacin
(Table 57.1). However, the level of these minor nutrients in wines is not nutritionally relevant, as the per-
centage of these nutrients with regard to the daily value per serving is too low. Nutritionally speaking,
the most interesting compounds in wine are polyphenols, the physiological properties attributed to these
compounds will be explained in more detail in subsequent sections.
Wine 741
TABLE 57.1
Compositional and Nutritional Characteristics of Red and White Wines (per Serving, 147 g Wine)
Nutrient Unit Red Wine White Wine
Water g 127 128
Energy kcal 125 121
Protein g 0.10 0.10
Lipid (fat) g 0.00 0.00
Minerals
Calcium mg 12 13
Iron mg 0.68 0.40
Magnesium mg 18 15
Phosphorus mg 34 26
Sodium mg 6.0 7.0
Zinc mg 0.21 0.18
Vitamins
Folate (DFE) µg 1.0 1.0
Niacin mg 0.329 0.159

Bioactive compounds are mainly nonnutrient elements that are typically found in small quantities in
foods. These compounds vary widely in chemical structure and function, and they can exert beneficial
effects on health. Wine represents a great source of bioactive compounds, mostly based on phenolic com-
pounds (Figure 57.1). White wines contain between 100 and 300 mg of gallic acid equivalents (GAE)/L
of total phenolics and red between 1000 and 3000 mg GAE/L. Many phenolic compounds have antioxi-
dant properties, which may play an important role in reducing cardiovascular risk. Table 57.2 shows the
content of the most representative wine polyphenols in both red and white wines [79]. As can be seen, the
main polyphenols found in red wine are anthocyanins, flavanols, and phenolic acids (hydroxycinnamic
and hydroxybenzoic acids), whereas white wines contain mainly flavanols and phenolic acids (hydroxy-
cinnamic and hydroxybenzoic acids).
The level of phenolic compounds, in particular wines, depends on numerous factors, such as grape
variety and maturity, environmental factors in the vineyards (climate, soil, and sanitary stage), and the
winemaking technology, as well as fermenting and aging conditions [7].
Polyphenols are present in wine in both free and bound forms conjugated to one or more sugar mole-
cules, such as glucose, galactose, sucrose, rhamnose, xylose, or arabinose. One exception is the hydroxy-
cinnamic acids that are only found bound to tartaric acid in esters such as caftaric (caffeoyltartaric),
p-coutaric (coumaroyltartaric), and fertaric (feruoyltartaric) esters. Polyphenols can also be found in
a polymerized form, including dimers to hexamers and some molecules with even higher degree of
polymerization such as the procyanidins and viniferines. The best known of the wine polyphenols are
the stilbenes, including resveratrol and piceid (resveratrol glucoside derivative).
O CO2H
OH HO
O H
CO2H
HO OH
OH HO H
Caftaric acid Resveratrol
OH OH
OH OH
HO O HO O
OH OH
OH OH O
Epicatechin Quercetin
CH3
O
OH
+
HO O O CH3
3
5
O
OH Glucose
Malvidin-3-glucoside
FIGURE 57.1 Chemical structures of common polyphenolics found in wine.
57.3.1 Antioxidant Activity of Wine Polyphenols

Phenolics compounds have been considered powerful antioxidants in vitro [8,9]. Their antioxidant
activity is based in their ability to donate a hydrogen or electron and their ability to delocalize the
unpaired electron within the aromatic structure. It has been demonstrated that wine polyphenols
exert great antioxidant activity in vitro as measured by 2,2′-azino-bis(3-ethylbenzothiazoline-6-
sulfonic acid) (ABTS), oxygen radical absorbance capacity (ORAC), and 2,2-diphenyl-1-picryl-
hydrazyl (DPPH) methods [10]. However, the effect of phenolics compounds in the antioxidant
activity in vitro cannot be simply extrapolated onto the redox balance in vivo due to biological
processes that modify their antioxidant capacity. This includes metabolism into smaller molecules,
methylation, or conjugation to glucuronides and sulfates metabolites. Plasma antioxidant capacity
(PAC) includes the activity of antioxidant compounds from the diet, but more importantly, endog-
enous compounds with key antioxidant roles, such as uric acid, albumin, bilirubin, and glutathione.
PAC has been proposed as a biomarker of antioxidant status [11] and has been used to evaluate real
antioxidant effects in plasma of polyphenols in intervention clinical studies including wine [12–15].
In a short-term clinical intervention study with red wine in humans, it was shown that PAC increased
with the wine dose (100, 200, and 300 mL), by 6.2%, 19.5%, and 28.9%, respectively [13]. In
another study, volunteers consumed 300 mL of red wine and the PAC was measured at different time
points (15, 30, 60, 120, and 240 min). PAC increased 16.9% using the ORAC or 5.5% using ferric-
reducing ability of plasma (FRAP) [15].
Although, red wine increased PAC after an acute ingestion, long-term consumption did not show any
effects [12,14]. In a clinical intervention study, healthy volunteers consumed red wine and dealcoholized
red wine daily for 6 weeks. Antioxidant activity was measured in serum and plasma at baseline and after
6 weeks. It was concluded that PAC was not affected by a regular consumption of red wine or dealcohol-
ized red wine [14].
Wine 743
TABLE 57.2
Polyphenols in Red and White Wines
Polyphenols Red Wine (mg/100 mL) White Wine (mg/100 mL)
Flavonoids
Flavanols
(+)-Catechin 1.4–39 0–4.6
(−)-Epicatechin 0–16.5 0–6
(+)-Gallocatechin 0.42–0.12 0–0.02
(−)-Epigallocatechin 0–0.3 —
(−)-Epicatechin-3-O-gallate 0–0.9 0–0.1
Procyanidin dimer B1 2.15–14 0–0.05
Procyanidin dimer B2 0.4–9 0–0.03
Procyanidin dimer B3 0–12 0–0.02
Procyanidin dimer B4 0.08–11.3 0–0.05
Procyanidin dimer B7 0.3 —
Prodelphinidin dimer B3 0.1 —
Procyanidin trimer C1 0.2–2.6 —
Procyanidin trimer T2 6.7 —
Flavanones
Naringenin 0.04–0.07 —
Hesperetin 0.05–0.06 —
Naringin 0.7–0.8 0.2–0.3
Flavonols
Kaempferol 0–0.3 0–0.3
Quercetin 0–3.1 0–0.8
Quercetin-3-O-glucoside 0.8–2.3 —
Quercetin 3-O-rhamnoside 0–1.8 —
Quercetin 3-O-rutinoside 0–3.2 0–0.9
Quercetin 3-O-arabinoside 0.4–0.5 0.1–0.3
Myricetin 0–1.8 —
Isorhamnetin 0.006–0.6 —
Kaempferol 3-O-glucoside 0.5–1 —
Isorhamnetin 3-O-glucoside 0.1–0.5 —
3,7-Dimethylquercetin — 0–0.02
Dihydroflavonols
Dihydroquercetin 3-O-rhamnoside 0.1–1.5 0.07–1.3
Dihydromyricetin 3-O-rhamnoside 4.5 0.3
Anthocyanins
Malvidin 3-O-(6″-p-coumaroyl-glucoside) 0.6–4.5 —
Cyanidin 3-O-glucoside 0.009–0.9 —
Peonidin 3-O-glucoside 0.15–6 —
Delphinidin 3-O-glucoside 0.2–2.5 —
Petunidin 3-O-glucoside 0.3–3.4 —
Malvidin 3-O-glucoside 0–38.2 —
Delphinidin 3-O-(6″-acetyl-glucoside) 0.06–1.2 —
Cyanidin 3-O-(6″-acetyl-glucoside) 0.05–0.3 —
Petunidin 3-O-(6″-acetyl-glucoside) 0.07–1.6 —
Malvidin 3-O-(6″-acetyl-glucoside) 0.5–11.3 —
Peonidin 3-O-(6″-acetyl-glucoside) 0.08–1.1 —
Peonidin 3-O-(6″-p-coumaroyl-glucoside) 0.02–1 —
Petunidin 3-O-(6″-p-coumaroyl-glucoside) 0.01–1.1 —
(Continued)

Polyphenols in Red and White Wines
Polyphenols Red Wine (mg/100 mL) White Wine (mg/100 mL)
Vitisin A 0.15–1 —
Delphinidin 3-O-(6″-p-coumaroyl-glucoside) 0.01–0.3 —
Pinotin A 0.01–1.8 —
Malvidin 3-O-(6″-caffeoyl-glucoside) 0.2 —
Phenolic Acids
Hydroxybenzoic acids
Protocatechuic acid 0–1 0.02–1.3
Gallic acid 0–12.6 0–1.1
Vanillic acid 0–0.75 0.02–0.1
Gentisic acid 0–0.8 0–2
4-Hydroxybenzoic acid 0–2.2 0–0.04
Syringic acid 0–2.3 0–0.02
2-Hydroxybenzoic acid 0–0.09 0.02–0.1
2,3-Dihydroxybenzoic acid 0–0.6 —
Gallic acid ethyl ester 1.4–1.7 —
Hydroxycinnamic acids
p-Coumaric acid 0–4 0–0.5
Caffeic aid 0–7.7 0–0.7
Ferulic acid 0–1 0.03–0.2
Caffeoyl tartaric acid 0.1–18 2.1–2.2
o-Coumaric acid 0.02–0.04 0–0.07
Sinapic acid 0–0.5 0–0.3
p-Coumaroyl tartaric acid 0.2–1.8 —
2,5-di-S-Glutathionyl caftaric acid 1.1–4.7 —
5-Caffeoylquinic acid — 0–0.3
Hydroxyphenylacetic acids
4-Hydroxyphenylacetic acid 0.1–0.2 0.07–0.1
Stilbenes
Stilbenes
trans-Resveratrol 0–1 0–0.08
trans-Resveratrol 3-O-glucoside 0–2.9 0.0005–0.4
Piceatannol 0–2.6 —
cis-Resveratrol 0–2.3 0–0.07
ε-Viniferin 0.01–0.4 0–0.02
δ-Viniferin 0–2.2 —
cis-Resveratrol 3-O-glucoside 0–1.5 0.002–0.2
Pallidol 0–0.2 0–0.03
Piceatannol 3-O-glucoside 0.6–1.3 0.1–0.8
Resveratrol 3-O-glucoside 0–4.4 0.002–0.6
Other Polyphenols
Hydroxybenzaldehydes
Syringaldehyde 0–4.4 —
Protocatechuic aldehyde 0–0.1 0.1
Tyrosols
Tyrosol 0.6–4.5 0.1–0.3
Hydroxytyrosol 0.05–1 0.1–0.3
Hydroxycoumarins
4-Hydroxycoumarin — 0.1–0.5
Source: Adapted from Phenol-Explorer, Phenol-Explorer 3.5: Database in Poylphenol Content in Foods, Published
online at: http://phenol-explorer.eu/ (acccessed January 14, 2015).
Wine 745
Therefore, although polyphenols have great antioxidant activity in vitro and they might be able to
modulate the redox balance in vivo after an acute ingestion. The aforementioned studies have shown that
the antioxidant activity of polyphenols and their metabolites is not the most important feature of their
recognized biological activity.
57.3.2 Anti-Inflammatory Activity of Wine Polyphenols

Polyphenols can modulate protein and enzyme expression by regulating the activity of nuclear factors,
inhibit cell signaling, and modulate activation of key proteins in cascade reactions [16–20]. For example,
in recent years, the role of dietary procyanidins as health-protective agents has become an important
area of human nutrition research, since procyanidins may modulate the anti-inflammatory response
by molecular mechanisms including the modulation of the arachidonic acid pathway by the inhibition
of cyclooxygenase (COX) and lysyl oxidase (LOX) enzymes, inhibition of gene transcription, protein
expression, and enzymatic activity of eicosanoid-generating enzymes, inhibition of the production and
secretion of inflammatory mediators (cytokines and nitric oxide), inhibition of mitogen-activated protein
kinase pathway activation, and modulation of the expression of the nuclear factor-kappa β (NF-κβ) [20].
Moreover, some studies suggest that the presence of galloyl moieties in the chemical structure may be
important for the inhibitory activity of COX and LOX enzymes [18,19].
Protocatechuic acid, the main metabolite of anthocyanidins and flavan-3-ols, has been proposed as
an antiatherogenic due to its vascular anti-inflammatory activity. It has been shown to inhibit monocyte
adhesion to tumor necrosis factor-α (TNF-α) in endothelial cells, and to reduce vascular cell adhe-
sion molecule-1 (VCAM-1), intracellular adhesion molecule-1 (ICAM-1) expression, and NF-κβ binding
activity [16].
Furthermore, quercetin metabolites have also shown anti-inflammatory activity. 3′-O-methyl-quercetin
and 4′-O-methyl-quercetin inhibited ICAM-1 expression at physiological concentrations in human
aortic endothelial cells. In contrast, other quercetin metabolites such as quercetin-3′-O-sulfate and
quercetin-3-O-glucuronide did not inhibit adhesion molecule expression [17]. However, in another study,
these metabolites showed to be able to inhibit VCAM-1 cell surface expression [21]. Therefore, quercetin
metabolites may inhibit the expression of key molecules involved in monocyte aggregation during the
early stages of atherosclerosis.
Although wine contains several polyphenols with biologic activity, such as resveratrol, at present, the
strongest evidence for the efficacy against chronic disease are reported in flavan-3-ol and flavan-3-ol-rich
food such as cocoa and wine [22].
57.3.3 Polyphenols and Platelet Function

Platelets play an important role in the development of atherosclerosis and are often found in arterial lesion
sites. They are associated with reactive oxygen species cell producers such as neutrophils, monocytes,
macrophages, and endotelial cells. Platelet adhesion, activation, and aggregation represent the first stage
of arterial thrombus formation. Adherent platelets are activated by several mediators, including collagen,
thromboxane, and thrombin. Once platelets are activated, they secrete chemotaxins, clotting factors, and
vasoconstrictors which promote thrombin generation and more platelet accumulation. Activated platelet
also change shape, which help to promote further aggregation and coagulation.
Current evidences suggest that polyphenols could inhibit platelet activation and related signal trans-
duction pathways, neutralize free radicals, enhance nitric oxide production, and block thromboxane
receptors like TxA2 [23]. In an ex vivo study, catechin and epicatechin were shown to reduce platelet
activity by inhibiting adenosine diphosphate (ADP)-induced expression of GP IIb-IIIa surface gly-
coproteins [24]. Recently, Ostetag et al. [25] showed that flavan-3-ols reduced ADP-induced platelet
aggregation and P-selectin expression in men and decreased thrombin receptor‒activating peptide-
induced platelet aggregation in women. Therefore, polyphenols may downregulate thromboxane A 2
receptors reducing platelet aggregation, block ADP and GIIb-IIIa receptors avoiding platelet aggrega-
tion, block collagen receptors preventing collagen-induced platelet aggregation, and lower the expres-
sion of P-selectin [23].
57.3.4 Mechanistic Effects of Polyphenols on Blood Pressure

Polyphenols may protect the cardiovascular system by improving endothelial function. The endothe-
lium controls vascular tone by releasing several vasorelaxing factors, such as nitric oxide (NO) and
endothelium-derived hyperpolarizing factor (EDHF) [26,27].
New evidence reveals that polyphenols cause NO-mediated endothelium-dependent relaxations and
increase endothelial formation of NO. Wine, grape juice, and grape skin extracts induce relaxation in rat
aortic rings with the endothelium in a concentration-dependent manner [28]. The grape-derived products
increased the synthesis of endothelial NO, which relaxed the vascular smooth muscle via the cyclic gua-
nosine monophosphate (cGMP)-mediated pathway. These endothelium-dependent relaxations are corre-
lated with the polyphenol concentrations in red wine and have been further observed in various types of
animal blood vessels [29,30]. Moreover, other food products rich in polyphenols, such as cocoa, tea, and
chokeberry have also been shown to induce endothelium-dependent NO-mediated relaxations in arter-
ies. Another pathway that polyphenols may decrease blood pressure is by increasing the intracellular free
calcium concentration, which activates endothelial NO synthase (eNOS) [31].
In summary, polyphenols have been associated with several health effects. Most of their pleiotropic
effects are rendered via several mechanisms, including antioxidant activity and chelation of metal ions,
reduction in platelet activation and aggregation, inhibition of LOX, COX, phospholipase A2, enhance-
ment of prostaglandins PGI2 release and anti-inflammatory action, and interaction with biomembranes.
57.4 Health Effects

57.4.1 Epidemiological Evidence
Scientific evidence suggests that wine compounds responsible for the health effects, attributed to mod-
erate wine consumption, are ethanol and polyphenols. According to the World Health Organization
(WHO), 3.3 million people die every year due to the harmful use of alcohol that represents 5.9% of all
deaths. However, regular or moderate alcohol consumption by healthy individuals is associated with
lower cardiovascular disease (CVD) and all-cause mortality, with a reduced risk of vascular dementia or
any dementia, and a decrease in CVD risk factors [32]. Nevertheless, epidemiological evidence cannot
demonstrate whether polyphenols are responsible for the difference between wine and other alcoholic
distillated (gin, vodka, etc.) or fermented drinks such as beer and cider [33]. For this reason, in order to
evaluate the different properties of ethanol and polyphenols, randomized intervention clinical trials that
evaluate ethanolic drinks with and without polyphenols are necessary. To date, the studies performed
have mainly focused on intermediate markers [34] and studies on hard endpoints as final variables are
still lacking. This fact may be explained since the duration of these studies is limited by the personal
difficulties in accomplishing long interventions [35].
Considering the effect of ethanol by itself, consumption of one-to-two standard drinks per day showed
a lower rate of events than either subjects abstaining from alcohol or high alcohol consumers, a relation-
ship represented by a J-shaped (or U-shaped) curve [34]. Moderate habitual alcohol use also appears
to be linked to lower risks of diabetes mellitus, stroke, heart failure, and total mortality [36]. Increase
of plasma high-density lipoprotein (HDL)-cholesterol and apolipoprotein-AI (Apo-AI) is the most
described effect of moderate alcohol consumption, and until recently, this was considered as the main
protective effect of alcohol.
Light-to-moderate alcohol intake has also been shown to improve outcomes in patients with estab-
lished CVD or dementia. In a meta-analysis of eight prospective studies involving 16,351 patients
with a history of CVD, the familiar J-shaped curve was observed, with a maximal protective effect of
alcohol at approximately 26 g/day (or about two drinks daily) [37]. This effect is observed after the
consumption of any alcoholic beverage in a dose-dependent manner, and accordingly, it is considered
to be due to ethanol itself [38]. Accumulating scientific evidences also suggest that light-to-moderate
alcohol intake may enhance insulin sensitivity, increase adiponectin levels, and improve endothelial
function [39]. However, the protective effect of moderate alcohol consumption also depends on the
Wine 747
pattern of alcohol use, whether the intake is concentrated at certain times, usually at the weekends, or
is regularly spread over the week during meals. Binge drinking, usually defined as episodic excessive
alcohol intake (≥5 drinks within a few hours), often with intent to become intoxicated, is associated
with a two-fold higher risk of mortality [40]. Even occasional binges attenuate the protection offered
by otherwise light-to-moderate consumption [41]. In order to evaluate the different properties of etha-
nol and polyphenols, randomized intervention clinical trials that evaluate alcoholic drinks with and
without polyphenols are necessary.
57.4.2 Clinical Intervention Trials

57.4.2.1 Wine Polyphenols and Oxidative Status
While alcohol by itself is known to induce oxidative stress, wine polyphenols seem to counteract
this effect [35]. Several clinical trials have shown the antioxidant effect of moderate red wine
consumption. The intake of wine polyphenols was found to increase PAC [42]. However, the total
antioxidant capacity assay is inconclusive, because it detects urate as the main contributor, and the
clinical significance of the increase in total antioxidant capacity due to urate concentration is unclear.
On the other hand, wine polyphenols apparently decrease plasma malondialdehyde [38], a measure-
ment of oxidative stress status, as well as inhibit low-density lipoprotein (LDL) cholesterol particles
oxidation and increase the activity of several antioxidant enzymes [43]. Despite a general consensus
attributing antioxidant benefits in healthy volunteers to sustained wine consumption, there is not
enough scientific evidence, at present, that sustained wine consumption provides antioxidant benefits
other than to counteract a possible pro-oxidative effect of the alcohol [38]. On the contrary, data on
the antioxidant protective effects of red wine in oxidative stress situations are promising. Thus, post-
prandial oxidative stress seems to be counteracted by the ingestion of red wine, effect being attributed
to wine polyphenols, despite the diversity of biomarkers used for its evaluation. It has been proposed
that polyphenols, including the nonabsorbable ones, could exert antioxidant and other cytoprotective
effects in the gastrointestinal tract, where they are present in high quantities [6]. On this basis, the
Mediterranean diet pattern that includes moderate wine consumption with meals would counteract
the pro-oxidant effect of food digestion.
57.4.2.2 Wine Polyphenols and Inflammation

There is growing evidence for the anti-inflammatory effects of sustained wine consumption [39],
even in low cardiovascular risk individuals [39]. Oxidative stress and inflammation are intertwined
processes [38]. Inflammation promotes oxidative stress and oxidative damage, and vice versa [44].
Red wine, but not gin, diminished C-reactive protein (CRP) in plasma in healthy subjects [39] and
favored anti-inflammatory interleukins [45]. Similar results were observed for cell adhesion and cyto-
kines [39], molecules that participate in the recruitment of circulating leukocytes to the vascular endo-
thelium, initiating the atherosclerotic process. However, when dealcoholized red wine was compared
with the same alcoholized wine, the phenolic content was found to be responsible for modulating
leukocyte adhesion molecules, whereas both ethanol and polyphenols of red wine may modulate soluble
inflammatory mediators in high-risk patients [45].
57.4.2.3 Wine Polyphenols and Blood Pressure

A linear correlation between alcohol intake and increased blood pressure has been reported in normal
subjects and hypertensive patients [46]. However, in moderate amounts it appears to exert neutral or even
beneficial effects on blood pressure, and red wine seems be superior to other alcoholic beverages in this
respect [35]. Thus, in a crossover feeding trial, moderate doses of dealcoholized red wine decreased
systolic and diastolic blood pressure while increasing plasma NO concentration. Red wine tended to
induce a similar pattern to dealcoholized red wine, although the changes did not achieve statistical
significance, while moderate gin consumption had no effect. Thus, these blood-pressure-lowering and
NO-raising effects should be attributed to red wine polyphenols and not to alcohol. Polyphenol effects
on blood pressure have also been demonstrated in the PREDIMED trial. The decrease in blood pressure
was correlated with an increase in plasma NO concentration, and wine was one of the main sources of
polyphenols in Mediterranean population [47].
57.4.2.4 Wine Polyphenols and Lipid Profile

Compared to other alcoholic beverages, red wine consumption may exert an additional protective effect
against CVD. Thus, in an intervention trial comparing the effects on wine, dealcoholized wine and gin,
plasma LDL-cholesterol and apolipoprotein-B (Apo-B) concentrations were significantly reduced after
wine and dealcoholized wine interventions, but not after gin, suggesting that this effect should be related
to nonalcoholic content of red wine, mainly polyphenols [34]. In addition, both cross-sectional and inter-
vention studies have shown that moderate wine intake reduces the plasma concentrations of in vivo
oxidized LDL, which has also been related to the polyphenolic content of red wine [48]. According
to Cartron et al. [12], the beneficial effect of red wine seems to mainly be explained by its action on
lipid and lipoprotein constants and not by its antioxidant capacity. Polyphenols in red wine also help to
improve the lipid profile.
57.4.2.5 Wine Polyphenols and Diabetes Mellitus

In some prospective studies, the inverse association between moderate alcohol intake and low diabe-
tes risk was most apparent in wine and beer drinkers compared to those who reported liquor intake
[49]. The results of the few randomized clinical studies that examined the effects of moderate alcohol
consumption on insulin sensitivity, an accurate measurement of the glucose metabolism, have been
inconsistent. Although some have reported null results [50], another study has concluded that red wine
exerts higher protective effects than other alcoholic beverages [51]. Thus, in a recent randomized clini-
cal trial that examined the effects of three interventions (red wine, dealcoholized red wine, and gin),
both red and dealcoholized wines decreased homeostasis model assessment–insulin resistance index,
a measurement of insulin sensitivity, by 30% and 22%, respectively [38], suggesting that polyphenols
contained in wine exert a positive effect on glucose metabolism. However, in another trial that included
51 postmenopausal women, consumption of 30 g/day of alcohol (ethanol in orange juice) was associated
with a 7.2% improvement in insulin sensitivity compared with 0.0 g/day, while consumption of 15 g/day
of alcohol had no effect. Therefore, considering all these results together, it seems that both ethanol and
polyphenols contained in alcoholic beverages are responsible for beneficial effect observed on glucose
metabolism.
Interestingly, adiponectin has been proposed as an important link between moderate alcohol con-
sumption and lower incidence of type-II diabetes. To address this question, in an open randomized
crossover intervention study, plasma adiponectin concentration significantly increased after moderate
consumption of wine, beer, and ethanol, but the changes after wine consumption (30% increase) was
higher than after ethanol (17%) or beer (16%). In this case, the additional effect of wine on adiponectin
concentration could not be attributed to their polyphenolic content since dealcoholized wine did not exert
any significant effect. Changes in sex hormones or dietary pattern on adiponectin concentrations may be
other potential explanations for these findings [52].
57.4.2.6 Wine Prebiotic Effects

In a study, 10 healthy males received red wine, dealcoholized red wine, or gin for 20 days; the results
showed that red wine polyphenols can inhibit nonbeneficial bacteria from the human microbiota and
potentiate the growth of probiotic bacteria such as bifidobacteria, which could be implicated in the reduc-
tion of CRP and cholesterol [53]. Red wine, but not gin or dealcoholized red wine, consumption seemed
to be able to lower plasma lipopolysaccharides in metabolic endotoxemia, by means of the modulation
of the composition of the gut microbiota, while acute red wine intake did not modify the postprandial,
probably because changing gut microbiota requires a longer stimulus than a meal [54].
Wine 749
57.4.2.7 Wine Polyphenols and Cancer

To evaluate the effect of wine on cancer, intervention clinical trials are very difficult to perform due to
ethical reasons and the length of the studies. However, when DNA damage markers are studied, it seems
that wine polyphenols may protect against cancer initiation, since DNA strand breaks are reduced after
6 weeks of daily consumption of red wine [14].
57.4.2.8 Red Wine and Dementia

There are still no papers published using wine in intervention clinical trials to evaluate the possible
protective effect of wine on dementia. However, polyphenols may benefit Alzheimer’s disease by modu-
lating multiple disease-modifying modalities, both β-amyloid-dependent and β-amyloid-independent
mechanisms [55].
57.5 Current Government Policies Affecting Wine Consumption

As explained earlier, effects of wine on the human body are complex. The consumption of alcoholic
beverages such as wine in heavy amounts is considered to be a risk factor for diseases such as certain
cancers, alcoholic cirrhosis of the liver, and alcoholic pancreatitis. Alcohol-related harm is also indepen-
dently considered to be a noncommunicable disease. Conversely, light-to-moderate consumption reduces
the risk of CVD, certain cancers, diabetes, and dementias such as Alzheimer’s disease [56]. From a
review of the literature, moderate consumption is generally defined as approximately 20–40 g alcohol
per day [57].
The J-shaped relationship between wine consumption and CVD (lowest risk at moderate consumption)
is currently acknowledged by WHO [58]. For example, the WHO’s global status report on alcohol [59]
includes the following statement: “The relationship between alcohol consumption and CVD is complex.
Light-to-moderate drinking can have a beneficial impact on morbidity and mortality for ischemic heart
disease and ischemic stroke. However, the beneficial cardioprotective effect of drinking disappears with
heavy drinking occasions.”
The relationship between wine and cancer per se is not necessarily J-shaped and consequently the 4th
edition of the WHO-related European code against cancer (2014) states that “If you drink alcohol of any
type, limit your intake. Not drinking alcohol is better for cancer prevention.” In contrast, the previous
3rd edition of this code stated that “If you drink alcohol, whether beer, wine, or spirits, moderate your
consumption to two drinks per day if you are a man or one drink per day if you are a woman” [60]. This
previous code is more consistent with conclusions of the European Prospective Investigation into Cancer
and Nutrition, which showed that although risk for cancer increases with alcohol consumption, the risk
of dying overall decreases with moderate drinking when compared with nondrinkers [61].
Few, if any, national alcohol drinking or dietary guidelines now take into account the potential ben-
eficial health effects of alcohol consumption; those of Australia, Canada, and the United Kingdom did
so previously. This is because the science behind the health effects of both moderate and heavy con-
sumption is not the only factor taken into consideration by governments when producing public health
policies and guidelines. The purpose of guidelines is to facilitate a change in behavior, and a popula-
tion’s prevailing drinking culture is also an important factor. Those countries with a Mediterranean-style
diet, lifestyle, and consumption patterns, such as drinking wine with daily meals, have traditionally had
higher recommended maximum levels for alcohol consumption than do other countries, especially those
with a culture of binge beer and spirits drinking [62]. Distinctions between countries’ consumption pat-
terns and recommendations, are disappearing, however, as beverage preferences have begun to converge
globally [62]. This can be seen in a trend toward binge drinking and intoxication among young adults
irrespective of country [63].
In addition, most governments are concerned with reducing the economic, health, and social conse-
quences of alcohol abuse and misuse. Their recommendations are aimed at the population groups that
are misusing alcohol or drinking cultures that are likely to lead to misuse. Indeed, of the 2 billion people
that consume alcoholic beverages worldwide, approximately 76.3 million or 3.8% have alcohol-related
problems due to alcohol abuse, and 1.8 million or 0.09% are estimated to likely die from alcohol-related
harmful effects [64]. Alcohol is a leading cause of the global burden of disease along with childhood and
maternal underweight, unsafe sex, hypertension, and tobacco use and is estimated to cause 20%–30%
of esophageal and liver cancers, cirrhosis of the liver, and epileptic seizures worldwide [65,66]. In this
context, the beneficial health effects of moderate wine consumption appear much less relevant. They are
much more likely to be taken into account where recommendations are aimed at the whole population
and are set in the context of the diet as a whole, as is the case with the current U.S. Dietary Guidelines
for Americans.
It may further be argued that in some cases, recommendations intended to change the behavior of
those misusing alcohol may conflict with those intended to maximize the potential beneficial health
effects in a population. For example, any advice to abstain from drinking on some days of the week is at
odds with the potential beneficial effects of regular, daily moderate drinking on the cardiovascular sys-
tem. This consumption pattern has been observed to prolong any acute and short-term beneficial effects
of alcohol and phenolic components on hemostasis [67,68] and also maintain or promote any long-term
beneficial effects, including those on blood pressure [69,70]. Similarly, recommendations that seek to
reduce overall alcohol consumption in a population may also reduce that of moderate consumers, which
may eventually be reflected in an increased incidence of CVD within a population. CVD remains a lead-
ing cause of mortality in the developed world, accounting for 25%–50% of all deaths, where its incidence
is increasing in developing countries.
When formulating recommendations on maximum levels of alcohol consumption, hopefully govern-
ments will recognize these potential problems and seek ways of resolving them. Continuing research will
also continue to change alcohol’s role as part of a healthy diet and lifestyle. It has already been estab-
lished that the extent of the beneficial health effects of moderate alcohol consumption is closely related
to background diet [71]. The most significant effects of drinking, for example, are found when alcohol is
consumed as part of a diet that is high in saturated fat [72]. Another important finding is that beneficial
health effects of moderate drinking are enhanced in diets that are already high in plant-derived phenolic
compounds, such as a Mediterranean-style diet. The findings on the potential beneficial effects of moder-
ate alcohol consumption on diabetes mellitus and obesity [73] are especially important in light of present
global trends in body weight. Similar future findings would make a strong case for the importance of
taking the positive outcomes of moderate consumption of alcohol into account when formulating both
recommended maximum levels of drinking and population dietary guidelines.

The historical legacy of wine and its cultural importance also means that the definition of wine is highly
regulated. For instance in the United States, the sale of “wine” is restricted to those beverages created
solely by the fermentation of grapes [74]. The addition of any other fruits as well as any flavorings is
strictly forbidden. The geographic origin of the fruit can be an important indicator of quality, and so
those terms are regulated worldwide in winegrowing regions. In many traditional European regions,
the grape varieties are specified, as well as the grape growing protocols and winemaking procedures.
Therefore, in this environment of tradition and regulation, innovation is either limited or results in prod-
ucts that are derived from wine but must be called something else.
While polyphenolics are considered to be very important to the health impact of wine, most regulatory
frameworks do not permit producers to inform consumers about the content of these substances. In the
United States, alcoholic beverages are not considered sources of nutrition, and thus producers are not
permitted to post the standard nutrition label found on packaged food. One producer, Willamette Valley
Vineyards, was able to obtain permission to declare the resveratrol content of one wine, but no subse-
quent permits have been allowed [75].
A general increase in the phenolic content of red wine is easily achieved by a thorough extraction
of the skins and seeds, for instance, by leaving the wine in contact with the pomace after fermentation
is complete, using a winemaking technique called extended maceration [76]. Grapes grown in cooler
Wine 751
regions contain high levels of phenolics and thus can yield high phenolic wine. However, tannins com-
prise about half of the phenolics that are extracted and such wines can be very unpalatable due to high
astringency and bitterness.
Modifying wines, for instance, in the manner described by Arnold of Villa Nueva, by using the wine
to extract the active components from herbal medicines, is rarely practiced today, and only as a home
remedy. The availability of high-strength alcohol, a much better extraction solvent than wine, reduces
interest in using wine for such extracts. There are a few traditional liqueurs that were created as medici-
nal herbal extracts, but today are sold as flavored or bitter liqueurs, for example, Chartreuse, Benedictine,
Drambuie, Cointreau, Vermouth, and Amaro.
The expectation that there will be wines with enhanced or new medicinal properties is limited by
the regulatory marketing constraints on producers [77]. Even if a company developed a truly effective
medicinal wine, the channels to market are very murky if not totally opaque, at least in the United States.
In fact, the U.S. wine industry has studiously avoided marketing wine based on its now fairly well-
established health benefits, in part due to regulations. Therefore, the prospects for new, functional bev-
erages based on wine are unlikely barring some major changes in the law. Instead, we expect new wines
to arise based on desirable flavor profiles that come from novel varieties or other production scenarios.
57.7 Conclusion
Historical claims that moderate consumption of wine is healthy have been substantiated in numerous
modern medical studies. The best documented areas of health benefit involve reducing CVD, but there
are other areas of distinct benefit. An important point to note is that regular, moderate wine consumers
have a lower overall mortality rate compared to nonconsumers. Explanations for these health benefits
are now becoming clear. For instance, recent studies show that wine polyphenolics and their metabolites
prevent inflammatory responses and this, for instance, can reduce blood clotting. However, these clear
health benefits must be offset by the social burden of alcohol abuse, and regulatory agencies struggle
to find ways to minimize this burden through outreach and regulation while still acknowledging the
health benefits to moderate consumers. The regulatory framework for wine sales worldwide greatly
limits innovation, especially those that might offer additional health benefits, because in most countries,
wine composition is strictly defined, and it is difficult if not impossible to market the health benefits of
their product.
REFERENCES
1. McGovern, P.E., Zhang, J.H., Tang, J.G., Zhang, Z.Q., Hall, G.R., Moreau, R.A., Nunez, A. et al.,
Fermented beverages of pre- and proto-historic china. Proc. Natl. Acad. Sci. U.S.A., 101, 17593–17598,
2004.
2. Sayon-Orea, C., Martinez-Gonzalez, M.A., and Bes-Rastrollo, M., Alcohol consumption and body
weight: A systematic review. Nutr. Rev., 69, 419–431, 2011.
3. Regueiro, J., Vallverdu-Queralt, A., Simal-Gandara, J., Estruch, R., and Lamuela-Raventos, R.,
Development of a lc-esi-ms/ms approach for the rapid quantification of main wine organic acids in
human urine. J. Agric. Food Chem., 61, 6763–6768, 2013.
4. Regueiro, J., Vallverdu-Queralt, A., Simal-Gandara, J., Estruch, R., and Lamuela-Raventos, R.M.,
Urinary tartaric acid as a potential biomarker for the dietary assessment of moderate wine consump-
tion: A randomised controlled trial. Br. J. Nutr., 111, 1680–1685, 2014.
5. Weisse, M.E., Eberly, B., and Person, D.A., Wine as a digestive aid: Comparative antimicrobial effects
of bismuth salicylate and red and white wine. Br. Med. J., 311, 1657–1660, 1995.
6. Kanner, J., Gorelik, S., Roman, S., and Kohen, R., Protection by polyphenols of postprandial human
plasma and low-density lipoprotein modification: The stomach as a bioreactor. J. Agric. Food Chem.,
60, 8790–8796, 2012.
7. Fang, F., Li, J.-M., Zhang, P., Tang, K., Wang, W., Pan, Q.-H., and Huang, W.-D., Effects of grape vari-
ety, harvest date, fermentation vessel and wine ageing on flavonoid concentration in red wines. Food
Res. Int., 41, 53–60, 2008.
8. Sato, M., Ramarathnam, N., Suzuki, Y., Ohkubo, T., Takeuchi, M., and Ochi, H., Varietal differences
in the phenolic content and superoxide radical scavenging potential of wines from different sources.
J. Agric. Food Chem., 44, 37–41, 1996.
9. Villaño, D., Fernández-Pachón, M.S., Troncoso, A.M., and García-Parrilla, M.C., Comparison of anti-
oxidant activity of wine phenolic compounds and metabolites in vitro. Anal. Chim. Acta, 538, 391–398,
2005.
10. Fernandez-Panchon, M.S., Villano, D., Troncoso, A.M., and Garcia-Parrilla, M.C., Antioxidant activ-
ity of phenolic compounds: From in vitro results to in vivo evidence. Crit. Rev. Food Sci. Nutr., 48,
649–671, 2008.
11. Ghiselli, A., Serafini, M., Natella, F., and Scaccini, C., Total antioxidant capacity as a tool to assess
redox status: Critical view and experimental data. Free Radic. Biol. Med., 29, 1106–1114, 2000.
12. Cartron, E., Fouret, G., Carbonneau, M.-A., Lauret, C., Michel, F., Monnier, L., Descomps, B., and
Leger, C.L., Red-wine beneficial long-term effect on lipids but not on antioxidant characteristics in
plasma in a study comparing three types of wine: Description of two o-methylated derivatives of gallic
acid in humans. Free Radic. Res., 37, 1021–1035, 2003.
13. Simonetti, P., Gardana, C., and Pietta, P., Plasma levels of caffeic acid and antioxidant status after red
wine intake. J. Agric. Food Chem., 49, 5964–5968, 2001.
14. Arendt, B.M., Ellinger, S., Kekic, K., Geus, L., Fimmers, R., Spengler, U., Müller, W.-U., and Goerlich,
R., Single and repeated moderate consumption of native or dealcoholized red wine show different
effects on antioxidant parameters in blood and DNA strand breaks in peripheral leukocytes in healthy
volunteers: A randomized controlled trial (isrctn68505294). Nutr. J., 4, 33, 2005.
15. Cao, G., Russell, R.M., Lischner, N., and Prior, R.L., Serum antioxidant capacity is increased by con-
sumption of strawberries, spinach, red wine or vitamin C in elderly women. J. Nutr., 128, 2383–2390,
1998.
16. Wang, D., Wei, X., Yan, X., Jin, T., and Ling, W., Protocatechuic acid, a metabolite of anthocyanins,
inhibits monocyte adhesion and reduces atherosclerosis in apolipoprotein e-deficient mice. J. Agric.
Food Chem., 58, 12722–12728, 2010.
17. Lotito, S.B., Zhang, W.-J., Yang, C.S., Crozier, A., and Frei, B., Metabolic conversion of dietary flavo-
noids alters their anti-inflammatory and antioxidant properties. Free Radic. Biol. Med., 51, 454–463,
2011.
18. Hou, D.X., Masuzaki, S., Hashimoto, F., Uto, T., Tanigawa, S., Fujii, M., and Sakata, Y., Green tea
proanthocyanidins inhibit cyclooxygenase-2 expression in lps-activated mouse macrophages: Molecular
mechanisms and structure-activity relationship. Arch. Biochem. Biophys., 460, 67–74, 2007.
19. Kundu, J.K., Na, H.K., Chun, K.S., Kim, Y.K., Lee, S.J., Lee, S.S., Lee, O.S., Sim, Y.C., and Surh, Y.J.,
Inhibition of phorbol ester-induced cox-2 expression by epigallocatechin gallate in mouse skin and cul-
tured human mammary epithelial cells. J. Nutr., 133, 3805S–3810S, 2003.
20. Martinez-Micaelo, N., Gonzalez-Abuin, N., Ardevol, A., Pinent, M., and Blay, M.T., Procyanidins and
inflammation: Molecular targets and health implications. BioFactors, 38, 257–265, 2012.
21. Tribolo, S., Lodi, F., Connor, C., Suri, S., Wilson, V.G., Taylor, M.A., Needs, P.W., Kroon, P.A., and
Hughes, D.A., Comparative effects of quercetin and its predominant human metabolites on adhesion
molecule expression in activated human vascular endothelial cells. Atherosclerosis, 197, 50–56, 2008.
22. Del Rio, D., Rodriguez-Mateos, A., Spencer, J.P.E., Tognolini, M., Borges, G., and Crozier, A., Dietary
(poly)phenolics in human health: Structures, bioavailability, and evidence of protective effects against
chronic diseases. Antioxid. Redox Sign., 18, 1818–1892, 2013.
23. Santhakumar, A.B., Bulmer, A.C., and Singh, I., A review of the mechanisms and effectiveness of
dietary polyphenols in reducing oxidative stress and thrombotic risk. J. Hum. Nutr. Diet., 27, 1–21, 2014.
24. Rein, D., Paglieroni, T.G., Wun, T., Pearson, D.A., Schmitz, H.H., Gosselin, R., and Keen, C.L., Cocoa
inhibits platelet activation and function. Am. J. Clin. Nutr., 72, 30–35, 2000.
25. Ostertag, L.M., Kroon, P.A., Wood, S., Horgan, G.W., Cienfuegos-Jovellanos, E., Saha, S., Duthie, G.G.,
and de Roos, B., Flavan-3-ol-enriched dark chocolate and white chocolate improve acute measures of
platelet function in a gender-specific way: A randomized-controlled human intervention trial. Mol. Nutr.
Food Res., 57, 191–202, 2013.
26. Taylor, S.G. and Weston, A.H., Endothelium-derived hyperpolarizing factor: A new endogenous inhibi-
tor from the vascular endothelium. Trends Pharmacol. Sci., 9, 272–274, 1988.
Wine 753
27. Palmer, R.M., Ferrige, A.G., and Moncada, S., Nitric oxide release accounts for the biological activity
of endothelium-derived relaxing factor. Nature, 327, 524–526, 1987.
28. Fitzpatrick, D.F., Hirschfield, S.L., and Coffey, R.G., Endothelium-dependent vasorelaxing activity of
wine and other grape products. Am. J. Physiol., 265, H774–H778, 1993.
29. Burns, J., Gardner, P.T., O’Neil, J., Crawford, S., Morecroft, I., McPhail, D.B., Lister, C. et al.,
Relationship among antioxidant activity, vasodilation capacity, and phenolic content of red wines.
J. Agric. Food Chem., 48, 220–230, 2000.
30. Fitzpatrick, D.F., Fleming, R.C., Bing, B., Maggi, D.A., and O’Malley, R.M., Isolation and characteriza-
tion of endothelium-dependent vasorelaxing compounds from grape seeds. J. Agric. Food Chem., 48,
6384–6390, 2000.
31. Martin, S., Andriambeloson, E., Takeda, K., and Andriantsitohaina, R., Red wine polyphenols increase
calcium in bovine aortic endothelial cells: A basis to elucidate signalling pathways leading to nitric
oxide production. Br. J. Pharmacol., 135, 1579–1587, 2002.
32. Ruitenberg, A., van Swieten, J.C., Witteman, J.C.M., Mehta, K.M., van Duijn, C.M., Hofman, A.,
and Breteler, M.M.B., Alcohol consumption and risk of dementia: The Rotterdam study. Lancet, 359,
281–286, 2002.
33. Poli, A., Marangoni, F., Avogaro, A., Barba, G., Bellentani, S., Bucci, M., Cambieri, R. et al., Moderate
alcohol use and health: A consensus document. Nutr. Metab. Cardiovasc. Dis., 23, 487–504, 2013.
34. Chiva-Blanch, G., Urpi-Sarda, M., Ros, E., Valderas-Martinez, P., Casas, R., Arranz, S., Guillén, M.
et al., Effects of red wine polyphenols and alcohol on glucose metabolism and the lipid profile: A ran-
domized clinical trial. Clin. Nutr., 32, 200–206, 2013.
35. Estruch, R. and Lamuela-Raventós, R., Wine, alcohol, polyphenols and cardiovascular disease. Nutr.
Aging, 2, 101–109, 2014.
36. O’Keefe, J.H., Bybee, K.A., and Lavie, C.J., Alcohol and cardiovascular health: The razor-sharp double-
edged sword. J. Am. Coll. Cardiol., 50, 1009–1014, 2007.
37. Costanzo, S., Di Castelnuovo, A., Donati, M.B., Iacoviello, L., and de Gaetano, G., Alcohol consump-
tion and mortality in patients with cardiovascular disease: A meta-analysis. J. Am. Coll. Cardiol., 55,
1339–1347, 2010.
38. Covas, M.I., Gambert, P., Fitó, M., and de la Torre, R., Wine and oxidative stress: Up-to-date evidence
of the effects of moderate wine consumption on oxidative damage in humans. Atherosclerosis, 208,
297–304, 2010.
39. Estruch, R., Sacanella, E., Badia, E., Antúnez, E., Nicolás, J.M., Fernández-Solá, J., Rotilio, D., de
Gaetano, G., Rubin, E., and Urbano-Márquez, A., Different effects of red wine and gin consumption on
inflammatory biomarkers of atherosclerosis: A prospective randomized crossover trial. Effects of wine
on inflammatory markers. Atherosclerosis, 175, 117–123, 2004.
40. Mukamal, K.J., Ascherio, A., Mittleman, M.A., Conigrave, K.M., Camargo, C.A., Kawachi, I., Stampfer,
M.J., Willett, W.C., and Rimm, E.B., Alcohol and risk for ischemic stroke in men: The role of drinking
patterns and usual beverage. Ann. Intern. Med., 142, 11–19, 2005.
41. O’Keefe, J.H., Bhatti, S.K., Bajwa, A., DiNicolantonio, J.J., and Lavie, C.J., Alcohol and cardiovascular
health: The dose makes the poison… or the remedy. Mayo Clin. Proc., 89, 382–393, 2014.
42. Micallef, M., Lexis, L., and Lewandowski, P., Red wine consumption increases antioxidant status and
decreases oxidative stress in the circulation of both young and old humans. Nutr. J., 6, 27, 2007.
43. Estruch, R., Sacanella, E., Mota, F., Chiva-Blanch, G., Antúnez, E., Casals, E., Deulofeu, R. et al.,
Moderate consumption of red wine, but not gin, decreases erythrocyte superoxide dismutase activity:
A randomised cross-over trial. Nutr. Metab. Cardiovasc. Dis., 21, 46–53, 2011.
44. Crowley, S.D., The cooperative roles of inflammation and oxidative stress in the pathogenesis of hyper-
tension. Antioxid. Redox Sign., 20, 102–120, 2014.
45. Chiva-Blanch, G., Urpi-Sarda, M., Llorach, R., Rotches-Ribalta, M., Guillén, M., Casas, R., Arranz, S.
et al., Differential effects of polyphenols and alcohol of red wine on the expression of adhesion mol-
ecules and inflammatory cytokines related to atherosclerosis: A randomized clinical trial. Am. J. Clin.
Nutr., 95, 326–334, 2012.
46. Fuchs, F.D., Chambless, L.E., Whelton, P.K., Nieto, F.J., and Heiss, G., Alcohol consumption and the
incidence of hypertension: The atherosclerosis risk in communities study. Hypertension, 37, 1242–1250,
2001.
47. Medina-Remón, A., Tresserra-Rimbau, A., Pons, A., Tur, J.A., Martorell, M., Ros, E., Buil-Cosiales, P.
et al., Effects of total dietary polyphenols on plasma nitric oxide and blood pressure in a high
cardiovascular risk cohort. The PREDIMED randomized trial. Nutr. Metab. Cardiovasc. Dis., 25,
60–67, 2014.
48. Schroder, H., Marrugat, J., Fíto, M., Weinbrenner, T., and Covas, M.-I., Alcohol consumption is directly
associated with circulating oxidized low-density lipoprotein. Free Radic. Biol. Med., 40, 1474–1481,
2006.
49. Wannamethee, S.G., Camargo, C.A., Manson, J.E., Willett, W.C., and Rimm, E.B., Alcohol drinking
patterns and risk of type 2 diabetes mellitus among younger women. Arch. Intern. Med., 163, 1329–1336,
2003.
50. Sierksma, A., Patel, H., Ouchi, N., Kihara, S., Funahashi, T., Heine, R.J., Grobbee, D.E., Kluft, C., and
Hendriks, H.F.J., Effect of moderate alcohol consumption on adiponectin, tumor necrosis factor-alpha,
and insulin sensitivity. Diabetes Care, 27, 184–189, 2004.
51. Napoli, R., Cozzolino, D., Guardasole, V., Angelini, V., Zarra, E., Matarazzo, M., Cittadini, A., Saccà,
L., and Torella, R., Red wine consumption improves insulin resistance but not endothelial function in
type 2 diabetic patients. Metabolism, 54, 306–313, 2005.
52. Imhof, A., Plamper, I., Maier, S., Trischler, G., and Koenig, W., Effect of drinking on adiponectin in
healthy men and women: A randomized intervention study of water, ethanol, red wine, and beer with or
without alcohol. Diabetes Care, 32, 1101–1103, 2009.
53. Queipo-Ortuño, M.I., Boto-Ordóñez, M., Murri, M., Gomez-Zumaquero, J.M., Clemente-Postigo, M.,
Estruch, R., Cardona Diaz, F. Andrés-Lacueva, C., and Tinahones, F.J., Influence of red wine polyphe-
nols and ethanol on the gut microbiota ecology and biochemical biomarkers. Am. J. Clin. Nutr., 95,
1323–1334, 2012.
54. Clemente-Postigo, M., Queipo-Ortuño, M.I., Boto-Ordoñez, M., Coin-Aragüez, L., Roca-Rodriguez,
M.D.M., Delgado-Lista, J., Cardona, F., andres-Lacueva, C., and Tinahones, F.J., Effect of acute and
chronic red wine consumption on lipopolysaccharide concentrations. Am. J. Clin. Nutr., 97, 1053–1061,
2013.
55. Pasinetti, G.M., Novel role of red wine-derived polyphenols in the prevention of alzheimer’s disease
dementia and brain pathology: Experimental approaches and clinical implications. Planta Med., 78,
1614–1619, 2012.
56. Ronksley, P.E., Brien, S.E., Turner, B.J., Mukamal, K.J., and Ghali, W.A., Association of alcohol
consumption with selected cardiovascular disease outcomes: A systematic review and meta-analysis.
Br. Med. J., 342, d671, 2011.
57. Klatsky, A.L. and Udaltsova, N., Alcohol drinking and total mortality risk. Ann. Epidemiol., 17,
S63–S67, 2007.
58. World Health Organization (WHO), Global Strategy to Reduce Harmful Use of Alcohol, WHO, Geneva,
Switzerland, 2010.
59. World Health Organization (WHO), Global Status Report on Alcohol and Health, WHO, Geneva,
Switzerland, 2011.
60. Boyle, P., Autier, P., Bartelink, H., Baselga, J., Boffetta, P., Burn, J., Burns, H.J.G. et al., European code
against cancer and scientific justification: Third version. Ann. Oncol., 14, 973–1005, 2003.
61. Bergmann, M.M., Rehm, J., Klipstein-Grobusch, K., Boeing, H., Schuetze, M., Drogan, D., Overvad, K.
et al., The association of pattern of lifetime alcohol use and cause of death in the European prospective
investigation into cancer and nutrition (epic) study. Int. J. Epidemiol., 42, 1772–1790, 2013.
62. Bloomfield, K., Stockwell, T., Gmel, G., and Rehn, N., International comparisons of alcohol consump-
tion. Alcohol Res. Health, 27, 95–109, 2003.
63. Britton, A., Nolte, E., White, I.R., Gronbaek, M., Powles, J., Cavallo, F., and McPherson, K., A compari-
son of the alcohol-attributable mortality in four European countries. Eur. J. Epidemiol., 18, 643–651,
2003.
64. World Health Organization (WHO), Global Status Report—Alcohol Policy, WHO, Geneva, Switzerland,
2004.
65. World Health Organization (WHO), The World Health Report 2002—Reducing Risks, Promoting
Healthy Life, WHO, Geneva, Switzerland, 2002.
66. World Health Organization (WHO), Global Health Risks: Mortality and Burden of Disease Attributable
to Selected Major Risks, WHO, Geneva, Switzerland, 2009.
Wine 755
67. Hendriks, H.F.J., Veenstra, J., Velthuistewierik, E.J.M., Schaafsma, G., and Kluft, C., Effect of moder-
ate dose of alcohol with evening meal on fibrinolytic factors. Br. Med. J., 308, 1003–1006, 1994.
68. Renaud, S. and de Lorgeril, M., Wine, alcohol, platelets, and the French paradox for coronary heart
disease. Lancet, 339, 1523–1526, 1992.
69. Klatsky, A.L., Blood pressure and alcohol intake, in Hypertension: Pathophysiology, Diagnosis and
Management, Laragh, J.H.M. and Brenner, B.M., Eds., Raven Press, New York, 1995, pp. 2649–2667.
70. Puddey, I.B., Beilin, L.J., Vandongen, R., Rouse, I.L., and Rogers, P., Evidence for a direct effect
of alcohol-consumption on blood-pressure in normotensive men—A randomized controlled trial.
Hypertension, 7, 707–713, 1985.
71. de Lorgeril, M. and Salen, P., Wine ethanol, platelets, and mediterranean diet. Lancet, 353, 1067–1067,
1999.
72. Mezzano, D., Leighton, F., Strobel, P., Martinez, C., Marshall, G., Cuevas, A., Castillo, O., Panes, O.,
Munoz, B., Rozowski, J., and Pereira, J., Mediterranean diet, but not red wine, is associated with benefi-
cial changes in primary haemostasis. Eur. J. Clin. Nutr., 57, 439–446, 2003.
73. Wannamethee, S.G., Shaper, A.G., Perry, I.J., and Alberti, K.G.M.M., Alcohol consumption and the
incidence of type II diabetes. J. Epidemiol. Commun. Health, 56, 542–548, 2002.
74. Code of Federal Regulations, Standards of Identity for Wine (Title 27, Volume 1, Part 4), Washington
State Legislature, Olympia, WA, 2014.
75. Willamette Valley Vineyards, Winery Receives Federal Label Approval on Antioxidant Content, PR
Newswire, 2004, Published online at: http://www.prnewswire.com/news-releases/winery-receives-
federal-label-approval-on-antioxidant-content-54143477.html (accessed January 15, 2015).
76. Busse-Valverde, N., Bautista-Ortin, A.B., Gomez-Plaza, E., Fernandez-Fernandez, J.I., and Gil-Munoz,
R., Influence of skin maceration time on the proanthocyanidin content of red wines. Eur. Food Res.
Technol., 235, 1117–1123, 2012.
77. U.S. Department of the Treasury, Alcohol and Tobacco and Trade Bureau, Health Claims and Other
Health-Related Statements in the Labeling and Advertising of Alcohol Beverages, Final Rule, Federal
Register (Volume 68, No 41, March 3, 2003), 2003, pp. 10076–10106, U.S. Department of the Treasury,
Washington, DC.
79. Phenol Explorer, Phenol-Explorer 3.5: Database in Polyphenol Content in Foods. Published online at:
http://phenol-explorer.eu/ (accessed January 14, 2015).
58
Maple Water
CONTENTS
58.1 Introduction....................................................................................................................................757
58.4 Health Effects.................................................................................................................................762
58.5 Novel Products/Formulation and Future Trends............................................................................762
58.6 Conclusion..................................................................................................................................... 763
Acknowledgment.................................................................................................................................... 763
References............................................................................................................................................... 763
58.1 Introduction
In an increasingly competitive and, arguably, oversaturated functional beverage market, there are few
naturally derived and minimally processed so-called “vegetable-derived waters” that are available com-
mercially to consumers. Coconut water, the liquid endosperm of the coconut (Cocos nucifera L.) fruit [1],
is a good example of such a product. Moreover, coconut water, which has been consumed for centuries
in many developing countries, mainly in the tropics, is rapidly growing in popularity among consumers
in modern cultures such as in the United States, Canada, the United Kingdom, and the rest of Europe.
Given that there are few commercially successful functional beverages in this “vegetable-derived and
natural waters” arena, there exists great potential for the development of new products.
The eastern North American region, primarily the province of Quebec (in Canada), leads the world’s
commercial production of maple syrup. This is a premium natural sweetener that contains sucrose as its
major sugar and is obtained by boiling the sap collected from certain maple species. The main maple
tree used for this purpose includes the sugar maple (Acer saccharum) species, which is native to this
region of the world. Maple sap, which is primarily of a watery consistency (~95%–99% water, hence also
referred to as “maple water”), is concentrated wherein ~40 L yields 1 L of maple syrup. Traditionally,
the indigenous people of North America consumed maple water by itself as a tonic for a variety of health
benefits. Even today, various cultures such as in South Korea collect and consume large quantities of
maple water from the maple tree known as gorosoe or “tree good for the bones” as a “treasured elixir”
that is used for osteoporosis due to the rich mineral (e.g., calcium) content of the sap and other health
benefits [2]. Moreover, every year, in Hadong in South Korea alone, over 1.2 million liters of maple
water is produced (and sold for ~$2–3 a liter) where it is widely consumed as a beverage. Given the well-
established commercial maple syrup industry in Quebec and other related areas in the eastern North
American region, with established technological ability for large-scale collection of sap, there exists
great potential for this beverage to be positioned as a new and emerging functional beverage in this part
of the world. In this light, herein, the chemical composition and biological effects of maple water in rela-
tion to its potential functional beverage applications are being discussed. Since maple water is a newly
emerging candidate in the functional beverage arena, and there is limited published data available on its
757
bioactive constituents, the majority of the data presented here have been generated by the research group
of the authors in collaboration with the Federation of Quebec Maple Syrup Producers (FPAQ; French:
Fédération des producteurs acéricoles du Québec, located in the province of Quebec in Canada), where
the vast majority of the world’s supply of maple syrup is produced.

Maple water is the xylem sap obtained by the tapping of maple trees in the late winter to early spring
when there is a combination of freeze–thaw cycles, namely, cold nights followed by warm days, that
allows the sap to flow freely. Being that this is a plant-derived natural product, it is not surprising
that the constituents of the natural maple water would include macronutrients, micronutrients, and
phytochemicals, and these substances have been well documented by extensive research over the
years [3–7].
Because maple water has been primarily consumed in our food chain in its concentrated form, namely,
as the natural sweetener, maple syrup, the nutritional characteristics of the latter substance is well estab-
lished and is available from reputable government websites such as the U.S. Department of Agriculture
(USDA) [8]. Notably, beyond sucrose, which is the major sugar present in maple syrup, this sweetener
also contains minerals (including calcium, iron, magnesium, potassium, sodium, and zinc) and vitamins
(including thiamin, riboflavin, and niacin), which are all listed on the aforementioned USDA Nutrient
Database [8]. It should be noted here that considering that maple syrup is technically “concentrated
maple sap/water without any additives,” it should not be surprising that many of the constituents found in
maple syrup are present originally in the sap, albeit at much lower quantities in the latter.
Given that maple water is a relatively new functional beverage and has recently emerged as a com-
mercial product on the market, its nutritional characteristics have not yet been established by the
aforementioned USDA Nutrient Database. In this chapter, we present data shown in Table 58.1 on the
compositional and nutritional characteristics of maple water (per 100 mL) as provided by the FPAQ [9].
Briefly, 100 mL of maple water (~95%–99% water) contains the following: energy = 7.57 kcal, lipid =
0.0 g, protein = 0.0 g, carbohydrate = 2.1 g of which sucrose = 2 g, glucose = 0.02 g, fructose = 0.01 g,
and complex sugars = 0.05 g. Other constituents including minerals and vitamins along with free amino
acids (arginine + threonine, histidine, leucine, and proline), organic acids (acetic, fumaric, gluconic,
malic, pyruvic, and succinic acid), and phytohormones (abscisic acid, phaseic acid, and their metabolites)
are also provided in Table 58.1.
In addition, the labeling of commercial maple water beverages includes “not a significant source of
saturated fat, trans fat, cholesterol, fiber, vitamin A, vitamin C or iron.” Moreover, the FPAQ has devel-
oped a natural, authentic, pure, sterile, and integral (NAPSI) certification for their commercial maple
water beverages with several claims allowed by Health Canada [10] as follows: low source of calories,
2.12 mg polyphenols per 500 mL serving, no sugar added, natural sweet 100%, pure and natural, no
preservatives or additives, 100% vegan, unprocessed, and great natural taste.
In collaboration with FPAQ, our laboratory has recently reported the levels of carbohydrate, minerals,
organic acids, and amino acids in maple sap samples both before (n = 34) and after sterilization (n = 14)
or pasteurization (n = 35) [11]. These thermal treatments are being used by FPAQ and their commercial
partners to eliminate/reduce microbial growth in maple sap in order for this beverage to be safe for
human consumption and for its large-scale commercial production. Our published data showed that the
constituents in maple water are being largely preserved after both types of thermal treatments [11].

Phenolic compounds constitute one of the most ubiquitous classes of phytochemicals present in human
diet. Moreover, an overwhelming body of studies has shown that phenolic phytochemicals have a wide
variety of biological properties [12].
Maple Water 759
TABLE 58.1
Compositional and Nutritional Characteristics of Maple Water (per 100 mL)
Nutrient Unit Quantity
Water g 95–99
Energy kcal 7.57
Protein g 0.0
Lipid (fat) g 0.0
Carbohydrate g 2.1
Sucrose g 2.0
Glucose g 0.02
Fructose g 0.01
Complex sugars g 0.05
Minerals
Calcium mg 6.19
Copper mg 0.30
Iron mg 0.41
Magnesium mg 1.31
Manganese mg 0.41
Phosphorus nd —
Potassium mg 7.89
Selenium mg —
Sodium mg 5.30
Zinc mg 0.26
Vitamins
Vitamin C mg —
Thiamin mg —
Niacin mg 0.003
Riboflavin mg 0.034
Pantothenic acid mg —
Vitamin B6 mg —
Folate (DFE) mg —
Free Amino Acids mg 2.17
Arginine + threonine mg 1.12
Histidine mg 0.03
Leucine mg 0.14
Proline mg 0.88
Organic Acids mg 32.6
Acetic mg 1.11
Fumaric mg 0.38
Gluconic mg 4.06
Malic mg 25.01
Pyruvic mg 0.84
Succinic mg 1.24
Phytohormones µg 6.15
Abscisic acid µg 0.27
Phaseic acid µg 3.44
Others ABA metabolites µg 2.44
Source: Adapted from the Federation of Quebec Maple Syrup Producers (FPAQ), Quebec Maple
Syrup, Published online at: http://www.siropderable.ca/home.aspx (accessed April 28, 2014).
In the past few years, in collaboration with FPAQ, our laboratory has been involved in a comprehen-
sive research program to isolate and identify bioactive compounds present in maple plant species and
maple food products, namely syrup and sap [6–7,13–16]. Interestingly, the majority of phytochemicals
present in maple water and maple syrup are found as phenolics as reported by our group [6–7,13] and
others [3,17–19]. However, data on the identification of phenolic compounds in maple water in relation to
its functional beverage applications is scarce. Because of this lack of data, our group recently evaluated
the effects of pasteurization (at 78°C) and sterilization (at 140°C) on the chemical composition (sugars,
free amino acids, organic acids, minerals, and phenolics) and antioxidant activities of maple water [11].
To the best of knowledge, this was the first report to characterize the chemical composition and evaluate
the antioxidant activities of maple water after undergoing thermal treatments. It should be noted that
these sterilization processes are necessary to reduce and/or eliminate microbial growth, which inevitably
happens in sap due to its largely watery nature [20]. Therefore, maple water has to be pasteurized and/
or sterilized to eliminate or reduce microbial growth before it would be safe and suitable for large-scale
human consumption as a commercial beverage.
As previously reported, we generated ethyl acetate extracts of the maple water samples, before and
after the thermal treatments, to evaluate their antioxidant activities by the 2,2′-diphenyl-1-picrylhydrazyl
(DPPH) radical scavenging assay [11]. The antioxidant values of maple water in their original, pasteur-
ized, and sterilized forms had IC50 values of 597, 567, and 511 μg/mL, respectively, proving that the
antioxidant activity was being maintained despite the thermal processes (Table 58.2). Notably, many
functional beverages include additives, such as ascorbic acid (vitamin C), to boost their antioxidant
effects beyond their natural constituents alone. Thus, it is impressive that the phenolic constituents pres-
ent in natural maple sap are exerting an antioxidant activity to the beverage without any antioxidant
additives such as vitamin C.
As mentioned earlier, we have previously isolated and elucidated the chemical structures using nuclear
magnetic resonance (NMR) of over 50 compounds from maple syrup, which are predominantly pheno-
lics and belong to various subclasses of lignans, flavonoids, coumarins, and stilbene [6–7,13]. Thus, using
a combination of high-performance liquid chromatography (HPLC) by comparison of retention time and
ultraviolet chromatograms to the previously isolated maple standards, as well as through further isolation
efforts and subsequent structure elucidation (by NMR), we were able to identify a total of 16 phenolic
compounds in maple water. The identities of these compounds are listed in Table 58.3, and their chemical
structures are shown in Figure 58.1. Notably, over 25 individual peaks, indicative of phenolic compounds
(at a detection wavelength of 280 nm), were observed in the maple water samples, but only 16 phenolics
were identified. Thus, there remain a large number of compounds yet to be identified in maple water.
The major compounds identified in maple water thus far also belong to the lignan subclass as previously
observed for maple syrup. Therefore, it is possible that several of the currently unidentified compounds in
maple water would have persisted in maple syrup during its production (in the intensive heating process),
and therefore, there could be considerable overlap in the presence of these compounds in maple water
and maple syrup. However, further research would be required to confirm this.
TABLE 58.2
Total Phenolic Content and Antioxidant Activity of Maple Water Extracts
Total Phenolic Content DPPH Radical Scavenging Activity
Samples (mg GAE/100 g)a (IC50 in μg/mL)b
Maple water 0.25 ± 0.09 597 ± 161
Pasteurized maple water 0.27 ± 0.06 567 ± 92
Sterilized maple water 0.27 ± 0.06 511 ± 92
Source: Adapted from Yuan, T. et al., J. Funct. Foods, 5, 1582, 2013. With permission.
Note: The data are presented as mean ± standard deviation.
a Assay conducted with neat samples of original maple water.
b Assay conducted with ethyl acetate extracts of maple water.
Abbreviations: GAE, gallic acid equivalents; DPPH, 2,2′-diphenyl-1-picrylhydrazyl.

Maple Water 761
TABLE 58.3
Phenolic Compounds Identified in Maple Water
No. Compound
1 C-Veratroylglycol
2 2,3-Dihydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-propanone
3 3-Hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)propan-1-one
4 Vanillin
5 Syringaldehyde
6 Threo-guaiacylglycerol-β-O-4′-dihydroconiferyl alcohol
7 Erythro-guaiacylglycerol-β-O-4′-dihydroconiferyl alcohol
8 3-[4-[(6-Deoxy-α-l-mannopyranosyl)oxy]-3-methoxyphenyl]methyl]-5-(3,4-dimethoxyphenyl)dihydro-3-hydroxy-
4-(hydroxymethyl)-2(3H)-furanone
9 5-(3″,4″-Dimethoxyphenyl)-3-hydroxy-3-(4′-hydroxy-3′-methoxybenzyl)-4-(hydroxymethyl)dihydrofuran-2-one
10 Erythro-1-(4-hydroxy-3-methoxyphenyl)-2-[4-(3-hydroxypropyl)-2,6-Dimethoxyphenoxy]-1,3-propanediol
11 Icariside E4
12 3′,4′,5′-Trihydroxyacetophenone
13 Dehydroconiferyl alcohol
14 Acernikol
15 2-[4-[2,3-Dihydro-3-(hydroxymethyl)-5-(3-hydroxypropyl)-7-methoxy-2-benzofuranyl]-2,6-dimethoxyphenoxy]-
1-(4-hydroxy-3-methoxyphenyl)-1,3-propanediol
16 3′,5′-Dimethoxy-4′-hydroxy-(2-hydroxy)acetophenone
Source: Adapted from Yuan, T. et al., J. Funct. Foods, 5, 1582, 2013. With permission.
OH OH
O O 8
7 R
H3CO R1
OH R2 O
R2 HO
HO HO OCH3 OH
R1 R3 H3CO
1 R1=H, R2 =OH 4 R1=R2=H , R3=OCH3 6 7,8-threo, R=H
2 R1=OCH3, R2 =OH 5 = =
R1 R3 OCH3, R2 =H 7 7,8-erythro, R=H
3 R1=OCH3, R2 =H 12 R1=R3=OH, R 2
=CH3 10 7,8-erythro, R=OCH3
H3CO
OCH3 OCH3 OCH3
OH
O HO O
RO 7 8
O
OH OH
HO
H3CO HO H3CO HO
11 R=Rham 14 7,8-erythro
13 R=H 15 7,8-threo
O
HO
O
H3CO O OCH3
H3CO OCH3
H3CO HO HO OR
OH
8 R=Rham 16
9 R=H
FIGURE 58.1 The chemical structures of phenolic compounds identified in maple water. (Adapted from Yuan, T. et al.,
J. Funct. Foods, 5, 1582, 2013. With permission.)
58.4 Health Effects

It is now widely accepted that the consumption of phytochemical-rich foods and beverages may prevent
and/or delay the onset of oxidative-stress and inflammatory-mediated diseases. This is supported by
scientific and epidemiological data, which has led to increased public and research interest in so-called
“functional foods and nutraceuticals” a fast-growing sector of the modern food industry.
Among the diverse chemical subclasses of phytochemicals, phenolics are arguably the most ubiqui-
tous of these compounds present in human diet. Moreover, phenolics are abundant in colorful fruit and
vegetables, grains, and plant-derived foods and beverages such as dark chocolate, fruit juices, tea, red
wine, and coffee.
Maple water is primarily consumed in our food chain in its concentrated form, namely, maple syrup,
a premium natural sweetener. Therefore, it is not surprising that more studies have been conducted on
maple syrup compared to maple water. While the chemical constituents and biological effects of maple
syrup have been recently reviewed [21], there are limited published data on the biological effects of
maple water.
Thériault et al. [18] have shown that phenolic-enriched extracts of maple sap, collected at different
periods in the season, exhibit antioxidant and antimutagenic activities. Interestingly, sap collected later
in the season had higher concentrations of phenolics, which increased from 16.51 (early season) to 24.6 g
gallic acid equivalents (GAE)/100 g (late season). Legault et al. [22] also investigated the antioxidant
activity, inhibition of nitric oxide overproduction in RAW 264.7 cells (to measure anti-inflammatory
activity), and human cancer cell antiproliferative effects of maple sap and maple syrup extracts collected
from 30 producers at different periods of harvest from three different regions in Quebec, Canada [22].
Similar to Thériault et al. [18], this study also showed that later collections of sap contained higher
levels of phenolics and as a result, displayed enhanced biological activity. Therefore, these studies sup-
port the hypothesis that phenolic compounds present in maple sap and syrup are contributing to their
observed biological effects. We have also recently reported on the anti-inflammatory effects, and under-
lying mechanisms of actions, of a phenolic-enriched maple syrup extract in the RAW 264.7 macrophage
model [23]. Despite these reports, the overall contribution of other substances present in maple sap and
maple syrup, including “nonphenolic” substances, to their observable biological effects should not be
excluded. Moreover, it has been shown that multiple constituents present in a whole food or extract exert
greater biological effects in toto than any single purified constituent alone due to additive, synergistic,
and/or complementary effects.
To date, there are no clinical studies to support the health benefits of maple water apart from anecdotal
reports arising out of the traditional medicinal uses of maple water as a tonic by the indigenous peoples
of North America and other cultures such as in South Korea. Considering that maple water is newly
emerging in the functional beverage market, it is possible that there may be future preclinical and clinical
studies to support the potential health benefits, if any, of this beverage. Given the dearth of data support-
ing the biological effects of natural compounds and the unique combination of macronutrients (including
complex polysaccharides), micronutrients (vitamins and minerals), and phytochemicals (including phe-
nolic and nonphenolic compounds) in maple water, it is possible that this beverage could impart health
effects when consumed by humans. However, further studies would be required to confirm this.

Maple water can be regarded as “vegetable-derived natural water” that has immense potential as a new
functional beverage. In this regard, it is important to discuss the integral role of the previously mentioned
organization, FPAQ, in the commercial development of this natural product in Canada with ~7400 maple
businesses across the province.
The maple growers and producers in Quebec have established and sophisticated technology for large-
scale collection and processing of maple water since they are the world’s leader in the commercial pro-
duction of maple syrup (~80%). Indeed, FPAQ has developed proprietary and patent pending methods
Maple Water 763
to develop sterilization techniques that preserve many of the original properties of maple water for
18 months at room temperature. To ensure authenticity and quality, these techniques are grouped with
an accompanying NAPSI certification seal. The harvesting methods and processes of NAPSI-certified
products meet strict standards that guarantee that the maple water is
• Natural: Harvested from maple trees.

• Authentic: It is the same sap that the trees themselves produce.
• Pure: No agents or extra ingredients are added.
• Sterile: Devoid of any microorganism.
• Integral: It is unrefined, not from concentrate, and contains all the original compounds pro-
vided by nature.
Thus far, there are already commercial maple water products available in Canada, and it is projected
that maple water will help promote economic growth through the added value generated by the NAPSI
process. In terms of future trends, it is projected that maple water, as a new product, will provide great
potential for growth opportunities in the Quebec maple syrup industry, which already contributes up to
three-quarters of a billion dollars to Canada’s GDP ($750 million).
58.6 Conclusion
Currently, maple water is primarily utilized in eastern North America to produce maple syrup, a pre-
mium natural sweetener. However, the great abundance and refreshing palatability of maple water make
its functional beverage applications very attractive from both a business and consumer perspective.
In addition, from a research perspective, the unique combination of macronutrients, micronutrients, and
phytochemicals in maple water, and the preservation of these constituents after pasteurization and ster-
ilization (in order to reduce microbial growth so it can be suitable for large-scale commercial production
for human consumption), supports it as an emerging and attractive functional beverage candidate for the
modern food industry. Ultimately, though, the commercial success of this functional beverage would
require considerable investment and commitment such as has already been started by FPAQ, in the prov-
ince of Quebec in Canada, where the vast majority of worldwide supply of maple syrup is commercially
produced.
Acknowledgment
The authors would like to acknowledge the Federation of Quebec Maple Syrup Producers (FPAQ) in
Canada for leading and supporting the research necessary for the large-scale commercial development
of maple water as a functional beverage as detailed in this chapter.
REFERENCES
1. Yong, J.W.H., Ge, G., Ng, Y.F., and Tan, N.S., The chemical composition and biological properties of
coconut (Cocos nucifera L.) water. Molecules, 14, 5144–5164, 2009.
2. The New York Times, South Korea, drinks are on the maple tree, March 2009, Published online at: http://
www.nytimes.com/2009/03/06/world/asia/06maple.html (accessed April 28, 2014).
3. Potter, T.L. and Fagerson, I.S., Phenolic compounds in maple sap, in Phenolic Compounds and Their
Effects on Health, part I, Ho, C.T., Lee, C.Y., and Huang, M.T., Eds., ACS Symposium Series 506,
American Chemical Society, Washington, DC, 1992.
4. Ball, D.W., The chemical composition of maple syrup. J. Chem. Educ., 84, 1647–1650, 2007.
5. Perkins, T.D. and van den Berg, A.K., Maple syrup-production, composition, chemistry, and sensory
characteristics. Adv. Food Nutr. Res., 56, 101–143, 2009.
6. Li, L. and Seeram, N.P., Maple syrup phytochemicals include lignans, coumarins, a stilbene and other
previously unreported antioxidant phenolic compounds. J. Agric. Food Chem., 58, 11673–11679, 2010.
7. Li, L. and Seeram, N.P., Further investigation into maple syrup yields three new lignans, a new phenyl-
propanoid, and twenty-six other phytochemicals. J. Agric. Food Chem., 59, 7708–7716, 2011.
9. Federation of Quebec Maple Syrup Producers (FPAQ), Quebec maple syrup, Published online at: http://
www.siropderable.ca/home.aspx (accessed April 28, 2014).
10. Health Canada, Canada’s food guide, February 2007. Published online at: http://www.hc-sc.gc.ca/
index-eng.php (accessed April 28, 2014).
11. Yuan, T., Li, L., Zhang, Y., and Seeram, N.P., Pasteurized and sterilized maple sap as functional bever-
ages: Chemical composition and antioxidant activities. J. Funct. Foods, 5, 1582–1590, 2013.
12. Shahidi, F. and Ho, C.-T., Phenolics in food and natural health products: An overview, in Phenolic
Compounds in Foods and Natural Health Products, Shahidi, F. and Ho, C.-T., Eds., ACS Symposium
Series 909, American Chemical Society, Washington, DC, 2005, pp. 1–8.
13. Li, L. and Seeram, N.P., Quebecol, a novel phenolic compound isolated from Canadian maple syrup.
J. Funct. Foods, 3, 125–128, 2011.
14. Yuan, T. Wan, C., Gonzalez-Sarrias, A., Kandhi, V., Cech, N.B., and Seeram, N.P., Phenolic glycosides
from sugar maple (Acer saccharum) bark. J. Nat. Prod., 74, 2472–2476, 2011.
15. Yuan, T., Wan, C., Liu, K., and Seeram, N.P., New maplexins F-I and phenolic glycosides from red
maple (Acer rubrum) bark. Tetrahedron, 68, 959–964, 2012.
16. Wan, C., Yuan, T., Li, L., Kandhi, V., Cech, N.B., Xie, M., and Seeram, N.P., Maplexins, α-glucosidase
inhibitors from red maple (Acer rubrum) stems. Bioorg. Med. Chem. Lett., 22, 597–600, 2012.
17. Kermasha, S., Goetghebeur, M., and Dumont, J., Determination of phenolic compound profiles in maple
products by high performance liquid chromatography. J. Agric. Food Chem., 43, 708–716, 1995.
18. Théirault, M., Caillet, S., Kermasha, S., and Lacroix, M., Antioxidant, antiradical and antimutagenic
activities of phenolic compounds present in maple products. Food Chem., 98, 490–501, 2006.
19. Abou-Zaid, M.M., Nozzolillo, C., Tonon, A., Coppens, M., and Lombardo, A.D.A., High performance
liquid chromatography characterization and identification of antioxidant polyphenols in maple syrup.
Pharm. Biol., 46, 117–125, 2008.
20. Filteau, M., Lagacé, L., Lapointe, G., and Roy, D., Maple sap predominant microbial contaminants are
correlated with the physicochemical and sensorial properties of maple syrup. Int. J. Food Microbiol.,
154, 30–36, 2012.
21. Li, L. and Seeram, N.P., Chemical composition and biological effects of maple syrup, in Emerging
Trends in Dietary Components for Preventing and Combating Disease, Patil, B.S., Jayaprakasha, G.K.,
Murthy, K.N.C., and Seeram, N.P., Eds., ACS Symposium Series 1093, American Chemical Society,
Washington, DC, 2012, pp. 323–333.
22. Legault, J., Girard-Lalancette, K., Grenon, C., Dussault, C., and Pichette, A., Antioxidant activity, inhi-
bition of nitric oxide overproduction, and in vitro antiproliferative effect of maple sap and syrup from
Acer saccharum. J. Med. Food, 13, 460–468, 2010.
23. Nahar, P.P., Driscoll, M.V., Li, L., Slitt, A.L., and Seeram, N.P., Phenolic mediated anti-inflammatory
effects of a maple syrup extract in RAW 264.7 murine macrophages. J. Funct. Foods, 6, 126–136, 2014.
Section VII
Fermented and Fortified

Functional Beverages
59
Fermented Functional Beverages
(Kefir, Koumiss, and Ayran)
Frank Sherkat, Kambiz Shamsi, and Amir Arjmand
CONTENTS
59.1 Introduction....................................................................................................................................767
59.2.1 Kefir.................................................................................................................................. 768
59.2.2 Koumiss............................................................................................................................ 769
59.2.3 Ayran................................................................................................................................ 769
59.3.1 Antihypertensive Peptides................................................................................................ 772
59.3.2 Immunomodulating Peptides........................................................................................... 772
59.3.3 Antitumor and Anticarcinogenic Peptides....................................................................... 773
59.3.4 Opioid Peptides................................................................................................................. 773
59.3.5 Antithrombotic Peptides................................................................................................... 773
59.3.6 Antioxidative Peptides...................................................................................................... 773
59.3.7 Mineral-Transporter Peptides........................................................................................... 773
59.3.8 Antimicrobial Peptides..................................................................................................... 773
59.4 Health Effects.................................................................................................................................774
59.4.1 Cardiovascular and Endothelial Health.............................................................................775
59.4.2 Digestive Health............................................................................................................... 776
59.4.3 Alleviation of Lactose Intolerance................................................................................... 776
59.4.4 Anticarcinogenic Effect.................................................................................................... 776
59.4.5 Protection from Osteoporosis........................................................................................... 777
59.4.6 Stimulation of the Immune System.................................................................................. 777
59.4.7 Antiallergy Effects........................................................................................................... 777
59.4.8 Lifestyle and Well-Being.................................................................................................. 777
59.4.9 Other Health Benefits....................................................................................................... 778
59.5.1 Novel Approaches to Traditional Fermented Dairy Beverages....................................... 778
59.5.2 Future Trends and Perspectives........................................................................................ 779
59.6 Conclusion..................................................................................................................................... 780
References............................................................................................................................................... 780
59.1 Introduction
Health-conscious consumers in modern societies routinely consume functional foods and drinks
believing that they promote better health and well-being, prevent the onset of chronic diseases, and
increase longevity. Fermented dairy products are on the top of the list of such products since the
origin of their consumption dates back to antiquity. Many types of traditional fermented milks have
originated in different geographical regions by spontaneous fermentation of the milk of at least
767
11 species of animals including cow, buffalo, goat, sheep, mare, donkey, camel, yak, and reindeer,
among others.
The spontaneous fermentation of milk was triggered by the action of native milk bacteria as well as
the adventitious lactic acid bacteria (LAB), yeast, and molds from the environment, the vessels (e.g.,
bags made of animal hides), or adding the leftovers of the previous day’s sour milk. Beverages such as
kefir and koumiss have been produced and consumed for millennia with proven health benefits, from the
Steppes of Central Asia to the Middle East, and from the Caucasus and Asia Minor to Eastern Europe,
for their refreshing qualities and nutritional and health benefits. The pioneering works of Metchnikoff in
the early twentieth century helped describe the lactic fermentation and paved the way for better under-
standing of the health benefits of fermented milk products. It is claimed that over 3,500 different types of
fermented foods and drinks are produced across the globe and are among the most commonly consumed
functional foods. This chapter highlights the nutritional and health benefits of three main traditional
dairy beverages (kefir, koumiss, and ayran) and describes the future trends in developing more stable
products and formulations to meet the ever-increasing demand for these beverages.

Nutritional profile of kefir, koumiss, and ayran is determined by the type and composition of the milk
used and the microbiological and biochemical changes the milk undergoes during fermentation.
59.2.1 Kefir
Kefir is a self-carbonated beverage made with milk of cow or goat, using kefir grains. It originated in
Caucasian mountains and eventually found its way to Mongolia [1]. Kefir has been consumed as a popu-
lar beverage associated with health and longevity from Inner Mongolia to Russia for thousands of years.
It has gained popularity in the Southwest Asia, Eastern and Northern Europe, North America, Japan,
Middle East, North Africa, Brazil, and Israel [2,3].
Kefir grain (Figure 59.1) is a complex colony of more than 30 different beneficial microorganisms
(including various species of Lactobacillus, Lactococcus, Leuconostocs, acetic acid bacteria, and
yeast species, both lactose fermenting and nonlactose fermenting) [4,5]. The grain is constituted of
FIGURE 59.1 Kefir grain.

Fermented Functional Beverages (Kefir, Koumiss, and Ayran) 769
108 –10 9 CFU/mL LAB, 105 –10 6 CFU/mL yeasts, and 105 –10 6 CFU/mL acetic acid bacteria held
symbiotically in a gelatinous polysaccharides biofilm matrix, known as kefiran, to form a protective
colony against outside pathogens [6,7]. The composition, flavor, and nutritional characteristics of kefir
vary considerably from region to region depending on the type of milk and the concentration of strains of
each group of microflora within the kefir grain (Table 59.1) [8–10]. The Codex standard for kefir compo-
sition is 2.80% milk protein, <10.0% milk fat, and a minimum of 0.6% titratable acidity (as lactic acid),
107 CFU/g LAB, and minimum 104 CFU/g of yeasts [11].
The enzymes produced by kefir microflora increase the digestibility and biological value of kefir pro-
teins. This makes kefir a good source of bioactive peptides, essential amino acids, and conjugated lin-
oleic acid (CLA) [12]. Amino acid profile of kefir increases during fermentation and storage with higher
levels of threonine, serine, alanine, and lysine than milk [12,13].
Kefir has lower lactose content than the milk from which it is made due to the alcoholic and lac-
tic fermentation processes, thus is better tolerated by lactose malabsorbers. Lactose digestion by the
action of β-galactosidase continues in the gastrointestinal tract (GIT) due to slower transit time of
fermented milks than milk [9]. Almost all of the lactic acid produced during kefir fermentation is L(+)
isomer, but D(−) may also be produced depending on the type of grains used. The L(+) is more readily
absorbed and converted to glycogen than D(−) lactic acid that is mostly excreted in the urine. Kefir also
contains acetaldehyde, acetoin, diacetyl, ethanol, CO2, vitamins, and amino acids [14]. It is an excel-
lent source of B vitamins especially biotin that aids body’s assimilation of other B vitamins such as
folic acid, pantothenic acid, and B12. The vitamin content of kefir is influenced by the type of milk as
well as by the supplementing flora [15]. Propionibacterium species are more effective in the synthesis
of B vitamins. Ewe’s milk kefir has been shown to contain more thiamin (B1) than that made from
cow’s milk [15]. The fermentation process increases the solubility and bioavailability of kefir minerals
such as calcium, iron, phosphorus, magnesium, potassium, sodium, copper, zinc, molybdenum, and
manganese [9,16–18].
59.2.2 Koumiss
Koumiss is a mildly alcoholic and lightly carbonated milk beverage, with a mild to sharp acidic taste that
does not coagulate [19–21]. It is made from equine (mare’s) milk by lactic and alcoholic fermentation and
is mainly consumed in Central Asia, Turkey, and Russia [22,23]. Mares’ milk and its products are widely
used in Russia and Mongolia, and a considerable interest has been reported regarding their use for human
consumption in Western Europe [24]. Malacarne et al. [25] reported the energy value of mare’s milk as
480 kcal/L with an average composition of 2.1% protein, 1.2% fat, 6.4% lactose, and 0.4% ash. With
higher level of immunoglobulins than cow’s milk and with a 1:1 ratio of casein to whey proteins, mare’s
milk is closer to human milk and is thus recognized as an important nutritional resource, especially for
the elderly, convalescents, and infants [26,27].
Koumiss is made by adding 10%–30% of previous day batch to boiled mare’s milk at 26°C–28°C.
Depending on the inoculum rate, incubation time and temperature, mild, medium, or strong koumiss
may be made, with pH ranging from 3.3 to 5.0 (Table 59.1). Koumiss microflora includes Lactobacillus
(L. delbrueckii subsp. bulgaricus, L. salivarius, L. buchneri, L. plantarum, L. casei, L. helveticus, and
L. fermentum) [28], lactose-fermenting yeasts (Saccharomyces lactis, Torula koumiss, Kluyveromyces
lactis), and nonlactose-fermenting yeasts (S. unisporus) [29,30]. Koumiss has low mineral content; how-
ever, with a calcium-to-potassium ratio of 2:1, it is quite suitable for human consumption [31]. Despite the
high lactose content of mare’s milk (6.2%), the strong koumiss may contain as low as 1.4% lactose and up
to 4.4% in mild variety. Koumiss also contains B vitamins (Table 59.1), a high ratio of polyunsaturated
fatty acids (PUFA), and low cholesterol content [31].
59.2.3 Ayran
Ayran is a refreshing beverage made simply by diluting 1/3 yogurt with 2/3 water (sometimes milk),
lightly salted, and topped with diced mint or other aromatic herbs and may or may not be carbonated.
It has a smooth and creamy mouthfeel and a mildly salty, and sour flavor with no added sugar and
TABLE 59.1
Compositional, Nutritional, and Physicochemical Characteristics of Kefir, Koumiss, and Ayran (per 100 mL)
Unit Kefir Koumiss Ayran References
Water g 87.5 88.7–89.4 84.4–96.4 [1,3,4,6,7,10,11,15,20,21]
Protein g 3.0–3.4 2.0–2.5 1.5–3.1
Fat (lipid) g 0.1–3.5 0.6–1.3 1.50–1.55
Carbohydrate (lactose) g 1.8–3.8 1.4–4.4 1.5–3.0
Ash g 0.5–0.7 0.4–1.0 0.7–1.0 [1,3,4,10]
pH 4.6 3.3–5.0 3.35–3.89
Minerals
Calcium mg 86–160 168 40–50 [1,3,4,15,35]
Cobalt µg 160 na na
Copper mg 0.73 0.15–0.16 0.10–0.20
Iron mg 0.05–2.0 0.05–2.0 0.10
Manganese mg 1.3 0.004 na
Magnesium mg 12–14.5 0.10–0.16 7.5–16.5
Molybdenum µg 330 na na
Phosphorus mg 100 86–137 11.5–18.8
Potassium mg 150–165 na 45–86
Sodium mg 50 0.45–1.0 47–98
Zinc mg 9.27 0.18 0.3–0.6
Vitamins
Biotin mg na na na [3,4,6,15,16,20,35]
Cobalamin (B12) mg 0.5 tr na
Folate (DFE) mg na na na
Niacin mg 0.09–0.3 1.06 na
Pantothenic acid (B5) mg 0. 3 tr 0.19
Pyridoxine mg 0.05 na tr
Riboflavin mg 0.13–0.17 0.03 0.16
Thiamin mg 0.04–1.0 0.02 na
Vitamin A (RAE) mg 0.06 0.01 na
Vitamin C mg 1.0 10–25 tr
Vitamin K mg na 1.25–2.60 na
Ethanol g Industrial: 0.2–0.3 0.7–2.5 na [5,13,23,29]
Authentic: 1.0–3.0
Lactic Acid g 0.6–3.7 0.6–1.2 0.45–0.61
Carbon Dioxide g 0.2 0.5–0.9 na
Amino Acids g 1.75 2.19 na
Organic Acids
Acetic acid mg/mL na 0.95 na [14,22,24,31]
Citric acid mg/mL 1760 0.91 na
Pyruvic acid mg/mL 18–68 0.07 na
Uric acid mg/mL na 0.01 na
Other Substances
Acetaldehyde µg/mL 5–21 na 7.6–10.9 [14,17,22,24,27,31]
Acetoin µg/mL 16–25 na na
Diacetyl µg/mL tr tr na
Abbreviations: DFE, dietary folate equivalents; RAE, retinol activity equivalents; tr, trace; na, not available.
is nonalcoholic. The best ayran is made from ewe’s milk yogurt and is best served chilled. Ayran is very
popular in Turkey, where it has recently been proclaimed the national summer drink. It is consumed in
many countries under different local names such as aryan in Bulgaria; ariani or ayráni in Greece; doogh
in Iran and Afghanistan; shinēna in Iraq; dew or do in Syria, Lebanon, Kazakhstan, Kyrgyzstan, and
across the Caucasus; eyran in Azerbaijan; tahna in Armenia; and dawe in Assyrian/Chaldean region.
Nutritionally, it provides many nutrients such as proteins and essential amino acids, short-chain fatty
acids, vitamins (riboflavin and B5), and most minerals (such as calcium, copper, iron, magnesium, potas-
sium, phosphorus, sodium, and zinc), including highly absorbable calcium in a low-energy beverage.
Milk and fermented milks, including kefir, koumiss, and ayran, are known to modulate the satiety,
thus controlling food intake and obesity-related metabolic disorders. This effect is measured by high
plasma concentrations of satiety factors such as amino acids, glucose-dependent insulinotropic polypep-
tide, glucagon-like peptide-1, and cholecystokinin (CCK) [32]. The mechanisms involved is proposed to
include slowing the GIT motility, perhaps via opioid receptors, and direct or indirect stimulation of gut
hormone receptors, including CCK and glucagon-like peptide-1 receptors [33].

The origin of bioactive peptides found in fermented dairy beverages can be traced back to three main
sources; milk proteins, the products of milk fermentation by starter and nonstarter bacteria, and the
bioactive compounds released from the lysed bacterial cells in the GIT.
Caseins and whey proteins in their native forms play important physiological roles in the body [34–36].
However, during fermentation, milk proteins undergo a series of catabolic changes by the action of
proteases synthesized by the LAB and liberate bioactive peptides, the cell wall peptidoglycan, and cyto-
plasmic fractions that further improve the nutritional and health-promoting functions of the beverage.
Milk bioactive peptides may function as exogenous regulatory substances with hormonelike activity on
different intestinal and peripheral target sites of the mammalian organism. The peptides released from
casein fermentation such as opiate, antiopiate, antihypertensive, mineral transporter, immunomodifier,
platelet aggregation inhibitor, and mitogenous peptides are further activated by the digestive enzymes
in human GIT [36,37]. These bioactive peptides demonstrate antimicrobial, antihypertensive, antioxida-
tive, anticytotoxic, immunomodulatory, opioid, and mineral-carrying functionalities that impart health-
promoting and disease-preventing properties to fermented milk products [38]. The type and amount
of these bioactive compounds are determined by the milk type, the type of the product made, types
and numbers of starter and nonstarter bacteria used, incubation time, and storage conditions. Some are
metabolized by microorganisms, such as enzymes, vitamins, antibiotics, and bacteriocins, while others
result from the breakdown of food matrix, such as peptides, organic acids, and carbonyl compounds,
each with specific function and health-promoting attribute [38].
Most starter and nonstarter cultures employed in the production of kefir, koumiss, and ayran possess
a complement of proteolytic enzymes that enable them to release bioactive peptides from milk proteins.
For example, L. lactis, L. helveticus, L. plantarum, L. rhamnosus, L. acidophilus, L. delbrueckii ssp.
Bulgaricus, and Streptococcus thermophilus possess a vast array of cell-wall-bound proteinases, trans-
port system, and intracellular peptidases including endopeptidases, aminopeptidases, tripeptidases, and
dipeptidases. The extracellular proteinases degrade caseins to long-chain oligopeptides, the transport
systems allow the internalization of some of the released oligopeptides, and the intracellular peptidases
further hydrolyze them into smaller peptides and amino acids to cater for bacterial nitrogen need [37,38].
Studies on LAB have found that less than 25% of casein-derived oligopeptides are transported inside the
bacterial cell to sustain growth, the nontransported peptides accumulating in the fermentation medium
to constitute a significant pool of biologically active peptides [39]. These oligopeptides having undergone
a number of proteolytic changes during fermentation become more prone to producing bioactive peptides
such as valyl-prolyl-proline (VPP) and isoleucyl-prolyl-proline (IPP) by the action of pepsin and trypsin
in human GIT [40].
Caseins (αs1-CN and β-CN) act as precursors for casomorphins (opioid antagonists), casokinins
(inhibitors of angiotensin I–converting enzyme [ACE]), phosphopeptides (mineral binding), and
immunopeptides (immunomodulatory), whereas κ-casein is the precursor for casoxins (opioid antagonist),
casoplatelins (antithrombotic), and glycomacropeptide. Hydrolysis of α-lactalbumin and β-lactoglobulin
results in lactorphins (opioid agonist) and lactokinins (ACE inhibitor), whereas serum albumin hydroly-
sis produces the opioid agonist serorphin [41].
The functionality of bioactive peptides is dependent on their inherent amino acid sequence that
may vary from 2 to 20 amino acid residues. Furthermore, many bioactive peptides are known to have
multifunctional properties [42]. For example, the antimicrobial peptide Casecidin 17 also exerts ACE-
inhibitory, immunomodulating, antioxidant, and antithrombotic activities, whereas β-CN f(59–68) also
called V-β-casomorphin-9 exhibits ACE-inhibitory, antioxidant, and opioid activity. The β-CN f(60–70)
shows immunostimulatory, opioid, and ACE-inhibitory activities [43]. The αs1-CN f(194–199) shows
immunomodulatory and ACE-inhibitory activities, whereas the opioid peptides α- and β-lactorphin also
exhibit ACE-inhibitory activity, and the calcium-binding phosphopeptides (caseinophosphopeptides
[CPPs]) also show immunomodulatory properties [42–44].
The extracellular proteases released by the LAB during milk fermentation are essential for the produc-
tion of multitude of bioactive peptides from caseins. Strains of L. helveticus are shown to release anti-
hypertensive peptides, especially ACE-inhibitory tripeptides IPP and VPP. Cell membrane proteinase is
believed to be the key enzyme capable of initiating the degradation of caseins to β-CN f(74–76), β-CN
f(84–86), and κ-CN f(108–110) [45]. Kefir is shown to have a higher rate of proteolysis, hence a poten-
tial source of bioactive peptides. Liu et al. [46] reported that both cow’s milk kefir and soy milk kefir
possessed significant antimutagenic and antioxidant activities. Ebner et al. [47] reported a 1.7–2.4-fold
increase in peptide content of kefir compared to unfermented milk. A total of 257 peptides were identi-
fied mainly released from β-CN (43%), followed by αs1- (22%), κ- (21%), and αs2-CN (14%). No bioactive
peptides from the major whey proteins were found. The majority of peptides (236) were unique to kefir,
with 16 described as being bioactive, showing ACE-inhibitory, antimicrobial, immunomodulating, opi-
oid, mineral-binding, antioxidant, and antithrombotic effects.
59.3.1 Antihypertensive Peptides

ACE is an important regulator of blood pressure. This enzyme converts the inactive angiotensin I into
angiotensin II, a strong vasoconstrictor that affects ion retention and fluid excretion, thus increasing
the blood pressure. ACE-inhibitory peptides of fermented milks block the conversion of angiotensin I
to angiotensin II and prevent the degradation of the vasodilator and immunostimulating bradykinin.
Thus far, 12 sequences among kefir peptides are reported to be ACE inhibitors [47]. It has been sug-
gested that peptides with hydrophobic amino acids at the C-terminal position could be the most likely
ACE inhibitors [48]. Therefore, food proteins with proline-rich regions on their sequence are potential
candidates for ACE inhibition [49]. Some peptides such as Tyr-Pro, purified from yogurt fermented
with L. helveticus CPN4 strain, showed significant antihypertensive activity at low dose without show-
ing strong ACE-inhibitory activity [49]. Different strains of L. helveticus have been isolated from kefir
grains and koumiss [50]. Sedlábek et al. [51] isolated six strains of L. helveticus from natural koumiss
made from mare’s milk in the central Mongolia. Quirós et al. [52] isolated 16 ACE-inhibitory peptides
including PYVRYL and LVYPFTGPIPN from goat’s milk kefir.
59.3.2 Immunomodulating Peptides

Bioactive peptides formed during the fermentation of milk and subsequent digestions in the GIT have
been shown to stimulate the immune system in animal models [37,47]. In a kefir feeding trial to young
and old rats, an enhanced mucosal immune response measured as significant increase in total nonspe-
cific blood immunoglobulin G (IgG) level was reported [53]. The authors concluded that this effect may
be produced by bacterial cell wall components. Furukawa et al. [54] showed that oral administration of
the water-soluble polysaccharide of kefiran may act as a modulator of the immune response. Clinical
studies have shown that exopolysaccharides can stimulate the mammalian immune system by activat-
ing macrophages, thereby increasing the T-cell cascade and enhancing the secretion of cytokines [9,55].
Fedechko et al. [26] reported a strong immune response in white mice and chickens after feeding with
cow’s milk koumiss. According to Ebner et al. [47], the ACE-inhibitory peptides of kefir could also be
potential immunomodulators.
59.3.3 Antitumor and Anticarcinogenic Peptides

Recent studies have shown that several nonnutrient components in ayran, kefir, and koumiss, such as
sphingolipids, CLA, and butyric acid, may play a role as anticancer agents. Murofushi et al. [56] reported
that the water-soluble polysaccharides fraction of kefiran inhibited pulmonary metastasis of Lewis lung
carcinoma. The water-insoluble kefiran-containing microorganisms is reported to significantly inhibit
metastasis of highly colonized B16 melanoma through an increased natural killer cell activity [57].
59.3.4 Opioid Peptides

Milk-derived peptides are capable of binding to opioid receptors, thus exerting agonistic or antagonistic
effects, resulting in antidiarrheal effects or amino acid transport modulation. Matar et al. [37] demon-
strated that trypto-peptic hydrolysis of milk fermented with L. rhamnosus GG released opioid peptides
from casein. The β-casein f(60–70) and the β-casomorphin (YPFPGPIPASL) exert a morphine-like
activity, whereas peptide VYPFPGPIPN f(59–68) identified in kefir shows ACE-inhibitory, antioxidant,
and opioid activity [58].
59.3.5 Antithrombotic Peptides

It has been shown that the κ-caseinoglycopeptide (f106–116) inhibits the binding of human fibrinogen
on the platelet receptors, thus enhancing fibrinolysis and inhibiting platelet aggregation and thrombus
formation in the blood. Fiat et al. [59] reported the antithrombotic activity of casopiastrin, a fragment
obtained by tryptic hydrolysis of κ-casein that inhibited fibrinogen binding on platelets. The antithrom-
botic and immune response modulation properties of casoplatelins were reported [42].
59.3.6 Antioxidative Peptides

Several vascular diseases are linked to the increased levels of reactive oxygen species (ROS) that can
damage lipids, proteins, and the DNA. The unsaturated fatty acids in the cholesterol esters and phospho-
lipids can be oxidized by the ROS to produce hydroperoxides that render low-density lipoprotein (LDL)
even more atherogenic. Natural fat-soluble antioxidants present in blood plasma prevent this oxidation.
However, this defense mechanism can be outstripped leading to elevated coronary heart disease (CHD)
risk. Tsopmo et al. [60] isolated two peptides (YGYTGA and ISELGW) from human milk after a two-
step gastric digestion followed by incubation in the presence of pancreatin and bile salts. These peptides
showed high-proxyl radical scavenging activity and inhibition of lipid hydroperoxides due to the presence
of tryptophan that serves as a hydrogen donor or free radical scavenger, thus breaking the radical chain
reactions. Grishina et al. [61] reported a significantly greater antioxidant activity in the supernatants
of kefir and ayran than in milk. This was linked to high levels of acetic and lactic acids that was dose-
dependent with ayran, whereas with kefir the effect was shown to be stronger than vitamin E.
59.3.7 Mineral-Transporter Peptides

CPPs, the phosphoseryl residues of αs1-, αs2-, and β-CNs, are powerful chelators of divalent ions such
as Ca2+, Mg2+, Zn2+, and Cu2+. One kefir peptide, the β-CN f(29–41), has been confirmed for calcium-
binding property, but other 62 identified kefir phosphopeptides may also have this property.
59.3.8 Antimicrobial Peptides

The antimicrobial effects of kefir and koumiss have been reported to be due to competition for nutri-
ents from their microflora against other microorganisms via the synthesis of metabolites such as poly-
saccharides, peptides, bacteriocins, organic acids, and free fatty acids (FFA) during fermentation.
Cevikbas et al. [62] determined the antibacterial effects of kefir and kefir grains against Staphylococcus
aureus, S. epidermidis, Pseudomonas aeruginosa, Proteus vulgaris, Klebsiella pneumoniae, and Bacillus
subtilis. The greatest antimicrobial activity of kefir was exhibited against Gram-positive microbes and
against strains of Candida, Torulopsis glabrata, Microsporum nanum, Trichophyton mentagrophytes,
and T. rubrum. However, no antifungal effect was observed against Cryptococcus neoformans and
Candida parapsilosis.
Garrote et al. [63] studied the inhibitory activity of kefir and its organic acids (lactic and acetic acids)
on Gram-negative microorganisms isolated from human fecal matter and Gram-positive bacteria i solated
from food samples using the agar spot test or the agar well diffusion assay. Diameters of the inhibi-
tion zones against Gram-positive species were nearly twice that of Gram-negative microorganisms. The
authors related the inhibitory effect to the undissociated forms of lactic and acetic acids produced during
fermentation. L. plantarum ST8KF, isolated from kefir, was shown to produce a 35 kDa bacteriocin with
a narrow spectrum of activity. Kakisu et al. [64] reported that the incubation of milk with Bacillus cereus
spores in the presence of 5% kefir grains prevented spore germination and growth of vegetative forms.
The antimicrobial peptides produced during fermentation selectively bind to the outer lipid membrane
of the bacterial cell and cause blisters or pores that eventually lead to cell lysis and death. Most LABs
found in kefir also produce hydrogen peroxide with antimicrobial properties. Sphingolipids and their
metabolites may exert antimicrobial effects either directly or upon digestion.
Studies show that kefir and koumiss have significantly high concentration of FFA with C18:1 (oleic),
C18:2 (linoleic), and C18:3 (α-linolenic), nearly 1/3 being linoleic and α-linolenic acids. The LAB has
been shown to produce CLA from linoleic acid [65]. The addition of linoleic acid (up to 0.1%) to nonfat
yogurt has been shown to significantly increase the (c9t11) CLA isomer content without adversely affect-
ing the sensory properties.
59.4 Health Effects

Milk, the main ingredient of fermented dairy beverages, is the source of essential nutrients and a
broad range of biologically active compounds. Specific health effects claimed on milk proteins include
improvement of physical performance, recovery after exercise, and prevention of muscular atrophy, sati-
ety and weight management, cardiovascular health, anticancer effect, wound care and repair, manage-
ment of microbial infections and mucosal inflammation, hypoallergenic infant nutrition, and healthy
aging (Table 59.2). Fermented milks, due to their additional bioactive peptides, especially antimicrobial
peptides, produced during fermentation, are believed to be more effective than unfermented milks in
health promotion and treatment of diseases.
Kefir has been used in the former Soviet Union hospitals and sanatoria for the treatment of metabolic
disorders, allergies, atherosclerosis, tuberculosis, and cancer as well as to relieve gastrointestinal disor-
ders, promote bowel movement, reduce flatulence, and establish a balanced inner ecosystem for optimum
health and longevity [18,19]. It was used to increase the essential fatty acids intake in patients suffering
from metabolic and GIT disorders and those with atherosclerosis, ischemic heart disease, obesity, peptic
ulcers, and liver and gallbladder pathologies. Kefir has also been used to help patients suffering from
AIDS, chronic fatigue syndrome, herpes, and cancer [1,3].
Animal studies have shown the antimicrobial, immunological, antitumoral, and hypocholesterolemic
effects of kefir consumption. Tryptophan, an abundant essential amino acids in kefir, has been shown to
exhibit a relaxing effect on the nervous system [16]. Guzel-Seydim et al. [10] reported kefir’s antimuta-
genic, anticarcinogenic, and antimicrobial properties, cholesterol metabolism, ACE-inhibitory activity,
stimulation of the immune system, and alleviation of lactose intolerance. Casecidin 17 is considered
of particular significance to kefir’s health-promoting effects due to its abundance and multifunctional
properties [47]. Regular consumption of kefir is shown to help achieve better gut health, antimicrobial
activity, cancer prevention, control of serum glucose and cholesterol levels, control of lactose intolerance,
and better immune system [18,66].
Koumiss enjoys a long-established image as a healthy beverage, preventing stomach upset and promot-
ing general health. The LAB strains of koumiss show acid and bile tolerance, cholesterol reduction ability,
TABLE 59.2
Bioactive Functions of Milk Proteins
Milk Proteins Functions References
Caseins (αs1, αs2, β, Ion carriers (Ca, PO4, Fe, Zn, and Cu), precursors of bioactive peptides, [33,36,38,40–44,58,59]
and κ) immune regulatory, and anticarcinogenic.
β-Lactoglobulin Vitamin carrier, potential antioxidant, precursor for bioactive peptides, [32,40–44]
and fatty acid binding capacity.
α-Lactalbumin Effector of lactose synthesis in mammary gland, a calcium carrier, [32,40–44]
immunomodulatory, precursor for bioactive peptides, and potentially
anticarcinogenic.
Immunoglobulins Function as antibodies (70%–80% of total protein content in colostrum [36–38,43,44]
(IgG, IgM, and but only 1%–2% in milk) and are able to prevent the adhesion of
IgA) microbes and inhibit their metabolism by agglutination, promote
phagocytosis, and neutralize toxins and viruses.
Glycomacropeptide Antimicrobial, antithrombotic, prebiotic, and gastric hormone [32,38,40,58]
regulator.
Lactoferrin Antimicrobial, antioxidative, anticarcinogenic, anti-inflammatory, iron [35,36,38,40]
transport, cell growth regulation, with immunomodulatory properties,
and a precursor of bioactive peptides and stimulates osteoblast
proliferation.
Lactoperoxidase Antimicrobial enzyme and has synergistic effect with [38,40]
immunoglobulins, lactoferrin, and lysozyme.
Lysozyme An antimicrobial enzyme and has synergistic effect with [32,38,40,43,44]
immunoglobulins, lactoferrin, and lactoperoxidase.
Serum albumin A precursor for bioactive peptides. [32,40,44,59]
Milk basic protein Stimulates osteoblast proliferation and suppresses bone resorption. [32,35,36,40,44]
Growth factors Stimulate cell growth, protect and repair intestinal cells, and regulate [35,40,41,44,70]
immune system.
and activity against molds and pathogens such as Escherichia coli and S. aureus [20,28]. Favorable influ-
ence of koumiss consumption on the alimentary canal’s activity, the circulatory and nervous systems,
blood-forming organs, kidney functions, endocrine glands, and the immune system have been reported
[26–29]. The suitability of koumiss for weight gain, increasing energy, and alleviation of cardiovascular,
hepatitis, chronic ulcer, and gynecological disease symptoms is reported. Russian scientists reported
that koumiss can reduce the blood sugar and enhance insulin secretion, thus helpful for diabetic patients
[21,67]. Koumiss therapy is a well-established practice in medical centers in Mongolia and Russia for the
treatment of tuberculosis, emphysema, and pneumonia [68].
The health benefits of ayran stems from the yogurt from which it is made and is linked to the presence
of S. thermophilus, L. delbrueckii ssp. bulgaricus and prebiotic Lactobacillus and Bifidobacteria
(in probiotic yogurt). Despite the fact that ayran is a diluted drinking yogurt, it is usually consumed in
larger doses than yogurt. The health effects of ayran include alleviation of inflammatory bowel disease
and constipation, enhanced immune system, and weight management when prepared from low-fat or
nonfat yogurt. Researchers at Harvard School of Public Health reported that intake of 1 oz (28.3 g)
yogurt per day (equivalent to one glass of ayran) for several weeks can lower the risk of type 2 diabetes
by 18% [69]. The high level of B vitamins in kefir as well as koumiss and ayran have many health benefits
ranging from regulation of the kidneys, liver, and nervous system to help relieve skin disorders, boost
energy, and promote longevity [3,15].
59.4.1 Cardiovascular and Endothelial Health

High blood pressure is considered a major independent risk factor for cardiovascular disease (CVD).
The monolayer cell lining of the endothelium mediates vascular homeostasis, releasing substances in
response to physical or chemical stimulus that control vasodilation and vasoconstriction. Every 1%
reduction in vasodilation correlates to 12% increase in the risk of CVD. Peptides found in kefir and kou-
miss are shown to enhance the immune system and lower the blood pressure in animal models. Regular
koumiss consumption is shown to reduce blood lipids and cholesterol content and blood clot formation.
Studies have shown that the probiotic bacteria in yogurt and ayran can decrease the absorption of choles-
terol and promote cardiovascular health. The α-lactorphin derived from α-lactalbumin has been shown
to possess antihypertensive properties. The presence of ACE-inhibitory peptides, for example, IPP and
VPP, in yogurt/ayran can significantly reduce the blood pressure [47–52].
59.4.2 Digestive Health

Kefir, koumiss, and ayran containing LAB and probiotics can play a beneficial role in restoring the
imbalance of microflora in antibiotic-related diarrhea or in acute rotavirus diarrhea in young children.
The positive effects of kefir intake in preventing Campylobacter jejuni colonization in chicken, improv-
ing systemic immune response in rats, and inducing mucosal resistance to gastrointestinal infection
in mice have been reported [53]. Animal studies have shown that the β-casomorphins present in these
products demonstrate a potent antisecretory effect on the intestinal mucosa and can prolong gastroin-
testinal transit time by slowing intestinal peristalsis and motility [70]. Bifidokefir containing 5 × 107
CFU/mL of Bifidobacterium bifidum is reported to positively normalize human intestinal Bifidobacteria/
Lactobacillus balance with concurrent reduction in pathogen numbers within 1–2 weeks. Kefir has been
used in the treatment of peptic ulcers of stomach and duodenum.
59.4.3 Alleviation of Lactose Intolerance

During the fermentation, up to 30% of milk lactose is converted to lactic acid by the action of bacterial
β-galactosidase (EC 3.2.1.23). The β-galactosidase activity is reported to be low in kefir and koumiss;
however, the alcoholic fermentation further reduces the lactose content, and the subsequent intraintesti-
nal hydrolysis of the residual lactose by β-galactosidase attenuates the body’s reaction and makes it better
tolerated by lactose maldigesters [66]. Guzel-Seydim et al. [10] reported that the consumption of kefiran
by lactose maldigesters significantly reduces the incidence of flatulence.
59.4.4 Anticarcinogenic Effect

Kefir was found to be effective in inhibiting tumor growth and inducing apoptosis of human T-cell
lymphotropic virus type 1–negative malignant T-lymphocytes [71]. Feeding kefir (2 g/kg body weight)
has been shown to be more effective than yogurt in inhibiting the proliferation of Lewis lung carcinoma
[54,57]. Peritoneal injection of liquid kefir to mice was shown to arrest the growth of fusiform cell sar-
comas and disappearance of tumor necrosis [62]. Most of these effects could be linked to the presence
of casein-derived fractions, such as αs1-casein f(90–95), αs1-casomorphin f(158–162), β-casomorphins
5 f(60–64), β-casomorphins 7 f(60–66), and morphiceptin f(60–63)-NH2, and fractions derived from
whey proteins including bovine lactoferricin (LfcinB) in fermented milks. On this basis, kefir, koumiss,
and ayran can be used as a part of probiotic-based therapies of cancer patients to alleviate symptoms of
mucositis, a painful side effect of chemotherapy.
Sieber et al. [65] reported the conversion of some of milk fatty acids into CLA by the LAB during
fermentation. Reported health benefits of the CLA include reduced risk of carcinogenesis, athero-
sclerosis, and obesity, improved hyperinsulinemia, and prevention of catabolic effects of the immune
system [16,65]. Nisin, a bacteriocin produced by the LAB during fermentation of yogurt, kefir, and
koumiss, has been shown to induce preferential apoptosis, cell cycle arrest, and cell proliferation reduc-
tion in some types of cancer. These anticancer effects could be related to the detoxification of ingested
carcinogens, cellular uptake of mutagenic compounds, alteration of intestinal microflora, production
of short-chain fatty acids (e.g., butyrate), and inhibition of carcinogen-producing enzymes by other
colonic microbes [72].
59.4.5 Protection from Osteoporosis

Senile osteoporosis affects the long bones in both men and women aged over 70 years, whereas
postmenopausal osteoporosis affects spongy bones in about 30% of women [9]. A three-pronged nutritional
intervention involving calcium, vitamin D, and proteins has been suggested to combat osteoporosis, all
three of which are present in kefir, koumiss, and ayran in various levels. The calcium in kefir, koumiss,
and ayran is more bioavailable than milk due to its increased solubility. The CPPs in fermented milks are
found to prevent calcium phosphate precipitation and to increase the concentration of soluble calcium,
hence be beneficial on dental remineralization and calcium resorption. Vitamin D functions as essential
prohormone in the phosphate and calcium metabolism and facilitates normal bone mineralization and
the intestinal absorption of calcium and phosphate. Milk proteins slow down the bone loss. It has been
reported that an intake of 1.0–1.5 g milk protein per kg of body weight increases the level of insulin-like
growth factor 1 in the bloodstream that attenuates the reduction in bone mineral density in the elderly.
Fermented milks provide up to 5 g per serving of protein with a very high biological value. The most likely
benefit of traditional dairy beverages is believed to be gained by direct oral ingestion on an empty stomach.
59.4.6 Stimulation of the Immune System

The presence of some 70% of the immune response in the guts indicates that consuming probiotic-rich
foods and maintaining healthy intestinal flora can lead to a healthy immune system. Some bioactive
peptides in fermented milks have been linked to potentiation of the immune system and inhibition of
tumor development [37]. The β-endorphin derived from whey proteins indirectly enhances lymphocyte
proliferation, NK cell activity, and neutrophil locomotion via modulation of opioid µ receptor on
T lymphocytes and human phagocytic leucocytes [41]. Peptides derived from peptic hydrolysis of whey
proteins are shown to enhance the phagocytic activity of human neutrophils [43].
59.4.7 Antiallergy Effects

Allergic infants have been reported to have more clostridia and fewer Bifidobacteria in their intestinal
microbiota. A number of studies suggest that probiotics could have a beneficial impact on reducing the
incidence of food allergies particularly in children [73]. A link is established between probiotics intake
during pregnancy and a 30% reduction in the instance of childhood eczema. The following mechanisms
have been suggested on the basis of animal and in vitro studies: (1) improved mucosal barrier func-
tion, (2) degradation of food antigens, (3) modulation of intestinal microbiota composition and activity,
(4) enhanced production of secretory immunoglobulin A (IgA), (5) change in mucus production, and
(6) direct immune modulation [73]. Direct modulation of the immune system by kefir and koumiss may
be through induction of anti-inflammatory cytokines or through secretion of IgA.
59.4.8 Lifestyle and Well-Being

An important group of nutritional products that are often overlooked are those that address lifestyle
issues, including health problems that reduce people’s quality of life but are not life-threatening diseases.
Sleeplessness is one such lifestyle issue that inflicts a significant percentage of consumers over the age
45. Melatonin is a hormone produced within the brain that helps to regulate our body clock (the circadian
rhythm). Melatonin levels in our bodies vary over the 24 h cycle and gradually decreases over the course
of our lives, thus leading to insomnia. It occurs naturally in milk and taking a glass of warm milk before
bed is a well-known remedy to overcome sleeplessness. This effect is even more noticeable by taking
some ayran, kefir, or koumiss, also due to their hypotensive functions.
Depression and anxiety afflict almost everyone in the workforce. Preclinical and clinical studies have
shown reduction in anxiety and depression from probiotic supplements, with reduction in inflammatory
cytokines as a likely mechanism [74]. In a recent study by Noori et al. [75], kefir was shown to be effec-
tive in the treatment of nicotine cessation‒induced depression, anxiety, and cognition impairment in
animal model. Another lifestyle factor directly affecting health and well-being is obesity. Many studies
have shown that fermented milks may be useful in treating obesity through the production of short-chain
fatty acids and the satiety factors that influence food intake.
59.4.9 Other Health Benefits

Numerous trials have shown that fermented milk products containing CPPs can reduce dental caries
and improve overall oral health by rebalancing the bacteria in the mouth. When the products come
into direct contact with the mouth, they also reduce dental plaque and gum disease. There is emerging
evidence that regular consumption of kefir, koumiss, and ayran can help prevent bad bacteria from invad-
ing the urinary tract. A lesser-known benefit of these beverages could be treating nasal congestion and
other sinus issues. Probiotic milk drinks are shown to decrease the levels of pathogenic bacteria in the
nasal passages, thus reducing the congestion and other symptoms associated with seasonal allergies. In
addition to taking probiotic beverages internally, there are a few reports that show promising results on
topical application of probiotic preparations for prophylaxis and therapy of cutaneous pro-inflammatory
immune reactions. Kefir and koumiss may prove effective in this regard.

The increasing demand by health-conscious consumers for traditional healthy foods along with prog-
ress in modern science and technology has resulted in many traditional fermented dairy products to be
produced at larger scales. The last two decades have seen a revolution in the production of fermented
functional dairy foods and beverages. These product lines employ sophisticated processing technologies
combined with the cutting edge scientific knowledge focused on product safety and health benefits by
incorporation of selected probiotics, prebiotics, and antioxidants from fruits and other plant sources.
These products are consumed either as refreshing beverages or as nutritious and therapeutic dietary
adjuncts. Many publications have reported the types and significance of fermented dairy products in
human nutrition [2].
59.5.1 Novel Approaches to Traditional Fermented Dairy Beverages

All three traditional dairy beverages listed in this chapter have distinct differences in terms of chemical
and sensory qualities. There are also variations among each group based on the type of milk used and the
region where they originated from. All these differences need to be addressed in developing industrial-
scale production plan.
Traditional kefir made from authentic kefir grains is currently produced on an industrial scale in bot-
tles and Tetra Pak containers for distribution in Russia, Eastern European, and Central Asian countries.
The so-called Russian-style industrial production of kefir involves two steps: the initial production of a
starter kefir using kefir grain and the use of the grain-free kefir at a rate of 1%–2% to produce the com-
mercial kefir [50]. Currently, a stirred method is employed in industrial kefir production where all stages
of the production from fermentation to agitation, ripening, and cooling occurs in one vessel.
Attempts have been made in some Western countries to formulate and package kefir-based products
with longer shelf life. However, due to continued growth of yeast and production of CO2 gas after pack-
aging, the package material should be flexible enough to retain the amount of gas produced or designed
in a manner to allow gas to escape and prevent swelling and bulging of the container. Major dairy culture
suppliers have developed a range of freeze-dried kefir cultures, composed mainly of homofermentative
Streptococcus, some citric-fermenting Streptococcus, a small proportion of Lactobacillus, and a very
low percentage of yeast.
From a regulatory point of view, the alcohol content of kefir and koumiss may reach up to 4 g per
100 mL, may be subject to custom excise levy in many countries [19]. The future success of the industrial
production of these beverages, therefore, relies on selected LAB to replace the kefir grain and to con-
trol the level of alcohol and CO2 gas produced during fermentation [19]. Many novel ideas have been
proposed, for example, using microencapsulated yeast to retain the yeast flavor or using immobilized
starters. A more recent approach is to eliminate yeast in the starter culture formulation by using a mix-
ture of Lactobacillus, Lactococcus, and S. thermophilus and a few species of probiotics. One starter
culture combination used in the United States consists of S. lactis, L. plantarum, S. cremoris, L. casei,
S. diacetylactis, Leuconostoc cremoris, and a small number of Saccharomyces florentinus [66]. These
developments have resulted in an unprecedented expansion of kefir and kefir-based products in the
marketplace, mostly in Europe and North America. Smaller kefir starter culture kits are also available
for home kefir production. Novel foods such as kefir ice cream, kefir cheese (paneer), and coconut kefir
have also been introduced in the U.S. market. A variant of kefir called water kefir (or Tibicos) is made
by growing kefir grain in water with added sugar and dry figs or lemon juice for a day or more at room
temperature. Soy milk kefir and probiotic kefir are reported to have enhanced antimicrobial and dietetic
properties.
Koumiss is mainly consumed locally in Mongolia where it has high social, ritual, and religious value
and is a national drink. It has not been commercialized in other parts of the world [25], but it is becom-
ing increasingly important in France, Italy, Hungary, and the Netherlands because of the special thera-
peutic properties claimed on equine milk due to its resemblance to human milk [20]. In the Netherlands
and Belgium, fermented equine milk is flavored with concentrated fruit extract [20]. There has been
an increasing interest in the manufacture of koumiss on an industrial scale for human nutrition and
health [21]. Attempts to replace mare’s milk with cow’s milk have necessitated the dilution of cow’s milk
with water and adding some sugar. Kücükçetin et al. [24] investigated the use of membrane technolo-
gies (ultrafiltration and nanofiltration) to alter the composition of cow’s milk for koumiss production.
Currently, cow’s milk is used in the industrial production of koumiss that differs from the specific flavor
and attributes of the traditional koumiss.
Despite the simple domestic preparation of ayran, there have been strong worldwide efforts for indus-
trial production and marketing of drinking yogurts. Drinking yogurt and buttermilk are currently pro-
duced industrially using selected strains of LAB and probiotic bacteria, with fruit juices added in some
products for improved antioxidant activity. Ayran is produced on an industrial scale in Turkey, Bulgaria,
and Iran either as still ayran (in Tetra Pak or sealed cups) or carbonated ayran (in crown-capped bottles),
sometimes blended with herbs or herbal flavors and carbonated with CO2 gas. Carbonation creates a
more refreshing and satisfying mouthfeel to the cost of reduced lactic flavor and decreased number of
S. thermophilus, but it has no effect on L. delbrueckii bulgaricus. Ayran is mostly marked packed in
clear or white opaque plastic bottles or single-serve disposable cups. Many brands of industrially pro-
duced ayran are marketed in Turkey, Iran, Bulgaria, and Balkan countries.
59.5.2 Future Trends and Perspectives

The future of fermented dairy products is predictably promising in light of recent and future increase
in scientific validation of their impact on our nutrition and health. While koumiss and kefir are used
in Mongolia as the first infant food, more developments will see the incorporation of these miracu-
lous foods in infant formulations outside Mongolia. Further research will enable to design kefir- and
koumiss-based products for the aging population of the twenty-first century; people on special diets
such as vegetarians or those with special dietary requirement, for example, lactose maldigesters; and
chemotherapy patients, among others. As the space flights become more frequent and longer in duration
and commercial space travels are predicted in this century, several nutritional and metabolic problem
such as the effects of weightlessness, nutrient intake change, stress, bone calcium loss, radiation damage,
and immune response change have been studied [76]. Fermented foods have been part of the human diet
for centuries and it may well be that they will become important in the diets of future space travellers.
Dried fermented dairy foods, including dried yogurt, kefir, and koumiss, may be helpful with the space
travel‒related nutritional challenges [9].
59.6 Conclusion
Fermented dairy beverages render tremendous potential as carriers of functional ingredients required for
health and wellness of the human beings. The traditional dairy beverages have been mostly consumed
locally for millennia with many proven health benefits. Kefir and koumiss market is still a niche market
compared with ayran, but market is growing throughout the Western world. There are several techno-
logical challenges that need to be addressed, for example, the selection and maintaining the viability of
starter cultures during manufacturing and marketing, defining and maintaining the physical, chemical,
and organoleptic properties of the products, and packaging design and development to maintain product
integrity during the shelf life. Further biotechnological developments are required to enhance the tradi-
tional fermented milk’s shelf life to expand their market share [8]. As better starter culture combinations
are developed and the technology to control gas production during storage becomes more sophisticated,
more expansion of the market share of these traditional health-promoting beverages will take place.
More research is needed for better identification and determination of the efficacy of specific bioactive
components that impart health benefits and validation of these benefits by in vitro and in vivo studies
including human trials.
REFERENCES
1. Jin, S.L., Ancient but new style milk beverage—Kefir. China Dairy Ind., 27, 18–23, 1999.
2. International Dairy Federation (IDF), Fermented milks—Science and technology. Bull. Int. Dairy Fed.,
227, 4–164, 1988.
3. Otles, S. and Cagindi, O., Kefir: A probiotic dairy—Composition, nutrition and therapeutic aspects.
Pak. J. Nutr., 2, 54–59, 2003.
4. Liut Kevicius, A. and Sarkinas, A., Studies on the growth conditions and composition of kefir grains—
As a food and forage biomass. Dairy Sci. Abst., 66, 903, 2004.
5. Pogačić, T., Šinko, S., Zamberlin, Š., and Samaržija, D., Microbiota of kefir grains. Mljekarstvo, 63,
3–14, 2013.
6. Arslan, S., A review: Chemical, microbiological and nutritional characteristics of kefir. CyTA J. Food,
12, 1–6, 2014.
7. Garrote, G.L., Abraham, A.G., and De Antoni, G.L., Chemical and microbiological characterisation of
kefir grains. J. Dairy Res., 68, 639–652, 2001.
8. Sarkar, S., Potential of kefir as a dietetic beverage—A review. Br. Food J., 109, 280–290, 2007.
9. Farnworth, R.E., Kefir—A complex probiotic. Food Sci. Technol. Bull: Funct. Foods, 2, 1–17, 2005.
10. Guzel-Seydim, Z.B., Tuğba, K.T., Annel, K.G., and Atif, C.S., Review: Functional properties of kefir.
Crit. Rev. Food Sci. Nutr., 51, 261–268, 2011.
11. FAO/WHO, Codex Standard for Fermented Milks #243, World Health Organization/Food and
Agriculture Organization of the United Nations, Rome, Italy, 2011.
12. Guzel-Seydim, Z.B., Greene, A.K., and Taş, T., Determination of antimutagenic properties of some
fermented milks including changes in the total fatty acid profiles including CLA. Int. J. Dairy Technol.,
59, 209–2015, 2006.
13. Guzel-Seydim, Z.B., Seydim, A.C., and Greene, A.K., Amino acid profile of kefir. Milchwissenschaft,
58, 158–160, 2003.
14. Guzel-Seydim, Z.B., Seydim, A.C., and Greene, A.K., Organic acids and volatile flavour components
evolved during refrigerated storage of kefir. J. Dairy Sci., 83, 275–277, 2000.
15. Kneifel, W. and Mayer, H.K., Vitamin profiles of kefirs made from milks of different species. Int. J.
Food Sci. Technol., 26, 423–428, 1991.
16. Lv, J.P. and Wang, L.M., Bioactive components in kefir and koumiss, in Bioactive Components in Milk
and Dairy Products, Park, Y.E., Ed., Wiley-Blackwell, Hoboken, NJ, 2009, pp. 251–262.
17. Irigoyen, A., Arana, I., Castiella, M., Torre, P., and Ibanez, F.C., Microbiological, physico-chemical ad
sensory characteristics of kefir during storage. Food Chem., 90, 613–620, 2005.
18. Ahmed, Z., Wang, Y., Ahmad, A., Khan, S.T., Nisa, M., Ahmad, H., and Afreen, A., Kefir and health:
A contemporary perspective, Crit. Rev. Food Sci. Nutr., 53, 422–434, 2013.
19. Wszolek, M., Kupiec-Teahan, B., Skov Guldager, H., and Tamime, A.Y., Production of kefir, koumiss
and other related products, in Fermented Milks, Tamime, A., Ed., Blackwell Science Ltd., Oxford, UK,
2008, pp. 174–216.
20. Uniacke-Lowe, T., Koumiss, in Encyclopedia of Dairy Science, 2nd ed., Fuquay, J.W., Fox, P.E., and
McSweeney, P.L.H., Eds., Academic Press, New York, 2011, pp. 512–517.
21. Mu, Z. and Bai, Y., Koumiss. J. Inn. Mong. Agric. Univ., 1, 116–120, 2003.
22. Kabak, B. and Dobson, A.D.W., An introduction to the traditional fermented foods and beverages of
Turkey. Crit. Rev. Food Sci. Nutr., 51, 248–260, 2011.
23. Akuzawa, R. and Surono, I.S., Fermented milks of Asia, in Encyclopedia of Dairy Science, Roginski,
H., Fuquay, J., and Fox, P.F., Eds., Academic Press Ltd., London, UK, 2002, pp. 1045–1049.
24. Küçükçetin, A., Yaygina, H., Hinrichsb, J., and Kulozikc, U., Adaptation of bovine milk towards mares’
milk composition by means of membrane technology for koumiss manufacture. Int. Dairy J., 13,
945–951, 2003.
25. Malacarne, M., Martuzzi, F., Summer, A., and Mariani, P., Protein and fat composition of mare’s milk:
Some nutritional remarks with reference to human and cow’s milk. Int. Dairy J., 12, 869–877, 2002.
26. Fedechko, I.M., Hrytsko, R.I., and Herasun, B.A., The anti-immunodepressive action of koumiss made
from cow’s milk. Likars’ka Sprava, 9, 104–106, 1995.
27. Guzel-Seydim, Z., Kök-Taş, T., and Greene, A.K., Kefir and koumiss: Microbiology and technology, in
Development and Manufacture of Yogurt and Other Functional Dairy Products, Yıldız, F., Ed., CRC
Press/Taylor & Francis Group, Boca Raton, FL, 2010, pp. 143–163.
28. Wang, J., Chen, X., Liu, W., Yang, M., Airidengcaicike, and Zhang, H., Identification of Lactobacillus
from koumiss by conventional and molecular methods. Eur. Food Res. Technol., 227, 1555–1561,
2008.
29. Tamime, A.Y., Muir, D.D., and Wszolek, M., Kefir, koumiss and kishk. Dairy Ind. Int., 64, 32–33, 1999.
30. Montanari, G., Zambonelli, C., Grazia, L., Kamesheva, G.K., and Shigaeva, M.K., Saccharomyces uni-
sporus as the principal alcoholic fermentation microorganism of the traditional koumiss. J. Dairy Res.,
2, 327–331, 1996.
31. Kinik, O., Akalin, S., and Gonc, S., A research on koumiss production and its properties. J. Food, 25,
379–384, 2000.
32. Luhovyy, B.L., Akhavan, T., and Anderson, G.H., Whey proteins in regulation of food intake and sati-
ety. J. Am. Coll. Nutr., 26(Suppl.), 704S–712S, 2007.
33. Daniel, H., Volwinkel, M., and Rehner, G., Effect of casein and betacasomorphins on gastrointestinal
motility in rats. J. Nutr., 120, 252–257, 1990.
34. Hayes, M., Stanton, C., Fitzgerald, G.F., and Ross, R.P., Putting microbes to work: Dairy fermentation,
cell factories and bioactive peptides, Part II: Bioactive peptide functions. Biotechnol. J., 2, 435–449,
2007.
35. Park, Y.W., Overview of bioactive components in milk and dairy products, in Bioactive Components in
Milk and Dairy Products, Park, Y.W., Ed., Wiley-Blackwell, Hoboken, NJ, 2009, pp. 3–12.
36. Korhonen, H., Bioactive components in bovine milk, in Bioactive Components in Milk and Dairy
Products, Park, Y.W., Ed., Wiley-Blackwell, Hoboken, NJ, 2009, pp. 15–42.
37. Matar, C., LeBlanc, J.G., Martin, L., and Perdigón, G., Biologically active peptides released in fer-
mented milk: Role and functions, in Handbook of Fermented Functional Foods, 2nd edn., Farnworth,
E.R., Ed., CRC Press/Taylor & Francis Group, Boca Raton, FL, 2008, pp. 177–201.
38. Korhonen, H. and Marnila, P., Bovine milk antibodies for protection against microbial human diseases,
in Nutraceutical Proteins and Peptides in Health and Disease, Mine, Y. and Shahidi, F., Eds., CRC
39. Foucaud, C. and Juillard, V., Accumulation of casein-derived peptides during growth of proteinase-
positive strains of L. lactis in milk: Their contribution to subsequent bacterial growth is impaired by
their internal transport. J. Dairy Res., 67, 233–240, 2000.
40. Pihlanto-Leppälä, A., Rokka, T., and Korhonen, H., Angiotensin I-converting enzyme inhibitory
peptides derived from bovine milk proteins. Int. Dairy J., 8, 325–331, 1998.
41. Clare, D.A. and Swasigood, H.E., Bioactive milk peptides: A prospectus. J. Dairy Sci., 83, 1187–1195,
2000.
42. Meisel, H. and FitzGerald, R.J., Biofunctional peptides from milk proteins: Mineral binding and cyto-
modulatory effects. Cur. Pharm. Des., 9, 1289–1295, 2003.
43. Migliore-Samour, D., Folch, F., and Jolles, P., Biologically active casein peptides implicated in immu-
nomodulation. J. Dairy Res., 56, 357, 1989.
44. Korhonen, H. and Pihlanto-Leppälä, A., Milk-derived bioactive peptides: Formation and prospects for
health promotion, in Handbook of Functional Dairy Products, Shortt, C. and O’Brien, J., Eds., CRC
45. Seppo, L., Jauhianien, T., Poussa, T., and Korpela, R., A fermented milk high in bioactive peptides has
a blood pressure-lowering effect in hypertensive subjects. Am. J. Clin. Nutr., 77, 326–330, 2003.
46. Liu, J.R., Chen, M.J., and Lin, C.W., Antimutagenic and antioxidant properties of milk-kefir and
soymilk-kefir. J. Agric. Food Chem., 53, 2467–274, 2005.
47. Ebner, J., Arslan, A.A., Fedorova, M., Hoffmann, R., Kücükcetin, A., and Pischetsrieder, M., Peptide
profiling of bovine kefir reveals 236 unique peptides released from caseins during its production by
starter culture or kefir grains. J. Proteom., 117, 41–57, 2015.
48. Ondetti, M.A. and Cushman, D.W., Angiotensin-converting enzyme inhibitors: Biochemical properties
and biological actions. Crit. Rev. Biochem., 16, 381–411, 1984.
49. Yamamoto, N., Maeno, M., and Takano, T., Purification and characterisation of an antihypertensive
peptide from yogurt-like product fermented by Lactobacillus helveticus CPN4. J. Dairy Sci., 2, 1388–
1393, 1999.
50. Simova, E., Beshkova, D., Angelov, A., Hristozova, T.S., Frengova, G., and Spasov, Z., Lactic acid bac-
teria and yeasts in kefir grains and kefir made from them. J. Ind. Micro. Biotechnol., 28, 1–6, 2002.
51. Sedlábek, I., Nováková, D., and Kvec, P., Ribotyping and biotyping of Lactobacillus helveticus from the
koumiss. Eur. Food Res. Technol., 230, 753–758, 2010.
52. Quirós, A., Hernández-Ledesma, B., Ramos, M., Amigo, L., and Rocio, I. Angiotensin-converting
enzyme inhibitory activity of peptides derived from caprine kefir. J. Dairy Sci., 88, 3480–3487, 2005.
53. Thoreux, K. and Schmucker, D.L., Kefir milk enhances intestinal immunity in young but not old rats.
J. Nutr., 131, 807–812, 2001.
54. Furukawa, N., Iiyama, R., Takahashi, T., and Yamanaka, Y., Effect of oral administration of water sol-
uble fraction from kefir grain on antibody production in mice. Anim. Sci. Technol. (Jpn.), 63, 428–436,
1992.
55. Kodama, N., Komuta, K., and Nanba, H., Effect of maitake (Grifola frondosa) D-fraction on the activa-
tion of NK cells in cancer patients. J. Med. Food, 6, 371–377, 2003.
56. Murofushi, M., Mizuguchi, J., Aibara, K., and Matuhasi, T., Immunopotentiative effect of polysaccha-
ride from kefir grain, KGF-C, administered orally in mice. Imm. Pharmacol., 12, 29–35, 1986.
57. Furukawa, N., Matsuoka, A., Takahashi, T., and Yamanaka, Y., Anti-metastatic effect of kefir grain
components on Lewis lung carcinoma and highly metastatic B16 melanoma in mice. J. Agric. Sci.,
Tokyo Nogyo Daigaku, 45, 62–70, 2000.
58. Jinsmaa, Y. and Yoshikawa, M., Enzymatic release of neocasomorphin and β-casomorphin from bovine
β-casein. Peptides, 20, 957–962, 1999.
59. Fiat, A.M., Migliore-Samour, D., Jollès, P., Crouet, L., Collier, C., and Caen, J., Biologically active
peptides from milk proteins with emphasis on two examples concerning antithrombotic and immuno-
modulating activities. J. Dairy Sci., 76, 301–310, 1993.
60. Tsopmo, A., Romanowski, A., Banda, L., Lovie, J.D., Jenssen, H., and Friel, J.K., Novel anti-oxidant
peptides from enzymatic digestion of human milk. Food Chem., 126, 1138–1143, 2011.
61. Grishina, A., Kulikova, I., Alieva, L., Dodson, A., Rowland, I., and Jin, J., Antigenotoxic effect of kefir
and ayran supernatants on fecal water-induced DNA damage in human colon cells. Nutr. Cancer, 63,
73–79, 2011.
62. Cevikbas, A., Yemni, E., Ezzedenn, F.W., Yardimici, T., Cevikbas, U., and Stohs, S.J., Antitumoural
antibacterial and antifungal activities of kefir and kefir grain. Phytother. Res., 8, 78–82, 1994.
63. Garrote, G.L., Abraham, A.G., and De Antoni, G.L., Inhibitory power of kefir: The role of organic acids.
J. Food Prot., 63, 364–369, 2000.
64. Kakisu, E.J., Abraham, A.G., Pérez, P.F., and De Antoni, G.L., Inhibition of Bacillus cereus in milk
fermented with kefir grains. J. Food Prot., 70, 2613–2616, 2007.
65. Sieber, R., Collomb, M., Aeschlimann, A., Jelen, P., and Eyer, H., Impact of microbial cultures on
conjugated linoleic acid in dairy products—A review. Int. Dairy J., 14, 1–15, 2004.
66. Hertzler, S.R. and Clancy, S.M., Kefir improves lactose digestion and tolerance in adults with lactose
maldigestion. J. Am. Diet. Assoc., 103, 582–587, 2003.
67. Jiang, B., Mu, W., Obiro, W., and Shi, J., Traditional Chinese functional foods, in Functional Foods of
the East, Shi, J., Ho, C.-T., and Shahidi, F., Eds., CRC Press/Taylor & Francis Group, Boca Raton, FL,
2010, pp. 13–49.
68. Zha, M., Kumiss Therapy, Inner Mongolian People’s Publishing House, Huhehaote, China, 1987.
69. Chen, M., Sun, Q., Giovannucci, E., Mozaffarian, D., Manson, J.E., Willett, W.C., and Hu, F.B., Dairy
consumption and risk of type 2 diabetes: Three cohorts of US adults and an updated meta-analysis.
BMC Med., 12, 215–229, 2014.
70. Defilippi, C. and Gomez, E., Effect of casein and casein hydrolysate on small bowel motility and
d-xylose absorption in dogs. Neurogastroenterol. Motil., 7, 229–234, 1995.
71. Maalouf, K., Baydoun, E., and Rizk, S., Kefir induces cell-cycle arrest and apoptosis in HTLV-1-negative
malignant T-lymphocytes. Cancer Manag. Res., 3, 39–47, 2011.
72. Ibeagha-Awemu, E.M., Liu, J.R., and Zhao, X., Bioactive components in yogurt products, in Bioactive
Components in Milk and Dairy Products, Park, Y.W. Ed., Wiley-Blackwell, Hoboken, NJ, 2009,
pp. 235–250.
73. Ouwehand, A.C., Antiallergic effects of probiotics: Effects of probiotics and prebiotics. J. Nutr.,
137(Suppl. 3), 794S–797S, 2007.
74. Dinan, T.G. and Quigley, E.M., Probiotics in the treatment of depression: Science or science fiction?
Aust. NZ. J. Psych., 45, 1023–1025, 2011.
75. Noori, N., Bangash, M.Y., Motaghinejad, M., Hosseini, P., and Noudoost, B., Kefir protective effects
against nicotine cessation-induced anxiety and cognition impairments in rats. Adv. Biomed. Res.,
3, 251–264, 2014.
76. Buckley, N.D., Champagne, C.P., Masotti, A.I., Wagar, L.E., Tomkins, T.A., and Green-Johnson, J.M.,
Harnessing functional food strategies for the health challenges of space travel—Fermented soy for
astronaut nutrition. Acta Astronaut., 68, 731–738, 2011.
60
Applications of Plant Sterols and
Stanols in Functional Beverages
Jerzy Zawistowski
CONTENTS
60.1 Introduction................................................................................................................................... 785
60.2 Plant Sterols.................................................................................................................................. 786
60.3 Plant Sterol‒Enriched Beverages and Health Effect.................................................................... 786
60.3.1 Hypocholesterolemic Effect of Plant Sterols.................................................................... 786
60.3.2 Plant Sterol Modulatory Effect on Other Lipids.............................................................. 789
60.3.3 Other Health Benefits of Plant Sterols............................................................................. 790
60.3.3.1 Antioxidant Efficacy......................................................................................... 790
60.3.3.2 Anti-Inflammatory Activity.............................................................................. 790
60.4 Mechanism of Action.................................................................................................................... 790
60.5 Products, Formulations, Challenges, and Future Trends...............................................................791
60.6 Regulations of Functional Beverages with Plant Sterols.............................................................. 793
60.6.1 European Union................................................................................................................ 793
60.6.2 Australia and New Zealand.............................................................................................. 793
60.6.3 Canada.............................................................................................................................. 793
60.6.4 United States of America................................................................................................. 795
60.6.5 Other Countries................................................................................................................ 796
60.7 Conclusion..................................................................................................................................... 796
Acknowledgment.................................................................................................................................... 796
References............................................................................................................................................... 796
60.1 Introduction
The health benefits of plant sterols have been recognized for more than six decades. Plant sterols and
stanols in ester and free forms have been the subject of over 200 clinical studies. It has been proven that
both types of sterols are highly effective in lowering total and low-density lipoprotein (LDL) cholesterols
and can be used in functional foods to reduce the risk of coronary heart disease (CHD). Clinically proven
hypocholesterolemic properties of plant sterols led to the worldwide regulatory approval of these com-
pounds for use in foods. In addition, many regulatory jurisdictions now allow the use of health claims
regarding foods that are enriched with plant sterols/stanols.
The hypocholesterolemic properties of plant sterols are associated with their chemical structures,
which resemble cholesterol with some differences in the side chain. Sterols occur in foods of plant origin.
The most significant concentration exists in nuts and unrefined vegetable oils such as soy and peanut oils.
However, their presence even in the vegetarian diet is below the efficacious level, ranging from 300 to
450 mg/day. The consumption of such a diet has little to no effect on blood cholesterol level. Therefore,
for the body to benefit from hypocholesterolemic properties, foods need to be enriched with plant sterols
up to 2 g per serving size and consumed daily. There are many functional food categories enriched with
plant sterols available in global markets. This includes vegetable fat spreads and other fat-based foods,
785
CH3
H3C H3C
CH3 CH3
H3C 24 H3C 24
CH3 H3C CH3

H3C H H
8 8
7 7
HO HO
Cholesterol β-Sitosterol
CH3
H3C H3C
CH3 CH3
H 3C 24 H3C 24
CH3 CH3
H3C H H3C H
8 8
7 7
HO HO
Campesterol β-Sitostanol
FIGURE 60.1 Chemical structures of most common sterols and stanols used in functional beverages.
dairy products including yogurts and cheese, breakfast cereals and cereal bars, and various drinks.
However, sterol-containing beverages are the most convenient and popular functional foods. This chap-
ter focuses on a review of plant sterol properties, their efficacy, and safety as well as associated formula-
tion challenges with a specific emphasis on their applications to functional beverages.
60.2 Plant Sterols

Plant sterols are lipid-like compounds that are naturally occurring in many foods of plant origin. This
includes vegetable oils, nuts, seeds, cereals, vegetables, and fruits. Vegetable oils such as olive, corn,
sesame, and soybean contain between 2 and 10 mg/kg of sterols [1]. On a commercial scale, plant sterols
are obtained as a by-product during the processing of coniferous trees and soybean oil.
There are about 200 sterols occurring in plant and marine sources [2], with the most common being
β-sitosterol, campesterol, and β-sitostanol (Figure 60.1). Sitostanol is a saturated counterpart of sitos-
terol. It is formed by the hydrogenation of the 5α position double bond in sitosterol. Stanols are less
abundant in plants than sterols, and they occur in oilseeds and wood pulp. Plant sterols are triterpenes
that are chemically closely related to cholesterol. However, the presence of a side-chain substitution of
a methyl (campesterol) or an ethyl (sitosterol) group on carbon 24 makes sterols more hydrophobic and
structurally slightly distinct from cholesterol (Figure 60.1). In nature, plant sterols exist as free alcohols,
steryl esters with the fatty acid moiety at C12–C18, steryl glucosides, and acetylated steryl glucosides [3].
Although plant sterols are a part of our diet, the consumption of sterols varies from 100 to 300 mg/day
(Western diet) to 450 mg/day (vegetarian and vegan diets) [4]. Thus, this amount is insufficient to produce
a hypocholesterolemic effect. However, inclusion of around 2 g plant sterols in a daily diet can provide
the optimal effect based as evidenced in numerous clinical studies.
60.3 Plant Sterol‒Enriched Beverages and Health Effect

60.3.1 Hypocholesterolemic Effect of Plant Sterols
The health effect of plant sterols is associated with risk reduction of CHD and has been studied since
the early 1950s. Between 1950 and 1980, Eli Lilly marketed Cytelin™, a drug derived from tall oil plant
sterols. The efficacy of Cytelin™ was determined in clinical studies by feeding people with 9–50 g/day
Applications of Plant Sterols and Stanols in Functional Beverages 787
of this product [5]. After a number of years, the company withdrew this product from the market due to a
lack of a proper delivery vehicle and large doses to achieve health benefits. In the early days, plant sterols
were prepared in liquid suspensions, capsules, tablets, and powder. However, due to low purity, its effi-
cacy and subsequently marketability was limited. Biscuits and candies were the very first foods enriched
with plant sterols used in clinical studies [6,7]. By far, the largest number of clinical studies is conducted
on plant sterols and stanol ester–enriched margarines and fat spreads. This is because the esterification
of plant sterols with polyunsaturated fatty acids (PUFA) renders them soluble in fat [8,9]. Law [8] con-
ducted meta-analysis of 14 randomized trials with sterol-enriched margarine and vegetable fat spreads.
It was shown that consumption of 2 g of sterols in fat-based food for 3–4 weeks reduced LDL cholesterol
between 10% and 13%. This was confirmed by various clinical studies with other fat-based foods con-
taining plant sterols: vegetable oils, butter, mayonnaise, dressings, and chocolate [10–12]. It is worthwhile
to emphasize that a dose of 2 g is an optimal, efficacious daily amount to be consumed for 3–4 weeks.
During the last decade, plant sterols have been formulated in low-fat and fat-free foods. Beverages
such as yogurt and yogurt drinks, milk, soy drinks, and juices play an important role as convenient
delivery vehicles for plant sterols. There have also been a number of clinical studies on the hypocho-
lesterolemic effect, particularly on the LDL cholesterol responses, of plant sterol‒enriched beverages
(Table 60.1) [13–37].
Dairy drinks such as a single-shot yogurt and milk have shown the highest efficacy among clini-
cally tested liquids. Clifton et al. [34] have demonstrated that the cholesterol-lowering effect of sterol-
containing foods depends on the matrix. For example, milk enriched with plant sterols is about three
times more efficacious than bread or cereals containing the same amount of sterols.
Consumption of yogurt drink enriched with 1.0–2.8 g of sterols once a day yielded 3.2%–15.6% LDL
cholesterol reduction. The differences in efficacy may be correlated to the dose, the genotype of the clini-
cal subject, and whether or not the drink is consumed with food [15]. Doornbos et al. [14] have shown
that consuming the same amount of sterols (2.8 g/day) for 28 days in a form of single-shot yogurt drink
reduces LDL by 9.3%–9.5% when yogurt is consumed with meals as compared to a 5.1%–6.9% LDL
reduction when yogurt is consumed without meals. Even more pronounced differences were reported by
Seppo et al. [15], showing a significant reduction of LDL cholesterol (by 11.8%) when sterol-containing
(2.0 g/day) single-shot yogurt was consumed with meals for 35 days in contrast to the consumption of
same yogurt for the same period of time but without meals (LDL, which was only reduced by 3.2%).
It has been postulated that food consumption, particularly energy and fat intake with meals, increases
bile flow and makes plant sterols more effective when consumed with meals [15].
In the past, it was a strong belief that sterol-enriched foods need to contain a high level of fat
(e.g., margarine and vegetable fat spreads) in order to be efficacious. However, recent clinical studies have
shown that low-fat dairy drinks as delivery vehicles are as effective as other matrices, including veg-
etable fat spreads, and are even more effective than bread and cereals [34]. Low-fat fermented milk with
plant sterols showed a high efficacy when given to subjects in multicenter, randomized, double-blinded,
placebo-controlled, and parallel studies. The subjects received a dose of 100 mL milk with 1.6 g of plant
sterols (as sterol esters) once a day for 42 days. The plasma LDL cholesterol levels decreased from base-
line by 10.1% after 3 weeks and 10.5% after 6 weeks. Total cholesterol levels were decreased in the range
of 6.7% to 7.8% [20]. These results show that low-fat dairy drinks provide effective matrices optimally
at 3 weeks. The results are similar to other studies conducted on low-fat fermented milk [19,26], low-fat
milk [23,27,30,35], and low-fat yogurt drink [18,31]. It is worthwhile to note that the frequency of con-
sumption of dairy drinks (once or twice a day) showed no significant difference in cholesterol-lowering
effects [18]. Furthermore, similar efficacy was shown for free sterols and sterol esters. However, at least
one study conducted with free sterol‒enriched fat-free beverages failed to decrease blood cholesterol
levels [37]. The efficacy of free sterols is dependent on the way they are incorporated into drinks. The
consumption of a reduced-calorie orange juice drink with plant sterols (1 g/240 mL) twice a day for 8
weeks lowered total and LDL cholesterols in 72 mildly cholesterolemic subjects by 5% and 9.4%, respec-
tively, as compared with the baseline, in a randomized clinical trial [25]. Using an emulsifying agent
such as lecithin increased the efficacy of free sterols when added to fat-free or low-fat foods and drinks
[33]. It has been demonstrated that plant sitostanol formulated with lecithin micelles was more effective
in reducing cholesterol absorption than sitostanol alone. A dose of 300 mg of sitostanol with lecithin
TABLE 60.1
Low-Density Lipoprotein Cholesterol Lowering Effect of Free and Esterified Plant Sterols and Stanols
When Consumed as Beverages
Relative
Changes (%)
Sterol/ Frequency Dose Duration as Compared
Beverage Stanol (Times/Day) (g/Day) (Days) with Placebo References
Yogurt drink Stanol ester 1 2.0 21 −10.8 [13]
Yogurt drink Sterol ester 1 (with meal) 2.8 28 −9.3 to 9.5 [14]
Yogurt drink Sterol ester 1 (without meal) 2.8 28 −5.1 to 6.9 [14]
Yogurt drink Stanol ester 1 (with meal) 2.0 35 −11.8 [15]
Yogurt drink Stanol ester 1 (without meal) 2.0 35 −3.2 [15]
Low-fat milk Stanol ester 2–3 2.0 35 −6.2 [15]
Yogurt drink Sterol ester 1 2.0 28 −4.5 [16]
Yogurt drink Sterol ester 1 1.0 28 −4.3 [17]
Low-fat yogurt drink Free sterol 1 1.0 28 −4.3 [18]
Low-fat yogurt drink Free sterol 2 2.0 56 −6.4 [18]
Low-fat fermented milk Sterol ester 1 1.6 21 −10.5 [19]
Soy drink Sterol ester 1 1.6 28 −7.2 [21]
Soy drink Sterol ester 1 1.6 56 −6.8 [21]
Vegetable and fruit Free sterol 1 0.8 28 −5.8 [22]
juice mix
juice mix
juice mix
juice mix
Low-fat milk Sterol ester 2 2.0 90 −9.9 [23]
Orange juice Free sterol 2 2.0 56 −12.4 [24]
Orange juice Free sterol 2 2.0 28 −9.4 [25]
Milk with healthy diet Sterol ester 2 2.0 90 −9.6 [27]
Milk with free diet Sterol ester 2 2.0 90 −7.0 [27]
Milk tea Sterol ester 2 1.5 35 −2.5 [28]
Milk tea Sterol ester 2 2.3 35 −3.4 [28]
Dairy beverage Sterol ester 2 0.4 28 −5.0 [29]
Dairy beverage Sterol ester 2 2.0 28 −8.9 [29]
Low-fat milk Free sterol 2 1.2 28 −7.1 [30]
Low-fat milk Free sterol 2 1.6 28 −9.6 [30]
Low-fat yogurt drink Free sterols 1 1.0 28 −11.1 [31]
Low-fat yogurt drink Free sterols 1 2.0 28 −15.6 [31]
Oat-based drink Stanol ester nr 8.8 70 −17.1 [32]
Lemonade-flavored Free stanol/ 3 1.9 28 −14.3 [33]
beverage lecithin
Milk Sterol ester 2 or more 1.6 21 −15.9 [34]
Low-fat milk Sterol ester 2 2.0 21 −7.9 [35]
Milk Free sterols 3 0.9 28 −7.4 [36]
Nonfat beverage Sterol/stanol 3 1.8 21 0 [37]
reduced cholesterol absorption by 34%. This is three-fold higher than the absorption achieved by using
sitostanol without any emulsifier [38]. A sitostanol/lecithin complex was used to formulate a fat-free
lemonade beverage [33]. Subsequently, lemonade (1.9 g/day of sitostanol) was a subject of clinical studies
conducted on 45 normal or mildly hypercholesterolemic males and females. During a 4-week treatment,
total cholesterol was reduced by 10.1%, while LDL was lowered by 14.3% [33]. Emulsification of plant
sterols with lecithin is an effective way to increase sterol efficacy and makes the suitable preparation for
uses in functional beverages. The addition of lecithin prevents crystallization of sterols and makes them
more soluble in the mixed micelles, the critical step to achieve efficacy [39].
60.3.2 Plant Sterol Modulatory Effect on Other Lipids

The consumption of functional foods with plant sterols typically does not affect plasma triacylglycerols
(TAG) and high-density lipoprotein (HDL) cholesterol. However, some current clinical studies have
shown that plant sterols may lower the TAG level in human blood. Plana et al. [20] conducted a large,
multicenter study with hypercholesterolemic subjects. In this randomized placebo-controlled parallel
study, 44 patients consumed plant sterol–fortified (1.6 g sterols/day) fermented milk for 6 weeks. TAG
levels were significantly reduced by 14% in addition to lowering total and LDL cholesterols. No effect
of HDL cholesterol was observed. In addition, it has also been shown that the consumption of low-fat
yogurt drinks, containing plant stanol esters (2 g/day), for 8 weeks by patients with dyslipidemic meta-
bolic syndrome lowered their blood TAG by 27.5% [40]. This was in contrast to no effect on TAG in
normolipidemic subjects in the same study. The authors suggested that this effect was due to the signifi-
cant reduction of hepatic large TAG-rich very-low-density lipoprotein (VLDL) particles in metabolic
syndrome subjects [40]. It is worthwhile to note that the lowering TAG effect of plant sterols is not
unique to a specific food matrix such as a beverage. A recent meta-analysis of 12 clinical studies, where
sterols were delivered in margarine and salad dressing, showed smaller but significant lowering effects
of plasma TAG. Results showed that consumption of 1.6–2.5 g/day of sterol esters (11 studies) and free
sterols (1 study) for 3–4 weeks by subjects with the normal to borderline high TAG levels lowered plasma
lipids by an average of 6%. The authors concluded that the hypolipidemic effect of sterols was dependent
on the initial baseline concentration of TAG [41], which is in line with the earlier research [40].
Meta-analysis of clinical studies conducted on the efficacy of plant sterols and stanols showed that
sterols had no effect on plasma HDL cholesterol levels [41,42] or had only a slight insignificant increase
in subjects with low baseline HDL levels [43]. However, a single-blind randomized crossover study con-
ducted on patients with polygenic hypercholesterolemia fed with plant sterol ester-enriched yogurt drink
(1.0 g of free sterol equivalent) for 4 weeks yielded a 4.3% increase of HDL cholesterol [17]. Interestingly,
an increase of the total HDL was the result of a significant increase in HDL3 (17.1%) and an insignifi-
cant reduction in HDL2 cholesterol subfractions [17]. Furthermore, Devaraj et al. [25] reported that the
consumption of an orange juice beverage enriched with sterols (1 g/240 mL) twice a day with meals for
8 weeks significantly increased the level of blood HDL cholesterol compared with baseline (6% increase)
in 72 healthy subjects participating in the clinical study. The levels of plasma HDL cholesterol were not
statistically different in the treatment and control groups after 8 weeks [25]. Although a few studies sug-
gest that sterols may affect the blood HDL level, it is inconclusive whether the food matrix (beverage
versus solid food) may contribute to this effect.
It has been generally accepted that physical activities such as aerobics, bicycling, or other forms of
exercise may decrease the risk of CHD by favorably modulating blood lipids. Meta-analysis conducted by
Wilmore [44] was shown that exercise training increased the HDL level and decreased TAG concentration
in subjects regardless of gender and age. It seems that it is beneficial to combine the consumption of ste-
rols that primarily lower plasma total and LDL concentrations and exercise intervention that may increase
HDL and TAG concentrations. Varady et al. [45] have reported that daily endurance training combined
with the consumption of 2 g/day of plant sterols for 8 weeks by 84 previously sedentary hypercholesterol-
emic subjects not only lowered the total and LDL levels but also significantly increased the HDL level (by
7.5%) and decreased TAG concentration by 13.3%. These results conflict with other clinical studies that
show consumption of stanol esters may attenuate the increase of plasma HDL levels generated by a rela-
tively vigorous (400 kcal expenditure/day for 5 days) treadmill exercise by healthy sedentary subjects [46].
60.3.3 Other Health Benefits of Plant Sterols

In addition to their cholesterol-lowering effect, plant sterols exhibit other activities that might contribute
to their efficacy in retarding the processes of atherosclerosis and CHD. The following sections summa-
rize some of the other effects of phytosterols.
60.3.3.1 Antioxidant Efficacy

Oxidative stress is caused by free radicals that are generated by molecules with unpaired electrons. Free
radicals may lead to membrane structural damage and dysfunction. They may also increase the risk of
myocardial ischemia due to decreased membrane fluidity and permeability. In contrast, some compounds
are able to accept/donate electrons and act as free radical scavengers that show antioxidant properties.
It has been reported that β-sitosterol has a protective effect against free radical damage in vivo due to its
ring structure with the acetyl group and the presence of oxygen that accept electrons [47]. The protective
effect was even more pronounced for β-sitosterol glycoside due to the presence of glucose [47]. The anti-
oxidant properties of plant sterols have also been suggested by other researchers. β-Sitosterol has some
activity in inhibiting LDL cholesterol oxidation in vitro. Some researchers have found that minor com-
pounds of olive oil including β-sitosterol inhibit free radical formation and may inhibit LDL cholesterol
oxidation [48,49]. It has also been reported that Δ5-avenasterol, a substantial sterol in oats, and fucosterol
from algae have antioxidant properties due to the presence of ethylidene group [3,50]. Furthermore, in the
ApoE-deficient mouse model, a mixture of plant sterols increased the activity of antioxidant enzymes,
glutathione reductase in red blood cells, and superoxide dismutase in plasma [51]. This study also showed
that plant sterols had a significantly better effect on the prevention of atherosclerotic lesions in mice than
probucol [51]. It has been postulated by Xie et al. [52] that increasing activity of antioxidant enzymes may
be a contributing factor in the atheroprotective effect in the ApoE-knockout mouse model.
Antioxidant properties of plant sterols have also been studied in various food matrices. It has been
shown that plant sterols inhibit the thermal oxidation of olive oil [53], soybean oil [54,55], and medium-
chain TAG oils [56] heated at 180°C. The antioxidant properties of sterols may also be related to antimi-
crobial activities shown by other antioxidants. We have shown in our studies that incorporating mixture
of coniferous sterols into milk extends its shelf life [57].
60.3.3.2 Anti-Inflammatory Activity

It is a common belief that inflammation may be a contributing factor in atherogenic plaque formation and
consequently the risk of CHD and stroke. Therefore, current clinical studies, in addition to investigating the
effect of hypocholesterolemic efficacy of plant sterols, are also focused on their anti-inflammatory properties.
In one clinical study, the consumption of orange juice enriched with plant sterols by mildly cholesterolemic
subjects resulted in a 12% reduction of C-reactive protein (CRP) [25]. CRP is recognized as an inflammation
biomarker. The consumption of a low-fat fermented drink enriched with plant sterols has also shown anti-
inflammatory properties; however, the decrease of CRP levels was not statistically significant [26]. In contrast,
the consumption of a portfolio diet composed of beverages and solid foods, combining bioactives such as
fiber, soy protein, and plant sterols, significantly reduced CRP levels in the range of 23.8% to 28% in clinical
subjects [58,59]. The consumption of plant sterols together with omega-3 fatty acids yielded an even more pro-
nounced reduction in CRP (39%) along with a reduction of other inflammatory markers such as interleukin-6,
tumor necrosis factor-α, and leukotriene B4 in clinical studies conducted on 60 hyperlipidemic adults [60].
60.4 Mechanism of Action

The consumption of plant sterols has been shown to inhibit the absorption of both dietary and biliary cho-
lesterol from the small intestine. The main mechanism underlying the hypocholesterolemic effect partly
involves the competition between plant sterols and cholesterol for micellar solubilization based on simi-
larities in their chemical structures [61,62]. Sterols have a relatively higher affinity for micelles than cho-
lesterol due to a bulkier hydrophobic side chain. Thus, cholesterol is partially prevented and/or displaced
Micelles Dietary cholesterol
Plant sterols
Absorption Bile salts
NPC1L1
Esterification
Intestinal mucosa
ABCG
LXR
Chylomicrons
Adipocytes
Liver
NPC1L1 – Niemann pick C1-like 1 protein
Biliary LXR – Liver X receptor
cholesterol ABCG (G5/G8) – ATP cassette protein-binding
transporters
FIGURE 60.2 Plant sterols mechanism of action.
from the mixed micelle and some of the dietary and biliary cholesterols, typically absorbed by the body
is blocked (Figure 60.2). The competition for micellar solubilization takes place between nonesterified
sterols and cholesterol. However, there is no significant difference between free sterols and sterol esters in
exerting this effect since the latter is hydrolyzed by pancreatic cholesterol esterase in the intestine.
Another mechanism explaining the reduction of cholesterol absorption is associated with poor solubil-
ity of plant sterols in the intestinal chyme, which interferes with the micellar solubility of cholesterol.
This results in the coprecipitation of cholesterol and plant sterols in the intestine lumen. The formed
mixed sterol crystals subsequently reduce cholesterol bioavailability.
Plant sterols may also compete with cholesterol for esterification in enterocytes; however, this step is
less effective since the rate of sterol esterification is 34%–89% in range of cholesterol [63]. Furthermore,
the mechanism of action may include the competition of plant sterols with cholesterol for transporters as
indicated by animal studies. Sterols inhibit the intestinal expression of Niemann-Pick C1-like 1 protein
(NPC1L1)—the cholesterol transporter located in the apical membrane of enterocytes [64]. Sterols may
act as agonists of the liver receptor (LXR) and activate transcriptional ABCA1 (ABC-binding cassette
transporter), adenosine triphosphate (ATP)-binding cassette transporters, and ABCG5/G8 in the intes-
tine, which regulates the efflux of cholesterol and sterols between lumen and the intestinal mucosa.
These additional mechanisms may also reduce cholesterol absorption into the bloodstream [64].
It is worthwhile to note that the reduction of cholesterol absorption is partially offset by the increase
of cholesterol synthesis, albeit this increase is not large enough to make a significant difference.
Consequently, the plasma circulating of total and LDL cholesterols are reduced. The levels of HDL and
TAG are typically not affected [61].
60.5 Products, Formulations, Challenges, and Future Trends

The formulation of beverages enriched with plant sterols presents a challenge. These bioactives are
insoluble in water. Incorporation of sterols in drinks that contain fat, such as dairy products, is also diffi-
cult because of poor solubility in fat. Due to their crystalline form, sterols may change sensory attributes
of beverages because of their waxiness and powderiness. Esterification of plant sterols significantly
increases their fat solubility and lowers the melting point, which makes them suitable for incorporation
into fat-based foods. Benecol margarine enriched with plant stanol esters was the very first functional
food introduced on the EU market in 1995 by Raisio Oy, a Finish Company, which also patented the pro-
cess of stanol esterification. Plant sterol and stanol esters are primarily used for fortification of vegetable
fat spreads, salad dressing, mayonnaise, and vegetable oils. However, a number of commercial dairy
beverages including milk and yogurt drinks are also enriched with plant sterol and stanol esters. In 2003,
Swiss Dairy Producer “Emmi” introduced a one-shot yogurt drink with Raiso stanol esters. A 2 g of sta-
nols (based on free sterols) in a small bottle of 65 mL of yogurt drink was deemed to be an optimal dose
for daily consumption. The marketing of this yogurt drink was a big success and encouraged many other
producers to introduce yogurt shots on the European market. These contain either free or esterified plant
sterols. The proven efficacy [35] and convenience of a single-shot drink has boosted the sale of yogurt
beverages [65]. Within 3 years, sale of these products reached 65% of revenue generated by well-estab-
lished fat spreads [65]. The majority of fat-free and low-fat sterol-containing beverages available on the
world market are formulated with free sterols. The formulation of plant sterols into drinks also presents
a significant challenge. Uniform distribution of sterols within liquid requires developing the stabilization
system, since free sterols tend to sediment or float within the matrix. Furthermore, the sterol particles
may aggregate forming nondispersible clusters. Thus, in order to stabilize dispersion, it is critical to use
plant sterols of a small particle size, preferably in the range of 15 to 20 µ. Smaller particles have more
surface area per unit volume, and therefore they can be mixed more efficiently with food ingredients. The
advantage of small particle size is an improvement of mouthfeel in the final beverage product. Particles
larger than 35 µ are perceived as having a gritty texture; therefore, in chocolate manufacturing, at least
80% of the cocoa powder particles have to be smaller than 22 µ to produce acceptable “mouthfeel” for
chocolate [66]. Size reduction can be achieved by mechanical grinding (e.g., dry milling and microni-
zation) as well as wet milling (e.g., colloidal/jet milling and microfluidization). Microfluidization, also
known as particle collision technology, allows for processing plant sterols in high-concentration slurries
and creates uniform dispersions comprising particles of less than 10 µ. This technique was successfully
used to formulate yogurt and soya drinks with plant sterols [67]. A novel technique for the preparation of
micron and submicron particle size for plant sterols is being investigated. This is a modified supercriti-
cal fluid CO2 extraction, depressurization of an expanded liquid organic solvent, using CO2-expanded
ethanol at low temperature [68].
Aqueous dispersions with free plant sterols can also be stabilized with lyophilic colloids (carrageenan,
alginate, guar gum, xanthan gum, etc.). This is also accomplished by using microcrystalline cellulose,
emulsifiers such as polysorbates, sucrose-fatty acid esters, lecithin [67], and protein [69]. An interesting
approach was developed by Ostlund et al. [70] who formulated plant stanols with lecithin in the form of
micelles for applications in fat-free beverages. This approach also allows using as small dose as 300 mg
of plant stanol to achieve the same or better efficacy as for the optimal dose of 2 g of stanol per day [70].
Another novel approach to increase aqueous dispersibility of plant sterols was recently proposed by
Japanese scientists. They developed a technique of making a sterol-egg yolk lipoprotein complex (PSY).
It has been shown that the PSY complex is readily dispersible in water due to emulsifying properties of
egg’s phospholipids. It is suitable for use in beverages [71]. There are a number of proprietary techniques
for the formulation of dispersible plant sterols for convenient beverage applications. Most of them include
admixing sterols with emulsifiers in different ratios and subjecting the mixture to homogenization.
Auriou and Ferreres [72] proposed to disperse free plant sterols in water in the presence of an emulsifier
with a higher hydrophilicity–lipophilicity balance (HLB) value than sterols, such as lysolecithin. Some
other techniques describe micellization (microencapsulation) of plant sterols with water surfactants fol-
lowed by spray drying [72,73]. Dispersible sterols can be conveniently used by food product manufactur-
ers or even consumers who can incorporate them into food prior consumption.
Nanotechnology is another emerging technique to make plant sterols dispersible, even in clear bev-
erages. This method relates to making plant sterol micelles of small size, preferably below 300 nm.
South Korean company, Eugene Science, has developed a technique, which includes thermally melting
a mixture of plant sterols and emulsifier and dispersing the resultant molten admixture in water with
high-speed stirring until micelles are formed [74]. It is claimed that nanosized sterols can be used in any
beverages regardless of pH. In addition, nanomicelles do not affect sensory attributes of drinks such as
taste, flavor, or appearance [74]. Dolhaine et al. [75] suggested another way of making nanosized plant
sterols in the 10–300 nm range. They used a rapid expansion of supercritical solutions of sterols and
sterol esters. The resulting sterols were not only readily dispersible in aqueous-based foods but were also
claimed to be more absorbable into the bloodstream upon consumption versus their conventional coun-
terparts [75]. The enhancement of the absorbability of plant sterols may present a safety problem. Some
authors suggest that absorbed plant sterols can increase the risk of CHD [76]. Although these findings are
not conclusive, increasing absorption of sterols to the bloodstream should be avoided, particularly since
this does not increase their efficacy.
Some examples of commercial beverages approved for the global market by various regulatory juris-
dictions are shown in Table 60.2. Plant sterol–containing beverages are surging onto the world market,
as they represent a good alternative to vegetable fat spreads and other fatty foods enriched with sterols
for the risk reduction of CHD.
60.6 Regulations of Functional Beverages with Plant Sterols

Functional foods enriched with plant sterols and stanols have been recognized by global regulatory
authorities, and they are approved for human consumption in many countries. The specific approval con-
ditions, food categories, and health claims that can be used on foods vary from country to country. This
section focuses on sterol-containing beverages. Table 60.2 shows regulatory venues and health claims on
some functional beverages approved by various regulatory jurisdictions.
60.6.1 European Union

A number of beverages that can be used as delivery vehicles for plant sterols have been approved by the
European Commission under the Novel Food Regulations—258/97 [77]. The approved drinks include
milk-based beverages, milk-type products with the addition of fruits and/or cereals, and fermented milk-
type beverages including yogurt drink, soy, and rice drinks [78,79]. Approved drinks must contain no
more than 3 g of sterols per one or three serving sizes [78]. It is worthwhile to note that fruit juices with
plant sterols are not approved for the EU market. Approved beverages are eligible to carry a health claim
label, which is authorized by the European Commission under Regulation (EC) No 1924/2006, article
14 [80]. Drinks that contain at least 2 g of plant sterols may carry the disease risk reduction claim:
“Phytosterols have been shown to lower/reduce blood cholesterol. High blood cholesterol is a risk factor
in the development of coronary heart disease.”
Recently, the European Commission also allowed informing consumers about the beneficial effects
(cholesterol reduction between 7% and 10%) of consumption of dairy drinks if enriched with plant ste-
rols in the range of 1.5 to 2.4 g and consumed daily for 2–3 weeks [81].
60.6.2 Australia and New Zealand

Plant sterols, stanols, and their esters are approved only for adding to beverages such as milk and yogurt
under the Code Standard 1.5.1 Novel Foods. Plant sterols in the range of no less than 3 g/L and no less
than 4 g/L may be added to milk that contains not more than 1.5% (w/w) total fat in accordance with
Standard 2.5.1–6. Sterols enrichment of yogurt must be carry on according with Standard 2.5.3–5.
These regulations allow total plant sterols content of no less than 0.8 g and no more than 1.0 g per pack-
age of yogurt that contains no more than 1.5% (w/w) of total fat, while a package should not be bigger
than 200 g [82].
60.6.3 Canada
Contrary to EU and Australia/New Zealand regulations, Canadian authorities allow use of plant ste-
rols for enrichment of vegetable and fruit juices in addition to yogurt drinks, under the Novel Food
TABLE 60.2
Examples of Commercial Beverages Enriched with Plant Sterols, Stanols, and Their Esters That Were
Approved under Various Regulations
Examples of
Regulatory Health Claim Examples of Commercial Foods
Country Venue Health Claim Wording Enriched with Phytosterols
USA GRAS Disease risk “May reduce risk Orange juice
Notification reduction of coronary heart Heart Wise (Minute Maid)
Self-GRAS disease.” Yogurt drink
Benecol (McNeil), Promise Active
(Unilever), and Healthy Heart
(Yoplait)
Rice beverage
Dream Heart Wise (Heinz Celestial)
Milk drink
Active Lifestyle (Kroger)
European Novel Food Disease risk “Plant sterols have Yogurt drink
Union Regulations reduction been shown to Emmi Benecol (Emmi AG), Benecol
lower/reduce (McNeil), Flora Pro-activ (Unilever
blood cholesterol. Bestfoods), Danacol (Danone),
Blood cholesterol Tesco Reducol (Tesco), Salutare
lowering may Reducol (Pingo Doce Portugal),
reduce the risk of Healthy Heart (Yoplait), and Optifit
coronary heart Cholest Drink (Aldi)
disease.” Milk
Flora Pro-activ (Unilever Bestfoods)
and Mimosa Benvital (Lactogal
Portugal)
Canada Novel Food Therapeutic claims “Plant sterols help Yogurt drink
Regulations reduce/lower BioBest (Parmalat), Danacol
cholesterol. High (Danone), and President’s Choice
cholesterol is a Blue Menu (Loblaws)
risk factor for Juice drink
heart disease.” Heart Wise Orange Juice (Coca Cola),
Oasis Heart Break Orange/Mango/
Cranberry, and Pineapple/Tropical
juices (A. Lassonde, Inc.)
Australia Novel Food Proposal P293— “With natural plant Milk
and New Standard, high-level health sterols that reduce Logicol (MeadowLea Foods)
Zealand Standard 1.5.1 claims. cholesterol
uptake.”
South Health Functional Disease risk “Phytosterols may Cholzero Green Tea, Coffee, and Milk
Korea Food Act reduction, reduce risk of Drinks (Eugene Science, Inc.)
structure function coronary heart
claim disease.”
Turkey Turkish Food — “Help to reduce Kalbim Benecol Yogurt Drink (Ülker)
Codex cholesterol.” Kalbim Benecol Chocolate Milk
Regulations (Ülker)
Taiwan Health Food Yes, approved by “Consumption of Plant Sterol Milk (Uni-President)
Control Act Department of the product may
Health help lower blood
total cholesterol.”
Malaysia Food Act and Nutrient function “Plant sterol or ActiCol Omega plus Milk (Nestle)
Regulation, Food claims stanol helps lower ActiCol Omega plus Soy Drink
Supplement or reduce (Nestle)
cholesterol.”
(Continued)

Examples of Commercial Beverages Enriched with Plant Sterols, Stanols, and Their Esters That Were
Approved under Various Regulations
Examples of
Regulatory Health Claim Examples of Commercial Foods
Country Venue Health Claim Wording Enriched with Phytosterols
Indonesia Drug and Food Disease risk “May reduce risk NesVita Proheart Milk (Nestle)
Control reduction of coronary heart Benecol Smoothes (Kalbe
Regulations disease.” Nutritionals)
Singapore Food Regulation No health claims “For people who ActiCol Omega plus Milk (Nestle)
Act, approval on permitted want to lower
case-by-case their blood
bases by Food cholesterol level.”
Control
Department of
Singapore
Thailand The Notification Nutrient function “Plant sterol may Benecol Yogurt Drink (Saha
for Foods for claims help lower Pathanapibul)
Special Dietary cholesterol.”
Uses
Philippines Food Fortification Structure function “This product NesVita Proheart Milk (Nestle)
Act claim contains natural NesVita Proheart Yogurt Drink
plant sterols that (Nestle)
help lower Benecol Smoothie (Kalbe)
cholesterol.”
Brazil Technical Health claims “Phytosterols help Milk Drink (Shefa Benecol)
regulation on in reducing the
procedures for absorption of
registration of cholesterol.”
foods and or new
ingredients
Chile Ministry of Health claims “Reduce Kaiku Benecol Yogurt Drink (Kaiku
Health Decree cholesterol.” Co. Alimentaria)
No. 977
Abbreviation: GRAS, generally recognized as safe.
Regulations. However, the use of sterols in milk is not allowed. The aforementioned drinks can be
enriched with up to 1 g of sterols per serving size to a total amount of 2 g as a daily dietary intake, which
is considered the optimal daily efficacious dose. Authorized drinks that are formulated to fulfill specific
prerequisites may bear a health claim label: “Plant sterols help reduce/lower cholesterol and high choles-
terol is a risk factor for heart disease” [83]. This claim is similar to the disease risk reduction claim for
plant sterols in the EU, albeit in Canada is classified as a therapeutic claim.
60.6.4 United States of America

The U.S. Food and Drug Administration (FDA) has approved plant sterols for use in foods including bev-
erages since 1998 under the generally recognized as safe (GRAS) notification and self-GRAS rulings.
In September 2000, the FDA issued an interim ruling that allows for a health claim that links the con-
sumption of plant sterol and stanol esters, and free sterols, to the risk reduction of CHD. This claim was
amended in 2010. The new ruling requires eligible foods to contain at least 0.5 g of plant sterols or sta-
nols or their esters (free sterol bases) per serving size to a total amount of 2 g as a daily dietary intake in
order to justify the CHD reduction claim [84]. The regulations do not restrict the use of sterols in formu-
lation of specific beverages such as in EU, Canada, or Australia/New Zealand. However, in order to bear
a health claim, drinks must comply with the requirements of the disqualifying nutrient levels rule (CFR
21.101.14 a 4), the low–saturated fat (21CFR 101.62 c 2), and low-cholesterol (21CFR 101.62 d 2) rules.
Furthermore, they must contain at least 10% of the daily value for one of six nutrients per reference
amount as is stipulated by the minimum nutrient contribution requirement (CFR 21.101.14 e 6) [84]. If a
beverage meets prerequisites and contains the acceptable level of plant sterols, it qualifies to bear a health
claim as per the FDA model claim: “Foods containing at least 0.5 g per serving of plant sterols eaten
with meals or snacks for a daily total intake of 2 g as a part of a diet low in saturated fat and cholesterol
may reduce the risk of heart disease. A serving of [name of the food] supplies X g of plant sterols” [84].
60.6.5 Other Countries

The use of plant sterols in foods, including beverages, was granted in non-EU countries such as
Switzerland, Norway, and Iceland. Approval in Asia includes Japan, South Korea, Hong Kong, Taiwan,
Malaysia, Indonesia, Singapore, Thailand, and the Philippines. This also applies to Brazil, Chile, Turkey,
and Israel. The examples of approved beverages are listed in Table 60.2.
60.7 Conclusion
Plant sterols represent one of the most intensely and actively researched groups of bioactives in the area of
cardiovascular disease (CVD). This group of compounds has been clinically proven to reduce serum cho-
lesterol. With nearly 100 million Americans and another 100 million Europeans experiencing elevated
cholesterol levels, plant sterols containing functional foods may offer preventive solutions that are safe
and effective alternatives that are complementary to medical strategies in lowering blood cholesterol.
Plant sterols play an important role in functional beverages. Proven efficacy combined with conve-
nience of a delivery vehicle such as drink makes beverages a good alternative to vegetable fat spreads
and other fatty foods enriched with sterols for risk reduction of CHD. There is no doubt that health is a
major selling point in the functional industry today; however, contrary to other food formats, beverages
are appealing to consumers because they are easier to incorporate in the daily diet.
Acknowledgment
I would like to thank my son Robert and my wife Urszula for inciting discussion and proofreading of
the manuscript.
REFERENCES
1. Moreau, R.A., Norton, R.A., and Hicks, K.B., Phytosterols and phytostanols lower cholesterol. Inform,
10, 572–577, 1999.
2. Piironen, V., Lindsay, D.G., Miettinen, T.A., Toivo, J., and Lampi, A.M., Plant sterols: Biosynthesis,
biological function and their importance to human nutrition. J. Sci. Food Agric., 80, 939–966, 2000.
3. Moreau, R.A., Whitaker, B.D., and Hicks, K.B., Phytosterols, phytostanols, and their conjugates in
foods: Structural diversity, quantitative analysis, and health-promoting uses. Prog. Lipid Res., 41,
457–500, 2002.
4. Nair, P.P., Turjman, N., Kessie, G., Calkins, B., Goodman, G.T., Davidovitz, H., and Nimmagadda, G.,
Diet, nutrition intake, and metabolism in populations at high and low risk for colon cancer. Dietary
cholesterol, beta-sitosterol, and stigmasterol. Am. J. Clin. Nutr., 40(Suppl. 4), 927–930, 1984.
5. Pollak, O.J. and Kritchevsky, D., Sitosterol (Monographs on Atherosclerosis), S. Karger AG, Basel,
Switzerland, 1981.
6. Barber, J.M. and Grant, A.P., The serum cholesterol and other lipids after administration of β-sitosterol.
Br. Heart J., 17, 296–298, 1955.
7. Fahrenbach, M.J., Lewry, H.V., Riccardi, B.A., Dunnett, C.W., Saunders, J.C., Lourie, E., Blodinger, J.,
Ruegsegger, J.M., Grant, W.C., and Jukes, T.H., Effect of β-sitosterol and unsaturated vegetable oil
combinations on human serum cholesterol levels. Circulation, 18, 491, 1958.
8. Law, M., Plant sterol and stanol margarines and health, Br. Med. J., 320, 861–864, 2000.
9. Neil, H.A.W. and Huxley, R.R., Efficacy and therapeutic potential of plant sterols. Atheroscler. Supp.,
3, 11–15, 2002.
10. Nestel, P., Cehun, M., Pomeroy, S., Abbey, M., and Weldon, G., Cholesterol-lowering effects of plant
sterol esters and non-esterified stanols in margarine, butter and low-fat foods. Eur. J. Clin. Nutr.,
12, 1084–1090, 2001.
11. de Graaf, J., de Sauvage-Nolting, P.R.W., van Dam, M., Belsey, E.M., Kastelein, J.J.P., Pritchard, P.H.,
and Stalenhoef, A.F.H., Consumption of tall oil-derived phytosterols in a chocolate matrix signifi-
cantly decreases plasma total and low-density lipoprotein-cholesterol levels. Br. J. Nutr., 88, 479–488,
2002.
12. Vanstone, C.A., Raeini-Sarjaz, M., Parsons, W.E., and Jones, P.J., Unesterified plant sterols and stanols
lower LDL-cholesterol concentrations equivalently in hypercholesterolemic persons. Am. J. Clin. Nutr.,
76, 1272–1278, 2002.
13. Algorta Pineda, J., Chinchetru Ranedo, M.J., Aguirre Anda, J., and Terreros, F.S., Hypocholesteremic
effectiveness of a yogurt containing plant stanol esters. Rev. Clin. Esp., 205, 63–66, 2005.
14. Doornbos, A.M., Meynen, E.M., Duchateau, G.S., van der Knaap, H.C., and Trautwein, E.A., Intake
occasion affects the serum cholesterol lowering of a plant sterol-enriched single-dose yoghurt drink in
mildly hyper-cholesterolemic subjects. Eur. J. Clin. Nutr., 60, 325–333, 2006.
15. Seppo, L., Jauhiainen, T., Nevala, R., Poussa, T., and Korpela, R., Plant stanol esters in low-fat milk
products lower serum total and LDL cholesterol. Eur. J. Nutr., 46, 111–117, 2007.
16. Khandelwal, S., Demonty, I., Jeemon, P., Lakshmy, R., Mukherjee, R., Gupta, R., Snehi, U. et al.,
Independent and interactive effects of plant sterols and fish oil n-3 long-chain polyunsaturated fatty
acids on the plasma lipid profile of mildly hyperlipidemic Indian adults. Br. J. Nutr., 102, 722–732,
2009.
17. Ruiu, G., Pinach, S., Veglia, F., Gambino, R., Marena, S., Uberti, B., Alemanno, N., Burt, D., Pagano, G.,
and Cassader, M., Phytosterol-enriched yogurt increases LDL affinity and reduces CD36 expression in
polygenic hypercholesterolemia. Lipids, 44, 153–160, 2009.
18. Niittynen, L.H., Jauhiainen, T.A., Poussa, T.A., and Korpela, R., Effects of yoghurt enriched with free
plant sterols on the levels of serum lipids and plant sterols in moderately hypercholesterolemic subjects
on a high-fat diet. Int. J. Food Sci. Nutr., 59, 357–367, 2008.
19. Mannarino, E., Pirro, M., Cortese, C., Lupattelli, G., Siepi, D., Mezzetti, A., Bertolini, S. et al., Effects
of a phytosterol-enriched dairy product on lipids, sterols and 8-isoprostane in hypercholesterolemic
patients: A multicenter Italian study. Nutr. Metab. Cardiovasc. Dis., 19, 84–90, 2009.
20. Plana, N., Nicolle, C., Ferre, R., Camps, J., Cos, R., Villoria, J., and Masana, L., Plant sterol-enriched
fermented milk enhances the attainment of LDL-cholesterol goal in hypercholesterolemic subjects.
Eur. J. Nutr., 47, 32–39, 2008.
21. Weidner, C., Krempf, M., Bard, J.M., Cazaubiel, M., and Bell, D., Cholesterol lowering effect of a soy
drink enriched with plant sterols in a French population with moderate hypercholesterolemia. Lipids
Health Dis., 7, 35, 1–8, 2008.
22. Hironaka, T., Shroya, N., Matsubara, H., Matsuoka, Y., and Itakura, H., Double-blind, placebo-
controlled study of effects of plant sterol enriched vegetable juice on serum cholesterol concentrations
in mildly hyper-cholesterolemic subjects and safety evaluation. J. Oleo Sci., 55, 593–606, 2006.
23. Banuls, C., Martinez-Triguero, M.L., López-Ruiz, A., Morillas, C., Lacomba, R., Victor, V.M., Rocha,
M., and Hernandez-Mijares, A., Evaluation of cardiovascular risk and oxidative stress parameters in
hypercholesterolemic subjects on a standard healthy diet including low-fat milk enriched with plant
sterols. J. Nutr. Biochem., 21, 881–886, 2010.
24. Devaraj, S., Jialal, I., and Vega-Lopez, S., Plant sterol-fortified orange juice effectively lowers choles-
terol levels in mildly hypercholesterolemic healthy individuals. Arterioscler. Thromb. Vasc. Biol., 24,
e25–e28, 2004.
25. Devaraj, S., Autret, B.C., and Jialal, I., Reduced-calorie orange juice beverage with plant sterols lowers
C-reactive protein concentrations and improves the lipid profile in human volunteers. Am. J. Clin. Nutr.,
84, 756–761, 2006.
26. Hansel, B., Nicolle, C., Lalanne, F., Tondu, F., Lassel, T., Donazzolo, Y., Ferrieres, J. et al., Effect of low-
fat, fermented milk enriched with plant sterols on serum lipid profile and oxidative stress in moderate
hypercholesterolemia. Am. J. Clin. Nutr., 86, 790–796, 2007.
27. Hernández-Mijares, A., Banuls, C., Rocha, M., Morillas, C., Martinez-Triguero, M.L., Victor, V.M.,
Lacomba, R. et al., Effects of phytosterol ester-enriched low-fat milk on serum lipoprotein profile in
mildly hypercholesterolemic patients are not related to dietary cholesterol or saturated fat intake. Br. J.
Nutr., 104, 1018–1025, 2010.
28. Li, N.Y., Li, K., Qi, Z., Demonty, I., Gordon, M., Francis, L., Molhuizen, H.O., and Neal, B.C., Plant
sterol-enriched milk tea decreases blood cholesterol concentrations in Chinese adults: A randomised
controlled trial. Br. J. Nutr., 98, 978–983, 2007.
29. Racette, S.B., Lin, X., Lefevre, M., Spearie, C.A., Most, M.M., and Ostlund, R.E.J., Dose effects of
dietary phytosterols on cholesterol metabolism: A controlled feeding study. Am. J. Clin. Nutr., 91,
32–38, 2010.
30. Thomsen, A.B., Hansen, H.B., Christiansen, C., Green, H., and Berger, A., Effect of free plant sterols in
low-fat milk on serum lipid profile in hypercholesterolemic subjects. Eur. J. Clin. Nutr., 58, 860–870, 2004.
31. Volpe, R., Niittynen, L., Korpela, R., Sirtori, C., Bucci, A., Fraone, N., and Pazzucconi, F., Effects of
yoghurt enriched with plant sterols on serum lipids in patients with moderate hypercholesterolemia.
Br. J. Nutr., 86, 233–239, 2001.
32. Gylling, H., Hallikainen, M., Nissinen, M.J., and Miettinen, T.A., The effect of a very high daily plant
stanol ester intake on serum lipids, carotenoids, and fat-soluble vitamins. Clin. Nutr., 29, 112–118, 2010.
33. Spilburg, C.A., Goldberg, A.C., McGill, J.B., Stenson, W.F., Racette, S.B., Bateman, J., McPherson,
T.B., and Ostlund, R.E.J., Fat-free foods supplemented with soy stanol-lecithin powder reduce choles-
terol absorption and LDL cholesterol. J. Am. Diet. Assoc., 103, 577–581, 2003.
34. Clifton, P.M., Noakes, M., Sullivan, D., Erichsen, N., Ross, D., Annison, G., Fassoulakis, A., Cehun, M.,
and Nestel, P., Cholesterol-lowering effects of plant sterol esters differ in milk, yoghurt, bread and
cereal. Eur. J. Clin. Nutr., 58, 503–509, 2004.
35. Noakes, M., Clifton, P.M., Doornbos, A.M., and Trautwein, E.A., Plant sterol ester-enriched milk and
yoghurt effectively reduce serum cholesterol in modestly hypercholesterolemic subjects. Eur. J. Nutr.,
44, 214–222, 2005.
36. Beer, M.U., Pritchard, H., Belsey, E.M., and Davidson, M., Effect of a milk drink enriched with increas-
ing doses of free tall oil phytosterols on plasma lipid levels of mildly hypercholesterolemic subjects.
Document submitted by Altus Foods to FDA concerning the interim final rule for Vegetable oil sterol
esters and coronary heart disease. Docket Nos. 00P-1275 and 00P-1276, 2001.
37. Jones, P.J.H., Vanstone, C.A., Raeini-Sarjaz, M., and St-Onge, M.P., Phytosterols in low- and nonfat
beverages as part of a controlled diet fail to lower plasma lipid levels. J. Lipid Res., 44, 1713–1719, 2003.
38. Ostlund, R.E., Spilburg, C.A., and Stenson, W.F., Sitostanol administered in lecithin micelles potently
reduces cholesterol absorption in humans. Am. J. Clin. Nutr., 70, 826–831, 1999.
39. Engel, R. and Schubert, H., Formulation of phytosterols in emulsions for increased dose response in
functional foods. Inn. Food Sci. Emer. Technol., 6, 233–237, 2005.
40. Plat, J. and Mensink, R.P., Plant stanol esters lower triacylglycerol concentrations via a reduced hepatic
VLDL-1 production. Lipids, 44, 1149–1153, 2009.
41. Demonty, I., Ras, R.T., van der Knaap, H.C.M., Meijer, L., Zock, P.L., Geleijnse, J.M., and Trautwein,
E.A., The effect of plant sterols on serum triglyceride concentrations is dependent on baseline concen-
trations: A pooled analysis of 12 randomised controlled trials. Eur. J. Nutr., 52, 253–260, 2013.
42. Morusi, K.G., Oosthuizen, W., and Opperman, A.M., Phytosterols/stanols lower cholesterol concentra-
tions in familial hypercholesterolemic subjects: A systematic review with meta-analysis. J. Am. Coll.
Nutr., 25, 41–48, 2006.
43. Demonty, I., Ras, R.T., van der Knaap, H.C.M., Duchateau, G.S.M.J.E., Meijer, L., Zock, P.L., Geleijnse,
J.M., and Trautwein, E.A., Continuous dose-response relationship of the LDL-cholesterol-lowering
effect of phytosterol intake. J. Nutr., 139, 271–284, 2009.
44. Wilmore, J.H., Dose-response: Variation with age, sex, and health status. Med. Sci. Sports Exerc.,
33(Suppl. 6), S622–S634, 2001.
45. Varady, K.A., Ebine, N., Vanstone, C.A., Parsons, W.E., and Jones, P.J.H., Plant sterols and endurance
training combine to favorably alter plasma lipid profiles in previous sedentary hypercholesterolemic
adults after 8 wk. Am. J. Clin. Nutr., 80, 1159–1166, 2004.
46. Alhassan, S., Reese, K.A., Mahurin, J., Plisance, E.P., Hilson, B.D., Graner, J.C., Wee, S.O., and
Grandjean, P.W., Blood lipid responses to plant stanol ester supplementation and aerobic exercise train-
ing. Metab. Clin. Exp., 55, 541–549, 2006.
47. Van Rensburg, S.J., Daniels, W.M., van Zyl, J.M., and Taljaard, J.J., A comparative study of the effects
of cholesterol, beta-sitosterol, beta-sitosterol glucoside, dehydroepiandrosterone sulphate and melatonin
on in vitro lipid peroxidation. Metab. Brain Dis., 15, 257–265, 2000.
48. Aviram, M. and Eias, K., Dietary olive oil reduces low density lipoprotein uptake by macrophages and
decreases the susceptibility of the lipoprotein to undergo lipid peroxidation. Ann. Nutr. Metab., 37,
75–84, 1993.
49. Moreno, J.J., Effect of olive oil minor components on oxidative stress and arachidonic acid mobilization
and metabolism by macrophages RAW 267.7. Free Radic. Biol. Med., 35, 1073–1081, 2003.
50. Boskou, D., Frying temperatures and minor constituents of oils and fats. Grasas y Aceites, 49, 326–330,
1998.
51. Moghadasian, M.H., McManus, B.M., Godin, D.V., Rodrigues, B., and Frohlich, J.J., Proatherogenic
and antiatherogenic effects of probucol and phytosterols in apolipoprotein E-deficient mice. Possible
mechanism of action. Circulation, 99, 1733–1739, 1999.
52. Xie, C., Kang, J., Burris, R., Ferguson, M.E., Schauss, A.G., Nagarajan, S., and Wu, X., Acai juice
attenuates atherosclerosis in ApoE deficient mice through antioxidant and anti-inflammatory activities.
Atherosclerosis, 216, 327–333, 2011.
53. Gordon, M.H. and Magos, P., The effect of sterols on the oxidation of edible oils. Food Chem., 10,
141–147, 1983.
54. White, P.J. and Armstrong, L.S., Effect of selected oat sterols on the deterioration of heated soybean oil.
J. Am. Oil Chem. Soc., 63, 525–529, 1986.
55. Winkler, J.K., Warner, K., and Glynn, M.T., Effect of deep-fat frying on phytosterol content in oils with
differing fatty acid composition. J. Am. Oil Chem. Soc., 84, 1023–1030, 2007.
56. Du, K. and Zawistowski, J., Oxidative stability of designer oil, presented at the Institute of Food
Technologists Annual Meeting and Food Expo, Anaheim, CA, June 15–19, 2002, Abstract 136–165.
57. Monu, E., Blank, G., Holley, R., and Zawistowski, J., Phytosterol effects on milk and yogurt microflora.
J. Food Sci., 73, 121–126, 2008.
58. Jenkins, D.J., Kendall, C.W., Marchie, A., Faulkner, D.A., Josse, A.R., Wong, J.M.W., de Souza, R.
et al., Direct comparison of dietary portfolio versus statin on C-reactive protein. Eur. J. Clin. Nutr., 59,
851–860, 2005.
59. Jenkins, D.J., Kendall, C.W., Nguyen, T.H., Marchie, A., Faulkner, D.A., Ireland, C., Josee, A.R. et al.,
Effect of plant sterols in combination with other cholesterol-lowering foods. Metabolism, 57, 130–139,
2008.
60. Micallef, M.A. and Garg, M.L., Anti-inflammatory and cardioprotective effects of n-3 polyunsaturated
fatty acids and plant sterols in hyperlipidemic individuals. Atherosclerosis, 204, 476–482, 2009.
61. Gylling, H. and Miettinen, T.A., Cholesterol reduction by different plant stanol mixtures and with
variable fat intake. Metabolism, 48, 575–580, 1999.
62. Hendriks, H.F.J., Weststrate, J.A., van Vliet, T., and Meijer, G.W., Spread enriched with three different
levels of vegetable oil sterols and the degree of cholesterol lowering in normocholesterolemic and mildly
hypercholesterolemic subjects. Eur. J. Clin. Nutr., 53, 319–327, 1999.
63. Moghadasian, M.H., Pharmacological properties of plant sterol in vivo and in vitro observations. Life
Sci., 67, 605–615, 2000.
64. Calpe-Berdiel. L., Escola-Gil. J.C., and Blanco-Vaca, F., Phytosterol-mediated inhibition of intestinal cho-
lesterol absorption is independent of ATP-binding cassette transporter A1. Br. J. Nutr., 95, 618–622, 2006.
65. Nielsen, A.C., Top 100 grocery brands, AC Nielsen Retail News, Croydon, UK, March 2006.
66. Rostagno, W., Chocolate particle size and its organoleptic influence. The Manufacturing Confectioner,
49, 81–86, 1969.
67. Zawistowski, J., Method of preparing microparticles of phytosterols or phytostanols, European Patent,
EP 1 148 793 B1, August 13, 2003.
68. Temelli, F., Cordoba, A., Elizondo, E., Cano-Sarabia, M., Veciana, J., and Ventosa, N., Phase behavior
of phytosterols and cholesterol in carbon dioxide-expanded ethanol. J. Supercrit. Fluids, 63, 59–68,
2012.
69. Moreau, R.A., Plant sterols in functional foods, in Phytosterols as Functional Food Components and
Nutraceuticals, Dutta, P.C., Ed., Marcel Dekker, New York, pp. 317–345, 2004.
70. Ostlund, R.E. Jr., Spilburg, C.A., and Stenson, W.F., Sitostanol administration in lecithin micelles
potently reduces cholesterol absorption in humans. Am. J. Clin. Nutr., 79, 826–831, 1999.
71. Matsuoka, R., Muto, A., Kimura, M., Hoshina, R., Wakamatsu, T., and Masuda, Y., Cholesterol-lowering
activity of plant sterol-egg yolk lipoprotein complex in rats. J. Oleo Sci., 57, 309–314, 2008.
72. Auriou, N. and Ferreres, V., Emulsions and aqueous dispersions of phytosterols, PCT patent application,
WO 02/065859 A1, August 29, 2002.
73. Burruano, B., Bruce, R.D., and Hoy, M.R., Method for producing water dispersible sterol formulations,
U.S. Patent, US 6,110,502, August 29, 2000.
74. Yoon, W.T., Kim, K.S., and Hong, H.P., Mixing powder of plant sterol and emulsifier, and method for
preparing the same, PCT patent application, WO 03/077680 A1, August 25, 2003.
75. Dolhaine, H.G., Kropf, C., Christophliemk, P., Biermann, M., and Schroeder, C., Use of nanoscale ste-
rols and sterol esters, US patent, US 6,352,737 B1, March 5, 2002.
76. Weingartner, O., Bohm, M., and Laufs, U., Controversial role of plant sterol esters in management of
hypercholesterolemia. Eur. Heart J., 30, 404–409, 2009.
77. European Commission, Regulation (EC) No 258/97 of the European Parliament and the Council of 27
January 1997 concerning novel foods and novel food ingredients. Off. J. Eur. Union, L 43/1, 1, February
14, 1997.
78. European Commission, Commission Decision 2004/333/EC. Off. J. Eur. Union, L 105/40, 1, April 14,
2004.
79. European Commission, Commission Decision 2008/36/EC. Off. J. Eur. Union, L 8/15, 1, January 11,
2008.
80. European Commission, Commission Regulation (EC) No. 983/2009 of 21 October 2009 on the authori-
sation and refusal of authorisation of certain health claims made on food and referring to the reduction
of disease risk and to children’s development and health. Off. J. Eur. Union, L 277/3, 1, October 22,
2009.
81. European Commission, Commission Regulation (EU) No. 384/2010 of 5 May 2010 on the authorisation
and refusal of authorisation of certain health claims made on foods and referring to the reduction of
disease risk and to children’s development and health. Off. J. Eur. Union, L 113/6, 1, May 6, 2010.
82. Food Standards Australia New Zealand, Standard 1.5.1. Novel foods standard, in: Australia New Zealand
Food Standards Code. Commonwealth of Australia Gazette, FSC 31, November 9, 2006.
83. Health Canada, Notice of Assessment of Certain Categories of Foods Containing Added Phytosterols,
Bureau of Nutritional Sciences Food Directorate, Health Products and Food Branch Health Canada,
Montreal, Quebec, Canada, May 2010.
84. U.S. Food and Drug Administration, Code of Federal Regulations, Food Labeling, Health Claim,
Phytosterols and risk of coronary heart disease, Propose Rule. Federal Reg., 75, 76526–76571,
December 08, 2010.
61
Beverages Fortified with Omega-3 Fatty Acids,
Dietary Fiber, Minerals, and Vitamins
Fereidoon Shahidi and Priyatharini Ambigaipalan
CONTENTS
61.1 Introduction................................................................................................................................... 801
61.2 Omega-3 Fatty Acids..................................................................................................................... 803
61.2.1 Nutritional Characteristics............................................................................................... 803
61.2.2 Applications in the Beverage Industry............................................................................. 804
61.2.2.1 Dairy Products.................................................................................................. 804
61.2.2.2 Other Beverages................................................................................................ 806
61.2.3 Health Effects of Omega-3 Fatty Acids........................................................................... 807
61.2.3.1 Cardiovascular Disease..................................................................................... 807
61.2.3.2 Cancer............................................................................................................... 807
61.2.3.3 Inflammatory Diseases..................................................................................... 808
61.2.3.4 Mental Health and Neural Function................................................................. 808
61.3 Dietary Fiber................................................................................................................................. 808
61.4 Minerals and Vitamins...................................................................................................................810
61.5 Conclusion......................................................................................................................................810
References................................................................................................................................................810
61.1 Introduction
Fortification of beverages with omega-3 (ω-3) fatty acids, dietary fiber, minerals, and vitamins has been
growing tremendously. For years, consumers have considered beverages only as thirst-quenching and
good-tasting products, while now they specifically look for their health benefits. A survey in 2006 by the
Institute of Food Technologists on the greatest promise as a delivery vehicle for nutraceuticals showed
that nearly 45% of the 327 respondents chose beverages and juices as the most promising delivery vehi-
cles for health-promoting ingredients [1]. It has been reported that 38% of consumers specifically look
for ω-3 fatty acids in the purchase of food and beverages [2]. Thus, today, ω-3 fatty acids have been
incorporated into commonly consumed food and beverage products. Making a new functional food and
beverage product without a significant decrease in the flavor and sensory acceptability has become a
challenge for researchers. However, recently, ω-3 fatty acids have been incorporated into dairy products,
beverages, infant nutrition products, breads and other baked goods, processed meats, cooking oils, and
various prepared foods using advanced technologies [3]. According to the Mintel Global New Products
database, between 2012 and 2013, more than 70 beverages were produced globally by fortification with
ω-3 fatty acids.
Figure 61.1 shows the global market share of ω-3 fortified products by categories with respect
to total market value of US$25.4 billion in 2011. Infant formula had the highest market value,
accounting for approximately 40% of the total, followed by fortified foods and beverages at 31% [3].
801
4%
5%
7%
40%
Infant formula
Food and beverage
13%
Nutritional supplements
Pharmaceutical
Clinical nutrition
Pet food, treats, and supplements
31%
FIGURE 61.1 Global market share for EPA/DHA omega-3 products, 2011. (Adapted from Packaged Facts, Global Market
for EPA/DHA Omega-3 Products, 2012, Published online at: http://www.packagedfacts.com/Global-EPA-DHA-7145087/,
accessed on November 3, 2014.)
5%
18%
North America
51% Europe
Asia-Pacific
Rest of World
26%
FIGURE 61.2 EPA/DHA omega-3 fortified food and beverages: global sale by region, 2011. (Adapted from Packaged
Facts, Global Market for EPA/DHA Omega-3 Products, 2012, Published online at: http://www.packagedfacts.com/Global-
EPA-DHA-7145087/, accessed on November 3, 2014.)
Nutritional supplements place third, followed by pharmaceutical products. The smallest of the product
categories was covered by the pet food, treat, supplement, and other fortified products (eicosapentaenoic
acid [EPA]/docosahexaenoic acid [DHA]). It has been predicted that the market value of EPA/DHA
packaged products would reach US$34.7 billion in 2016, representing a compound annual growth rate
of 6.4%. Omega-3 fatty acids in the beverage market are predominantly found in refrigerated juices and
milk products [3]. North America had the largest share (51%) of the EPA/DHA fortified foods and bever-
ages category at US$4.0 billion in 2011, followed by Europe at US$2.05 billion (26%) and Asia-Pacific
at US$1.4 billion (18%) (Figure 61.2) [3]. This chapter summarizes the nutritional characteristics and
health effects of omega-3 fatty acids, dietary fiber, minerals, and vitamins as well as their application in
beverage industry.
Beverages Fortified with Omega-3 Fatty Acids, Dietary Fiber, Minerals, and Vitamins 803
61.2 Omega-3 Fatty Acids

61.2.1 Nutritional Characteristics
The omega-3 polyunsaturated fatty acids (ω-3 PUFA) exert a wide range of beneficial health effects. The
ω-3 PUFA, such as EPA (20:5ω-3), DHA (22:6ω-3), and to a lesser extent docosapentaenoic acid (DPA;
22:5ω-3), as well as alpha-linolenic acid (ALA; 18:3ω-3) have been shown to exhibit health benefits.
Chemical structures of these fatty acids are presented in Figure 61.3. The ω-3 fatty acids are mainly
derived from marine and plant sources, typically fish, marine mammals, algae, or flax. Traditional sources
of ω-3 fatty acids are fish oils from anchovy or cod liver, but recently, ω-3 fatty acids are also extracted
OH
OH
O
O
Alpha-linolenic acid Eicosapentaenoic acid

ALA (18:3n-3) EPA (20:5n-3)
O
OH
OH
Docosapentaenoic acid Docasahexaenoicacid

DPA (22:5n-3) DHA (22:6n-3)
FIGURE 61.3 Chemical structures of omega-3 fatty acids.

from krill, squid (or calamari), and algae, among others [3]. However, the main sources of fish oils are the
pelagic species caught in large quantities, particularly those with oily flesh, such as salmon, tuna, mack-
erel, and herring or small fish such as anchovy and capelin [4]. The ALA is found in walnut, flaxseed,
chia seed, soybean, and canola oils. Humans can synthesize up to approximately 5% long chain PUFA
(LC-PUFA) through desaturation and elongation from dietary ALA [5]. DHA is the most abundant
LC-PUFA in the gray matter of the brain and in the retina of the eye [6]. Hoffmann et al. [7] reported
that DHA occurs naturally in breast milk and is essential for normal development of infant brain and eye.
61.2.2 Applications in the Beverage Industry

Several studies have shown that ω-3 fatty acids influence blood lipid profiles, cardiovascular health,
membrane lipid composition, eicosanoid biosynthesis, cell signaling cascades, and gene expression [8].
Generally, the intake of ω-3 fatty acids is too low among population living away from coastal areas.
National health and nutrition examination survey revealed that average LC-PUFA consumption in the
United States is less than 185 mg/day, while the International Society for the Study of Fatty Acids and
Lipids recommends 500 mg/day [9]. Deficiency of ω-3 fatty acids has been associated with sluggish
memory and mental performance, tingling sensation of the nerves, poor vision, increased tendency of
blood clots, diminished immune function, increased levels of triacylglycerols (TAG) and low-density
lipoprotein (LDL), impaired membrane function, hypertension, arrhythmia, learning disorders, meno-
pausal discomfort, and growth retardation in infants and children [3,9].
Food processing/technology have now advanced to the point where, in theory, any product with an
oily or fatty component can be enriched with ω-3 PUFA [10]. In Europe and Japan, fish oils are found
in variety of foods, including baked goods, margarines, infant formulas, dietary supplements, and
beverages [11]. Bread is considered to be an ideal medium for ω-3 PUFA fortification, because the car-
bon dioxide released in the proofing and baking process may act favorably to exclude oxygen and prevent
oxidation while bread is at high temperatures [12]. The most commonly observed adverse effects of ω-3
PUFA supplementation are nausea, gastrointestinal upset, and “fishy” burp [13]. However, a large meta-
analysis by Mozaffarian and Rimm [14] also showed the favorable risk-to-benefit ratio (1:400) associated
with a high consumption of fish.
Oxidation of marine ω-3 fatty acids in beverages could cause fishy odors, and improper stabilization
could also result in fishy tastes. Fish and other marine organism–derived oils, similar to other edible
oils, are subjected to different processing steps. Shahidi [15] reported that it is important to treat the
resultant refined, bleached, and deodorized oils with appropriate antioxidants in order to enhance their
oxidative stability. Encapsulating marine lipids for food and beverage applications, referred to as micro-
encapsulation, could produce dry powders or liquid emulsion products [16]. Jin et al. [16] reported that
encapsulation and microencapsulation highly unsaturated oils increase the body demand for vitamin
E. Generally, to improve the shelf life of the oil, tocopherol is added. Hansen et al. [17] suggested that
encapsulated oil particles must be smaller than 100 μm to avoid mouth detection and alteration of food
texture.
61.2.2.1 Dairy Products

Fortification of dairy products with vitamins/minerals, probiotics, or ω-3 fatty acids has become quite
popular these days. As an example, milk and other dairy products have been fortified with vitamin D
since 1933, and consumers are aware of fortification of dairy products [3]. In addition, dairy products,
including drinks, are believed to supply over 20% of the daily requirement for calcium and vitamins A,
D, B12, and riboflavin (vitamin B2) as well as minerals such as magnesium and potassium [18]. Dairy-
based beverages are considered as an ideal delivery vehicle for ω-3 fatty acids, antioxidants as well as
proteins due to their shorter shelf life and need of refrigeration for storage. Currently, available ω-3
fortified dairy products are yogurt (spoonable or drinkable), milk drinks, fresh and ultrahigh tempera-
ture (UHT) processed milk, and products such as margarines and spreads. It has been reported that the
increase in consumption of sweetened beverages decreased the per capita total fluid milk consumption
from 36.4 gallon (~138 L)/person in the 1950s to 22.6 gallon (~86 L)/person in 2000 [19].
Parmalat Finanziaria S.P.A. introduced the ω-3 fortified milk in 1998 [16], which contained 80 mg
of ω-3 per 100 mL serving. The increase in the DHA content in cow milk by natural enrichment has
also occurred [20]. Cows were fed with DHA-enriched feed, which allowed the production of milk with
increased DHA content [20]. They suggested that DHA in the feed is less susceptible to biohydrogena-
tion than ALA and other PUFA, and ALA could pass through the gut without any changes and can be
converted to DHA. Neilson® Dairy produces naturally fortified milk to the Canadian market, which
contains 10 mg of DHA per serving of 250 mL. According to the Agriculture and Agri-Food Canada
[21], ω-3 in most of the dairy products are vegetable-based ALA from flax oil, which is added to milk
either at the dairy or in milk produced by cattle fed a diet rich in flax and flavors may also be added to
milk. The report reveals that the consumption of ω-3 oils has been rising since 2004 as dairy processors
launched ω-3 milk, ice cream, cheese, and yogurt. Examples of current functional dairy foods in Canada
are presented in Table 61.1 [21,22].
Visioli et al. [23] observed that the daily consumption of 500 mL of a commercially produced EPA +
DHA-enriched long-life milk (Parmalat Plus Omega 3®) decreased plasma TAG (~19%) and increased
high-density lipoprotein (HDL) (~19%) after a 6-week period. In another study, a similar observation
was made and the authors concluded that replacing semiskimmed long-life milk with enriched milk
was an effective way to decrease cardiovascular disease (CVD) risk factors [24]. Furthermore, flaxseed-
enriched milk-based beverage to deliver ω-3 fatty acids was shown to be an acceptable product, despite
sensory and compositional differences compared to chocolate milk [1]. In addition, commercially avail-
able functional dairy beverages by Dairy Crest, which milk is enriched with ω-3, Munch Bunch Drinky
Plus is aimed at children in the United Kingdom and Lactalis’s Primevé and Coralis’s Arilait milk
enriched with ω-3 in France [21]. Dawczynski et al. [25] reported that 3 g of ω-3 fatty acids per day in a
dairy drink was associated with improvements in a range of cardiovascular risk factors, including total
cholesterol and TAG levels, and the ratio of arachidonic acid (AA) to EPA.
TABLE 61.1
Commercially Available Functional Dairy Foods in Canada
Company Brand Name Products Functional Components
Agropur Natrel Pro Natrel Milk Calcium
Probiotics
Omega-3
Probiotics
Parmalat Astro Bio-Best Yogurt Omega-3
Parmalat Beatrice Vitalité Premium milk Calcium
Parmalat Beatrice Chocolate milk Omega-3
Calcium
Vitamins
Saputo Dairyland Premium milk Calcium
Probiotics
Omega-3
Vitamins and calcium extra
Saputo Milk 2 Go Premium milk Calcium
Probiotics
Omega-3
Vitamins and calcium extra
Neilson Dairy Ltd. Neilson Dairy Oh! Premium milk DHA
Omega-3
GlenOaks Farms GlanOaks Farms Drinkable yogurt Omega-3
Fibre
Calcium
Abbreviation: DHA, docosahexaenoic acid.
61.2.2.2 Other Beverages

Functional food beverages could be prepared by using fish oil in liquid (30% EPA and DHA) or powder-
microencapsulated (10%) form. Kolanowski [26] suggested that a low-pH water-based product (fruit
beverages) facilitates hydrolysis and oxidation of added fish oil, thereby strongly decreasing the shelf
life of the enriched products. The addition of α-tocopherol (vitamin E; 150 mg/L), β-carotene (20 mg/L),
and ascorbic acid (vitamin C; 200 mg/L) to decrease the oxidation of enriched beverages (in the case of
0.25% level of fish oil addition) is recommended [27]. Food enrichment with fish oil at a significant level
of up to 0.1%, which provided 0.03% EPA and DHA, was found to be not suitable for liquid products of
low flavors and sweetness intensity such as milk or vegetable juices, due to the decreased palatability
[26]. It has been reported that the ω-3 ingredient arena has seen an increased demand for vegan or veg-
etarian diet [28]. Hence, Royal Dutch-based multinational life sciences and materials sciences company
(named DSM) in the Netherlands added Martek’s life DHA (now DSM), a vegetarian DHA sourced from
algae to its ω-3 product [28].
Commercially available beverages fortified with ω-3 are summarized in Table 61.2. Market leaders
in the fortified juice category in the United States include Minute Maid Enhanced Pom Blueberry and
Tropicana Pure Premium Health Heart Orange Juice [3]. ProSure® fortified with EPA is mainly used
as part of an overall cancer-care program for patients with cancer-associated malnutrition and unin-
tended weight loss [29]. Hasbrouck Heights Zymes delivers solubilizing and stabilizing technology for
TABLE 61.2
Commercially Available Omega-3 Fortified Beverages
Company Brand Name Products Form of Omega-3
Oceans Omega ΩmegaActiv from Nutrigrity
® Sodas, waters, fortified Menhaden oil: consists of
Life’s DHA™ from DSM waters, energy drinks, balanced level of EPA, DHA,
Nutritional Products juices, RTD teas, liquid and DPA.
nutritional shots, gels, Fish free DHA from algae.
and gelatins
The Wright Group O3 Complete™ Sports drink Microencapsulated omega-3.
Lifeaid Beverage GolferAID Energy drink Omega-3 and Vitamin B.
Co. FitAID
PartyAID
Minute Maid Minute Maid® Enhanced Fruit juice DHA, choline, vitamins B12, C,
Pomegranate Blueberry and E.
Tropicana Tropicana Pure Premium Health Orange juice EPA, DHA, Vitamin C, folic
Heart acid, and potassium.
Indian River Grape Fruit Juice Fruit juice Omega-3 from fish oil and fish
gelatin, calcium, and dietary
fibre.
Genesis Today Omega Orange Fruit juices Algal oil.
Acaí Berry Classic Juice Omegas-3, -6 and -9, vitamins
B12 and C.
A. Lassonde Oasis Concord Grape-Blueberry Fruit juices Microencapsulated fish oil or
Oasis Premium Orange with linseed oil.
Omega-3
Oasis Health Break Strawberry
and Kiwi with Omega-3
Oasis Fruit Fusion
Big Shotz Vi+amineral™ Super-vitamin health Omega-3, pre-biotic, vitamins B,
drink C, D, E, K, and β-carotene.
Ross ProSure® Nutritional beverage EPA from sardine oil, pre-biotics,
and vitamins C and E.
Smartfish Recharge Fresh juice-based drink Marine EPA and DHA.
for children and adults
Abbreviations: EPA, eicosapentaenoic acid; DHA, docosahexaenoic acid; DPA, docosapentaenoic acid.
ω-3 ingredients. For example, the water-soluble Ubisol-Aqua technology from Hasbrouck Heights Zymes
offers a shelf-stable ω-3 ingredient in clear beverages [28]. Zymes beverage also contains vitamins C, E,
B3, B6, B5, and B12 and is naturally sweetened with Stevia with zero calories. The beverage is currently
available in two flavors, Clearberry and lemon-lime (lemon-lime contains electrolytes). Norwegian ω-3
specialist Smartfish produced a juice-based beverage enriched with ω-3 drink containing DHA and EPA
(110 mg), vitamin D3 (10 μg), and protein (8 g) and was sold in pharmacies, hospitals, and rest homes
as a medical food targeting the malnourished elderly [30,31]. Nestlé has also launched a nutrient drink,
Resource SeniorActiv in Switzerland targeting the malnourished elderly in 2009, is enriched with EPA,
DHA, vitamin D, calcium, and protein [31].
61.2.3 Health Effects of Omega-3 Fatty Acids

61.2.3.1 Cardiovascular Disease
Numerous prospective and retrospective trials from many countries have shown that moderate fish oil con-
sumption decreases the risk of major cardiovascular events, such as myocardial infarction, sudden cardiac
death (SCD), coronary heart disease (CHD), atrial fibrillation, and most recently, death in patients with
heart failure [13,32]. Current research suggests that ω-3 PUFA may prevent fatal arrhythmias via their abil-
ity to inhibit fast, voltage-dependent sodium channels, and L-type calcium channels [33]. Kris-Etherton
et al. [34] reported that EPA and DHA specifically reduce plasma TAG concentration and thereby the risk
of CVD. A recent systematic review summarized 47 trials and found that fish oil supplementation was
effective in reducing TAG concentration by 0.34 mmol/L over an average treatment period of 24 weeks in
participants with hypertriglyceridemia [35]. Isocaloric replacement of about 5% of energy from saturated
fatty acids with oleic acid (or PUFA) has been estimated to reduce CHD risk by 20%–40% mainly via
LDL cholesterol reduction [34]. Although fatty acids are classically observed as an energy substrate, they
are also endogenous ligands for peroxisome proliferator-activated receptors and regulate the expression of
genes encoding key proteins controlling fatty acid uptake and metabolism and the formation of very-low-
density lipoprotein (VLDL) carrying TAG in the liver [36]. Harris [37] reported that ω-3 PUFA confer
cardiovascular benefits largely through EPA and DHA enrichment of membrane phospholipids. In addi-
tion, ω-3 PUFA render vasodilation, reduce blood pressure, improve arterial and endothelial function, and
reduce platelet aggregation [38]. One of the important suggested mechanisms for the potential beneficial
effect of fish consumption on CVD is that LC ω-3 PUFA can prevent cardiac death through inhibition of
malignant cardiac arrhythmias, a primary cause of sudden death [39]. Goel et al. [40] demonstrated that
specific fatty acids can significantly inhibit the activity of the cardiac sarcolemmal Na+/H+ exchanger,
which induces arrhythmias, cell damage, and eventually cell death. The American Heart Association has
endorsed the use of ω-3 PUFA at a dose of approximately 1 g/day of combined EPA and DHA, either in
the form of fatty fish or fish oil supplements (in capsules or liquid form) in patients with documented CHD
[34]. vonSchacky and Harris [41] proposed the ω-3 index (percentage of EPA + DHA of total fatty acids in
red blood cells) as a risk factor for SCD and concluded that this index should be higher than 8%.
61.2.3.2 Cancer
Epidemiological studies have indicated that a high intake of saturated fat and/or animal fat increases the
risk of colon and breast cancer [42]. Several studies have pointed out that ω-3 fatty acid consumption
is associated with a decreased risk of several cancers such as breast, prostate, colon, and kidney [43].
Whether this effect is mediated by the actual EPA and DHA or by the EPA- and DHA-derived eicosanoids
and docosanoids has not yet been clarified [44]. A meta-analysis by Fay et al. [45] showed that LC ω-3
PUFA had only a small protective effect, while ω-6 PUFA had a strong tumor-enhancing effect. Recently,
a therapeutic study in breast cancer patients where DHA was combined with the chemotherapeutic drugs
epirubicin, cyclophosphamide, and 5-fluorouracil emphasized that an interindividual uptake and incor-
poration of DHA can alter the treatment response [46]. The high incorporating group was characterized
by delayed time to tumor progression and longer overall survival compared to the low incorporating
group. Cyclooxygenase (COX) II was increased in a variety of human cancers, including colon, breast,
esophagus, hepatocellular, cervical, bladder, and pancreas and may suppress apoptosis [47]. Stehr and
Heller [47] reported that ω-3 fatty acids downregulate COX II expression, alter its metabolites, and con-
sequently avoid prostaglandin E2 (PGE2)-induced increase of estrogen in breast cancer and prostaglandin
H2 (PGH2)-induced DNA damage. Animal studies on DHA supplementation as cancer prevention have
shown that DHA-enriched or fish oil-enriched diets can inhibit the formation of papillomas and of mam-
mary carcinogenesis, carcinogenesis of the large and small intestine, and lungs [48]. Sharma et al. [49]
demonstrated that ω-3 fatty acids have variable antiproliferative effects on a selected population of ovar-
ian cancer cell lines. It has been reported that excess ω-6 fatty acids may increase the risk of breast cancer
metastasis, while ω-3 fatty acids had the opposite action. Thus, the ratio of ω-3/ω-6 fatty acids is of prime
importance [50]. A case control study of nonmenopausal women found that the risk of developing breast
cancer was 50% lower in women with the lowest ω-3/ω-6 ratio than in the group with the highest ratio [51].
61.2.3.3 Inflammatory Diseases

The common denominator that might explain the beneficial effects of ω-3 fatty acids and DHA in
particular, in this great variety of diseases and syndromes, is their anti-inflammatory property [44].
Omega-3 fatty acids, if adequately protected from oxidation, are involved in alleviating atherosclerosis,
chronic hepatitis, inflammatory bowel diseases, psoriasis, and rheumatoid arthritis [52]. There is exten-
sive documentation in the rheumatology, ophthalmology, and cardiovascular literature on the beneficial
anti-inflammatory effects of a high-dose fish oil in the reduction of joint pain from rheumatoid and osteo-
arthritis, improvement in dry eyes and macular degeneration, and also major positive effects on lipid
profile, plaque formation arrhythmias, and reduction in infarction from coronary arthrosclerosis, which
is now considered an inflammatory disease [53]. Several studies have reported that the supplementation
of ω-3 PUFA to patients with rheumatoid arthritis results in a significant reduction of symptoms in all
studies (13 double-blind and placebo-controlled studies), including reduced duration of morning stiff-
ness, number of tender or swollen joints, joint pain, and time to fatigue and increased grip strength [54].
61.2.3.4 Mental Health and Neural Function

Research suggests that essential fatty acids are involved in neurodevelopment of cognitive and emotional
function and that supplementation of essential fatty acids, particularly EPA, can improve mood and
alleviate depression [55]. LC ω-3 fatty acids have now been investigated in epidemiological, prospec-
tive cohort, cross-sectional, and clinical trials in bipolar disorder, schizophrenia, depression, dementia
and Alzheimer’s disease, and developmental disorders including attention deficit hyperactivity disorder,
dyslexia, dyspraxia, and the autistic spectrum disorders [56], with some conflicting and some positive
indications for their use as a supplementary treatment for symptoms [57]. EPA and DHA have also been
related to many functions important for neural activity including myelination, membrane fluidity, neuro-
transmission, ion channel and enzyme regulation, and gene expression [57]. Horrobin [58] reported that
DHA and AA were present in high concentrations in the grey matter of the cerebral cortex of the brain,
specifically in the membranes of neuronal synapses, approximately 6%–8% of its nonaqueous weight.
Recently, Jordan [59] reported that adequate ω-3 intake in pregnant and nursing women and young chil-
dren was acknowledged for its positive role in increasing gestation and birth weight and its effects on
neonatal cognitive and visual function.
61.3 Dietary Fiber

Dietary fiber is of growing interest because of its beneficial effect on human health such as weight con-
trol, cholesterol level, diabetes, and in the prevention of CVD, diverticulitis, varicose veins, hiatus of the
hernia, and colon cancer [60]. Generally, dietary fiber exists as soluble and insoluble forms. Akad et al.
[60] reported that insoluble dietary fibers are difficult to maintain in proper suspension or dispersion,
due to accumulation in the bottom of containers of drinks and beverages. In addition, insoluble dietary
fibers could affect the taste of beverages as well as texture of the liquid foodstuffs. Thus, soluble fiber
is widely used for beverage fortification. Daily intake of dietary fiber recommended by the American
Dietetic Association is 38 g for men and 25 g for women of 50 years and younger, and 30 and 21 g for
men and women over 50, respectively [61].
Generally, gums and starches are considered as healthy sources of soluble dietary fiber. Gums are
widely used in beverage formulation due to their ability to prevent sensory properties such as taste and
smell as well as other benefits, such as stabilization of milk proteins at low pH, emulsion stabilization
in drinks, pleasant mouthfeel, good suspension, and desirable levels of cloud in beverages [62]. In addi-
tion, fructose polymers inulin and oligofructose and polydextrose are used as a source of dietary fiber.
Functional soluble dietary fibers are also produced from novel plant sources such as apple pomace,
orange peel, and chicory. A beverage comprising soluble dietary fiber supplement (gum Arabic and
pectin) with a fiber level of at least 2 g per 8 fluid oz (~237 mL) beverage has been successfully prepared
without alteration in sensory characteristics [60]. Sáyago-Ayerdi et al. [63] reported that a beverage of
Hibiscus sabdariffa flowers, widely consumed in Mexico, contained dietary fiber as the largest compo-
nent (33.9%) and soluble dietary fiber of 0.66 g/L in beverage.
Blecker et al. [64] reported that inulin improves body and mouthfeel in cheese analogues or ice cream,
and reduces synaeresis in yogurt and other fermented milk products. However, there are concerns of
potential disadvantages of the formulation of the beverage with dietary fiber, due to its ability to decrease
the bioavailability of calcium provided in the milk. Propylene glycol alginate (PGA) is used in place
of alginates in beverages, where the carboxylic acid groups of alginates have been esterified to prevent
gelation and precipitation at low pH. Thus, fruit drinks containing pulp are fortified with medium esteri-
fied PGA at 0.1% with high calcium to prevent sedimentation, which forms a cross-linked network with
higher viscosity [65]. Beristain et al. [61] reported that the addition of 0.4%–0.5% (w/w) of carboxymethyl
cellulose or xanthan gum to a noncentrifuged cloudy apple juice showed stable turbidity for extended
periods of storage. Examples of dietary fibers used in the beverage industry are listed in Table 61.3.
TABLE 61.3
Examples of Dietary Fibers Used by the Beverage Industry and Their Purpose
Dietary Fiber Purpose
Polypropylene glycol alginate Increase viscosity.
Polypropylene glycol alginate and Particle suspension, calcium fortification, and viscosity control.
xanthan gum
High-methoxy pectins Viscosity control.
Xanthan gum Improves the mouthfeel of citrus and fruit-flavored beverages and shear thinning.
Microcrystalline cellulose or Aids in suspending, thickening, and stabilizing.
cellulose gel, and sodium
carboxymethyl cellulose
Guar gum Provides medium viscosity without excessive gumminess, enhances mouthfeel, and
helps to evenly suspend particulates, emulsifying flavor oils in beverage
emulsions, providing a stable cloud in the drink and boosting the cloud from
added fruit pulps and juices.
Gum arabic and gum tragacanth Used to reduce the viscosity without reducing emulsion stability, resulting in a
thin, pourable consistency.
Carrageenan Give good body to control to beverages while pouring, without causing gummy or
slimy coatings that linger in the mouth, used to control syneresis and to enhance
mouthfeel in cultured dairy beverages.
Locust bean gum Used in the formulation of milkshakes at concentrations less than 0.1% w/w to
thicken and impart a creamier mouthfeel to the product.
Modified tapioca starches Enhance mouthfeel and texture of liquid foods and beverages, including solutions
too low in viscosity for starch granules to remain suspended have been developed.
Polydextrose Used in transparent beverages, low glycemic, and can be used in sugar-free
beverages suitable for diabetics.
Inulin and fructooligosaccharides Act as soluble fiber and both substances contribute fewer calories than sugar or
starch, so that they can be used in the formulation of low-carbohydrate beverages.
61.4 Minerals and Vitamins

Tables 61.1 and 61.2 show some commercially available functional dairy foods and beverages enriched with
minerals and vitamins. The major consideration in formulating beverages with added minerals and vitamins
and is their stability and solubility [61]. For example, vitamins A, B, and C tend to break down when exposed
to sugar, oxygen, and various levels of pH. Beristain et al. [61] reported that large doses of water-soluble
vitamins are quite safe, while doses above 50 mg/day of vitamin B6 could affect the sensory nerve function.
Additionally, fat soluble vitamins (A, D, E, and K) are toxic at higher dosage levels. Energy drinks are widely
formulated with antioxidants such as vitamins A, C, and E. Major vitamins and minerals desired by consum-
ers for supplementation of beverages are calcium, iron, Vitamin C, B complex vitamins, and folic acid [66].
Fortification with minerals could result in solubility problems. For example, calcium could affect that
solubility of protein. Calcium and iron are widely used, as well as chromium, selenium, and zinc, which
are used in small quantities. Calcium is widely used to fortify fruit juices due to its ability to maintain
bone particularly in postmenopausal women and young children. In addition, bioavailability of minerals
is another important factor to be considered when formulating beverages with minerals. It has been men-
tioned that organic calcium salts, such as calcium citrate, calcium lactate, calcium lactate gluconate, and
calcium gluconate, generally provide more bioavailability and more solubility than inorganic calcium
sources [61]. Salts are important ingredients of isotonic and sports drinks due to their ability to replace
the electrolytes (sodium and potassium) that the body loses through sweat.
Presently, cod liver oil is dominant in the market and contains high levels of vitamins A and D. Shahidi
[4] reported that although cod as well as halibut liver oils are rich in ω-3 PUFA, they were originally used
primarily as a source of vitamins A and D and hence may often be diluted with vegetable oils. Vitamin D
plays a key role in immunity development, reducing inflammation and also promotes bone and joint health.
Calcium is known to be an essential element required for numerous functions in our body including
the strengthening of teeth and bones, nerve function, and many enzymatic reactions that require calcium
as a cofactor [67]. Larsen et al. [68] reported that consumption of whole small fish is nutritionally ben-
eficial providing with a rich source of calcium and shown that calcium in fish could be absorbed to the
body as tested in vivo. A vitamin- and mineral-fortified beverage that is stable and contains vitamin A in
the form of encapsulated β-carotene, vitamin C, and riboflavin was developed earlier [69]. Chandler et al.
[70] produced a liquid beverage consisting of water, calcium glycerophosphate as a source of calcium, a
vitamin D emulsion using a gum selected from gum Arabic, gum tragacanth, and xanthan gum.
61.5 Conclusion
Several studies have shown that ω-3 fatty acids, dietary fiber, minerals and vitamins are effective in
lowering cholesterol and blood pressure, preventing heart disease, reducing the risk of stroke and certain
cancers, controlling diabetes and inflammatory diseases, aiding in weight loss, as well as enhancing
calcium assimilation, skin healing, and mental clarity [3]. Global market for ω-3, dietary fiber, miner-
als, and vitamins-enriched beverages are growing steadily. Presently, technological advances such as
microencapsulation masks fishy taste and smell of the ω-3 oils. This provides impetus for manufacturers
to produce more diversified ω-3 fortified beverages. Overall, these studies suggest that the formulation
and usage of nutrition-enriched milk beverages and juices are feasible tools to increase ω-3 fatty acids,
dietary fiber, minerals, and vitamins intake.
REFERENCES
1. Lau, C.S., Formulation and physical, chemical, and sensory analysis of a novel flaxseed-enriched milk-
based beverage to deliver Omega-3 fatty acids, PhD thesis, Virginia Polytechnic Institute and State
University, Blacksburg, VA, 2007.
2. Oceans Omega, Oceans Omega market research, 2014, Published online at: http://www.oceansomega.
com/our-products-services-2 (accessed November 3, 2014).
3. Packaged Facts, Global market for EPA/DHA Omega-3 products, 2012. Published online at: http://
www.packagedfacts.com/Global-EPA-DHA-7145087/ (accessed November 3, 2014).
4. Shahidi, F., Omega-3 oils: Sources, applications, and health effects, in Marine Nutraceuticals and
Functional Foods, Barrow, C. and Shahidi, F., Eds., CRC Press/Taylor & Francis Group, Boca Raton,
FL, 2008, pp. 23–61.
5. Hulshof, K.F.A.M., van Erp-Baart, M.A., Anttolainen, M., Becker, W., Church, S.M., Couet, C.,
Hermann-Kunz, E. et al., Intake of fatty acids in western Europe with emphasis on trans fatty acids:
The TRANSFAIR study. Eur. J. Clin. Nutr., 53, 143–157, 1999.
6. Salem Jr., N., Omega-3 fatty acids: Molecular and biochemical aspects, in New Protective Roles for
Selected Nutrients, Spiller, G.A. and Scala, J., Eds., Alan R. Liss, Inc., New York, 1989, pp. 109–228.
7. Hoffmann, D.R., Boettcher, J.A., and Diersen-Schade, D.A., Toward optimizing vision and cognition
in term infants by dietary docosahexaenoic acid and arachidonic acid supplementation: A review of
randomized controlled trials. Prostaglandins Leukot. Essent. Fatty Acids, 81, 151–158, 2009.
8. Shahidi, F. and Miraliakbari, H., Marine oils: Compositional characteristics and health effects, in
Nutraceutical and Speciality Lipids and Their Co-Products, Shahidi, F. Ed., CRC Press/Taylor &
9. Jacobsen, J., Counting the benefits of omega-3 fatty acids. Beverage Ind., 104, 54–55, 2013.
10. Alasalvar, C., Shahidi, F., and Quantick, P., Food and health applications of marine nutraceuticals: A
review, in Seafoods-Quality, Technology and Nutraceutical Applications, Alasalvar, C. and Taylor, T.,
Eds., Springer, Heidelberg, Germany, 2002, pp. 175–204.
11. Nice, D.J. and Little, A.D., Overview of marine nutraceuticals: Opportunities and considerations for new
food products. Paper presented at the Annual Meeting of the Institute of Food Technologists, Orlando,
FL, June 14–18, 1997, abstract no. (pp. 150).
12. Newton, I. and Snyder, D., Nutritional aspects of long-chain omega-3 fatty acids and their use in bread
enrichment. Cereal Foods World, 42, 126–131, 1997.
13. Lavie, C.J., Milani, R.V., Mehra, M.R., and Ventura, H.O., Omega-3 polyunsaturated fatty acids and
cardiovascular diseases. J. Am. Coll. Cardiol., 54, 585–594, 2009.
14. Mozaffarian, D. and Rimm, E.B., Fish intake, contaminants, and human health: Evaluating the risks
and the benefits. J. Am. Med. Assoc., 296, 1885–1899, 2006.
15. Shahidi, F., Marine oils from marine by-products, in Maximising the Value of Marine By-Products,
Shahidi, F., Ed., Woodhead Publishing, Cambridge, UK, 2007, pp. 258–278.
16. Jin, Y., Perrie, C., Zhang, W., Diepen, C.V., Curtis, J., and Barrow, C.J., Microencapsulation of marine
lipids as a vehicle for functional food delivery, in Marine Nutraceuticals and Functional Foods, Barrow,
C. and Shahidi, F., Eds., CRC Press/Taylor & Francis Group, Boca Raton, FL, 2007, pp. 115–155.
17. Hansen, L.T., Allan-Wojtas, P.M., Jin, Y.L., and Paulson, A.T., Survival of Ca-alginate microencapsulated
Bifidobacterium spp. in milk and simulated gastrointestinal conditions. Food Microbiol., 19, 35–45, 2002.
18. Fortitech, Inc., New innovations in functional beverages, 2011, Published online at: https://www.
fortitechpremixes.com/wp-content/uploads/2012/06/Functional_Beverages_FINAL_ENG.pdf
(accessed November 5, 2014).
19. USDA, Agriculture Fact Book 2001–2002, USDA, Washington, DC, 2003.
20. Holub, B.J., McBride, B., and Wright, T.C., U.S. Patent No. 5,932,257, U.S. Patent and Trademark Office,
21. Functional Foods-Dairy, Agriculture and Agri-Food Canada, 2007, Published online at: http://www4.
agr.gc.ca/resources/prod/doc/misb/fb-ba/nutra/pdf/ffd2007_e.pdf (accessed November 6, 2014).
22. Euromonitor International Ltd., Report on Global Fortified/Functional Foods & Beverages, London,
UK, 2006.
23. Visioli, F., Rise, P., Plasmati, E., Pazzucconi, F., Sirtori, C.R., and Galli, C., Very low intakes of n-3
fatty acids incorporated into bovine milk reduce plasma triacylglycerol and increase HDL-cholesterol
concentrations in healthy subjects. Pharmacol. Res., 41, 571–576, 2000.
24. Carrero, J.J., Baró, L., Fonollá, J., Gonzalez-Santiago, M., Martinez-Ferez, A., Castillo, R., Jimenez, J.,
Boza, J.J., and Lopez-Huertas, E., Cardiovascular effects of milk enriched with ω-3 polyunsaturated
fatty acids, oleic acid, folic acid, and vitamins E and B6 in volunteers with milk hyperlipidemia.
Nutrition, 20, 521–527, 2004.
25. Dawczynski, C., Martin, L., Wagner, A., and Jahreis, G., n-3 LC-PUFA-enriched dairy products are able
to reduce cardiovascular risk factors: A double-blind, cross-over study. Clin. Nutr., 29, 592–599, 2010.
26. Kolanowski, W., Possibilities of fish oil application for food products enrichment with omega-3 PUFA.
Int. J. Food Sci. Nutr., 50, 39–49, 1999.
27. Hoffman La Roche, Bringing the Nutritional Gap: n-3 PUFA Food Enrichment, Hoffman La Roche
Ltd., Vitamins and Fine Chemicals Division, Basel, Switzerland, 1996.
28. Jacobsen, J., Omega-3 carving its niche. Beverage Ind., 102, 82, 2011.
29. ProSure, Key features of ProSure, 2014. Published online at: http://prosure.com/product (accessed
November 6, 2014).
30. Starling, S., Scandinavians target elderly with 1000 mg + omega-3 drink. NutraIngredients, August 26,
2010.
31. Hernandez, E.M., Jong, L.D., and Hosokawa, M., Applications of omega-3 fats in foods, in Omega-3
Oils: Applications in Functional Foods, Hernandez, E.M. and Hosokawa, M., Eds., AOCS Press,
Urbana, IL, 2011, pp. 151–176.
32. Yokoyama, M., Origasa, H., Matsuzaki, M., Matsuzawa, Y., Saito, Y., Ishikawa, Y., Oikawa, S. et al.,
Japan EPA lipid intervention study (JELIS) investigators. Effects of eicosapentaenoic acid on major
coronary events in hypercholesterolaemic patients (JELIS): A randomised open-label, blinded endpoint
analysis. Lancet, 369, 1090–1098, 2007 (published correction appears in Lancet, 370, 220, 2007).
33. Anand, R.G., Alkadri, M., Lavie, C.J., and Milani, R.V., The role of fish oil in arrhythmia prevention.
J. Cardiopulm. Rehabil. Prev., 28, 92–98, 2008.
34. Kris-Etherton, P.M., Monounsaturated fatty acids and risk of cardiovascular disease. Circulation, 100,
1253–1258, 1999.
35. Eslick, G.D., Howe, P.R., Smith, C., Priest, R., and Bensoussan, A., Benefits of fish oil supplementation
in hyperlipidemia: A systematic review and meta-analysis. Int. J. Cardiol., 136, 4–16, 2008.
36. Berger, J. and Moller, D.E., The mechanisms of action of PPARs. Annu. Rev. Med., 53, 409–435, 2002.
37. Harris, W.S., Omega-3 fatty acids and cardiovascular disease a case for omega-3 index as a new risk
factor. Pharmacol. Res., 55, 217–223, 2007.
38. O’Keefe Jr, J.H., Abuissa, H., Sastre, A., Steinhaus, D.M., and Harris, W.S., Effects of omega-3 fatty
acids on resting heart rate, heart rate recovery after exercise, and heart rate viability in men with healed
myocardial infarctions and depressed ejection fractions. Am. J. Cardiol., 97, 1127–1130, 2006.
39. Huikuri, H.V., Castellanos, A., and Myerburg, R.J., Sudden death due to cardiac arrhythmias. New Eng.
J. Med., 345, 1473–1482, 2001.
40. Goel, D.P., Maddaford, T.G., and Pierce, G.N., Effects of omega-3 polyunsaturated fatty acids on cardiac
sarcolemmal Na+/H+ exchange. Am. J. Physiol. Heart Circ. Physiol., 283, H1688–H1694, 2002.
41. vonSchacky, C. and Harris, W.S., Cardiovascular benefits of omega-3 fatty acids. Cardiol. Res., 73,
310–315, 2007.
42. Hardman, W.E., Omega-3 fatty acids to augment cancer therapy. J. Nutr., 132(Suppl.), 3508S–3512S,
2002.
43. Thiebaut, A.C.M., Chajes, V., Gerber, M., Boutron-Ruault, M.-C., Joulin, V., Lenoir, G., Berrino, F.,
Riboli, E., Bénichou, J., and Clavel-Chapelon, F., Dietary intakes of ω-6 and ω-3 polyunsaturated fatty
acids and the risk of breast cancer. Int. J. Cancer, 124, 924–931, 2009.
44. Gleissman, H., Johnsen, J.I., and Kogner, P., Omega-3 fatty acids in cancer, the protectors of good and
the killers of evil. Exp. Cell Res., 316, 1365–1373, 2010.
45. Fay, M., Freedman, L., Clifford, C., and Midthune, D., Effect of different types and amounts of fat on
the development of mammary tumours in rodents: A review. Cancer Res., 57, 1379–1388, 1997.
46. Bougnoux, P., n-3 Polyunsaturatedfatty acids and cancer. Curr. Opin. Clin. Nutr. Metab. Care, 2,
121–126, 1999.
47. Stehr, S.N. and Heller, A.R., Omega-3 fatty acids effects on biochemical indices following cancer sur-
gery. Clin. Chim. Acta, 373, 1–8, 2006.
48. Toriyama-Baba, H., Iigo, M., Asamoto, M., Iwahori, Y., Park, C.B., Han, B.S., Takasuka, N. et al.,
Organotropic chemopreventive effects of n-3 unsaturated fatty acids in a rat multi-organ carcinogenesis
model. Jpn. J. Cancer Res., 92, 1175–1183, 2001.
49. Sharma, A., Belna, J., Espat, J., Rodriguez, G., Cannon, V.T., and Hurteau, J.A., Effects of omega-3 fatty
acids on components of the transforming growth factor beta-1 pathway: Implication for dietary modifi-
cation and prevention in ovarian cancer. Am. J. Obstet. Gynaecol., 200, 516e1–516e6, 2009.
50. Chajes, V. and Bougnoux, P., Omega-6/omega-3 polyunsaturated fatty acid ratio and cancer. World Rev.
Nutr. Diet., 92, 133–151, 2003.
51. Goodstine, S.L., Zheng, T., Holford, T.R., Ward, B.A., Carter, D., Owens, P.H., and Mayne, S.T., Dietary
(n-3)/(n-6) fatty acid ratio: Possible relationship to premenopausal but not postmenopausal breast cancer
risk in U.S. women. J. Nutr., 133, 1409–1414, 2003.
52. Gil, A., Polyunsaturated fatty acids and inflammatory diseases. Biomed. Pharmacother., 56, 388–396,
2002.
53. Maroon, C.J. and Bost, J.W., Omega-3 fatty acids (fish oil) as an anti-inflammatory: An alternative to
nonsteroidal anti-inflammatory drugs for discogenic pain. Surg. Neurol., 65, 326–331, 2006.
54. Calder, P.C., Polyunsaturated fatty acids, inflammation and immunity. Lipids, 36, 1007–1024, 2001.
55. Fontani, G., Corradeschi, F., Felici, A., Alfatti, F., Migliorini, S., and Lodi, L., Cognitive and physi-
ological effects of omega-3 polyunsaturated fatty acid supplementation in healthy subjects. Eur. J. Clin.
Invest., 35, 691–699, 2005.
56. Richardson, A.J., Omega-3 fatty acids in ADHD and related neurodevelopmental disorders. Int. Rev.
Psychiatr., 18, 155–172, 2006.
57. Sin, N. and Howe, P.R.C., Mental health benefits of omega-3 fatty acids may be mediated by improve-
ments in cerebral vascular function. Biosci. Hypoth., 1, 103–108, 2008.
58. Horrobin, D., The Madness of Adam and Eve: How Schizophrenia Shaped Humanity, Bantam Press,
London, UK, 2001.
59. Jordan, R.G., Prenatal omega-3 fatty acids: Review and recommendations. J. Midwifery Wom. Health,
55, 520–528, 2010.
60. Akad, M.E., Bolles, A.D., Dougan, D.G., and Hoersten, K.P., U.S. Patent No. 4,988,530, U.S. Patent and
Trademark Office, Washington, DC, 1991.
61. Beristain, C.I., Cruz-Sosa, F., Lobato-Calleros, C., Pedroza-Islas, R., Rodríguez-Huezo, M.E., and
Verde-Calvo, J.R., Applications of soluble dietary fibers in beverages. Rev. Mex. Ing. Quim., 5, 81–95,
2006.
62. Saunders, L., Beverage creation, design elements, food product design, 1994, Published online at: http://
www.foodproductdesign.com/archive/1994/0494DE.html (accessed November 21, 2014).
63. Sáyago-Ayerdi, S.G., Arranz, S., Serrano, J., and Goñi, I., Dietary fiber content and associated anti-
oxidant compounds in roselle flower (Hibiscus sabdariffa L.) beverage. J. Agric. Food Chem., 55,
7886–7890, 2007.
64. Blecker, C., Chevalier, J.P., Van Herck, J.C., Fougnies, C., Deroanne, C., and Paquot, M., Inulin: Its
physicochemical properties and technological functionality. Rec. Res. Dev. Agric. Food Chem., 5,
125–131, 2001.
65. Clare, K., Algin, in Industrial Gums: Polysaccharides and Their Derivatives, 3rd ed., BeMiller, J.N.
and Whistler, R.L., Eds., Academic Press, Inc., San Diego, CA, 1993, pp. 105–143.
66. Liebrecht, J.W. and Phillips, K.M., U.S. Patent No. 6,106,874, U.S. Patent and Trademark Office,
67. Jung, W.K., Shahidi, F., and Kim, S.K., Calcium from fish bone and other marine resources, in Marine
Nutraceuticals and Functional foods, Barrow, C. and Shahidi, F., Eds., CRC Press/Taylor & Francis
Group, Boca Raton, FL, 2008, pp. 419–429.
68. Larsen, T., Thilsted, S.H., Kongsbak, K., and Hansen, M., Whole small fish as a rich calcium source.
Br. J. Nutr., 83, 191–196, 2000.
69. Heckert, D.C., Hughes, D.L., Mehansho, H., and Nakel, G.M., U.S. Patent No. 4,992,282, U.S. Patent
and Trademark Office, Washington, DC, 1991.
70. Chandler, M.A., Dewille, N.T., Geraghty, M.E., Johnson, C.D., Mazer, T.B., Ragan, R.J., and Snowden,
G.A., U.S. Patent No. 5,597,595, U.S. Patent and Trademark Office, Washington, DC, 1997.
62
Probiotics and Prebiotics in Functional Beverages
Koen Venema
CONTENTS
62.1 Introduction....................................................................................................................................815
62.3.1 Probiotics...........................................................................................................................817
62.3.2 Prebiotics...........................................................................................................................817
62.3.3 Antioxidants and Bioactives Responsible for Antioxidant Activity................................. 820
62.4 Health Effects.................................................................................................................................821
62.6 Conclusion..................................................................................................................................... 824
References............................................................................................................................................... 824
62.1 Introduction
Probiotics are “live microorganisms that, when administered in adequate amounts, confer a health ben-
efit to the host” [1]. The two main genera from which species and strains are selected as probiotic bacte-
ria are Lactobacillus and Bifidobacterium. Especially, lactobacilli (and other lactic acid bacteria) have
historically been used to produce fermented dairy products and are considered generally recognized as
safe (GRAS) microorganisms. As consumers’ understanding of the role of probiotics in health grows, so
does the popularity of foods containing them, and hence certain strains of both genera are increasingly
utilized to formulate functional foods, also beyond dairy products.
Prebiotics are defined as “nondigestible food ingredients that beneficially affects the host by selec-
tively stimulating the growth and/or activity of one or a limited number of bacteria in the colon, and thus
improve host health” [2]. This definition was later refined by the same authors to include other bodily
areas of the host that may benefit from selective targeting of particular microorganisms [3]. A more
recently adopted definition by the International Scientific Association of Probiotics and Prebiotics is “a
selectively fermented ingredient that results in specific changes in the composition and/or activity of the
gastrointestinal microbiota, thus conferring benefit(s) upon host health” [4], in which the term “gastro-
intestinal (GI)” reappeared, indicating that the jury is still out on non-GI prebiotics. Even though other
areas of the body are being targeted by prebiotics, this is usually done by giving prebiotics orally. Hence,
the number of products containing prebiotics has also increased substantially over the past decades.
The result of the aforementioned phenomena is a large increase in the number of probiotic- and
prebiotic-containing foods available for public consumption, including a rapidly emerging variety of
probiotic- and/or prebiotic-containing (nondairy) beverages, which provide a convenient way to improve
and maintain health. This contribution focuses on functional beverages containing probiotics and/or pre-
biotics. Previously, probiotics have been primarily added to dairy products. The composition of nondairy
probiotic beverages can pose specific challenges to the survival of probiotics. To overcome these chal-
lenges, strain selection and protection techniques play an integral part in formulating a stable product
to protect the viable cells. One way of accomplishing this has been the addition of prebiotics, where the
815
prebiotics (usually carbohydrates) are thought to protect the probiotic cells in the product. In addition to
protecting the cells, prebiotics are also considered a “lunch box” for the bacteria as they travel through
the hostile GI tract, allowing them to survive better [2]. This chapter highlights recent developments in
functional beverages containing probiotic and/or prebiotics.

Many probiotic microorganisms were originally isolated from fermented dairy foods and are now
increasingly incorporated into dairy and nondairy products. Fermentation increases the nutritional con-
tent, improves sensory characteristics, and preserves many nonalcoholic, cereal-based beverages [5].
A variety of traditional non-dairy-fermented beverages are produced around the world including Boza,
Bushera, Mahewu, Pozol, Togwa, and Hardaliye [5]. Traditional food fermentation utilized to produce
these nondairy probiotic beverages is usually performed to prevent spoilage, and hence provides a means
for obtaining a safe, yet nutritious beverage consumed in developing countries, where starvation and
undernutrition are common challenges. At the same time, the Western world, which is battling modern
diseases associated with obesity and impaired immunity, is taking notice of probiotic benefits in prevent-
ing and treating a number of health conditions. Increasing health consciousness and growing awareness
of the benefits of probiotic yogurts and fermented milks or other beverages, combined with vast use of
probiotic supplements, are main contributors of the growth of probiotic market. The nondairy probiotic
beverages commercially available are mostly refrigerated juice and juice drinks containing nonstarter
but viable cultures of probiotics. As stated above, more and more, these probiotic beverages are being
supplemented with prebiotics. As an example, one of the nondairy beverages is a 100% juice smoothie
containing Bifidobacterium lactis HN019, fructooligosaccharides, and a blend of juices. The formulation
contains fructooligosaccharides, which contribute to the sensory experience and nutritional composi-
tion of the juice and possibly change its physicochemical properties through water activity reduction.
The latter possibly increases the stability of the probiotic bacteria [6]. Benefits of formulating products
with prebiotics extend beyond just product applications. Products with both probiotics and prebiotics
are considered synbiotics, where both functional components individually may have their own benefit
for the host [2]. Another important aspect of products with probiotics and prebiotics is their nutritional
profile. Dietary guidelines in numerous countries around the globe recommend making healthier food
choices to reduce the prevalence of overweight and obesity, which have taken global epidemic propor-
tions in Western countries. The guidelines recommend among others increasing consumption of veg-
etables, fruits, whole grains, milk, and milk products to obtain more potassium, dietary fiber, calcium,
and vitamin D and to consume less sodium, saturated and trans fats, added sugars, and refined grains.
Many ingredients belonging to the recommended healthy food groups have been shown to have prebiotic
effects on various strains of bacteria and to serve as substrates during the fermentation process, as well
as protecting viable cells from exposure to acid, digestive enzymes, and bile during passage through the
GI tract [7].
Modern lifestyles demand convenience, and therefore consumers continue to look for “portable”
products consumed on-the-go and requiring minimum preparation time. Shelf-stable formulated
beverages with probiotics and/or prebiotics provide a convenient way to supplement daily diets and
to improve digestive health and immunity, which are the two main claims that are carried by such
products. Therefore, formulated (nondairy) probiotic and prebiotic beverages can serve as a success-
ful means to deliver health-promoting benefits, nutrition, great taste, and convenience in today’s
demanding world. Such products with probiotics also fulfill a novel role for an efficient cell factory
for the production of functional biomolecules and food ingredients to enhance the quality of func-
tional beverages [8]. Fermented foods encompass approximately 25% of the European diet and 60%
of that in developing countries. Lactic acid bacteria fermentation is a safe, economical, and tradi-
tional method of food preservation as referred to in the beginning of this contribution. Such fermen-
tations have the additional benefits of flavor, texture, and nutritional improvement, besides rendering
the beverages safe, in a chemical-free, consumer-friendly manner, which is more and more preferred
by the health-conscious consumer.
Probiotics and Prebiotics in Functional Beverages 817

Recent reviews have described the development of nondairy probiotic beverages and cereal-based fer-
mented products [5,8–12]. Some of the functional aspects discussed in these reviews are highlighted
here, in addition to other recent studies.
62.3.1 Probiotics
In addition to their acidification (and hence preservation) capacity, lactic acid bacteria also have the abil-
ity to contribute to the production of several important functional molecules through fermentation. These
range from substances that are important for aroma and flavor as well as texture and other organoleptic
features to nutraceuticals [8]. For instance, traditional foods/beverages made from cereal grains can be
lacking flavor, aroma, and body, whereas cereal fermentation leads to the production of several com-
pounds, which contribute to a complex blend of flavors in these products, such as amino acids, acetoin,
diacetyl, ethyl esters of acids (fruity flavor), and other volatile compounds, reviewed more extensively
by Waters et al. [8]. A recent example is a novel malt beverage fermented with B. breve NCIMB 702257,
a strain of human origin [13]. Eight volatile compounds were produced through the metabolic activity
of the bifidobacterial strain, among which acetic acid, maltol, pyranone, 2-(5H)-furanmethanol, and
3-furanmethanol. The latter four are Maillard-derived products with organoleptic characteristics that
together provide the fermented cereal beverage with mild sweet and malty notes.
On the other hand, microbially produced exopolysaccharides contribute to texture [14]. For instance,
Weissella cibaria MG1 was shown to produce an exopolysaccharides (a dextran) and glucooligosaccha-
rides during sucrose-supplemented barley-malt-derived worth fermentation [15]. The exopolysaccharide
contributed significantly to the rheological profile of the resulting fermented product. Small amounts of
organic acids were formed, and ethanol remained below 0.5%, the threshold for a potential health claim
designation.
Recent examples of fermented beverage products that contain probiotics are listed in Table 62.1
[16–36]. In most cases, these products used the (potential) probiotic strain as a starter culture for fer-
mentation. In very few cases, the probiotic is added after the fermentation phase. Mostly, Lactobacillus
plantarum has been used.
To increase survival of probiotics in these new functional beverages, encapsulation technologies have
been applied. This is elaborated upon by Gawkowski and Chikindas [5]. However, encapsulation does
not always lead to an enhanced survival. Sohail et al. [37] showed that encapsulation in alginate did not
result in a different survival of either of the two probiotic strains L. rhamnosus GG or L. acidophilus
NCFM. Encapsulation of these two bacteria, however, reduced acidification of orange juice at 4°C and
25°C. In agreement with this, encapsulated L. rhamnosus GG also reduced acidification in pear and
peach fruit-based beverages at 25°C, but not significantly (P > 0.05) at 4°C.
62.3.2 Prebiotics
Usually, the first prebiotic encountered in life is human breast milk [38,39]. The composition of human
breast milk oligosaccharides is very diverse and complex [39]. The prebiotic activity and beneficial
effects of human milk oligosaccharides [40] have been mimicked by infant formula producers by add-
ing fructooligosaccharides and galactooligosaccharides (Figure 62.1) to infant formula [41]. Other
than infant formula, there has been relatively little research on the addition of prebiotics to func-
tional beverages, either alone or in combination with probiotics. A few examples are discussed in the
succeeding text.
Ramnani et al. [42] have described the prebiotic effects of fruit and vegetable shots containing inulin
(Figure 62.1) from the Jerusalem artichoke. The shots were composed of either pear–carrot–sea buckthorn
(PCS) or plum–pear–beetroot (PPB) and contained inulin at a dose of 5 g/day. The effects were com-
pared to placebo, but unfortunately, the PCS and PPB without added inulin were not tested. The authors
concluded that both products had a prebiotic effect (elaborated upon in the section on health benefits),
TABLE 62.1
Recent Examples of Products Containing (Potential) Probiotics
Product Category Product Probiotic Strain References
Cereal based Mixed cereal substrate-fermented L. plantarum NCIMB 8826 [16]
beverage L. acidophilus NCIMB 8821
An ancient wheat (emmer)-based L. rhamnosus SP1 [17]
beverage
A yogurt-like product containing L. plantarum LP09 [18]
fermented oat flakes
Oat-based products L. plantarum B28 [19]
L. reuteri ATCC 55730 [20]
L. plantarum ATCC 8014 [21]
A product based on a mixture of L. plantarum 6E, LB1, POM1, M6, PR1, [22]
cereals, soy, and grape must and 1LC5
L. rossiae LB5
Weissella cibaria POM9, M1, and 3XLC3
Pediococcus pentosaceus SWE5
Vegetable based Several products of vegetable origin Several different probiotic strains [23]
Cabbage juice L. plantarum C3 [24]
L. casei A4
L. delbrueckii subsp. bulgaricus D7
Beet juice L. acidophilus LA 39 [25]
L. casei A4
L. delbrueckii subsp. bulgaricus D7
L. plantarum C3
Fruit-based juices Cashew juice Leuconostoc mesenteroides NRRL B512F [26]
Cantaloupe juice L. casei NRRL B-442 [27]
Cashew apple juice L. casei NRRL B-442 [28]
Tomato juice L. plantarum POM1, POM8, POM27, [29]
POM35, and POM43
L. brevis POM2
Enterococcus faecium POM3
W. cibaria POM11
P. pentosaceus POM10
Lactobacillus sp. POM44
Black currant juice L. plantarum 299v [30]
Grape juice L. plantarum DSM19463 [31]
Pineapple juice L. casei NRRL B442 [32]
Other A herbal extract-containing L. acidophilus ATCC 4356 [33]
nutraceutical beverage
Peanut–soy milk P. acidilactici UFLA BFFCX 27.1 [34]
L. lactis CCT 0360
L. rhamnosus LR 32
An extract from quinoa with soy L. casei LC-1, with added [35]
fructooligosaccharides
Whey-based beverages L. acidophilus CRL 636 [36]
L. delbrueckii subsp. bulgaricus CRL 656
Streptococcus thermophilus CRL 804
although the data did not substantiate that this was solely caused by inulin [42]. It might have been that
other components in the fruit and vegetable shots (e.g., pectin) contributed to the observed effects.
Yeo and Liong [43] studied the effect of the addition of prebiotics on the growth of several probiotics in
soy milk. Despite that the title of this report states that viability was investigated, this was not the case.
Only the growth of six bacterial strains was studied in the fermented product. The growth of L. FTDC
2113 (species not reported), L. acidophilus FTDC 8033, L. acidophilus ATCC 4356, L. casei ATCC
OH
O O OH
HO
O
OH OH
O HO
O
OH
O
HO
OH
O
n
Gluco-oligosaccharides
O OH
HO OH
OH
O
O OH
OH
O OH
n
Fructo-oligosaccharides; n = 2–10
OH
O
O
OH
O
HO
O OH
OH
O
HO
O
OH
HO O
OH
Galacto-oligosaccharides
O OH
OH
HO
O OH
n
OH O OH
HO O OH
HO
OH
OH O
Inulin; n = 10–60
FIGURE 62.1 Chemical structures of the most used prebiotics.

393, B. FTDC 8943 (species not reported), and B. longum FTDC 8643 was significantly enhanced by
supplementation with maltodextrin (glucooligosaccharides; Figure 62.1), pectin, mannitol, and fructooli-
gosaccharides, but not by supplementation with inulin. It was observed that supplementation with fruc-
tooligosaccharides and maltodextrin led to enhanced hydrolysis and utilization of soy oligosaccharides
and simpler sugars such as fructose and glucose in soy milk [43].
Bernat et al. [44] developed a new fermented almond “milk” that combined the properties of both
almonds and probiotics. Almond milk was fermented with L. reuteri ATCC 55730 and Streptococcus
thermophilus CECT 986. The authors added glucose, fructose, and inulin to the almond milk as carbon
source for the probiotics. The authors concluded that survival of the probiotics was high due to the pres-
ence of inulin, although almond milk without inulin was not tested. Fermentation increased the viscosity
of the product. In addition, mannitol was detected in the fermented almond milk, presumably produced
by L. reuteri, which the authors concluded was beneficial, as mannitol has been described as a nonmeta-
bolic sweetener with antioxidant properties [44].
Pimentel et al. [45] described the organoleptic properties and the acceptability of clarified apple juice
in which sugar was substituted with oligofructose (also known as fructooligosaccharides) or sucralose,
and which was fermented with L. paracasei ssp. paracasei LC-01. There were no differences (P > 0.05)
in the acceptance of the appearance, aroma, or texture, even though the probiotic increased the turbid-
ity of the juice. Juices with oligofructose showed less sweetness than those with sucrose. The authors
concluded that it was possible to develop a synbiotic functional beverage by adding L. paracasei ssp.
paracasei LC-01 as a probiotic culture and oligofructose as a sugar substitute and prebiotic [45]. To the
best of our knowledge, the health effect of this product on the host has not been studied.
To study the potential addition of probiotics to carbonated drinks, Walsh et al. [46] have studied the
effects of carbonation on probiotic survivability, physicochemical, and sensory properties of milk-based
synbiotic beverages. Pomegranate and vanilla yogurt beverages were formulated, containing inulin as
a prebiotic, along with the probiotic bacteria, L. acidophilus LA-5 and B. animalis subsp. lactis BB-12.
Carbonation of both yogurt types did not affect probiotic survival and showed the potential for probiotics
to be used in carbonated beverages [46].
62.3.3 Antioxidants and Bioactives Responsible for Antioxidant Activity

Cereal fermentation represents an important tool for increasing the extractability of bioactive compounds
such as antioxidants from various cereal grains or releasing nutraceuticals due to the bacterial metabo-
lism. However, the type of raw material (cereal, pseudocereals, and legumes) used is of key importance
in the process to ensure optimal release of bioactive compounds for human nutrition [8]. Among the
bioactive compounds, phenolic acids, primarily located on the outer layers (aleurone and pericarp) of
cereals, can act as antioxidants through the so-called chain-breaking mechanisms, which include hydro-
gen donation and radical interception. Cereal fermentation by lactic acid bacteria represents an efficient
means to increase the level of phenolic compounds, provided that appropriate fermentation conditions
are applied [8]. Increases of up to 90% of free phenolic acids upon fermentation of cereals have been
reported.
Antioxidants have also been used to increase the stability of probiotics in juices [47]. The authors
tested L. rhamnosus HN001, B. lactis HN001, and L. paracasei LPC 37 in a model juice with or without
various vitamins or antioxidants, namely, white grape seed extract, green tea extract, vitamin B2, vitamin
B3, vitamin B6, vitamin C, and vitamin E. Probiotic bacteria did not survive well in the harsh environ-
ment of the model fruit juice. However, the addition of either vitamin C or grape extract or green tea
extract led to a better survival of the probiotic bacteria, with counts of 4.3–7.4 log CFU/mL at the end of
a 6-week storage period, depending on the supplement added [47].
Some phenolic antioxidant compounds may be inhibiting bacteria. Therefore, Servili et al. [48]
tested whether antioxidant phenolic compounds extracted from olive vegetable water were inhibitory to
γ-amino butyric acid (GABA)-producing L. plantarum C48 and L. paracasei 15N. Addition of either
100 or 200 mg/L extracted from olive vegetable water did not affect the survival of either probiotic
strain. The resulting fermented milk beverages contained antioxidants as well as viable probiotics, in
addition to GABA, which was produced by one of the probiotic strains [48].
Kullisaar et al. [49] described an antioxidant in a goat milk fermented product. The goat milk was fer-
mented with the antioxidant probiotic strain L. fermentum ME-3, which has been well-characterized by
the same group. This strain possesses substantial antioxidative activity, expresses manganese superoxide
dismutase, eliminates hydroxyl radicals, and contains reduced glutathione, a potent cellular antioxidant.
The health effects of this product are described in the next section.
62.4 Health Effects

The health effects of probiotics and prebiotics have been dealt with in numerous reviews [50], and several
health benefits have been claimed for probiotics and prebiotics that go beyond the scope of this contribu-
tion to discuss them. Table 62.2 describes some of these benefits and some of the approved claims by
the European Food Safety Authority (EFSA), Food and Drug Administration (FDA), Food for Specified
Health Uses (FOSHU), or other relevant regulatory agencies. As can be observed from Table 62.2, very
few, although scientifically well documented, products containing probiotics or prebiotics may carry a
claim.
The definition of probiotics requires that the term “probiotic” only be applied to live microbes having
a substantiated beneficial effect on the host [51], although preparations of dead cells and cell components
may also exert some health-promoting physiological effects [52]. The term “probiotic” has been loosely
used in the examples discussed earlier. However, in very few cases, an actual probiotic, with clinical
proof of benefits for the host, as mentioned in the definition of probiotics, has been used. In most cases, a
starter culture that was able to ferment the product under study (fruit, vegetable, or grain) was used, and
these strains, although capable of providing the product with beneficial attributes, are not necessarily
probiotics. In a few instances, true probiotics have been used, and these products are likely to have a ben-
efit for the host. However, because of a change of food matrix and changes due to different fermentation
and/or product environmental conditions, it would be highly advisable that these supposedly functional
beverages are actually tested in clinical trials.
As far as we are aware, only two of the functional beverages described in more detail in the preceding
text have been tested in a clinical trial: the fruit and vegetable smoothie with added inulin [42] and the
fermented goat’s milk [49].
Although the controls of the smoothies without the inulin were not tested, Ramnani et al. [42] showed
that the smoothies with added inulin exhibited a prebiotic effect compared to placebo using fluores-
cent in situ hybridization in healthy human volunteers. Bifidobacteria were significantly increased by
more than half a log. A small but significant increase was also seen with the probe that recognized
Lactobacillus/Enterococcus. Total bacteria, Bacteroides, Clostridium perfringens/histolyticum sub-
group, Eubacterium rectale/Clostridium coccoides group, Atopobium spp., Faecalibacterium prausnit-
zii, and Propionibacterium, remained unaffected [42].
The consumption of fermented goats’ milk improved antiatherogenicity in healthy subjects [49].
It prolonged resistance of the lipoprotein fraction to oxidation, lowered levels of peroxidized lipoproteins,
oxidized low-density lipoprotein (ox-LDL), 8-isoprostanes and glutathione redox ratio, and enhanced
total antioxidative activity. The consumption of fermented goats’ milk also altered both the prevalence
and proportion of lactic acid bacteria species in the gut microbiota of the healthy subjects. The authors
concluded that this functional beverage with the special antioxidative strain L. fermentum ME-3 exhibits
antiatherogenic effects in humans [49].
It has proven to be very difficult to procure claims approved for products containing probiotics or pre-
biotics. Particularly, the EFSA in the European Union and the FDA in the United States have set the bar
extremely high for probiotic producers to prepare a dossier that contains sufficient evidence that meets
the criteria of these regulatory agencies [53–58]. Surprisingly, some claims that were widely used in the
past and approved by several national health agencies have been rejected, such as those for probiotics and
prebiotics. This has led to lively debates, even in the scientific literature [58–60]. This has been slightly
easier in Japan with the FOSHU [61]. Japanese health regulatory officials, using their FOSHU system,
have approved human health claims for numerous probiotic products. In addition, Health Canada is less
strict and has allowed a few general claims for probiotics (Table 62.2) [62].
TABLE 62.2
Proposed Benefits of Products Containing Probiotics and Prebiotics and Claims Allowed
by Regulatory Agents
Region/Country
(Regulatory Body) Claim Product/Probiotic
European Union No claims have been approved by EFSA for probiotics Live yogurt
(EFSA) or prebiotics. The only claim on products with lactic
acid bacteria that is allowed is for live yogurt, which
contains the starter microorganisms L. delbrueckii
subsp. bulgaricus and S. thermophilus on improving
lactose digestion.
Switzerland The Federal FSVO allowed the following structure B. lactis HN019
(FSVO) function claim: supporting digestion by reducing
transit time.
Japan (FOSHU) General approved claims in Japan for products B. longum BB536
containing the probiotics that are listed: B. breve Yakult
• Reaches the intestines alive. B. lactis Bb-12
• Increases the intestinal lactobacilli/bifidobacteria. B. lactis FK120
• Promotes the maintenance of a good intestinal B. lactis LKM512
environment. L. delbrueckii subsp. bulgaricus 2038
• Regulates and helps maintain a good GI condition. L. helveticus CK60
• Maintains the intestine in good health. L. rhamnosus GG
• Helps balance the intestinal microbiota.a L. johnsonii LC-1
• Reduces harmful bacteria. L. casei Shirota (YIT9029)
L. casei NY1301
L. acidophilus CK92
S. salivarius subsp. thermophilus 1131
B. subtilis K-2
The following can be claimed for a multitude of >500 products
products containing prebiotics and fibers:
• Suitable for improving the regulation and control of
the intestines.
• Improves bowel movement.
• Or something similar.
Korea (MFDS) The product containing the two strains can carry the L. rhamnosus GR-1 plus L. reuteri
claim: RC-14
• Can help vaginal health by increasing lactic acid
bacteria.
Canada (Health Non-strain-specific claims can be made on eligible B. adolescentis
Canada) species: B. animalis subsp. animalis
• Probiotic that naturally forms part of the gut B. animalis subsp. lactis
microbiota.a B. bifidum
• Provides live microorganisms that naturally form B. breve
part of the gut microbiota.a B. longum subsp. infantis
• Probiotic that contributes to healthy gut B. longum subsp. longum
microbiota.a L. acidophilus
• Provides live microorganisms that contribute to L. casei
healthy gut microbiota.a L. fermentum
A serving of a product should contain a minimum level L. gasseri
of 1.0 × 109 colony forming units of one or more of the L. johnsonii
eligible microorganism(s) that is (are) the subject of L. paracasei
the claim. L. plantarum
L. rhamnosus
L. salivarius
(Continued)

Proposed Benefits of Products Containing Probiotics and Prebiotics and Claims Allowed
by Regulatory Agents
Region/Country
(Regulatory Body) Claim Product/Probiotic
Strain-specific claims are claims about the health No products
benefits or effects of specific strains of probiotics. At
the present time, no strain-specific claims have been
accepted by Health Canada.
United States The U.S. FDA has not approved any health claims for No products
(FDA) probiotics. Nor does the agency have a regulatory
definition of probiotics. Probiotics are regulated based
on the product category into which they fall, for
example, food, food additive, cosmetic, dietary
supplement, or drug. FDA does not have a central
office that deals specifically with probiotics.
New Zealand and The FSANZ has issued a new standard (Standard 1.2.7 Not regulated, except when novel food
Australia Nutrition, Health, and Related Claims) to regulate
(FSANZ) nutrition content and health claims in 2013. FSANZ
has not attempted to specifically regulate health claims
about probiotics.
Brazil (ANVISA) In Brazil, claims are allowed by the ANVISA for 10 L. acidophilus
probiotic species. The claim is phrased as follows: L. casei
• The probiotic contributes to the balance of the L. casei subsp. rhamnosus
intestinal microbiota. Its consumption should be L. casei subsp. defensis
associated with a balanced diet and healthy L. paracasei
lifestyle.a L. lactis
B. bifidum
B. animalis
B. lactis
B. longum
E. faecium
For prebiotics and dietary fiber, the following claim is Many products
allowed provided that 3 g of fiber is present if the food
is solid or 1.5 g fiber if the food is liquid:
• Dietary fibers assist in the functioning of the
intestine. Its consumption should be associated with
a balanced diet and healthy lifestyle.
a Original claim contains the outdated term “flora” rather than “microbiota.”
Abbreviations: EFSA, European Food Safety Authority; FSVO, Food Safety and Veterinary Office; FOSHU, Food for
Specified Health Uses; MFDS, Ministry of Food and Drug Safety; FDA, Food and Drug Administration;
FSANZ, Food Standards Australia New Zealand; ANVISA, Agência Nacional de Vigilância Sanitária.

Despite being mostly associated with fermented dairy foods and drinks, probiotic bacteria are increas-
ingly incorporated into fermented nondairy beverages and continue to be well received by consumers
looking for a convenient way to boost their health. Researchers have begun to focus on investigating
methods to use well-studied strains in novel functional beverages to achieve structure/function claims
related to digestive health and immunity. Probiotic survival during storage in the functional beverages
can be increased by careful selection of the efficacious strain in use or by encapsulating bacterial cells.
Currently available probiotic beverages must be stored at cold temperatures and have a short shelf life,
which limits convenience and availability. Novel concepts and technologies that allow these products
to be stored at ambient temperature without loss of probiotic viability are required. One of the ways to
achieve higher probiotic survival is the addition of (well-selected) prebiotics in synbiotic products. The
research in this field is still in its infancy, as little understanding is available about the proper combina-
tion of probiotics and prebiotics and about dose of each bioactive. However, the trend toward the devel-
opment of functional beverages with probiotics and prebiotics is likely to take off in the near future,
due to the convenience of these products. Nevertheless, before successful products can be marketed,
a significant amount of research still has to be performed to ensure optimal product formulation and
consumer acceptance.
62.6 Conclusion
Based on traditional food fermentations, several new functional beverages are being developed. Thus far,
fermentation is mostly carried out with strains that are optimal for the fermentation process, in terms of
acidification, safety, and organoleptic and structural features, but those are not yet established probiot-
ics. However, it is not entirely unlikely that these strains have probiotic properties, because several new
probiotic strains have already been isolated from traditional food fermentations. However, this has to
be studied in carefully designed clinical trials. The addition of prebiotics to beverages, either with or
without probiotics, may show more promise or faster commercial benefit, as the functionality of a pre-
biotic is not expected to change in different food matrices. For the consumer, an explosion of functional
beverages is expected.
REFERENCES
1. FAO/WHO, Guidelines for the evaluation of probiotics in food. Joint FAO/WHO Working Group
Meeting, London, UK, 2002.
2. Gibson, G.R. and Roberfroid, M.B., Dietary modulation of the human colonic microbiota: Introducing
the concept of prebiotics. J. Nutr., 125, 1401–1412, 1995.
3. Gibson, G.R., Probert, H.M., Loo, J.V., Rastall, R.A., and Roberfroid, M.B., Dietary modulation of the
human colonic microbiota: Updating the concept of prebiotics. Nutr. Res. Rev., 17, 259–275, 2004.
4. Blatchford, P., Ansell, J., de Godoy, M.R.C., Fahey, G., Garcia-Mazcorro, J.F., Gibson, G.R., Goh, Y.J.
et al., Prebiotic mechanisms, functions and applications. Int. J. Probiotics Prebiotics, 8, 109–132, 2013.
5. Gawkowski, D. and Chikindas, M.L., Non-dairy probiotic beverages: The next step into human health.
Benef. Microbes, 4, 127–142, 2013.
6. Crittenden, R.G. and Playne, M.J., Production, properties and applications of food-grade oligosaccha-
rides. Trends Food Sci. Technol., 7, 353–361, 1996.
7. Rivera-Espinoza, Y. and Gallardo-Navarro, Y., Non-dairy probiotic products. Food Microbiol., 27,
1–11, 2010.
8. Waters, D.M., Mauch, A., Coffey, A., Arendt, E.K., and Zannini, E., Lactic acid bacteria as a cell factory
for the delivery of functional biomolecules and ingredients in cereal-based beverages: A review. Crit.
Rev. Food Sci. Nutr., 55, 503–520, 2015.
9. Blandino, A., Al-Aseeri, M.E., Pandiella, S.S., Cantero, D., and Webb, C., Cereal-based fermented
foods and beverages. Food Res. Int., 36, 527–543, 2003.
10. Corbo, M.S., Bevilacqua, A., Petruzzi, L., Casanova, F.P., and Sinigaglia, M., Functional beverages: The
emerging side of functional foods—Commercial trends, research, and health implications. Comp. Rev.
Food Sci. Food Saf., 13, 1192–1206, 2013.
11. Granato, D., Branco, G.F., Nazzaro, F., Cruz, A.G., and Faria, J.A.F., Functional foods and nondairy pro-
biotic food development: Trends, concepts, and products. Comp. Rev. Food Sci. Food Saf., 9, 292–302,
2010.
12. Prado, F.C., Parada, J.L., Pandey, A., and Soccol, C.R., Trends in non-dairy probiotic beverages. Food
Res. Int., 41, 111–123, 2008.
13. Salmeron, I., Rozada, R., Thomas, K., Ortega-Rivas, E., and Pandiella, S.S., Sensory characteristics and
volatile composition of a cereal beverage fermented with Bifidobacterium breve NCIMB 702257. Food
Sci. Technol. Int., 20, 205–213, 2014.
14. Ganzle, M.G., From gene to function: Metabolic traits of starter cultures for improved quality of cereal
foods. Int. J. Food Microbiol., 134, 29–36, 2009.
15. Zannini, E., Mauch, A., Galle, S., Ganzle, M., Coffey, A., Arendt, E.K., Taylor, J.P., and Waters, D.M.,
Barley malt wort fermentation by exopolysaccharide-forming Weissella cibaria MG1 for the production
of a novel beverage. J. Appl. Microbiol., 115, 1379–1387, 2013.
16. Rathore, S., Salmeron, I., and Pandiella, S.S., Production of potentially probiotic beverages using single
and mixed cereal substrates fermented with lactic acid bacteria cultures. Food Microbiol., 30, 239–244,
2012.
17. Coda, R., Rizzello, C.G., Trani, A., and Gobbetti, M., Manufacture and characterization of functional
emmer beverages fermented by selected lactic acid bacteria. Food Microbiol., 28, 526–536, 2011.
18. Luana, N., Rossana, C., Curiel, J.A., Kaisa, P., Marco, G., and Rizzello, C.G., Manufacture and char-
acterization of a yogurt-like beverage made with oat flakes fermented by selected lactic acid bacteria.
Int. J. Food Microbiol., 185, 17–26, 2014.
19. Angelov, A., Gotcheva, V., Kuncheva, R., and Hristozova, T., Development of a new oat-based probiotic
drink. Int. J. Food Microbiol., 112, 75–80, 2006.
20. Bernat, N., Chafer, M., Gonzalez-Martinez, C., Rodriguez-Garcia, J., and Chiralt, A., Optimisation of
oat milk formulation to obtain fermented derivatives by using probiotic Lactobacillus reuteri microor-
ganisms. Food Sci. Technol. Int., 21, 145–157, 2015.
21. Gupta, S., Cox, S., and Abu-Ghannam, N., Process optimization for the development of a functional
beverage based on lactic acid fermentation of oats. Biochem. Eng. J., 52, 199–204, 2010.
22. Coda, R., Lanera, A., Trani, A., Gobbetti, M., and Di Cagno, R., Yogurt-like beverages made of a mix-
ture of cereals, soy and grape must: Microbiology, texture, nutritional and sensory properties. Int. J.
Food Microbiol., 155, 120–127, 2012.
23. Furtado Martins, E.M., Ramos, A.M., Lago Vanzela, E.S., Stringheta, P.S., de Oliveira Pinto, C.L., and
Martins, J.M., Products of vegetable origin: A new alternative for the consumption of probiotic bacteria.
Food Res. Int., 51, 764–770, 2013.
24. Yoon, K.Y., Woodams, E.E., and Hang, Y.D., Production of probiotic cabbage juice by lactic acid bacte-
ria. Bioresour. Technol., 97, 1427–1430, 2006.
25. Yoon, K.Y., Woodams, E.E., and Hang, Y.D., Fermentation of beet juice by beneficial lactic acid bacte-
ria. Lebensm. Wiss. Technol., 38, 73–75, 2005.
26. da Silva, I.M., Rabelo, M.C., and Rodrigues, S., Cashew juice containing prebiotic oligosaccharides.
J. Food Sci. Technol., 51, 2078–2084, 2014.
27. Fonteles, T.V., Costa, M.G.M., de Jesus, A.L.T., and Rodrigues, S., Optimization of the fermentation of
cantaloupe juice by Lactobacillus casei NRRL B-442. Food Bioprocess. Technol., 2011, doi: 10.1007/
s11947–011–0600.
28. Pereira, A.L.F., Maciel, T.C., and Rodrigues, S., Probiotic beverage from cashew apple juice fermented
with Lactobacillus casei. Food Res. Int., 44, 1276–1283, 2011.
29. Di Cagno, R., Surico, R.F., Paradiso, A., De Angelis, M., Salmon, J.C., Buchin, S., De Gara, L., and
Gobbetti, M., Effect of autochthonous lactic acid bacteria starters on health-promoting and sensory
properties of tomato juices. Int. J. Food Microbiol., 128, 473–483, 2009.
30. Luckow, T. and Delahunty, C., Which juice is ‘healthier’? A consumer study of probiotic non-dairy juice
drinks. Food Qual. Pref., 15, 751–759, 2004.
31. Di Cagno, R., Mazzacane, F., Rizzello, C.G., De Angelis, M., Giuliani, G., Meloni, M., De Servi, B., and
Gobbetti, M., Synthesis of gamma-aminobutyric acid (GABA) by Lactobacillus plantarum DSM19463:
Functional grape must beverage and dermatological applications. Appl. Microbiol. Biotechnol., 86,
731–741, 2010.
32. Costa, M.G., Fonteles, T.V., de Jesus, A.L., and Rodrigues, S., Sonicated pineapple juice as substrate
for L. casei cultivation for probiotic beverage development: Process optimisation and product stability.
Food Chem., 139, 261–266, 2013.
33. Lima, I.F., De Dea Lindner, J., Soccol, V.T., Parada, J.L., and Soccol, C.R., Development of an innova-
tive nutraceutical fermented beverage from herbal mate (Ilex paraguariensis A.St.-Hil.) extract. Int. J.
Mol. Sci., 13, 788–800, 2012.
34. Santos, C.C., Libeck Bda, S., and Schwan, R.F., Co-culture fermentation of peanut-soy milk for the
development of a novel functional beverage. Int. J. Food Microbiol., 186, 32–41, 2014.
35. Bianchi, F., Rossi, E., Gomes, R., and Sivieri, K., Potentially synbiotic fermented beverage with aqueous
extracts of quinoa (Chenopodium quinoa Willd) and soy. Food Sci. Technol. Int., 21, 403–415, 2015.
36. Pescuma, M., Hebert, E.M., Mozzi, F., and de Valdez, G.F., Functional fermented whey-based beverage
using lactic acid bacteria. Int. J. Food Microbiol., 141, 73–81, 2010.
37. Sohail, A., Turner, M.S., Prabawati, E.K., Coombes, A.G., and Bhandari, B., Evaluation of Lactobacillus
rhamnosus GG and Lactobacillus acidophilus NCFM encapsulated using a novel impinging aerosol
method in fruit food products. Int. J. Food Microbiol., 157, 162–166, 2012.
38. Gura, T., Nature’s first functional food. Science, 345, 747–749, 2014.
39. Hennet, T., Weiss, A., and Borsig, L., Decoding breast milk oligosaccharides. Swiss Med. Wkly., 144,
w13927, 2014.
40. Musilova, S., Rada, V., Vlkova, E., and Bunesova, V., Beneficial effects of human milk oligosaccharides
on gut microbiota. Benef. Microbes, 5, 273–283, 2014.
41. Fanaro, S., Boehm, G., Garssen, J., Knol, J., Mosca, F., Stahl, B., and Vigi, V., Galacto-oligosaccharides
and long-chain fructo-oligosaccharides as prebiotics in infant formulas: A review. Acta Paediatr. Suppl.,
94, 22–26, 2005.
42. Ramnani, P., Gaudier, E., Bingham, M., van Bruggen, P., Tuohy, K.M., and Gibson, G.R., Prebiotic
effect of fruit and vegetable shots containing Jerusalem artichoke inulin: A human intervention study.
Br. J. Nutr., 104, 233–240, 2010.
43. Yeo, S.K. and Liong, M.T., Effect of prebiotics on viability and growth characteristics of probiotics in
soymilk. J. Sci. Food Agric., 90, 267–275, 2010.
44. Bernat, N., Chafer, M., Chiralt, A., and Gonzalez-Martinez, C., Development of a non-dairy probiotic
fermented product based on almond milk and inulin. Food Sci. Technol. Int., 21, 440–453, 2015.
45. Pimentel, T.C., Madrona, G.S., and Prudencio, S.H., Probiotic clarified apple juice with oligofructose or
sucralose as sugar substitutes: Sensory profile and acceptability. LWT Food Sci. Technol., 62, 838–846,
2015.
46. Walsh, H., Cheng, J., and Guo, M., Effects of carbonation on probiotic survivability, physicochemical,
and sensory properties of milk-based symbiotic beverages. J. Food Sci., 79, M604–M613, 2014.
47. Shah, N.P., Ding, W.K., Fallourd, M.J., and Leyer, G., Improving the stability of probiotic bacteria in
model fruit juices using vitamins and antioxidants. J. Food Sci., 75, M278–M282, 2010.
48. Servili, M., Rizzello, C.G., Taticchi, A., Esposto, S., Urbani, S., Mazzacane, F., Di Maio, I., Selvaggini,
R., Gobbetti, M., and Di Cagno, R., Functional milk beverage fortified with phenolic compounds
extracted from olive vegetation water, and fermented with functional lactic acid bacteria. Int. J. Food
Microbiol., 147, 45–52, 2011.
49. Kullisaar, T., Songisepp, E., Mikelsaar, M., Zilmer, K., Vihalemm, T., and Zilmer, M., Antioxidative
probiotic fermented goats’ milk decreases oxidative stress-mediated atherogenicity in human subjects.
Br. J. Nutr., 90, 449–456, 2003.
50. Hill, C., Guarner, F., Reid, G., Gibson, G.R., Merenstein, D.J., Pot, B., Morelli, L. et al., Expert consen-
sus document. The International Scientific Association for Probiotics and Prebiotics consensus state-
ment on the scope and appropriate use of the term probiotic. Nat. Rev. Gastroenterol. Hepatol., 11,
506–514, 2014.
51. Reid, G., Sanders, M.E., Gaskins, H.R., Gibson, G.R., Mercenier, A., Rastall, R., Roberfroid, M.,
Rowland, I., Cherbut, C., and Klaenhammer, T.R., New scientific paradigms for probiotics and prebiot-
ics. J. Clin. Gastroenterol., 37, 105–118, 2003.
52. Adams, C.A., The probiotic paradox: Live and dead cells are biological response modifiers. Nutr. Res.
Rev., 23, 37–46, 2010.
53. Saxelin, M., Probiotic formulations and applications, the current probiotics market, and changes in the
marketplace: A European perspective. Clin. Infect. Dis., 46(Suppl. 2), S76–S79; discussion S144–S151,
2008.
54. Donovan, S.M., Schneeman, B., Gibson, G.R., and Sanders, M.E., Establishing and evaluating health
claims for probiotics. Adv. Nutr., 3, 723–725, 2012.
55. Farnworth, E.R., The evidence to support health claims for probiotics. J. Nutr., 138, 1250S–1254S,
2008.
56. Hoffmann, D.E., Fraser, C.M., Palumbo, F.B., Ravel, J., Rothenberg, K., Rowthorn, V., and Schwartz, J.,
Science and regulation. Probiotics: Finding the right regulatory balance. Science, 342, 314–315, 2013.
57. Salminen, S. and van Loveren, H., Probiotics and prebiotics: Health claim substantiation. Microb. Ecol.
Health Dis., 23, 2012.
58. Sanders, M.E., Heimbach, J.T., Pot, B., Tancredi, D.J., Lenoir-Wijnkoop, I., Lahteenmaki-Uutela, A.,
Gueimonde, M., and Banares, S., Health claims substantiation for probiotic and prebiotic products. Gut
Microbes, 2, 127–133, 2011.
59. Reid, G., Opinion paper: Quo vadis—EFSA? Benef. Microbes, 2, 177–181, 2011.
60. Katan, M.B., Why the European Food Safety Authority was right to reject health claims for probiotics.
Benef. Microbes, 3, 85–89, 2012.
61. Amagase, H., Current marketplace for probiotics: A Japanese perspective. Clin. Infect. Dis., 46(Suppl. 2),
S73–S75; discussion S144–S151, 2008.
62. Canadian Food Inspection Agency, Probiotic health claims, 2014. Published online at: http://www.
inspection.gc.ca/food/labelling/food-labelling-for-industry/health-claims/eng/1392834838383/1392834887794?
chap=9 (accessed November 28, 2014).
63
Functional Beverages in Weight Management
Debasis Bagchi, Anand Swaroop, and Manashi Bagchi
CONTENTS
63.1 Introduction................................................................................................................................... 829
63.2 Designing a Functional Beverage................................................................................................. 830
63.3 Nutraceutical and Functional Food Ingredients........................................................................... 830
63.3.1 Garcinia cambogia Extract and (–)-Hydroxycitric Acid............................................... 830
63.3.2 Chromium (III) Ingredients............................................................................................831
63.3.3 Green Coffee Bean Extract.............................................................................................831
63.3.4 Green Tea Extract and Oolong Tea.................................................................................831
63.3.5 Conjugated Linoleic Acid................................................................................................831
63.3.6 Trigonella foenum-graecum (Fenugreek) Extract.......................................................... 832
63.3.7 Chitosan.......................................................................................................................... 832
63.3.8 Glucomannan................................................................................................................. 832
63.3.9 5-Hydroxytryptophan..................................................................................................... 832
63.3.10 Berries and Selected Fruits............................................................................................ 832
63.3.11 Calcium.......................................................................................................................... 833
63.3.12 Caffeine and Ephedrine.................................................................................................. 833
63.3.13 Soluble Dietary Fiber, Thickeners, Anticoagulants, and Carbohydrates....................... 833
63.4 Salient Features of a Functional Beverage.................................................................................... 833
63.5 Novel Products/Formulation and Future Trends........................................................................... 834
63.6 Conclusion..................................................................................................................................... 835
References............................................................................................................................................... 835
63.1 Introduction
A recent survey by the World Health Organization (WHO) demonstrated that globally there are more
than 1 billion overweight adults and approximately 300 million of whom are obese [1]. It is important
to emphasize that wrestler and heavyweight champions are overweight due to healthy muscle and may
have little or no fat [2]. However, accumulation of excessive body fat due to increased consumption of
energy-dense foods rich in saturated fats and sugar, advancing age, compromised metabolism, sedentary
lifestyle, and lack of physical exercise may lead to obesity in conjunction with a series of degenerative
diseases and disorders [2]. Health professionals are exploring unique technologies to combat obesity,
which will ultimately help to combat a wide array of health dysfunctions.
Functional foods are foods and food components that provide proven health benefits beyond their basic
nutritional function. These include conventional food; fortified, enriched, or enhanced foods; nutraceuti-
cals; and dietary supplements [3]. A significant number of nutraceuticals and functional foods have been
introduced into the market, which demonstrated to cause a significant number of adverse effects includ-
ing toxicity and lack of efficacy. Therefore, researchers have assessed a number of products to introduce
safe and efficacious functional beverages to target obesity [2]. This chapter demonstrates the salient
features to design functional beverages with functional ingredients that demonstrated significant efficacy
in weight loss and weight maintenance, as well as stable and safe for long-term use.
829
63.2 Designing a Functional Beverage

Designing a functional beverage is an art, and novel formulations should be structured in a way so that
the safety, stability, and efficacy are not compromised under storage conditions. The following salient
features need to be considered:
• Structural integrity of the constituents: The structural integrity of the constituents should not
be altered on storage; however, minor degradation may occur, which should not exceed more
than 1%–3%. The formulation needs to be accomplished in such a way so that the combination
of multiple constituent(s) should work synergistically. The other important factor is that there
should not be yellowing of the beverage or precipitation on aging and storage. So, appropriate
antioxidants, preservatives, and stabilizers should be incorporated in requisite amounts.
• Safety and generally recognized as safe (GRAS) status: Extensive safety studies need to be con-
ducted to demonstrate the broad spectrum safety of the constituent(s) and the final formulation.
It is always recommended to use GRAS-affirmed constituents in the beverage formulations [4].
• Storage, pH, and temperature stability: The beverage samples need to be stored for at least
next to 1–2 years, so that the pH, stability, taste, and color should remain unchanged. Keeping
in mind that the beverages will be transported to various locations and exposed to diverse
environmental conditions of fluctuating temperatures (extreme low to high), sunlight, and
humidity, and the formulations must ascertain to maintain the integrity of the constituents
and the beverage. There should also be no precipitation on storage. Accelerated stability
studies are recommended at elevated temperatures. The formulation should be accomplished
using proper additives, antioxidants/preservatives, and stabilizers, so that the integrity of the
formulations remains unchanged.
• Light sensitivity: The beverage should not degrade, change its color remarkably, or precipitate
out to maintain its aesthetic appeal, while stored in shelves of the supermarket or convenience
stores at the temperatures recommended.
• Functionality and efficacy: The functionality and efficacy of the ingredients should not alter
under storage conditions.
• Aesthetic appeal, palatability, and sensory tests: The ingredient used in the formulation should
not alter the aesthetic appeal and taste of the beverage. Palatability and sensory tests need to be
performed carefully for market success.
63.3 Nutraceutical and Functional Food Ingredients

Over the last several years, beverages designed and formulated with structurally diverse nutraceutical
and functional food ingredients have gained significant popularity in the marketplace. People especially
younger generation are very careful, and they prefer functional beverages over regular cola drinks because
of their nutritional values. On the contrary, cola drinks are loaded with sugar and major causes for obesity
and diabetes. Although diet cola beverages were introduced about a couple of decades earlier, and in a
very recent study, diet cola drinks have claimed to exhibit appetite suppression and weight loss [5].
Following are the nutraceutical and functional food ingredients that are used in the beverage
formulations.
63.3.1 Garcinia cambogia Extract and (–)-Hydroxycitric Acid

(–)-Hydroxycitric acid (HCA) is derived from the fruits of Garcinia cambogia, which is known as Malabar
tamarind, brindal berry, and gorikapuli, and extensively used for culinary purposes as well as for pickling
fish in Southeast Asia [6]. It is important to mention that G. cambogia fruit is included in the United States
Functional Beverages in Weight Management 831
Department of Agriculture’s (USDA) inventory of perennial edible fruits of the tropics. The calcium–
potassium salt of HCA is freely soluble in water, while the other salts are reported to have 70%–99% water
solubility [6]. A significant number of animal and clinical studies repeatedly demonstrated its efficacy in
weight management without having any significant adverse events. Appetite suppression, mood alleviation,
and fat oxidation were demonstrated as the major mechanisms of weight management [6]. Several HCA-
based beverages including Fuze Slenderize [7–9], Skinny Water [10], and SoBe Lifewater and Lean [11]
are available in the United States, Europe, Japan, Korea, and Australia. It is important to mention that Fuze
beverages are integral part of the Coca-Cola Company (Atlanta, Georgia) [7,8], while SoBe Lifewater and
Lean products are manufactured by South Beach Beverage Company (Norwalk, Connecticut).
63.3.2 Chromium (III) Ingredients

Chromium (III) supplements are novel micronutrients that serve as a critical cofactor in the action of
insulin in human nutrition, promote lean body mass, and improve impaired glucose tolerance and insulin
sensitivity [12]. Several clinical and safety studies on chromium polynicotinate (commercially known as
ChromeMate), chromium picolinate (commercially known as Chromax), chromium histidinate, and chro-
mium dinicocysteinate (commercially known as Zychrome) have demonstrated the safety and efficacy
of chromium (III) supplements [12–14]. Most of these chromium (III) salts are readily soluble in water
and widely used in beverage formulations around the world. Several of these chromium (III) supple-
ments including ChromeMate [7–10] and Chromax [12,14] are being extensively used in commercially
available beverages including Fuze Slenderize (Coca-Cola Company), Skinny Water [8,10], and SoBe
Lifewater and Lean [11].
63.3.3 Green Coffee Bean Extract

Green coffee bean extract is a highly soluble nutraceutical, which is becoming increasingly popular as a
weight-loss supplement worldwide. Clinical studies have demonstrated the efficacy of green coffee bean
extracts, containing 50% total chlorogenic acid, in weight loss [15,16]. Ochiai et al. [17] demonstrated the effi-
cacy of a novel, green coffee bean beverage in improving vasoreactivity and hypertension in human subjects,
which is mainly caused by overweight and obesity. A novel, patent-pending, water-soluble, and low-caffeine
green coffee bean extract GCB-70 demonstrated remarkable weight-loss efficacy in animal and humans [18].
Test marketing of this product, alone and in combination with Trigonella foenum-graecum extract, is ongoing
by reputed beverage companies in the United States for weight management and metabolic syndrome.
63.3.4 Green Tea Extract and Oolong Tea

Green and oolong teas are very popular drinks. Caffeine and catechins contained in green tea may have
a thermogenic effect, favoring weight and body fat loss [19–22]. Several mechanistic pathways including
its antiangiogenic efficacy and inhibition of catechol O-methyl-transferase and phosphodiesterase have
been demonstrated to prevent the development of obesity. Baladia et al. [19] conducted a systemic review
and meta-analysis and concluded that green tea extract intake exerts no statistically significant effect in
obese adults. The authors indicated that there is a small effect on the decrease in the percentage of fat
mass, but it is not clinically relevant. On the contrary, clinical studies on oolong tea using a daily dosage
of 8 g/day demonstrated a decrease in body fat and body weight through improvement of lipid metabo-
lism [19]. Thus, chronic consumption of oolong tea may prevent against obesity [20]. Several popular
green tea beverages are available singly and in combination mostly with blueberry, raspberry, passion
fruit, and ginseng on the market by Snapple [23], Lipton [24], and Arizona Beverage Company [25].
63.3.5 Conjugated Linoleic Acid

Conjugated linoleic acid (CLA) is a popular weight-loss ingredient used in several formulations including
beverages. It reduces body weight, maintains lean muscle mass, and improves body composition [26].
The popular CLA ingredient Tonalin, derived from safflower oil, is well known to target stubborn belly
fat and reduces overall body fat [27]. It has been incorporated in both capsule and beverage formulations.
The key ingredients in Tonalin beverage formulations include CLA (3.5 g) and chromium picolinate
(120 μg) in a total volume of 8 fl. oz. (~237 mL) [27]. Another popular CLA-based weight management
beverage “Evolve” was introduced by CytoSport, Inc. (Benicia, CA) for weight management [28].
63.3.6 Trigonella foenum-graecum (Fenugreek) Extract

Fenugreek demonstrated significant antidiabetic and neuroprotective efficacy, probably mediated through
a decrease in hyperglycemia and oxidative stress, thereby ameliorating the incidences of diabetic com-
plications [29,30]. A novel, water-soluble, patented, and fenugreek extract (Fenfuro) has demonstrated
significant efficacy in marginal weight loss and potent antidiabetic efficacy [29,30]. Test marketing of this
product is currently underway by reputed beverage companies in the United States.
63.3.7 Chitosan
Chitin is a cellulose-like polymer found in the exoskeletons of marine invertebrates and arthropods such
as shrimp, crabs, and lobsters [31,32]. Chitin is composed of long chains of acetylated glucosamine, and
chitosan results from the deacetylation of chitin [31]. It is relatively insoluble in water but can be easily
dissolved by dilute acids, which makes it a highly viscous dietary fiber [31,32]. A number of clinical
studies demonstrated the safety and efficacy of chitosan in weight management [31–33]. Several model
beverage formulations have been discussed by Luo and Wang [34] to demonstrate weight management
and antioxidant properties.
63.3.8 Glucomannan
Glucomannan is a water-soluble dietary fiber enriched in polysaccharide and is used extensively for
achieving weight loss in overweight and moderately obese subjects. Clinical studies on glucomannan in
conjunction with psyllium husk or G. cambogia extract exhibited synergistic efficacy in weight manage-
ment [35,36]. Several mechanistic pathways including dilution of energy density, promotion of satiety,
and fecal energy loss have been demonstrated as the pathway of weight management by glucomannan
[35,36]. Soyum, a patented, high-fiber healthy sliming beverage containing glucomannan konjac fiber,
is marketed by Shanghai Huanging International Trading Co Ltd., Shanghai, China [37]. LuraLean,
another soluble and flavorless glucomannan from AHD International (Atlanta, GA), is available for mak-
ing weight-loss beverage in the United States and around the world [38].
63.3.9 5-Hydroxytryptophan
Oral administration of 5-hydroxytryptophan (5-HTP) (900 mg/day) was demonstrated to cause anorexia,
decreased food intake, and significant weight loss in obese subjects. Research demonstrated a reduc-
tion in carbohydrate intake and satiety in these subjects, and no adverse events were reported [39–41].
Fortitech (Schenectady, NY) premixes containing 5-HTP is recommended for preparing a beverage for
weight management in overweight and obese adults [42].
63.3.10 Berries and Selected Fruits

Berries have gained increasing global popularity due to their numerous health benefits, including weight
management. Several berry-based juices and beverages are already available in the marketplace. A novel
combination of six edible berries, including wild blueberry, bilberry, elderberry, strawberry, and rasp-
berry seeds, demonstrated potent antiangiogenic properties, including weight loss in animals [43]. Animal
studies showed that raspberry ketone prevents increase in overall body fat and visceral fat [44,45]. There
are several berry drinks in the U.S. marketplace [46,47]; however, an Ultra Malibu Miracle mixed berry
beverage for weight management by Dream Quest Nutraceuticals and Nature’s Plus (Melville, NY) is
very popular among young customers [46]. This Malibu Miracle beverage contains concentrated noni,
goji, pomegranate, açai, and mangosteen fruit juices (6 g of each berry juice) in conjunction with (–)-HCA
(1 g) in a total volume of 7 fl. oz. (~237 mL), which is recommended to be consumed within the next
3 h [47]. Another popular weight-loss berry drink is known as Aqua Slenda, marketed by Nature’s Way
(Lehi, UT), which contains a combination of mixed berries, including blueberry, strawberry, elderberry,
and raspberry, with demonstrated efficacy for satiety and appetite suppression.
63.3.11 Calcium
Various calcium supplements and drinks in conjunction with probiotics demonstrate significant benefits
in human health including attenuation of bone health and some benefits in weight management [48,49].
Several calcium salts including calcium carbonate, calcium chloride, tricalcium citrate, calcium lactate,
calcium lactate gluconate, and calcium gluconate are extensively used in the U.S. and E.U. beverage
markets. Several clear and cloudy calcium beverages in conjunction with juices of apple, cranberry,
orange, tangerine, and grapefruit are popular in both U.S. and E.U. markets [50,51].
63.3.12 Caffeine and Ephedrine

In most weight-loss beverage formulations, caffeine is an integral constituent. Although there are some
popularities in caffeine-free beverages, studies have demonstrated significant efficacy of caffeine and
ephedrine in weight loss [52,53]. Although, there are safety issues, it is very important to indicate that
ephedrine is now a banned substance by the Food and Drug Administration (FDA) [54]. The natural
form of ephedrine, Ma Huang, was very popular in the United States until early 2000; however, due to
its potential toxicity and danger, its use was abandoned [52].
63.3.13 Soluble Dietary Fiber, Thickeners, Anticoagulants, and Carbohydrates

Soluble dietary fiber, anticoagulants, and carbohydrates including fructose, psyllium husk, and barley
flakes are integral part of functional beverages as additives, sweeteners, and thickeners [55]. Fructose
is naturally available in beet, sugarcane, honey, and diverse fruits and vegetables. It is the sweetest of
all naturally occurring carbohydrates, approximately 1.73-fold as sweet as sucrose and exhibit a low
glycemic index (GI) [56]. Studies demonstrated that fructose produces a marginal rise in plasma glucose
level as compared to sucrose, glucose, potato starch, or wheat starch when consumed with a meal [57].
However, studies demonstrated that high fructose induces metabolic syndrome [57]. Therefore, it is very
important to regulate the amount of fructose in novel beverage formulations.
Psyllium husk (Plantago ovata), a viscous, water-soluble fiber, contains significant amount of hemi-
cellulose, composed of a xylan backbone linked with arabinose, rhamnose, and galacturonic acid units
(arabinoxylans) [58]. Studies demonstrated that psyllium husk exhibits significant benefits on body
weight reduction, satiety, cholesterol, and triacylglycerols (TAG), fasting glycemia, and blood pressure,
which suggests a potential role in the treatment of metabolic syndrome [58,59]. It is a common practice
to include psyllium husk in most beverage formulations as a thickener [55].
Barley flakes are good sources of soluble fiber and β-glucan. They significantly reduce glycemic responses
[60]. Pearled barley and cracked barley have lower GI as compared to sweet corn, rolled barley, and white
rice. Barley flakes may be recommended in maintaining blood glucose level and weight control [60,61].
63.4 Salient Features of a Functional Beverage

It is very important that nutraceutical and functional food ingredients exhibit the following properties for
incorporation in functional beverages.
• Water solubility or high dispersibility: The ingredients should preferably be water soluble or
have high water dispersibility. It is a common practice to use emulsifiers and additives in the
formulation. It is also very important to know that pH plays a vital role in solubility.
• Storage stability: The beverage formulations should have high storage stability. The ingredients
used in a beverage should not antagonistically interact with each other. The formulation should
be designed in such a way that they can withstand and maintain their individual integrity in the
beverage under storage conditions. It is important to remember that appropriate facilities may
not be available during transportation, storage, and handling. The accelerated storage stability
tests must be conducted for checking the stability up to 2–3 years.
• Color stability: There should be no significant change in the beverage color throughout the
storage. Primarily, the ingredients must be soluble or significantly compatible with water and
if multiple ingredients are chosen, then these ingredients should not interact antagonistically in
the beverage formulations or alter the pH or color. Color stabilizers are recommended to retain
the color.
• Flavor: It is very important that beverages should have an appealing flavor (taste and aroma).
Some of the nutraceutical ingredients may have bitter or unpleasant taste or strong smell.
Masking agents are added to the functional beverages to make them palatable.

A remarkable trend has been shown in the global market on the escalating share of weight-loss bever-
ages in the marketplace. The global beverage market has been forecasted to increase at a compound
annual growth rate of 4.6% over the next 5 years, to reach a value of US$1347 billion by 2017 [62],
while the global obesity was worth US$265 billion in 2012 [63]. Interestingly, nutraceutical food and
beverage market reached US$93 billion in 2011 [64]. There is a significant increasing trend in the mar-
keting of functional cola beverages during the last 2–3 decades; however, the new generation is looking
TABLE 63.1
Effect of Beverages on Weight Loss, BMI, and Energy Intake
Study Subjects Experimental Outline Observation References
Female, normal body 400 mL of drinking water Drinking water increased feelings of [65]
weight (n = 8) during fixed intake meal. satiety and diminished hunger. No
sustained effect after meal.
Female (n = 1,136) Cross sectional; diet Water intake from foods, but not [66]
questionnaire (150 food beverages, associated with lower
and beverages). BMI and waist circumference.
Female, overweight Mixed models used for Consuming ≥1 L water/day associated [67]
(n = 173), 12 months analysis (secondary analysis with greater weight loss (2 kg)
was carried out). versus water intake of <1 L/day.
Female (n = 51,603), 4-year Prospective cohort, sweetened Diet soft drinks help in weight loss. [68]
period beverage consumption with
weight gain.
Male (n = 21) Diet with 1150 g/day of soda Reduced energy intake and body [69]
Female (n = 9), normal sweetened with aspartame, weight with aspartame in males.
body weight high-fructose corn syrup, or
no drinks for 3 weeks.
Male (n = 569) Cross-sectional study on Tea drinkers (≥1 drink/week regularly [70]
Female (n = 641) habitual tea consumption. >10 years) have reduced body fat
and waist–hip ratio than nonhabitual
tea drinkers.
Male and female (n = 434) Low/no calorie sweetened Lost ≥13.6 kg and maintained weight [71]
beverages were used by the loss for >1 year.
subjects.
Abbreviation: BMI, body mass index.
for functional beverages that can provide certain health benefits. Some of these beverages are designed
based on novel supplements backed by scientific research studies. However, most of the claims for these
beverages are not appropriately substantiated. There are also only a few studies available on drinking
water, diet soft drinks, aspartame, high-fructose corn syrup, and tea, the results of which are summa-
rized in Table 63.1 [65–71].
63.6 Conclusion
A significant number of nutraceutical and functional food-based beverages targeted for weight manage-
ment are globally available in the marketplace. These are becoming increasingly popular worldwide.
Some of these beverages are backed by good scientific evidence on safety, efficacy, and stability, while
others are not. Credible research including safety, efficacy, stability, palatability, and sensory studies of
these beverages are required to demonstrate their efficacy in weight management in order to reduce the
risk of cardiovascular disease (CVD), diabetes, hypertension, and arthritis. Furthermore, reduction in
sugar content in beverage formulations is also recommended to keep a lower GI.
REFERENCES
1. World Health Organization (WHO), Obesity and overweight, May 2014, Published online at: http://
www.who.int/mediacentre/factsheets/fs311/en/ (accessed June 28, 2014).
2. Bagchi, D. and Preuss, H.G., Obesity: Epidemiology, Pathophysiology, and Prevention, 2nd edn., CRC
Press/Taylor & Francis Group, Boca Raton, FL, 2013.
3. Gyles, C.L., Lenoir-Wijnkoop, I., Carlberg, J.G., Senanayake, V., Gutierrez-Ibarluzea, I., Poley, M.J.,
Dubois, D., and Jones, P.J., Health economics and nutrition: A review of published evidence. Nutr. Rev.,
70, 693–708, 2012.
4. US Food and Drug Administration (US FDA), Generally recognized as safe (GRAS), February 2014,
Published online at: http://www.fda.gov/food/ingredientspackaginglabeling/gras/default.htm (accessed
June 28, 2014).
5. CNN Health, Diet soda helps weight loss, industry-funded study finds, May 2014, Published online at:
http://www.cnn.com/2014/05/27/health/diet-soda-weight-loss/ (accessed June 28, 2014).
6. Bagchi, D., Zafra-Stone, S., Bagchi, M., and Preuss, H.G., Overview of (−)-hydroxycitric acid in weight
management, in Obesity: Epidemiology, Pathophysiology, and Prevention, 2nd edn., Bagchi, D. and
Preuss, H.G., Eds., CRC Press/Taylor & Francis Group, Boca Raton, FL, 2013, pp. 511–531.
7. Wright, R., Coca-cola acquires Fuze beverages, Nutraceuticals World, February (Newsletter), 2007.
8. Fuze Slenderize Review, 2013, Published online at: http://www.dietspotlight.com/fuze-slenderize-
review/ (accessed November 13, 2014).
9. Fuze Slenderize, 2012, Published online at: http://diet-supplements.com/fuze-slenderize/ (accessed
November 13, 2014).
10. Skinny Water, 2011, Published online at: http://www.slideshare.net/SkinnyWater/skinny-power-
point-presentation (accessed November 13, 2014).
11. Sobe Lean Energy Beverage, 2013, Published online at: http://www.directionsforme.org/index.php/
directions/product/ISOTONIC/00739510000216 (accessed November 18, 2014).
12. Bagchi, D., Bagchi, M., Zafra-Stone, S., and Preuss, H.G., Chromium (III) in promoting weight loss and
lean body mass, in Obesity: Epidemiology, Pathophysiology, and Prevention, 2nd edn., Bagchi, D. and
Preuss, H.G., Eds., CRC Press/Taylor & Francis Group, Boca Raton, FL, 2013, pp. 501–510.
13. Sreejayan, N., Marone, P.A., Lau, F.C., Yasmin, T., Bagchi, M., and Bagchi, D., Safety and toxicological
evaluation of a novel chromium (III) dinicocysteinate complex. Toxicol. Mech. Methods, 20, 321–333, 2010.
14. Tian, H., Guo, X., Wang, X., He, Z., Sun, R., Ge, S., and Zhang, Z., Chromium picolinate supple-
mentation for overweight or obese adults. Cochrane Database Syst. Rev., 11, CD010063, 2013. doi:
10.1002/14651858.CD010063.pub2.
15. Flanagan, J., Billy, A., Rolland, Y., and Roller, M., Lipolytic activity of svetol, a decaffeinated green
coffee bean extract. Phtother. Res., 28, 946–948, 2014.
16. Vinson, J.A., Burnham, B.R., and Nagendran, M.V., Randomized, double-blind, placebo-controlled,
linear dose, cross over study to evaluate the efficacy and safety of a green coffee bean extract in over-
weight subjects. Diabetes Metab. Syndr. Obes., 5, 21–27, 2012.
17. Ochiai, R., Jokura, H., Suzuki, A., Tokimitsu, I., Ohishi, M., Komai, N., Rakugi, H., and Ogihara, T.,
Green coffee bean extract improves human vasoreactivity. Hypertens. Res., 27, 731–737, 2004.
18. Swaroop, A., Bagchi, M., Kumar, P., and Bagchi, D., Efficacy of a novel green coffee bean extract (GCB-70)
in weight management, presented at Experimental Biology 2014, San Diego, CA, April 26–30, 2014, A515
Abstract 1107.2.
19. Baladia, E., Basulto, J., Manera, M., Martinez, R., and Calbert, D., Effect of green tea or green tea
extract consumption on body weight and body composition; systematic review and meta-analysis. Nutr.
Hosp., 29, 479–490, 2014.
20. He, R.R., Chen, L., Lin, B.H., Matsui, Y., Yao, X.S., and Kurihara, H., Beneficial effects of oolong
tea consumption on diet-induced overweight and obese subjects. Chin. J. Integr. Med., 15, 34–41,
2009.
21. Hursel, R. and Westerterp-Plantenga, M.S., Catechin- and caffeine-rich teas for control of body weight
in humans. Am. J. Clin. Nutr., 98(Suppl. 6), 1682S–1693S, 2013.
22. Jurgens, T.M., Whelan, A.M., Killian, L., Doucettes, S., Kirk, S., and Foy, E., Green tea for weight loss
and weight management in overweight and obese adults. Cochrane Database Syst. Rev., 12, CD008650,
2012. doi: 10.1002/14651858.CD008650.pub2.
23. Diet snapple to help weight loss, Published online at: http://forum.bodybuilding.com/showthread.
php?t=142869831 (accessed November 17, 2014).
24. Sudipa, Lipton tea for weight loss, 2014, Published online at: http://fawesome.ifood.tv/diet/338783-
lipton-tea-for-weight-loss (accessed November 17, 2014).
25. Arizone Beverage Company, Arizona products, 2015, Published online at: www.drinkarizona.com/
productcart/ (accessed October 12, 2015).
26. Kong, R.Z.L., Antiobesity effects of conjugated linoleic acid, in Obesity: Epidemiology, Pathophysiology,
and Prevention, 2nd edn., Bagchi, D. and Preuss, H.G., Eds., CRC Press/Taylor & Francis Group, Boca
Raton, FL, 2013, pp. 555–575.
27. Tonalin, Conjugated linoleic acid and beverage, 2015, Published online at: www.tonalin.com/about.htm
(accessed October 12, 2015).
28. Congugated linoleic acid and evolve, 2015, Published online at: www.cytosport.com/our-story (accessed
October 12, 2015).
29. Swaroop, A., Bagchi, M., Kumar, P., Preuss, H.G., Tiwari, K., Marone, P.A., and Bagchi, D., Safety,
efficacy and toxicological evaluation of a novel, patented anti-diabetic extract of Trigonella foenum-
graecum seed extract (Fenfuro). Toxicol. Mech. Meth., 21, 1–25, 2014.
30. Kumar, P., Kale, P.K., and Baquer, N.Z., Antihyperglycemic and protective effects of Trigonella foenum-
graecum seed powder on biochemical alterations in alloxan diabetic rats. Eur. Rev. Med. Pharmacol.
Sci., 16(Suppl. 3), 18–27, 2012.
31. Preuss, H.G., Bagchi, D., and Kaats, G.R., Laboratory and clinical studies of chitosan, in Obesity:
Epidemiology, Pathophysiology, and Prevention, 2nd edn., Bagchi, D. and Preuss, H.G., Eds., CRC
32. Tan, S., Gao, B., Tao, Y., Guo, J., and Su, Z.O., Antiobese effects of capsaicin-chitosan microsphere
(CCMS) in obese rats induced by high-fat diet. J. Agric. Food. Chem., 62, 1866–1874, 2014.
33. Gavhane, Y.N., Gurav, A.S., and Yadav, A.V., Chitosan and its applications: A review of literature. Int.
J. Res. Pharm. Biomed. Sci., 4, 312–331, 2013.
34. Luo, Y. and Wang, Q., Recent advances of chitosan and its derivatives for novel applications in food
science. J. Food Process. Bever., 1, 1–13, 2013.
35. Keithley, J.K., Swanson, B., Mikolaitis, S.L., DeMeo, M., Zeller, J.M., Fogg, L., and Adamji, J., Safety
and efficacy of glucomannan for weight loss in overweight and moderately obese adults. J. Obes., 2013,
610908. doi: 10.1155/2013/610908. Epub December 30, 2013.
36. Swanson, B. and Keithley, J.K., Glucomannan in weight loss, in Obesity: Epidemiology, Pathophysiology,
and Prevention, 2nd edn., Bagchi, D. and Preuss, H.G., Eds., CRC Press/Taylor & Francis Group, Boca
Raton, FL, 2013, pp. 627–636.
37. Sun, G. (Inventor), Glucomannan high fiber beverage, U.S. Patent Pub. No. US 2003/0138545 A1,
July 24, 2003.
38. LuraLean, Dietary supplement, functional food and beverage ingredient, 2008, Published online at:
http://resources.nutraconference.com/forms/pdfs/AHDFinal.pdf (accessed November 17, 2014).
39. Cangiano, C., Ceci, F., Cascino, A., Del Ben, M., Laviano, A., Muscaritoli, M., Antonucci, F., and Rossi
Fanelli, F., Eating behavior and adherence to dietary prescriptions in obese adult subjects treated with
5-hydroxytryptophan. Am. J. Clin. Nutr., 56, 863–867, 1992.
40. Ceci, F., Cangiano, C., Cairella, M., Cascino, A., Del Ben, M., Muscaritoli, M., Sibilia, L., and Rossi
Fanelli, F., The effects of oral 5-hroxytryptophan administration on feeding behavior in obese adult
female subjects. J. Neural Transm., 76, 109–117, 1989.
41. Das, Y.T., Bagchi, M., Bagchi, D., and Preuss, H.G., Safety of 5-hydroxy-l-tryptophan. Toxicol. Lett.,
150, 111–122, 2004.
42. Research Applications, 5-Hydroxytryptophan, 2015, Published online at: http://www.fortitechpremixes.com/
glossary/5-hydroxytryptophan-5-htp/ (accessed October 12, 2015).
43. Zafra-Stone, S., Yasmin, T., Bagchi, M., Chatterjee, A., Vinson, J.A., and Bagchi, D., Berry antho-
cyanins as novel antioxidants in human health and disease prevention. Mol. Nutr. Food Res., 51,
675–683, 2007.
44. Morimoto, C., Satoh, Y., Hara, M., Inoue, S., Tsujita, T., and Okuda, H., Anti-obese action of raspberry
ketone. Life Sci., 77, 194–204, 2005.
45. Park, K.S., Raspberry ketones increases both lipolysis and fatty acid oxidation in 3T3-L1 adipocytes.
Planta Med., 76, 1654–1658, 2010.
46. Miller, M., Robard Corporation: Weight Management and Weight Loss Tips for Professionals and Dieters,
Product spot light: Berry drinks, 2014, Published online at: http://blog.robard.com/post/2014/07/29/
Product-Spotlight-Berry-Drinks.aspx (accessed November 17, 2014).
47. Weight loss and diet plans, Acai berry juice, 2005, Published online at: http://www.webmd.com/diet/
acai-berries-and-acai-berry-juice-what-are-the-health-benefits (accessed November 17, 2014).
48. Brackbauer, A. and Zemel, M.B., Dairy foods, calcium and weight management, in Obesity:
Epidemiology, Pathophysiology, and Prevention, 2nd edn., Bagchi, D. and Preuss, H.G., Eds., CRC
49. Larsen, S.C., Angquist, L., Ahluwalia, T.S., Skaaby, T., Roswall, N., Tjonneland, A., Halkjaer, J. et al.,
Interaction between genetic predisposition to obesity and dietary calcium in relation to subsequent
change in body weight and waist circumference. Am. J. Clin. Nutr., 99, 957–965, 2014.
50. Gerstner, G., How to fortify beverages with calcium by Dr. Gerhard Gerstner Food Marketing and
Technology, 2003, Published online at: http://www.jungbunzlauer.com/fileadmin/content/_PDF/How_
to_fortify_beverages_ with_Calcium_Oct03.pdf (accessed November 17, 2014).
51. Dairy Ingredients Market, Calcium fortification in beverages analyzed in new research reports, 2014,
Published online at: http://www.prnewswire.co.uk/news-releases/dairy-ingredients-market-calcium-forti-
fication-in-beverages-analyzed-in-new-research-reports-257643411.html (accessed November 28, 2014).
52. Greenway, F.L. and Bray, G.A., Treatment of hypothalamic obesity with caffeine and ephedrine. Endocr.
Pract., 14, 697–703, 2008.
53. Liu, A.G., Smith, S.R., Fujioka, K., and Greenway, F.L., The effect of leptin, caffeine/ephedrine, and
their combination upon visceral fat mass and weight loss. Obesity (Silver Spring), 21(10), 1991–1996,
2013. doi: 10.1002/oby.20416. Epub June 11, 2013.
54. Palamar, J., How ephedrine escaped regulation in the United States: A historical review of misuse and
associated policy. Health Policy, 99, 1–9, 2011.
55. Our Program, Step one foods and food ingredients, 2013, Published online at: http://www.steponefoods.
com/glossary.aspx (accessed November 17, 2014).
56. Cozma, A.I, Sievenpiper, J.L., de Souza, R.J., Chiavaroli, L., Ha, V., Wang, D.D., Mirrahimi, A. et al.,
Effect of fructose on glycemic control in diabetes: A systematic review and meta-analysis of controlled
feeding trials. Diabetes Care, 35, 1611–1620, 2012.
57. Park, D.Y., Ahn, Y.T., Huh, C.S., McGregor, R.A., and Choi, M.S., Dual probiotic strains suppress high
fructose-induced metabolic syndrome. World J. Gastroenterol., 19, 274–283, 2013.
58. Giacosa, A. and Rondanelli, M., The right fiber for the right disease: An update on the Psyllium seed
husk and the metabolic syndrome. J. Clin. Gastroenterol., 44(Suppl. 1), S58–S60, 2010.
59. Ziai, S.A., Larijani, B., Akhoondzadeh, S., Fakhrzadeh, H., Dastpak, A., Bandarian, F., Rezai, A.,
Badi, H.N., and Emami, T., Psyllium decreased serum glucose and glycosylated hemoglobin significantly
in diabetic outpatients. J. Ethnopharmacol., 102, 202–207, 2005.
60. Behall, M., Scholfield, D.J., and Hallfrisch, J., Comparison of hormone and glucose responses of
overweight women to barley and oats. J. Am. Coll. Nutr., 24, 182–188, 2005.
61. Granfeldt, Y., Eliasson, A.C., and Bjorck, I., An examination of the possibility of lowering the glycemic
index of oat and barley flakes by minimal processing. J. Nutr., 130, 2207–2214, 2000.
62. All-time Favorite Dishes, Food programs, 2015, Published online at: http://www.bbc.co.uk/food/
programmes/b01qm4t3 (accessed October 12, 2015).
63. Weight Loss & Obesity Management Market, Meal replacements, slimming centers, nutrition & psycho-
logical consultancy, treadmill, ellipticals, strength training, gastric bypass, intragastric balloon system,
stomaphyx—Global forecasts to 2017, 2009, Published online at: http://www.marketsandmarkets.com/
Market-Reports/weight-loss-obesity-management-market-1152.html (accessed August 21, 2014).
64. Nutraceuticals World, Nutraceuticals market to reach $204.8 billion by 2017, May 2014, Published
online at: http://www.nutraceuticalsworld.com/issues/2013–09/view_industry-news/nutraceuticals-
market-to-reach-2048-billion-by-2017/ (accessed June 28, 2014).
65. Lappalainen, R., Mennen, L., van Weert, L., and Mykkanen, H., Drinking water with a meal: A simple
method of coping with feelings of hunger, satiety and desire to eat. Eur. J. Clin. Nutr., 47, 815–819, 1993.
66. Murakami, K., Sasaki, S., Takahashi, Y., and Uenishi, K., Intake of water from foods, but not water
from beverages, is related to lower body mass index and waist circumference in free-living humans.
Nutrition, 24, 925–932, 2008.
67. Stookey, J.D., Constant, F., Popkin, B.M., and Gardner, C.D., Drinking water is associated with
weight loss in overweight dieting women independent of diet and activity. Obesity (Silver Spring),
16, 2481–2488, 2008.
68. Schulze, M.B., Manson, J.E., Ludwig, D.S., Colditz, G.A., Stampfer, M.J., Willett, W.C., and Hu, F.B.,
Sugar-sweetened beverages, weight gain, and incidence of type 2 diabetes in young and middle-aged
women. JAMA, 292, 927–934, 2004.
69. Tordoff, M.G. and Alleva, A.M., Effect of drinking soda sweetened with aspartame or high-fructose
corn syrup on food intake and body weight. Am. J. Clin. Nutr., 51, 963–969, 1990.
70. Wu, A.H., Tseng, C.C., Van Den Berg, D., and Yu, M.C., Tea intake, COMT genotype, and breast cancer
in Asian-American women. Cancer Res., 63, 7526–7529, 2003.
71. Catenacci, V.A., Pan, Z., Thomas, J.G., Ogden, L.G., Roberts, S.A., Wyatt, H.R., Wing, R.R., and
Hill, J.O., Low/no calorie sweetened beverage consumption in the national weight control registry.
Obesity, 22, 2244–2251, 2014.
64
Fortified Sports Drinks
Hernan Brice Kenmogne-Domguia and Cesarettin Alasalvar
CONTENTS
64.1 Introduction................................................................................................................................... 839
64.2 Sports Drinks................................................................................................................................ 840
64.2.1 Definition, Specification, and Use.................................................................................... 840
64.2.2 Adverse Effects of Sports Drinks..................................................................................... 840
64.3 Ingredients Used in Fortified Sports Drinks................................................................................. 841
64.3.1 Bioactives.......................................................................................................................... 841
64.3.2 Polysaccharides................................................................................................................ 842
64.3.3 Proteins............................................................................................................................. 843
64.3.4 Peptides............................................................................................................................. 847
64.3.5 Free Amino Acids............................................................................................................ 847
64.4 Factors Governing the Fortification of Sports Drinks.................................................................. 848
64.5 Conclusion..................................................................................................................................... 849
References............................................................................................................................................... 849
64.1 Introduction
Sports drinks are fortified, thirst-quenching beverages whose stated purpose is to help athletes
engaged in active training and competition. Sports drinks act by preventing dehydration and enhance
rehydration, by supplying carbohydrates to augment the available energy and glycogen replacement,
and by providing electrolytes to replace losses due to perspiration and sweating, leading to a better
exercise performance [1]. According to some sport regulatory authorities, they are generally made
up of carbohydrates, minerals, electrolytes, some vitamins, and flavoring [2,3] to improve their
palatability, which is one of the critical parameters that should be taken into account during their
designing [4]. Several studies have been conducted to clarify the physiological changes occurring
in athletes during exercise and define the appropriate nutritional needs for their activity. These stud-
ies show that the composition of the sports drinks and the timing of their consumption are critical
factors for fatigue reduction and performance improvement during training and exercise, and also
for the maximization of muscular recovery after exercise. Using this information and results from
sport nutrition studies, many commercially available sports drinks have been designed (Gatorade,
Powerade, Powerade Advance, Vitargo, Jxdrink, and Accelerade, etc.) [5].
Today, sports drinks are marketed not only to athletes but also to a broader consumer cross section
that does not necessarily engage in physical activity. For a few years now, misuse of sports drinks and
their interest are rising among those who are not engaged in physical activity. This could lead to health
outcomes (enamel erosion and weight gain) [6] that must be taken into account during their design
processes. Rising interest of nonathletic people for the sports drinks leads to an increase in the produc-
tion and the selling of many sports drinks on the market, which have the specificity to be fortified in
ingredients that enable these disadvantages to be overcome or have additional benefits for athletes. These
contain, other than the basic ingredients of sports drinks and according to the sports legislation, ingre-
dients such as proteins [7–9], polysaccharides [10–12], peptides [13–16,24], free amino acids [17,18], and
839
some bioactives [19–23] that had been found to have many positive functions and properties according
to the sports category. The sports benefits of these fortified drinks are related to improving glycogen
recovery, muscular protein synthesis, reduction of soreness and fatigue delay, as well as rehydration
[10,17,18,24–27]. Sports drink fortification can be achieved in two ways: a direct addition and/or a partial
replacement of carbohydrate by fortifying ingredients. Depending on the timing of ingestion in relation
to exercise, three types of sports drinks (preexercise drink, during exercise drink, and postexercise
drink) are differentiated based on their different compositions and functionalities [23,28]. This chapter
reviews recent research on the fortification of sports drinks, their benefits, and the functional behavior of
ingredients used in this fortification.
64.2 Sports Drinks

64.2.1 Definition, Specification, and Use
Sports drinks are liquid substances, containing a variety of nutrients and other substances dissolved
in water, which are consumed in association with sports or exercise either in preparation for exercise,
during exercise itself, or as a recovery drink after exercise. During exercise and depending on their
intensity and the exercise conditions, athletes could lose about 1%–8% of their body weight through
perspiration and sweating. This dehydration, which is a consequence of a body fluid loss that exceeds
fluid intake [28], is well documented to significantly impair aerobic exercise performance when it occurs
before or during exercise. For dehydration of 1%–2% of body weight, physiologic function begins to be
compromised and has a negative influence on performance. These adverse effects are further increased
for values greater than 3% from which physiological functions are further disturbed and could lead to
an increase the risk of exertional heat illness in athletes [29]. It is, therefore, recommended for athletes
participating in exercise to use sports drinks that help prevent dehydration and produce better hydra-
tion than water [30]. However, for an exercise time less than 1 h, some studies show that there is no
need for sports drinks and that water seems to be sufficient for hydration and lost electrolyte restora-
tion. Thus, some authors have come to the conclusion that for most athletes and individuals engaged in
physical activity, the use of sports drinks does not provide a benefit over water [2]. Many studies lead
to their effectiveness for improving sport performance [7,14,23,31–36]. The potential benefits of sports
drinks are tightly linked to their composition and ingested quantities, to the time it takes for the drink
to be emptied from the stomach and absorbed from the intestine, to whether the drink attenuates exo/
endogenous carbohydrate oxidation and protein damage, and to the nature and intensity of the exercise.
Regarding the composition, commercial sports drinks are generally made of carbohydrates (CHO)
and/or sugar (maltodextrins and/or fructose, glucose, and sucrose: 48–84 g/L) and electrolytes (sodium,
potassium, and chloride: 96–440 mg/L, 116–640 mg/L, 4–320 mg/L, respectively) [1–3]. In addition to
these nutrients, some commercially available sports drinks have numerous other nontraditional ingredients
including carbohydrate derivatives (fiber, polysaccharides, pyruvate, and lactate), protein (PRO) and pro-
tein derivatives (intact protein, branched-chain amino acids, free amino acids, keto-analogues, creatine, and
carnitine), fats (glycerol, medium chain triacylglycerols [TAG], and choline), micronutrients (B vitamins,
antioxidant vitamins, chromium, vanadium, and oxygenated fluids), and nonnutritive ingredients (caffeine,
bicarbonate buffers, herbs, ginseng, ciwujia, ginkgo biloba, and hydroxycitric acid, among others).
64.2.2 Adverse Effects of Sports Drinks

It is well known that consuming commercially available sports drinks improves sport performance [3,5],
but their use is not recommended for those who are not engaged in sports activity because it represents
an additional source of calories and can induce dental erosion.
During a human in situ study with five commercially available sports drinks, Hooper et al. [6] found
that these drinks caused significantly more dental erosion than water, and even if the degree of erosion
varied greatly between different subjects, it was considerable in some of them. Numerous studies have
reported that due to its acid pH (3–4), sports drinks can induce solubilization of tooth enamel resulting
Fortified Sports Drinks 841
in genesis of tooth decay among consumers, mainly in children and adolescents. Moreover, it is often
advocated to consume sports drinks when the oral cavity may be dehydrated, but in this condition, an
increase in the degree of erosion is likely to be observed [6].
If it is accepted that the consumption of sports drinks does not provide enough calories according to
those spent in the intensive sports activity among target consumers (athletes), these additional calories
are excessive in many consumers. In fact, today, most of the frequent sports drink consumers are chil-
dren and adolescents who participate in little or no physical activity. For this population, in which the
taste and the resultant functional properties of sports drinks are very attractive, its consumption is an
additional risk for weight gain and some degree of obesity.
64.3 Ingredients Used in Fortified Sports Drinks

During the last 15 years, to respond to the growing demand of athletes regarding the improvement of
their sport performance, many studies on athletes have been carried out to investigate the effects of well-
designed water and nutrient supply during exercise. The best way to do so is to use sports drinks. Today,
almost all commercially available sports drinks contain carbohydrates and electrolytes as main fortified
nutrients and thus are not frequently equivalent to beverages investigated in sports and nutrition studies.
However, the results of these studies provide insight into what properties of sports drinks may lead to
performance benefits.
64.3.1 Bioactives
Even if it is common for bioactives to be used in energy drinks, their use in sports drinks is scarce.
Among these compounds, creatine, caffeine, and taurine, which act as ergogenic aid, have been used
by athletes [28]. Creatine, most widely used to build muscle and enhance recovery, has been shown to
be effective in repeated short bursts of high-intensity activity in sports such as sprinting and weight
lifting but not for endurance sports such as distance running. Caffeine is used according to its role as a
central nervous system stimulant leading to a decrease perception of effort (Figure 64.1). Even though
considered safe for healthy adults, creatine supplementation has many adverse effects such as weight
gain, cramping, nausea, and diarrhea as well as dehydration, muscle strains/tears, and kidney damage.
Regarding caffeine, although the World Anti-Doping Agency has removed caffeine from the restricted
list of its Monitoring Programme in 2004, it is still a restricted substance by the National Collegiate
Athletic Association, where a positive doping test would be a caffeine level greater than 15 µg/mL of
urine. New evidence shows that caffeine, when used in moderation, does not cause dehydration, water–
electrolyte imbalance, hyperthermia, or reduced exercise-heat tolerance [37,38]. However, when rapid
hydration is necessary, athletes should rely on noncaffeinated beverages. Taurine is commonly used in
energy drinks at a higher level than 4000 mg/L where its purpose is to potentiate some effects or act in
combination with caffeine at the level of the heart and the brain (Figure 64.2). It also has an increasing
effect on postexercise cardiac muscle contractility similar to that of caffeine, and like caffeine, it affects
the intracellular calcium concentration in smooth muscles that may cause coronary vasospasm even if an
improving maximal performance and lowering heart rate at submaximal working intensity could also be
shown [39,40]. Thus, creatine, caffeine, and taurine are not recommended to be used before and during
exercise and are avoided in sports drinks.
O CH3
N
H3C N N
O N
H3C
FIGURE 64.1 Chemical structure of caffeine.

S OH
H2N O
FIGURE 64.2 Chemical structure of taurine.
Recent research has been carried out to study the effects of moderated caffeine-fortified sports drinks
on sports performance. In 2007, Cureton et al. [32] showed that supplemental sports drinks with caf-
feine (195 mg/L) led to an enhancement in cycling performance and a reduction of strength loss as
well as a reduction in fatigue and perception of effort, discomfort, and pain. More recently, it appears
that fortification of a CHO-PRO drink with caffeine (2600 mg/L) improved both cycling power output
and auditory response time following 2 h of moderate and high-intensity interval cycling compared
to a commercial shot containing a large caffeine dose [36]. These observations were in accordance
with those of Kammerer et al. [41] who found that when young adults consume drinks containing only
caffeine (80 mg in 250 mL) or taurine (1000 mg in 250 mL) or their combination during exercise, no
increase in physical and cognitive ability was observed. These studies may allow one to hypothesize
that caffeine has a synergistic ergogenic effect that depends tightly on other ingredients present in sports
drinks. In fact, caffeine fortification of a complex carbohydrate sports drinks leads to increase running
performance (2%), decrease fatigue and maintain glycemia [35], attenuate the muscle fatigue, and mini-
mize the drop in physical performance and to higher glycemia [23,42].
Over these common bioactives, other compounds with different biological function were recently used
as fortifying ingredients. It has been shown that when γ-aminobutyric acid–containing sports drinks
(5 g/L) are consumed during rest and exercise at high temperature, it could induced a larger decrease of the
body core temperature as well as the total heat production, which could be of great interest in improving
sports performance [19,22]. For several years, studies on the ergogenic potential of betaine in endurance
and resistance exercise resulted in nonsignificant trends before the study of Lee et al. [21]. They found that
the consumption of sports drinks fortified with betaine (4.2 g/L) by recreationally active men with at least
3 months of resistance training experience increases and maintains their physical performance accord-
ing to power and force measurements after a resistance exercise testing protocols. Fortification of sports
drinks could also have the aim of improving the health of athlete with no interest with sport performance.
It is the case of a recent fortification strategy developed to inhibit the enamel dental erosion induced by
several sports drinks. In fact, it has been shown in vitro that adding nanosized hydroxyapatite (5.5 g/L) to
Powerade might prevent dental erosion [43]. This latter study has opened the way to fortification of sports
drinks with ingredients/bioactives with healthy, but no sport performance, benefits.
64.3.2 Polysaccharides
It is well known that carbohydrate feeding during exercise improves prolonged endurance performance
by maintaining high rates of carbohydrate oxidation and preventing hypoglycemia [5,44]. The optimum
type and concentration of sugars to be added to drinks is of high importance in the sports drinks. For
example, Powerade containing 8% CHO (fructose, glucose, and maltodextrin) was found to yield run
performance, which was 8% faster compared to that of Gatorade containing lower CHO (6%, fructose
and glucose) [45]. However, high carbohydrate concentrations can delay gastric emptying and thus
reduce fluid absorption. Very high concentrations result in secretion of water into the intestine and thus
increase the dehydration [1]. Perhaps because of this effect, high sugar concentrations (>10%) may result
in an increased risk of gastrointestinal distress. Where there is a need to supply an energy source during
exercise, however, increasing the sugar content of drinks can increase the delivery of carbohydrate to
the site of absorption in the small intestine, which can benefit from the sport performance. Increasing
the carbohydrate content of sports drinks without the adverse effect of the high sugar content could be
achieved by adding high-molecular-weight polysaccharides instead of sugar or maltodextrin. One of the
first studies of the potential benefits of high-molecular-weight carbohydrates (HMW CHO) was carried
out by Piehl Aulin et al. [10], who studied the rate of muscle glycogen synthesis during 2 and 4 h of
recovery after depletion by exercise. They found that consumption of a carbohydrate solution contain-
ing glucose polymers (500–700 kDa) with a low osmolality could restore muscle glycogen at a faster
rate, compared to an energy-equivalent solution containing monomers (500 Da) with a high osmolality
during the first 2 h after exercise when muscle glycogen content is low [10]. The possible mechanisms
mentioned were more rapid gastric emptying and glucose delivery to the intestine and a postexercise
stimulated non-insulin-dependent glucose uptake into the muscle cell. If HMW CHO-consumed postex-
ercise show this effect, there is no effect of HMW CHO-fortified drinks during exercise when compared
to low-molecular-weight carbohydrate (LMW CHO). In fact, HMW CHO or LMW CHO ingested as
fortified drinks at the same rate during cycling exercise had the same effect on exogenous and endog-
enous substrate oxidation rates [11]. If this study failed to show a beneficial effect of consumption
of HMW CHO-fortified drinks during exercise, in a more recent study, it has been shown that a novel
288 kDa HMW CHO-fortified sports drink consumed during exercise could significantly enhance body
blood glucose concentration and delay fatigue with athletic performances nearly the same as those from
the subjects who consumed the Vitargo sports drink containing 500–700 kDa carbohydrates [12].
64.3.3 Proteins
One of the ingredients commonly used for fortification in sports drinks in relation to sport performance
are proteins. Regarding the efficacy of the fortification on the sport performance, several parameters
should be taken into account. First, is the type of fortification process, which could consist of either the
addition of 24%–53% of the carbohydrate fraction [7,25,46–49] or partial replacement of the carbo-
hydrate content (20%–26%) of the sports drinks [8,31,34,50]. Partial replacement of the carbohydrate
fraction should be preferred to the protein addition, because in this way, drinks could have lower caloric
content while improving the sport performance.
The second parameter that must be taken into account during fortification is the nature of the fortifying
protein, which is of great importance in sport performance. Recently, it has been suggested that the effects
of protein supplementation are highly dependent upon protein composition and digestion rate [49]. In fact,
by assessing the effect of protein composition on muscular performance after prolonged resistance train-
ing, Babault et al. [49] found that consumption of soluble whey protein-fortified sports drinks appeared
to significantly reduce muscle fatigue induced by intense resistance exercise than micellar casein. This
effect seems to be linked to the rate of digestion of these proteins and their subsequent amino acid absorp-
tion, which was previously found to be higher for whey protein than for casein when they were ingested
alone [51]. Under these conditions, dietary amino acids absorption is faster with whey protein (fast digest-
ible) than with casein, leading to a more important oxidation and stimulation of postprandial protein syn-
thesis (68%) than for casein (31%). From these findings, the main proteins used in drinks are whey protein
isolates or concentrates at concentration range from 7.5 to 44.3 g/L (Table 64.1). It has, thus, been found
that the addition of 1.4%–1.5% protein to a 6% carbohydrate Accelerade® drinks could increase rehydra-
tion and fluid retention after dehydration [25] and minimize muscle damage [47] when ingested after
cycling or preskiing, during skiing, and postskiing exercise, respectively. Many other benefits of ingestion
of whey protein-supplemented sports drinks are an improvement of sport performance and enhancement
of O2 capacity [7,8] of athletes. Moreover, ingestion of such drinks could also increase muscle glycogen
resynthesis and recovery process and improve exercise and aerobic endurance [31,34], as well as time to
fatigue during cycling [46]. Virtually all protein-fortified sports drinks contain whey protein, but a recent
study showed that in addition to an appropriate training, fortification of sports drinks with pea protein pro-
moted comparable increase of muscle thickness to whey protein [9] showing that such vegetable proteins
could be used as an alternative to whey protein in sports drinks. Interestingly, pea protein has an essential
branched-chain amino acids (BCAA; such as leucine, isoleucine, and valine) (Figure 64.3) content lower
than that of whey protein known to play an important role in muscle protein synthesis. From all these
studies, it, therefore, seems that the type of protein is tightly linked to the potential sports benefits related
to protein fortification of sports drinks. A hypothesis was previously corroborated by Tang et al. [26] who
studied the effect of consumption of sports drinks fortified with either micellar casein or soy protein dur-
ing recovery time. In fact, they found that postexercise muscle protein synthesis was greater for soy protein
(rapidly digestible) than for casein (slowly digestible).
844
TABLE 64.1
Sources and Bioactives in Fortified Sports Drinks
Bioactive/Supplemented Substance Expected Functional Characteristics Drinks and Timing of Use Content (g/L) References
High-Molecular-Weight Polysaccharides
Polysaccharide (500–700 kDa) Increases glycogen synthesis rate. PU 24–002a 150 [10]
Amylopectin waxy starch (500–750 kDa) Leads to same exo/endogenous oxidation than low-molecular-weight Vitargo drinkb 113 [11]
maltodextrin (8 kDa).
Jxsac polysaccharide (288 kDa) versus Jxsac polysaccharide enhances glycemia and delays fatigue with Jxdrink versus Vitargo drinkb 150 [12]
higher-molecular-weight polysaccharide performances near those obtained with Vitargo.
(500–700 kDa)
Protein Isolate and/or Concentrate
Whey protein isolate Improve sport performance. Gatoradeb 20 [7]
Whey protein Adding protein to low-carbohydrate drink may enhance O2 capacity. Experimental drinkb 11.5 versus 7.5 [8]
Nonspecified protein (see Accelerade®) Protein supplementation increase rehydration and fluid retention after Accelerade®a 15 [25]
dehydration.
Whey protein Protein coingestion exerts an ergogenic benefit leading to enhancement Experimental drinkb,c 21 [48]
of endurance capacity and reduction of ratings of perceived exertion.
Milk + whey protein isolate Increase muscle glycogen resynthesis/recovery process and improve Experimental drinka 67.8 [31]
exercise endurance.
Protein (NS) Protein supplementation minimizes muscle damage. Acceleradea,b 13.9 [47]
Whey protein Improves time to fatigue and reduce muscle damage. Experimental drinkb 18 [46]
Whey protein isolate versus micellar Fast-digestible whey protein reduces muscle fatigue with high intensity Experimental drinka,b 50 [49]
casein than micellar casein.
Whey protein isolate Reduces muscle soreness. Experimental drinka 23 [50]
Pea protein isolate versus whey protein Lower BCAA pea protein promotes comparable increase of muscle Experimental drinka,c 46.3 versus 44.3 [9]
concentrate thickness than whey protein.
Whey protein isolate Replacement of carbohydrate by protein in sports drink improves Reconstituted drinksb 12 [34]
aerobic endurance.
Protein Hydrolysate
Whey protein hydrolysate Promotes recovery and muscle protein accretion. Experimental drinka 40 [16]
Casein protein hydrolysate Reduces muscle soreness and enhance late-exercise time-trial Experimental drinka,b 18 [14]
performance.
(Continued)
Silk amino acid hydrolysate Peptides rich in alanine, glycine, serine, valine, and threonine, Carborade drink ®c 2 [52]
combined with an extensive and rigorous training, increase aerobic
Fortified Sports Drinks
endurance.
Whey protein hydrolysate versus micellar Whey protein hydrolysate stimulates muscle protein synthesis to a Experimental drinka 86 versus 88 [26]
casein and soy protein isolate greater extent than casein or soy. Type of protein, through amino acids
content and digestion rate, affects postexercise muscle anabolism.
Whey protein versus whey protein/fish Fish protein hydrolysate affects exercise metabolism during endurance Reconstituted drinksb 18.9 versus [53]
protein hydrolysate cycling but did not provide an ergogenic benefit. 15.3/3.3
Free Amino Acids
Leucine Supplementation increase muscular protein synthesis. Reconstituted drinksa 5.6 versus 3.0 [18]
Valine/leucine/isoleucine/arginine Reduce muscle soreness and fatigue sensation. Experimental drinka,b,c 2/4/2/2 [17]
Isoleucine/leucine/valine Attenuate muscle damage during prolonged endurance exercise. Musasha drinkb 0.5/1.2/0.73 [54]
Isoleucine/leucine/valine/green tea powder Show anti-delayed-onset muscle soreness effect and suppressed the Experimental drinkc 2/4.6/2.4/2 [27]
muscle force decrease (to 80%).
Isoleucine/leucine/valine/arginine Addition to carbohydrate beverage reduces muscle damage, decreases Experimental drinka,b,c 2/4/2/2 [33]
fatigue, and maintains exercise performance after consecutive days of
exercise.
Protein Hydrolysate + Free Amino Acids
Casein hydrolysate versus casein Protein hydrolysate with or without free leucine supplementation Experimental drinka 12.5 versus [62]
hydrolysate/leucine increases insulin secretion. 12.5/3.1
Wheat–gluten protein hydrolysate/free Peptide and free amino acids supplementation of carbohydrate drink Experimental drinka 57.1 [24]
leucine and phenylalanine stimulates glycogen synthesis.
(Whey, wheat, and pea) protein Supplementation of carbohydrate drink with free amino acids with or Experimental drink 57.1 [13]
hydrolysate, casein, arginine, leucine, without wheat peptides results in an insulinotropic effect, which is
phenylalanine, and glutamine 100% greater.
Caseinate, milk protein isolate, whey After intense activity, consumption of a complex protein beverage may VPX protein rusha 79.6 [15]
proteins, micellar casein, casein peptides, favorably influence subsequent physical performance better than an
di-/tripeptides, and whey peptides isocaloric carbohydrate drink.
(Continued)
845
846

Other Bioactives and Complex Ingredients Mix
γ-Amino butyric acid Downregulation of core temperature. Experimental drinkc 5 [19,22]
Hydroxyapatite Prevents dental enamel erosion. Powerade 2.5 [43]
Betaine Increases and maintains power and force in performance Gatorade©c 4.17 [21]
measurements.
Caffeine/taurine Caffeine, taurine, or their combination does not increase the physical Experimental drinka 0.32/4 [41]
and cognitive ability.
Caffeine/taurine/carnitine Caffeine leads to enhancement of cycling performance, reduction of Powerade Advance®a,b,c 0.195/0.192/0.046 [32]
strength loss of fatigue, perception of effort, discomfort, and pain.
Taurine/caffeine/glucuronolactone Bioactives mix has positive effects on mental performance and mood. Experimental drinkc 4/0.32/2.4 [20]
Taurine/caffeine/niacin versus proteins/ Proteins and caffeine improve cycling power output and auditory 5 Hour Energy® Body Glove 52/2.6 versus [36]
caffeine response time. Surge®b 8.4/3.6
Multiple Ingredients/Bioactives
Caffeine/vitamins B + BCAA/caffeine/ Consumption of well-designed sports drinks according to exercise Prematch drinkc + match 0.14/0.0014 + [23]
vitamins C, E, B + protein/lipids/ timing attenuates the muscle fatigue and minimizes the drop in drinkb + postmatch drinka 4/0.075/0.015,
vitamins C, E, B physical performance. 0.005, 0.0101 +
40/7.2/0.12, 0.02,
0.0849
Caffeine + BCAA + proteins/lipids (NS) Consumption of well-designed sports drinks according to exercise Preexercisec + during 0.14 + 4 + 40/7.2 [42]
timing leads to perceive less fatigue, to play with more intensity, and exerciseb + postexercisea
to higher glycemia.
BCAAs/caffeine Combination of carbohydrates to BCAA and caffeine increases running Nutratleticb,c 4/0.075 [35]
performance (2%), decreases fatigue, and maintains glycemia.
Whey protein/ vitamin E/vitamin C Protein and antioxidants supplementation attenuated postexercise Accelerade®b 18/0.08/0.50 [63]
muscle damage. Endurox®a 39/0.55/1.95
Recommended timing for sports drinks use: a postexercise/training, b during exercise/training, and c preexercise/training.
Abbreviations: BCAA, branched-chain amino acids; NS, nonspecified.
O CH3
HO CH3
NH2
Valine
O
CH3
HO
H2N CH3
Leucine
H3C
O
CH3
HO
NH2
Isoleucine
FIGURE 64.3 Chemical structures of branched chain amino acids.
64.3.4 Peptides
The rate of protein digestion and/or their postprandial rate of amino acid absorption are of great inter-
est for some beneficial effects of sports drinks. To make use of this finding, many recent studies have
supplemented these drinks with protein hydrolysates. Tang et al. [26] found that in comparison to slowly
(micellar casein) and rapidly (soy) digestible protein, whey protein hydrolysate ingestion after resistance
exercise in 90 g/L sports drinks resulted in a larger increase in blood essential and BCAA concen-
trations. Moreover, after exercise, muscle protein synthesis following whey protein hydrolysate drink
consumption was about 122% greater than casein and 31% greater than soy, showing that fortification
of sports drinks with protein hydrolysates could have more sport-related benefits compared to crude
proteins. These studies as well as many others show that consumption of peptide (protein hydrolysates)
fortified sports drinks during and after exercise could have an ergogenic benefit. Ingestion of sports
drinks containing a lower amount of peptides, from whey (40 g/L) or casein (18 g/L) than that of whey
protein hydrolysate (90 g/L) [26], either promotes recovery and muscle protein accretion [16] or reduces
muscle soreness and enhances late-exercise, time-trial performance [14], respectively. Peptides from
unusual sources of protein can also be used in sports drinks. Ingestion of silk amino acid hydrolysate-
supplemented Carborade® drink combined with an extensive and rigorous training schedule shows its
efficacy to increase an aerobic endurance [52]. These peptides, with a length of 18–19 amino acids, can
exert a large range of physiological effects including antidiabetic, antioxidant, and antitumor properties.
Moreover, the partial replacement (18%) of whey protein in sports drinks by fish protein hydrolysate was
shown to have significant effects on exercise metabolism during endurance cycling even if no ergogenic
benefit was observed [53].
64.3.5 Free Amino Acids

Until 2009, amino acids, branched-chain amino acids, and carnitine were considered ineffective on
sport performance enhancement [28]. However, several researchers have reported their positive effects
in fortified sports drinks. In contrast to proteins and peptides, a lower amount of supplemented amino
acids is needed in sports drinks to induce an effect, from 2.5 to 10 g/L (Table 64.1), but the amount
of added amino acids could affect the extent of the benefits observed. In sports drinks, other than the
concentration of amino acids, their composition is also important. In a recent study, the increase of the
concentration (3 vs. 5.6 g/L), and thus the ingested amount, of only leucine in a sports drinks consumed
during steady-state exercise elicited a greater muscle protein synthesis response during recovery [18].
This study supported the known assumption that some amino acids could induce ergogenic effects when
ingested in their free state and that muscle protein synthesis reaches maximal stimulation after the con-
sumption of 10 g essential amino acids. Leucine belongs to the BCAA group (leucine, isoleucine, and
valine), and it is usually used in sports drinks as a mix. BCAA effects could be explained by accelerated
transport of amino acids into muscle cells, resulting in higher intracellular amino acid concentrations
after exercise when BCAA ingestion is combined with prolonged endurance exercise. At a low con-
centration (~2.5 g/L) in sports drinks, BCAA mix (isoleucine, leucine, and valine) before and during
exercise attenuates muscle damage during prolonged endurance exercise [54]. At a higher concentra-
tion (8–9 g/L), ingestion of BCAA-fortified sports drinks could reduce muscle damage and soreness,
decreased fatigue, and maintained exercise performance after consecutive days of exercise when there
are ingested before, during, and after exercise [17,33]. It could also decrease to about 80% of the muscle
force [27] when they are ingested before the exercise.
64.4 Factors Governing the Fortification of Sports Drinks

The evolution of the taste of sports drinks during exercise phases is a characteristic of great importance
in the design of sports drinks. In fact, the main aim of sports drinks is to rehydrate the body and replace
electrolyte losses through sweat. It has been shown that during exercise, athletes could lose 1%–8%
of their body weight, mainly of water, depending on the exercise conditions. Some studies have dem-
onstrated that electrolyte concentration of beverages affects their palatability, which is important for
promoting their consumption, and also confirm that moderately high electrolyte content is essential if
the ingested fluid is to be retained in the body [55,56]. One of the difficulties that could be encountered
during evaluation of the palatability of sports drinks is the change of the perception of the taste of sports
drinks during exercise. For example, by recording sensory perceptions of sports drink formulations
when consumed before, at various points during and following exercise, Ali et al. [57] found that the rat-
ings of sweetness for the formulation tested, which also increased with duration of exercise, were higher
during exercise relative to pre- and postexercise conditions. This is in accordance with other study show-
ing that repeated exposure of sports drinks resulted in hedonic adjustment, and postexposure preferences
could not have been predicted with a traditional consumer preference test [4]. From this, care should be
taken into account to make easier the ingestion of high quantity of beverage needed for hydration during
exercise. Since long time, it is well known that fluid retention and palatability of beverages can be linked
to their composition (sodium content and carbonation). For example, a suitable sodium concentration in
a beverage (25 and 50 mM) can increase both its ingested volume and the subsequent whole-body hydra-
tion status after 3 h when compared to flavored water [56].
One of the important features of sports drinks that are increasingly taken into account during their
formulation is their time of consumption according to the preexercise, during exercise, and postexercise
states. In fact, during these states, the physiological and metabolic mechanisms are sought, and the nutri-
tional needs are different. Thus, according to the composition of the sports drinks, different sport benefits
could be observed [6]. Recently, Galloway et al. [58] found that ingestion of a 6.4% carbohydrate/electrolyte
sports drink at 30 min instead of 120 min before exercise produced a greater exercise capacity. Similarly,
another study showed that ingestion of protein supplement directly after resistance exercise, instead of
120 min later, could lead to a higher enhancement of total muscle mass as well as hypertrophy of single
muscle fibers in elderly humans [59]. Hence, many researchers focus on the design of suitable sports
drinks with regard to their time of consumption related to exercise. It was found that consumption of
well-designed sports drinks, according to exercise timing, attenuates the muscle fatigue, reduces fatigue
perception, increases physical performance, minimizes its drop, and leads to higher glycemia [23,42].
Moreover, it appears that other parameters need to be kept in mind for sports drinks fortification. It is the
case of fortification with free amino acids for which the maximum quantity that can be used in drinks
is limited by their bitter taste [60] and if it is possible to mask it. The presence of other ingredients in
sports drinks could also play a role in their final properties. It appears from some studies that the ben-
eficial effects of ingredient added to sports drinks could be linked to a synergistic effect. This is, for
example, the case for caffeine and amino acids [41], and of the proprietary ingredients in SOmaxP drink
with creatine, whey protein, and carbohydrate. In fact, this drink significantly improves strength, muscle
endurance, lean muscle mass, and percentage of body fat versus a counterpart with identical quantities
of creatine, whey protein, and carbohydrate [61].
64.5 Conclusion
It is well established that using sports drinks by athletes and those who are engaged in sports improves
exercise performance and recovery after exercise. If the composition of these drinks was limited to
carbohydrates and electrolytes during long period of time, it appears clear that fortification of sports
drinks with other nutrients (polysaccharides, proteins, peptides, and free amino acids) and/or bioactives
(caffeine, taurine, betaine, and γ-amino butyric acid) could raise their sports-related benefits. These
benefits are tightly linked to the nature and the content of fortifying ingredients, as well as the time of
consumption according to preexercise, during exercise, and postexercise states. Even if studies on forti-
fication of sports drinks rose in the functional food and sports research areas over the last 15 years, less
commercially available sports drinks make use of its findings. Producers of sports drinks should make
use of research findings to develop well-designed sports products according to their composition and
subsequent use in preexercise, during exercise, and postexercise. Until now, all ingredients and bioac-
tives used in sports drinks are connected to sport performance and after sport recovery. However, from
the fact that exercise could induce inflammatory and oxidative stress, it is easy to consider that other
health-related ingredients, such as those with anti-inflammatory or antioxidant properties (sterols, poly-
unsaturated fatty acids, polyphenols), could also be used in fortified sports drinks.
REFERENCES
1. Shirreffs, S.M., The optimal sports drink. Schweiz. Z. Sportmed. Sporttraumatol., 51, 25–29, 2003.
2. Coombes, J.S., Sports drinks and dental erosion. Am. J. Dent., 18, 101–104, 2005.
3. Committee on Nutrition and the Council on Sports Medicine and Fitness, Sports drinks and energy
drinks for children and adolescents: Are they appropriate? Pediatrics, 127, 1182–1189, 2011.
4. Kinnear, M. and de Kock, H.L., Would repeated consumption of sports drinks with different acidulants
lead to hedonic adjustment? Food Qual. Prefer., 22, 340–345, 2011.
5. Coombes, J.S. and Hamilton, K.L., The effectiveness of commercially available sports drinks. Sports
Med., 29, 181–209, 2000.
6. Hooper, S.M., Hughes, J.A., Newcombe, R.G., Addy, M., and West, N.X., A methodology for testing the
erosive potential of sports drinks. J. Dent., 33, 343–348, 2005.
7. van Essen, M. and Gibala, M.J., Failure of protein to improve time trial performance when added to a
sports drink. Med. Sci. Sports Exerc., 38, 1476–1483, 2006.
8. Martínez-Lagunas, V., Ding, Z., Bernard, J.R., Wang, B., and Ivy, J.L., Added protein maintains efficacy
of a low-carbohydrate sports drink. J. Strength Cond. Res., 24, 48–59, 2010.
9. Babault, N., Païzis, C., Deley, G., Guérin-Deremaux, L., Saniez, M.H., Lefranc-Millot, C., and Allaert,
F.A., Pea proteins oral supplementation promotes muscle thickness gains during resistance training: A
double-blind, randomized, placebo-controlled clinical trial vs. whey protein. J. Int. Soc. Sports Nutr.,
12, 3, 2015.
10. Piehl Aulin, K., Söderlund, K., and Hultman, E., Muscle glycogen resynthesis rate in humans after
supplementation of drinks containing carbohydrates with low and high molecular masses. Eur. J. Appl.
Physiol., 81, 346–51, 2000.
11. Rowlands, D.S., Wallis, G.A., Shaw, C., Jentjens, R.L., and Jeukendrup, A.E., Glucose polymer molecu-
lar weight does not affect exogenous carbohydrate oxidation. Med. Sci. Sports Exerc., 37, 1510–1516,
2005.
12. Hao, L., Chen, Q., Lu, J., Li, Z., Guo, C., Qian, P., Yu, J., and Xing, X., A novel hypotonic sports drink
containing a high molecular weight polysaccharide. Food Funct., 5, 961–965, 2014.
13. van Loon, L.J., Saris, W.H., Verhagen, H., and Wagenmakers, A.J., Plasma insulin responses after
ingestion of different amino acid or protein mixtures with carbohydrate. Am. J. Clin. Nutr., 72, 96–105,
2000.
14. Saunders, M.J., Moore, R.W., Kies, A.K., Luden, N.D., and Pratt, C.A., Carbohydrate and protein
hydrolysate coingestions improvement of late-exercise time-trial performance. Int. J. Sport Nutr. Exerc.
Metab., 19, 136–49, 2009.
15. Lynch, S., The differential effects of a complex protein drink versus isocaloric carbohydrate drink on
performance indices following high-intensity resistance training: A two arm crossover design. J. Int.
Soc. Sports Nutr., 10, 31, 2013.
16. Alghannam, A.F., Tsintzas, K., Thompson, D., Bilzon, J., and Betts, J.A., Post-exercise protein trial:
Interactions between diet and exercise (PEPTIDE): Study protocol for randomized controlled trial.
Trials, 15, 459, 2014.
17. Matsumoto, K., Koba, T., Hamada, K., Sakurai, M., Higuchi, T., and Miyata, H., Branched-chain amino
acid supplementation attenuates muscle soreness, muscle damage and inflammation during an intensive
training program. J. Sports Med. Phys. Fitness., 49, 424–431, 2009.
18. Pasiakos, S.M., McClung, H.L., McClung, J.P., Margolis, L.M., Andersen, N.E., Cloutier, G.J., Pikosky,
M.A., Rood, J.C., Fielding, R.A., and Young, A.J., Leucine-enriched essential amino acid supplementa-
tion during moderate steady state exercise enhances postexercise muscle protein synthesis. Am. J. Clin.
Nutr., 94, 809–818, 2011.
19. Miyazawa, T., Kawabata, T., Suzuki, T., Imai, D., Hamamoto, T., Yoshikawa, T., and Miyagawa, T.,
Effect of oral administration of GABA on temperature regulation in humans during rest and exercise at
high ambient temperature. Osaka City Med. J., 55, 99–108, 2009.
20. Seidl, R., Peyrl, A., Nicham, R., and Hauser, E., A taurine and caffeine-containing drink stimulates
cognitive performance and well-being. Amino Acids, 19, 635–642, 2000.
21. Lee, E.C., Maresh, C.M., Kraemer, W.J., Yamamoto, L.M., Hatfield, D.L., Bailey, B.L., Armstrong,
L.E., Volek, J.S., McDermott, B.P., and Craig, S.A., Ergogenic effects of betaine supplementation on
strength and power performance. J. Int. Soc. Sports Nutr., 7, 27, 2010.
22. Miyazawa, T., Kawabata, T., Okazaki, K., Suzuki, T., Imai, D., Hamamoto, T., Matsumura, S., and
Miyagawa, T., Oral administration of γ-aminobutyric acid affects heat production in a hot environment
in resting humans. J. Physiol. Anthropol., 31, 3, 2012.
23. Brink-Elfegoun, T., Ratel, S., Leprêtre, P.M., Metz, L., Ennequin, G., Doré, E., Martin, V. et al., Effects
of sports drinks on the maintenance of physical performance during 3 tennis matches: A randomized
controlled study. J. Int. Soc. Sports Nutr., 11, 46, 2014.
24. van Loon, L.J., Saris, W.H., Kruijshoop, M., and Wagenmakers, A.J., Maximizing postexercise muscle
glycogen synthesis: Carbohydrate supplementation and the application of amino acid or protein hydro-
lysate mixtures. Am. J. Clin. Nutr., 72, 106–111, 2000.
25. Seifert, J., Harmon, J., and DeClercq, P., Protein added to a sports drink improves fluid retention. Int. J.
Sport Nutr. Exerc. Metab., 16, 420–429, 2006.
26. Tang, J.E., Moore, D.R., Kujbida, G.W., Tarnopolsky, M.A., and Phillips, S.M., Ingestion of whey hydro-
lysate, casein, or soy protein isolate: Effects on mixed muscle protein synthesis at rest and following
resistance exercise in young men. J. Appl. Physiol., 107, 987–992, 2009.
27. Shimomura, Y., Inaguma, A., Watanabe, S., Yamamoto, Y., Muramatsu, Y., Bajotto, G., Sato, J.,
Shimomura, N., Kobayashi, H., and Mawatari, K., Branched-chain amino acid supplementation before
squat exercise and delayed-onset muscle soreness. Int. J. Sport Nutr. Exerc. Metab., 20, 236–244, 2010.
28. American Dietetic Association, Dietitians of Canada, and American College of Sports Medicine,
Nutrition and athletic performance. Med. Sci. Sports Exerc., 41, 709–731, 2009.
29. Casa, D.J., Armstrong, L.E., Hillman, S.K., Montain, S.J., Reiff, R.V., Rich, B.S., Roberts, W.O., and
Stone, J.A., National athletic trainers’ association position statement: Fluid replacement for athletes.
J. Athl. Train., 35, 212–224, 2000.
30. Higgins, J.P., Tuttle, T.D., and Higgins, C.L., Energy beverages: Content and safety. Mayo Clin. Proc.,
85, 1033–1041, 2010.
31. Niles, E.S., Lachowetz, T., Garfi, J., Sullivan, W., Smith, J.C., Leyh, B.P., and Headley, S.A.,
Carbohydrate-protein drink improves time to exhaustion after recovery from endurance exercise.
J. Exerc. Physiol. Online, 4, 45–52, 2001.
32. Cureton, K.J., Warren, G.L., Millard-Stafford, M.L., Wingo, J.E., Trilk, J., and Buyckx, M., Caffeinated
sports drink: Ergogenic effects and possible mechanisms. Int. J. Sport Nutr. Exerc. Metab., 17, 35–55,
2007.
33. Skillen, R.A., Testa, M., Applegate, E.A., Heiden, E.A., Fascetti, A.J., and Casazza, G.A., Effects of
an amino acid carbohydrate drink on exercise performance after consecutive-day exercise bouts. Int. J.
Sport Nutr. Exerc. Metab., 18, 473–492, 2008.
34. Ferguson-Stegall, L., McCleave, E.L., Ding, Z., Kammer, L.M., Wang, B., Doerner, P.G., Liu, Y., and
Ivy, J.L., The effect of a low carbohydrate beverage with added protein on cycling endurance perfor-
mance in trained athletes. J. Strength Cond. Res., 24, 2577–2586, 2010.
35. Peltier, S.L., Vincent, L., Millet, G.Y., Sirvent, P., Morin, J.B., Guerraz, M., Geyssant, A., Lescuyer,
J.F., Feasson, L., and Messonnier, L., Effects of carbohydrates-BCAAs-caffeine ingestion on perfor-
mance and neuromuscular function during a 2-h treadmill run: A randomized, double-blind, cross-over
placebo-controlled study. J. Int. Soc. Sports Nutr., 8, 22, 2011.
36. Seifert, J.G. and Connor, D.A., The influence of commercial energy shots on response time and power
output in recreational cyclists. J. Int. Soc. Sports Nutr., 11, 56, 2014.
37. Armstrong, L.E., Pumerantz, A.C., Roti, M.W., Judelson, D.A., Watson, G., Dias, J.C., Sokmen, B. et al.,
Fluid, electrolyte, and renal indices of hydration during 11 days of controlled caffeine consumption.
Int. J. Sport Nutr. Exerc. Metab., 15, 252–265, 2005.
38. Armstrong, L.E., Casa, D.J., Maresh, C.M., and Ganio, M.S., Caffeine, fluid-electrolyte balance, tem-
perature regulation, and exercise-heat tolerance. Exerc. Sport Sci. Rev., 35, 135–140, 2007.
39. Baum, M. and Weiss, M., The influence of a taurine containing drink on cardiac parameters before and
after exercise measured by echocardiography. Amino Acids, 20, 75–82, 2001.
40. Berger, A.J. and Alford, K., Cardiac arrest in a young man following excess consumption of caffeinated
“energy drinks.” Med. J. Aust., 190, 41–43, 2009.
41. Kammerer, M., Jaramillo, J.A., García, A., Calderón, J.C., and Valbuena, L.H., Effects of energy drink
major bioactive compounds on the performance of young adults in fitness and cognitive tests: A random-
ized controlled trial. J. Int. Soc. Sports Nutr., 11, 44, 2014.
42. Peltier, S.L., Leprêtre, P.M., Metz, L., Ennequin, G., Aubineau, N., Lescuyer, J.F., Duclos, M., Brink, T.,
and Sirvent, P., Effects of pre-exercise, endurance, and recovery designer sports drinks on performance
during tennis tournament simulation. J. Strength Cond. Res., 27, 3076–3083, 2013.
43. Min, J.H., Kwon, H.K., and Kim, B.I., The addition of nano-sized hydroxyapatite to a sports drink to
inhibit dental erosion: In vitro study using bovine enamel. J. Dent., 39, 629–635, 2011.
44. Jeukendrup, A.E., Carbohydrate intake during exercise and performance. Nutrition, 20, 669–677, 2004.
45. Millard-Stafford, M.L., Sparling, P.B., Rosskopf, L.B., and Snow, T.K., Should carbohydrate concentra-
tion of a sport drink be less than 8% during exercise in the heat? Int. J. Sport Nutr. Exerc. Metab., 15,
117–130, 2005.
46. Saunders, M.J., Kane, M.D., and Todd, M.K., Effects of a carbohydrate-protein beverage on cycling
endurance and muscle damage. Med. Sci. Sports Exerc., 36, 1233–1238, 2004.
47. Seifert, J.G., Kipp, R.W., Amann, M., and Gazal, O., Muscle damage, fluid ingestion, and energy supple-
mentation during recreational alpine skiing. Int. J. Sport Nutr. Exerc. Metab., 15, 528–536, 2005.
48. Alghannam, A.F., Carbohydrate-protein ingestion improves subsequent running capacity towards the
end of a football-specific intermittent exercise. Appl. Physiol. Nutr. Metab., 36, 748–757, 2011.
49. Babault, N., Deley, G., Le Ruyet, P., Morgan, F., and Allaert, F.A., Effects of soluble milk protein
or casein supplementation on muscle fatigue following resistance training program: A randomized,
double-blind, and placebo-controlled study. J. Int. Soc. Sports Nutr., 11, 36, 2014.
50. Millard-Stafford, M.L., Warren, G.L., Thomas, L.M., Doyle, J.A., Snow, T.K., and Hitchcock, K.,
Recovery from run training: Efficacy of a carbohydrate-protein beverage? Int. J. Sport Nutr. Exerc.
Metab., 15, 610–624, 2005.
51. Boirie, Y., Dangin, M., Gachon, P., Vasson, M.P., Maubois, J.L., and Beaufrère, B., Slow and fast dietary
proteins differently modulate postprandial protein accretion. Proc. Natl. Acad. Sci. U.S.A., 94, 14930–
14935, 1997.
52. Zubrzycki, I.Z., Ossowski, Z., Przybylski, S., Wiacek, M., Clarke, A., and Trabka, B., Supplementation
with silk amino acids improves physiological parameters defining stamina in elite fin-swimmers. J. Int.
Soc. Sports Nutr., 11, 57, 2014.
53. Siegler, J.C., Page, R., Turner, M., Mitchell, N., and Midgely, A.W., The effect of carbohydrate and
marine peptide hydrolysate co-ingestion on endurance exercise metabolism and performance. J. Int.
Soc. Sports. Nutr., 10, 29, 2013.
54. Greer, B.K., Woodard, J.L., White, J.P., Arguello, E.M., and Haymes, E.M., Branched-chain amino acid
supplementation and indicators of muscle damage after endurance exercise. Int. J. Sport. Nutr. Exerc.
Metab., 17, 595–607, 2007.
55. Maughan, R.J. and Leiper, J.B., Post-exercise rehydration in man: Effects of voluntary intake of four
different beverages. Med. Sci. Sports Exerc., 25, S2, 1993.
56. Wemple, R.D., Morocco, T.S., and Mack, G.W., Influence of sodium replacement on fluid ingestion
following exercise-induced dehydration. Int. J. Sport Nutr., 7, 104–116, 1997.
57. Ali, A., Duizer, L., Foster, K., Grigor, J., and Wei, W., Changes in sensory perception of sport drinks
when consumed pre, during, and post exercise. Physiol. Behav., 102, 437–443, 2011.
58. Galloway, S.D., Lott, M.J., and Toulouse, L.C., Pre-exercise carbohydrate feeding and high-intensity
exercise capacity: Effects of timing of intake and carbohydrate concentration. Int. J. Sport Nutr. Exerc.
Metab., 24, 258–266, 2014.
59. Esmarck, B., Andersen, J.L., Olsen, S., Richter, E.A., Mizuno, M., and Kjaer, M., Timing of post-
exercise protein intake is important for muscle hypertrophy with resistance training in elderly humans.
J. Physiol., 535, 301–311, 2001.
60. Suzuki, H., Kajimoto, Y., and Kumagai, H., Improvement of the bitter taste of amino acids through
the transpeptidation reaction of bacterial gamma-glutamyltranspeptidase. J. Agric. Food Chem., 50,
313–318, 2002.
61. Schmitz, S.M., Hofheins, J.E., and Lemieux, R., Nine weeks of supplementation with a multi-nutrient
product augments gains in lean mass, strength, and muscular performance in resistance trained men.
J. Int. Soc. Sports Nutr., 16, 40, 2010.
62. Kaastra, B., Manders, R.J., Van Breda, E., Kies, A., Jeukendrup, A.E., Keizer, H.A., Kuipers, H., and
Van Loon, L.J., Effects of increasing insulin secretion on acute postexercise blood glucose disposal.
Med. Sci. Sports Exerc., 38, 268–275, 2006.
63. Romano-Ely, B.C., Todd, M.K., Saunders, M.J., and Laurent, T.S., Effect of an isocaloric carbohydrate-
protein-antioxidant drink on cycling performance. Med. Sci. Sports Exerc., 38, 1608–1616, 2006.
65
Peptide-Enriched Functional Beverages
Kenji Sato and Tamami Kiyono
CONTENTS
65.1 Introduction................................................................................................................................... 853
65.2 Peptides as Applied to Sports Drinks........................................................................................... 854
65.2.1 Beneficial Activity of Peptide for Athletes....................................................................... 854
65.2.2 Other Factors for Peptide-Enriched Sports Drinks.......................................................... 854
65.3 Peptide-Enriched Beverages for Other Applications.................................................................... 855
65.3.1 Peptide-Enriched Beverages for Foods for Specified Health Use.................................... 855
65.3.2 New Types of Peptide-Enriched Beverages..................................................................... 855
65.4 Fermented Beverages Containing Peptides.................................................................................. 855
65.4.1 Peptides in Sour Milk....................................................................................................... 855
65.4.2 Peptides in Fermented Alcoholic Beverages.................................................................... 856
65.5 Conclusion..................................................................................................................................... 858
References............................................................................................................................................... 858
65.1 Introduction
Proteins and peptides have conventionally been considered as a source of amino acids in nutrition sci-
ence [1]. Recent studies have, however, demonstrated that the ingestion of peptides exerts higher ben-
eficial effects than their parent proteins in sports nutrition areas [2–4]. Peptides also generally exhibit
higher solubility and stability than proteins, and, therefore, sports drinks containing peptides have been
developed for decades. In addition to conventional nutritional aspects, one pioneering group at Kyoto
University (Japan) has explored the specific biological functions of food-derived peptides beyond the
conventional nutritional value [5,6]. They have demonstrated that some peptides exert antihypertension
activity upon ingestion, which has triggered further investigation into these functional foods. In recent
years, knowledge about the numerous biological activities of peptides has been published [7,8]. Protein
hydrolysates containing active peptides have been introduced into many food products such as energy
bars, soups, and beverages, as well as tablets.
Peptides are also generated by fermentation processes. Some traditionally fermented liquid foods,
such as fermented milk, contain biologically active peptides, and some of these fermented milk–based
beverages are used to moderate mild hypertension [9–11]. In addition to fermented milk, some traditional
alcoholic beverages also contain bioactive peptides. Japanese rice wine known as Sake, for example,
has been shown to contain some unique bioactive peptides. The aim of this chapter is not to provide a
comprehensive review of peptide functions in beverages, which can be obtained from other sources [7,8],
rather, it aims to provide an introduction to beverages containing biologically active peptides available
on the market and discusses the recent trends.
853
65.2 Peptides as Applied to Sports Drinks

65.2.1 Beneficial Activity of Peptide for Athletes
Protein supplementation just after exercise or physical activity can moderate exercise-induced muscle
pain and help increase muscle mass [12–14]. Milk whey protein and soy protein isolate are frequently
used for this purpose, with milk whey protein preferred most often. These proteins are usually avail-
able in the form of powder and are consumed by mixing in water, which may yield a turbid suspen-
sion. Recent animal studies have demonstrated that protein hydrolysates prepared enzymatically exert
greater efficacy than the parent proteins [15–17]. These facts suggest that some specific peptides, rather
than amino acids generated from proteins, may be responsible for the increased activity. Some specific
peptides generated by enzymatic digestion might act on intracellular receptors, such as the mammalian
target of rapamycin (mTOR) to increase muscle protein synthesis [18]. In addition, ingestion of soy pro-
tein hydrolysate has been shown to suppress the expression of fatty acid synthetase and hydroxymethyl-
glutaryl-CoA reductase, which is the rate-limiting enzyme for cholesterol synthesis [19]. However, there
is limited data available on peptides that can be absorbed into the bloodstream and reach the target cells
in organs, such as in the muscle and liver, in large amounts. As such, the mechanism underlying the
enhanced effect observed with the peptide, as compared to the protein, remains unclear.
65.2.2 Other Factors for Peptide-Enriched Sports Drinks

Table 65.1 summarizes peptide-based beverages for athletes both in liquid and powder types in the
Japanese market. The powder type is consumed after mixing with water. In the case of ready-to-use type,
demand for transparency of the final product has been on the rise. Recently, some peptides have been
isolated from the sediment derived from the soy protein hydrolysate of a model beverage that was stored
chilled [20]. All peptides in the sediment had a relatively large molecular mass of 2700–3800 Da and
were rich in hydrophobic amino acids. Therefore, these peptides could be removed by ultrafiltration.
In fact, the filtrate, which can be obtained using a membrane filter (Amicon YM3 > 3000 Da) did not
form any sediment under the aforementioned conditions.
Peptides with hydrophobic amino acids residues on the amino and carboxyl terminals often present
bitter or odd taste, which interferes with the application of peptides into beverages. Removal of such
hydrophobic residues from the terminal with peptidase can improve the flavor. For this purpose, fungal
protease preparations with significant exopeptidase activity are used along with bacterial protease prepa-
rations that are rich endoproteinase.
TABLE 65.1
Sources and Contents of Peptides in Beverages Designed for Athletes in the Japanese Market
Source of Peptide Form Peptide Content Total Volume/Weight
Soy protein hydrolysate Liquid 2000 mg 500 mL
Soy protein hydrolysate Powder 10.5 g 21 g
Soy protein and its hydrolysate Powder 14.1 g 21 g
Casein hydrolysate Powder 63.8 g 100 g
Casein hydrolysate Powder 2g 4.3 g
Milk whey and egg protein hydrolysates Powder 3.8 g 6.3 g
Milk hydrolysate Powder 5g 8g
Milk whey and gluten hydrolysates Powder 19 g 20 g
Source: Adapted from suppliers.
Note: Powder types are consumed by mixing the same in water.
Peptide-Enriched Functional Beverages 855
TABLE 65.2
Source and Content of Active Peptides in Peptide-Enriched Beverages as Foods for Specified Health Uses in
Japan
Content of Volume
Health Claim Source of Peptide Active Peptide Active Peptide (mL) References
Foods related to Sardine meat hydrolysate VY 0.4 mg 100 [21]
blood pressure
Sesame protein hydrolysate LVY 0.16 mg 350 [22]
Royal jelly protein hydrolysate VY, IY, and IVY 0.84 mg as IY 100 [23]
Mushroom extract IY 10.8 μg 120 [24]
Foods related to TAG Porcine hemoglobin hydrolysate VVYP 6 mg 50 [25]
Abbreviations: TAG, triacylglycerols; VY, valyltyrosine; LVY, leucylvalyltyrosine; IY, isoleucyltyrosine; IVY, isoleucyl-
valyltyrosine; VVYP, valylvalyltyrosylproline.
65.3 Peptide-Enriched Beverages for Other Applications

65.3.1 Peptide-Enriched Beverages for Foods for Specified Health Use
In Japan, some peptide-enriched beverages are classified as Foods for Specified Health Use (FOSHU),
which have the approval of the Japanese government to make certain health claims, such as “foods
related to blood pressure” and “foods related to triacylglycerols (TAG),” which can be stated on the
product. As summarized in Table 65.2, naturally occurring peptides in mushroom and enzymatic hydro-
lysates of sardine meat, sesame protein, royal jelly protein, and porcine hemoglobin are used [21–25].
65.3.2 New Types of Peptide-Enriched Beverages

In addition to the FOSHU products, collagen peptide–enriched beverages have also become prevalent in
Japan. While these products have not been approved by the Japanese government to make certain health
claims, there are many cases suggesting that consumption of collagen peptide-based products results in
improvement of skin conditions and moderation of joint pain. Although the academia has questioned the
suggested beneficial effects of collagen peptides, consumption of collagen peptide-based products has
increased in Japan, China, and elsewhere. Collagen peptides were originally provided in the powder form
in Japan. Since then, collagen peptide-enriched beverages (30–100 mL containing 1–10 g collagen peptide)
have been developed. Furthermore, since collagen peptides do not present a bitter taste, high concentra-
tions of the same product can be incorporated into beverages. The total annual sale of collagen peptide in
Japan in 2013 was approximately 70 billion Japanese Yen (nearly US$700 million), with beverage products
accounting for 40% of total sales. Recent placebo-controlled and double-blind human studies have dem-
onstrated the beneficial effects of collagen peptides for skin conditions [26,27]. In addition, the presence
of food-derived collagen peptides, such as prolylhydroxyproline (Pro-Hyp) or hydroxyprolylglycine (Hyp-
Gly) in human blood [28,29], which can enhance the growth of fibroblasts [29,30], has been reported. These
facts support the efficacy of collagen peptide-based products. The commercial success of collagen peptide-
enriched beverages and the subsequent researches describing mechanisms underlying the efficacy of col-
lagen peptides has helped establish new field for functional foods, including cosmetic or beautifying foods.
65.4 Fermented Beverages Containing Peptides

65.4.1 Peptides in Sour Milk
Fermented beverages contain significant amounts of peptides. The most commonly consumed fermented
beverage is liquid yogurt, which is produced by fermentation with Lactobacillus delbrueckii ssp. bulgaricus
and Streptococcus thermophilus. However, yogurt in a narrow sense contains relatively low levels of
peptides. On the other hand, various types of sour milk produced by fermentation with L. helveticus or
TABLE 65.3
Structures of Some Peptides with Inhibitory Activity against the Angiotensin Converting Enzyme (ACE)
Identified in Fermented Milk
Microorganisms Used Peptide Sequence Parent Protein References
Lactobacillus helveticus and Val-Pro-Pro, Ileu-Pro-Pro β-Casein and [31,32]
Saccharomyces cerevisiae κ-casein
Lactobacillus helveticus Trp-Pro Whey proteins [33]
Lactobacillus delbrueckii subsp. bulgaricus Ser-Lys-Val-Tyr-Pro-Pro-Gly-Pro-Ile β-Casein [34]
Enterococcus faecalis Leu-His-Leu-Pro-Leu-Pro β-Casein [35]
Enterococcus faecalis contain higher contents of peptides with inhibitory activity against the angioten-
sin converting enzyme (ACE), a key enzyme that induces hypertension, than yogurt. It has been shown
that sour milk produced by fermentation with L. helveticus and E. faecalis can moderate hypertension
[9–11,31–35]. ACE inhibitory peptides have been isolated from these sour milks; the structures of some of
the ACE-inhibitory peptides from sour milks are summarized in Table 65.3. These peptides are assumed to
be responsible for hypotensive effects on subjects with mild hypertension. A sour milk beverage produced
by fermentation with L. helveticus has been approved as a FOSHU product, with claims of it being a “food
related to blood pressure.” The lactotripeptides, isoleucylprolylproline (Ile-Pro-Pro) and leucylprolylproline
(Leu-Pro-Pro), have been suggested to be responsible for the hypotensive activity of this product. However,
the bioavailability of the proposed ACE inhibitory peptide is very low (nM level in plasma) [36]. Therefore,
it is possible that peptides in the sour milk may exert hypotensive activity via another mechanism.
65.4.2 Peptides in Fermented Alcoholic Beverages

Fermentation is also used to produce alcoholic beverages. Alcohol is produced from sugars by yeast
(Saccharomyces cerevisiae). Other microorganisms, such as lactic acid bacteria and fungi, are also used
depending on the type of alcoholic beverage. Fungi, especially Aspergillus oryzae, secrete proteases into
the extracellular medium, whereas yeast and lactic acid bacteria do not. Fungi are also used for saccharifica-
tion of starch from grain. Japanese rice wine, Sake, is one of the typical alcoholic beverages produced using
fungi (A. oryzae) for the saccharification process. ACE inhibitory peptides, histidyltyrosine (His-Tyr), valyl-
tyrosine (Val-Tyr), and tyrosylglycylglycyltyrosine (Tyr-Gly-Gly-Tyr) have been identified from Sake [37].
In addition to these peptides, Sake contains significant amounts of the N-blocked peptide (Figure 65.1).
25
Amino acid/peptide content (mM)
20
15
10
0
N-blocked Peptide Free amino acid
peptide
FIGURE 65.1 N-blocked peptide, peptide, and free amino acid content in two types of Japanese rice wine, Sake. The
N-blocked peptide was evaluated by amino acid analysis of HCl hydrolysate of the nonabsorbed fraction from a column
packed with a strong cation exchanger. (Unpublished data.)
O H2N CONHCHR1CONHCHR2CO........
H2N Peptide with N-terminal Gln
O N CONHCHR1CONHCHR2CO........
Pyroglutamyl peptide
FIGURE 65.2 Formation of pyroglutamyl peptide from peptides with amino terminal glutaminyl residue.
TABLE 65.4
Structure of Pyroglutamyl Peptides Identified in
Japanese Rice Wine (Sake)
Peptide Sequence
pyroGlu-Asn-Ile-Asp-Asn-Pro
pyropGlu-Asn-Ile
pyroGlu-Val
pyroGlu-Leu-Trp
pyroGlu-Val-Ala
pyroGlu-Val-Pro
pyroGlu-Val-Val
pyroGlu-Asn-Phe
pyroGlu-Leu
pyroGlu-Gln
pyroGlu-Ser-Gln
pyroGlu-Met
pyroGlu-Gly-Gln
pyroGlu-Tyr
pyroGlu-Phe
pyroGlu-Asn
pyroGlu-Ser
pyroGlu-Gly
pyroGly-Ala
Source: Adapted from Kiyono, T. et al., J. Agric. Food Chem.,
61, 11660, 2013. With permission.
Most of these peptides are pyroglutamyl peptides. A pyroglutamyl peptide is generated from peptides with
glutaminyl residues at their N-termini as shown in Figure 65.2. This reaction is irreversible and proceeds
even at low temperatures and is accelerated by heating. Recently, pyroglutamyl peptides in Sake have been
identified; the structure of these peptides is summarized in Table 65.4 [38]. Of these peptides, pyroglutamyl-
leucine (pyroGlu-Leu) is the major constituent and is present in Sake at approximately 40–60 μM, depending
on the final product [38]. It has been shown that oral administration of pyroGlu-Leu moderates galactos-
amine-induced hepatitis in rats [39] and dextran sulfate sodium (DSS)-induced colitis in mice [40]. These
effects exerted by pyroGlu-Leu were due to administration of relatively low doses of 20 and 0.1 mg/kg body
weight for hepatitis and colitis, respectively. The colonic microbiota of mice was changed due to colitis,
with a decrease in Bacteroidetes and increase in Firmicutes. Surprisingly, the administration of pyroGlu-
Leu at 0.1 mg/kg normalized the colonic microbiota in mice with DSS-induced colitis [40]. PyroGlu-Leu is
produced by proteolytic degradation of rice protein and not by synthesis from Leu. However, S. cerevisiae
is not directly involved in the production of pyroGlu-Leu [38]. Therefore, pyroGlu-Leu can be produced by
fermentation of steamed rice in acidified water. Nonalcoholic, sweet beverages containing approximately
100 μM of pyroGlu-Leu can be prepared using this procedure. Furthermore, it has been reported that abnor-
mal colonic microbiota is closely related to chronic diseases characterized by chronic inflammation [41].
Therefore, sweet beverages rich in pyroglutamyl peptides have the potential to serve as novel prebiotics,
which can moderate inflammation by normalizing colonic microbiota. However, the available data on the
effects of pyroglutamyl peptides on microbiota is very limited. Further studies on the structure and efficacy
of pyroglutamyl peptide in fermented beverages and better understanding of the underlying mechanism are
necessary.
65.5 Conclusion
Peptide-enriched functional beverages have been used for sports drinks and specific health applications.
In such cases, controlling the cost, taste, flavor, and appearance is critical for commercial success. In
addition to these applications, the use of collagen peptide-enriched beverages for improving skin condi-
tions has been growing and has helped established a new field of cosmetic or beautifying food prod-
ucts. Further studies for elucidating the mechanism underlying the improvement of skin conditions by
administration of collagen peptide are warranted. Some biologically active peptides have been identified
in fermented foods and beverages. Recently, the presence of modified peptides, such as pyroglutamyl
peptides in Japanese rice wine Sake, has been reported. Some of the pyroglutamyl peptides in Sake exert
anti-inflammatory activity at relatively low doses. Nonalcoholic beverages containing these pyroglu-
tamyl peptides can be prepared by fermenting steamed rice with fungi. A new type of sweet beverage
with anti-inflammatory activity may, therefore, be developed in the near future using this method.
REFERENCES
1. Grimble, G.K., The significance of peptides in clinical nutrition. Annu. Rev. Nutr., 14, 419–447, 1994.
2. Maebuchi, M., Samoto, M., Kohno, M., Ito, R., Koikeda, T., Hirotsuka, M., and Nakabou, Y., Improvement
in the intestinal absorption of soy protein by enzymatic digestion to oligopeptide in healthy adult men.
Food Sci. Technol. Res., 13, 45–53, 2007.
3. Boza, J.J., Moenno, D., Vuichoud, J., Gaudard-de-Weck, A.R.D., and Ballevre, O., Protein hydrolysate vs
free amino acid-based diets on the nutritional recovery of the starved rat. Eur. J. Nutr., 39, 237–243, 2000.
4. Boza, J., Martinez, O., Baro, L., Suarez, M.D., and Gil, A., Influence of casein and casein hydrolysate
diets on nutritional recovery of starved rats. J. Parenter. Enteral Nutr., 19, 216–221, 1995.
5. Yokoyama, K., Chiba, H., and Yoshikawa, M., Peptide inhibitors for angiotensin I-converting enzyme
from thermolysin digest of dried bonito. Biosci. Biotechnol. Biochem., 56, 1541–1545, 1992.
6. Yoshikawa, M., Fujita, H., Matoba, N., Takenaka, Y., Yamamoto, T., Yamauchia, R., Tsuruki, H.,
and Takahata, K., Bioactive peptides derived from food proteins preventing lifestyle-related diseases.
BioFactors, 12, 143–146, 2000.
7. Mine, Y. and Shahidi, F., Nutraceutical proteins and peptides in health and disease: An overview, in
Nutraceutical Proteins and Peptides in Health and Disease, Mine, Y. and Shahidi, F., Eds., CRC Press/
Taylor & Francis Group, Boca Raton, FL, 2005, pp. 3–9.
8. Kannan, A., Hettiarachchy, N.S., and Marshall, M.R., Food proteins and peptides as bioactive agents, in
Bioactive Food Proteins and Peptides, Hettiarachchy, N.S., Sato, K., Marshall, M.R., and Kannan, A.,
Eds., CRC Press/Taylor & Francis Group, Boca Raton, FL, 2012, pp. 1–28.
9. Hata, Y., Yamamoto, M., Ohni, M., Nakajima, K., Nakamura, Y., and Takano, T., A placebo-controlled study
of the effect of sour milk on blood pressure in hypertensive subjects. Am. J. Clin. Nutr., 64, 767–771, 1996.
10. Takano, T., Anti-hypertensive activity of fermented dairy products containing biogenic peptides.
Antonie van Leeuwenhoek, 82, 333–340, 2002.
11. Seppo, L., Jauhiainen, T., Poussa, T., and Korpela, R., A fermented milk high in bioactive peptides has
a blood pressure-lowering effect in hypertensive subjects. Am. J. Clin. Nutr., 77, 326–330, 2003.
12. Rasmussen, B.B., Tipton, K.D., Miller, S.L., Wolf, S.E., and Wolfe, R.R., An oral essential amino acid-
carbohydrate supplement enhances muscle protein anabolism after resistance exercise. J. Appl. Physiol.,
88, 386–392, 2000.
13. Hartman, J.W., Tang, J.E., Wilkinson, S.B., Tarnopolsky, M.A., Lawrence, R.L., Fullerton, A.V., and
Phillips, S.M., Consumption of fat-free fluid milk after resistance exercise promotes greater lean mass
accretion than does consumption of soy or carbohydrate in young, novice, male weightlifters. Am. J.
Clin. Nutr., 86, 373–381, 2007.
14. Flakoll, P.J., Judy, T., Flinn, K., Carr, C., and Flinn, S., Postexercise protein supplementation improves
health and muscle soreness during basic military training in marine recruits. J. Appl. Physiol., 96,
951–956, 2004.
15. Manninen, A.H., Protein hydrolysates in sports nutrition. Nutr. Metab., 6, 38, 2009.
16. Buckley, J.D., Thomson, R.L., Coates, A.M., Howe, P.R.C., DeNichilo, M.O., and Rowney, M.K.,
Supplementation with a whey protein hydrolysate enhances recovery of muscle force-generating capac-
ity following eccentric exercise. J. Sci. Med. Sport, 13, 178–181, 2010.
17. Morifuji, M., Kanda, A., Koga, J., Kawanaka, K., and Higuchi, M., Post-exercise carbohydrate plus
whey protein hydrolysates supplementation increases skeletal muscle glycogen level in rats. Amino
Acids, 38, 1109–1115, 2010.
18. Kanda, A., Nakayama, K., Fukasawa, T., Koga, J., Kanegae, M., Kawanaka, K., and Higuchi, M., Post-
exercise whey protein hydrolysate supplementation induces a greater increase in muscle protein synthe-
sis than its constituent amino acid content. Br. J. Nutr., 110, 981–987, 2013.
19. Inoue, N., Nagao, K., Sakata, K., Yamano, N., Gunawardena, P.E.R., Han, S.-Y., Matsui, T. et al.,
Screening of soy protein-derived hypotriglyceridemic di-peptides in vitro and in vivo. Lipids Health
Dis., 10, 85, 2011.
20. Nakamori, T., Nagai, M., Maebuchi, M., Furuta, H., Park, E.-Y., Nakamura, Y., and Sato, K., Identification
of peptides in sediments derived from acidic enzymatic soy protein hydrolysate solution. Food Sci.
Tech. Res., 20, 301–307, 2014.
21. Matsufuji, H., Matsui, T., Seki, E., Osajima, K., Nakashima, M., and Osajima, Y., Angiotensin
I-converting enzyme inhibitory peptides in an alkaline protease hydrolyzate derived from sardine mus-
cle. Biosci. Biotechnol. Biochem., 58, 2244–2245, 1994.
22. Nakano, D., Ogura, K., Miyakoshi, M., Ishii, F., Kawanishi, H., Kurumazuka, D., Kwak, C. et al.,
Antihypertensive effect of angiotensin I-converting enzyme inhibitory peptides from a sesame protein
hydrolysate in spontaneously hypertensive rats. Biosci. Biotechnol. Biochem., 70, 1118–1126, 2006.
23. Tokunaga, K., Yoshida, C., Suzuki, K., Maruyama, H., Futamura, Y., Araki, Y., and Mishima, S.,
Antihypertensive effect of peptides from royal jelly in spontaneously hypertensive rats. Biol. Pharm.
Bull., 27, 189–192, 2004.
24. Mashiko, K., Hiratsuka, H., Samejima, K., and Sato, T., Antihypertensive effect of an aqueous extract
from Mycoveptodonoides aitchisonii in high-normal and mild hypertensive (low-medium risk) group.
Jpn. Pharmacol. Ther., 31, 239–246, 2003.
25. Kagawa, K., Matsutaka, H., Fukuhama, C., Fujino, H., and Okuda, H., Suppressive effect of globin
digest on postprandial hyperlipidemia in male volunteers. J. Nutr., 128, 56–60, 1998.
26. Proksch, E., Segger, D., Degwert, J., Schunck, M., Zague, V., and Oesser, S., Oral supplementation of
specific collagen peptides has beneficial effects on human skin physiology: A double-blind, placebo-
controlled study. Skin Pharmacol. Physiol., 27, 47–55, 2014.
27. Proksch, E., Schunck, M., Zague, V., Segger, D., Degwert, J., and Oesser, S., Oral intake of specific bio-
active collagen peptides reduces skin wrinkles and increases dermal matrix synthesis. Skin Pharmacol.
Physiol., 27, 113–119, 2014.
28. Iwai, K., Hasegawa, T., Taguchi, Y., Morimatsu, F., Sato, K., Nakamura, Y., Higashi, A., Kido, Y.,
Nakabo, Y., and Ohtsuki, K., Identification of food-derived collagen peptides in human blood after oral
ingestion of gelatin hydrolysates. J. Agric. Food Chem., 53, 6531–6536, 2005.
29. Shigemura, Y., Akaba, S., Kawashima, E., Park, E.-Y. Nakamura, Y., and Sato, K., Identification of a
novel food-derived collagen peptide, hydroxyprolyl-glycine, in human peripheral blood by pre-column
derivatisation with phenyl isothiocyanate. Food Chem., 129, 1019–1024, 2011.
30. Shigemura, Y., Iwai, K., Morimatsu, F., Iwamoto, T., Mori, T., Oda, C., Taira, T., Park, E.-Y.,
Nakamura, Y., and Sato, K., Effect of prolyl-hydroxyproline (Pro-Hyp), a food-derived collagen peptide
in human blood, on growth of fibroblasts from mouse skin. J. Agric. Food Chem., 57, 444–449, 2009.
31. Nakamura, Y., Yamamoto, N., Sakai, K., Okubo, A., Yamazaki, S., and Takano, T., Purification and
characterization of angiotensin I-converting enzyme inhibitors from sour milk. J. Dairy Sci., 78,
777–783, 1995.
32. Nakamura, Y., Yamamoto, N., Sakai, K., and Takano, T., Antihypertensive effect of sour milk and
peptides isolated from it that are inhibitors to angiotensin I-converting enzyme. J. Dairy Sci., 78,
1253–1257, 1995.
33. Yamamoto, N., Maeno, M., and Takano, T., Purification and characterization of an antihypertensive
peptide from a yogurt-like product fermented by Lactobacillus helveticus CPN4. J. Dairy Sci., 82,
1388–1393, 1999.
34. Ashar, M.N. and Chand, R., Fermented milk containing ACE-inhibitory peptides reduces blood pres-
sure in middle aged hypertensive subjects. Milchwissenschaft, 59, 363–366, 2004.
35. Miguel, M., Recio, I., Ramos, M., Delgado, M.A., and Aleixandre, M.A., Antihypertensive effect of
peptides obtained from Enterococcus faecalis-fermented milk in rats. J. Dairy Sci., 89, 3352–3359,
2006.
36. Foltz, M., Meynen, E.M., Bianco, V., van Platerink, C., Koning, T.M., and Kloek, J., Angiotensin con-
verting enzyme inhibitory peptides from a lactotripeptide-enriched milk beverage are absorbed intact
into the circulation. J. Nutr., 137, 953–958, 2007.
37. Saito, Y., Wanezaki, K., Kawato, A., and Imayasu, S., Structure and activity of angiotensin I converting
enzyme inhibitory peptides from Sake and Sake lees. Biosci. Biotechnol. Biochem., 58, 1767–1771, 1994.
38. Kiyono, T., Hirooka, K., Yamamoto, Y., Kuniishi, S., Ohtsuka, M., Kimura, S., Park, E.-Y., Nakamura, Y.,
and Sato, K., Identification of pyroglutamyl peptides in Japanese rice wine, Sake: Presence of hepato-
protective pyroGlu-Leu. J. Agric. Food Chem., 61, 11660–11667, 2013.
39. Sato, K., Egashira, Y., Ono, S., Mochizuki, S., Shimmura, Y., Suzuki, Y., Nagata, M. et al., Identification
of a hepatoprotective peptide in wheat gluten hydrolysate against d−galactosamine-induced acute hepa-
titis in rats. J. Agric. Food Chem., 61, 6304–6310, 2013.
40. Wada, S., Sato, K., Ohta, R., Wada, E., Bou, Y., Fujiwara, M., Kiyono, T., Park, E.-Y., Aoi, W., Takagi, T.,
Naito, Y., and Yoshikawa, T., Ingestion of low dose pyroglutamyl leucine improves dextran sulfate
sodium-induced colitis and intestinal microbiota in mice. J. Agric. Food Chem., 61, 8807–8813, 2013.
41. Ott, S.J., Musfeldt, M., Wenderoth, D.F., Hampe, J., Brant, O., Folsch, U.R., Timmis, K.N., and
Schreiber, S., Reduction in diversity of the colonic mucosa associated bacterial microflora in patients
with active inflammatory bowel disease. Gut, 53, 685–693, 2004.
Food Science and Technology
Handbook of Functional
Nutraceutical
11
Science
and
Technology
Nutraceutical Science
and Technology
Series Editor: Fereidoon Shahidi
11
Beverages and Human Health
Handbook of
Handbook of Functional Beverages

Handbook of Functional Beverages and Human Health provides potential appli-
cations and new developments in functional beverages, nutraceuticals, and health
foods. In addition to serving as a reference manual, it summarizes the current state
of knowledge in key research areas and contains novel ideas for future research and Functional
Beverages and
development. Additionally, it provides an easy-to-read text suitable for teaching senior
undergraduate and postgraduate students in the relevant areas.
and Human Health

Human Health
The book is divided into seven major sections: Section I covers market trends,
global regulations, flavor challenges, chemistry, and health with specific reference
to cancer chemoprevention and the prevention of postprandial metabolic stress due
to the consumption of functional beverages. Section II, by far the largest part of
the book, has 39 chapters on the most popular fruit juices (apple juice, lemon juice,
pomegranate juice, watermelon juice, etc.). Section III reports on herbal and vegetable
juices (carrot juice, Chinese medicinal herbs and root-based beverages, tomato juice,
and vegetable-containing juices).
Edited by
Section IV details caffeinated beverages, including different varieties of tea (green,
black, oolong, and herbal teas), coffee (coffee and beverages from green coffee beans), Fereidoon Shahidi
and cocoa and chocolate. Section V is on dairy and soy beverages, while Section VI is
on alcoholic beverages (wine) and water (maple water). Finally, Section VII describes
fermented (kefir, koumiss, and ayran) and fortified functional beverages (applications
of plant sterols and stanols in functional beverages, beverages fortified with omega-3
fatty acids, dietary fiber, minerals and vitamins, probiotics and prebiotics in functional
beverages, functional beverages in weight management, fortified sports drinks, and
peptide-enriched functional beverages).
Shahidi
Alasalvar
K20775

Handbook of PDF

Uploaded by

Copyright:

Available Formats

You might also like

Handbook of PDF

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Handbook of PDF

Uploaded by

Copyright:

Available Formats

Food Science and Technology

Handbook of Functional Beverages

and Human Health

1. Phytosterols as Functional Food Components and Nutraceuticals

Boca Raton London New York

CRC Press is an imprint of the

No claim to original U.S. Government works

International Standard Book Number-13: 978-1-4665-9642-9 (eBook - PDF)

Section I Market Trends, Regulations, Chemistry, and Health Aspects

2. Global Nutraceutical Regulations for Functional Beverages.......................................................17

3. Flavor Challenges in Functional Beverages.................................................................................. 27

4. Chemistry of Functional Beverages.............................................................................................. 35

5. Cancer Chemopreventive Effects of Selected Fruit Juices......................................................... 47

6. Fruit Juices and the Prevention of Postprandial Metabolic Stress in Humans........................ 69

Section II Fruit Juices

9. Apricot Juice/Nectar..................................................................................................................... 107

10. Aronia Juice....................................................................................................................................119

11. Blackberry Juice.............................................................................................................................135

12. Black Currant Juice.......................................................................................................................147

13. Blueberry Juice...............................................................................................................................163

14. Cherry Juice...................................................................................................................................175

15. Cherry Laurel Syrup (Pekmez)....................................................................................................187

16. Coconut Juice................................................................................................................................. 193

17. Cranberry Juice............................................................................................................................ 205

18. Date Syrup......................................................................................................................................217

19. Dragon Fruit Juice.........................................................................................................................231

20. Goji Berry Juice............................................................................................................................ 239

22. Grape Juice.................................................................................................................................... 271

23. Grapefruit Juice............................................................................................................................ 281

24. Guava Juice.................................................................................................................................... 297

25. Hawthorn Juice..............................................................................................................................311

26. Indian Gooseberry (Amla) Juice..................................................................................................321

27. Kiwifruit Juice................................................................................................................................331

28. Lemon Juice................................................................................................................................... 339

29. Lime Juice...................................................................................................................................... 349

30. Mango Juice................................................................................................................................... 359

31. Mangosteen Juice.......................................................................................................................... 373

32. Melon Juice.................................................................................................................................... 385

33. Mulberry Juice.............................................................................................................................. 399

34. Noni Fruit Juice............................................................................................................................. 409

35. Orange Juice.................................................................................................................................. 423

36. Papaya Juice................................................................................................................................... 439

37. Passion Fruit Juice.........................................................................................................................455

38. Peach Juice..................................................................................................................................... 463

39. Pear Juice........................................................................................................................................475

40. Pineapple Juice.............................................................................................................................. 489

41. Plum, Prune, and Ume Juices...................................................................................................... 501

42. Pomegranate Juice.........................................................................................................................513

43. Raspberry Juice............................................................................................................................. 527

44. Strawberry Juice............................................................................................................................541

45. Watermelon Juice...........................................................................................................................553

Section III Herbal and Vegetable Juices

47. Chinese Medicinal Herbs and Root-Based Beverages............................................................... 583

48. Tomato Juice.................................................................................................................................. 593

49. Vegetable-Containing Juices (Carrot, Kale, and Sprout)......................................................... 609

Section IV Caffeinated Beverages: Tea, Coffee, and Cocoa/Chocolate

51. Herbal Teas.................................................................................................................................... 645

53. Beverages from Green Coffee Beans........................................................................................... 677

1.3 Consumer-Oriented New Product Development

2.5 Regulatory Requirements for Ingredients in Beverages and