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Shabnum Shaheen · Sehrish Ramzan
Farah Khan · Mushtaq Ahmad

Adulteration
in Herbal
Drugs: A
Burning Issue
Adulteration in Herbal Drugs: A Burning Issue
Shabnum Shaheen • Sehrish Ramzan
Farah Khan • Mushtaq Ahmad

Adulteration in Herbal
Drugs: A Burning Issue
Shabnum Shaheen Sehrish Ramzan
Department of Plant Sciences Lahore College for Women University
Lahore College for Women University Lahore, Pakistan
Lahore, Pakistan
Mushtaq Ahmad
Farah Khan Quaid-i-Azam University
Lahore College for Women University Islamabad, Pakistan
Lahore, Pakistan

ISBN 978-3-030-28033-8    ISBN 978-3-030-28034-5 (eBook)


https://doi.org/10.1007/978-3-030-28034-5

© Springer Nature Switzerland AG 2019


This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of
the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation,
broadcasting, reproduction on microfilms or in any other physical way, and transmission or information
storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology
now known or hereafter developed.
The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication
does not imply, even in the absence of a specific statement, that such names are exempt from the relevant
protective laws and regulations and therefore free for general use.
The publisher, the authors, and the editors are safe to assume that the advice and information in this book
are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the
editors give a warranty, express or implied, with respect to the material contained herein or for any errors
or omissions that may have been made. The publisher remains neutral with regard to jurisdictional claims
in published maps and institutional affiliations.

This Springer imprint is published by the registered company Springer Nature Switzerland AG
The registered company address is: Gewerbestrasse 11, 6330 Cham, Switzerland
Dedicated
to
My beloved parents

Every journey begins with a single step


You made me to learn to take this step
I promise to make the journey completed for you
Foreword

Adulteration is a legal term meaning that a food or herbal product fails to meet the
legal standards. Adulteration is basically found in two major fundamentals of life,
i.e., in food items and in herbal drugs. In recent years, adulteration has become an
increasing problem globally in the food as well as in herbal industry. Since food
adulteration is a huge concern in all parts of the globe, there is voluminous literature
on various aspects of food adulteration including its detection. In fact, methodologi-
cal procedures of food adulteration detection have been reviewed extensively.
Adulteration and fraudulent labeling of a foodstuff’s origin costs the global food
industry an estimated $10 to $15 billion per year. Adulteration usually refers to mix-
ing other matter of an inferior and sometimes harmful quality with food or drink
intended to be sold. As a result of adulteration, food or drink becomes impure and
unfit for human consumption.
Adulteration in herbal drugs means “a practice of substituting original crude
drug partially or whole with other similar looking substances but the latter is either
free from or inferior in chemical and therapeutic properties.” The adulteration and
substitution of the herbal drugs is the burning problem in herbal industry, and it
has caused a major threat in the research on commercial natural products. The
deforestation and extinction of many species and incorrect identification of many
plants have resulted in adulteration and substitution of raw drugs. So the present
book provides a detailed textual as well as pictorial data of adulteration particu-
larly with the reference of herbal drugs which is now a burning issue in the world
as most of the world rely on herbal drugs. As per data available, over three-quar-
ters of the world population relies mainly on plants and plant extracts for their
healthcare needs.

Lahore, Pakistan Shabnum Shaheen

vii
Preface

Herbal plants can be defined as the plants which are used to treat or prevent a
specific disease in human beings. For millions of years, human population is using
herbal plants for multiple health issues. It has been widely admitted that human
body is just like a machine which needs maintenance from time to time; and a
diseased body is like a broken-down machine which needs to be fixed. Man has
been using herbal plants for treating the various diseases from thousands of years.
According to the WHO (World Health Organization), most of the human beings
depend upon the traditional mode of treatment for their physical health. Rural
areas of various developed and under developing countries still depend upon the
herbal drugs in order to resolve their health issues because these traditional medi-
cines are comparatively cheaper and safer than the modern formulated medicines.
People native to the areas where such herbal plants grow have come to know by
their personal experience that these traditional therapies are very useful to main-
tain the health of both man and animals. However, mostly, they remain unaware
of the effects of these medicines in the human body. Knowledge about herbal
plants provides new approaches for the improvement of modern drugs. Various
causes like no side-effect treatment, easy accessibility, and low-cost were the rea-
sons behind their popularity. About 25% of drugs are originated either directly or
indirectly from the plants. Moreover, they are also a source of raw materials for
pharmaceutical companies and represent a large proportion in worldwide drug
market. Inhabitants of rural areas also depend on the plant-based remedy in all the
developing countries. Man used plants as well as animals in order to complete his
needs. Nowadays, millions of people, for their survival, depend on the flora and/
or fauna products even in the developed countries.
From all over the world, more than 422,000 flowering plant species exist, from
which almost 5000 species are used for medicinal purposes. Statistics say that the
value of herbal drug trade has increased annually about 12–15%. Above 80% of the
world population rely on these traditional plants for their various diseases. Herbal
products are traded globally and are major sources of global economy, and its
demand has been increasing both in developing and in developed countries. Through
the literature review, it becomes evident that above 1000 companies are now engaged

ix
x Preface

in synthesizing the herbal products and their annual revenue may exceed US$60
billion. Statistics indicate the rapid growth in herbal markets, approximately at 15%
per annum, and the number of consumers who are interested in herbal medicines
from various countries is increasing. Pakistan is a rich country in flora because of
appropriate soil condition, diverse climate, and many ecological regions. Traditional
medication system is dominant in Pakistan. Around 50,000 Hakims use herbal
plants and run their own clinics in urban and rural areas all over Pakistan. People
prefer traditional medication due to the several reasons like absence of allopathic
doctors and fear of harmful effects of modern medicine. Pakistan has a vast history
of research on herbal plants. The local communities of Pakistan have traditional
knowledge and practices of most plants belonging to the region. The traditional
knowledge about plants has been transferred from one generation to another by oral
demonstration or personal experience. The local inhabitants use native herbal plants
for their primary healthcare.
The term adulteration means mixing or replacement of original herbal drugs with
its resembled, less inferior plant which has different chemical or therapeutic proper-
ties. Substitution or adulteration in traded herbal raw material is a common practice.
Adulteration or substitution in herbal medicine is a burning issue of the herbal
industry. The reasons behind this fact are extinction of required species, deforesta-
tion, or maybe incorrect taxonomical identification. Adulteration may be accidental
or intentional. Faith in herbal method of treatment has lessened down due to adul-
teration. Even adulteration in herbal samples is major drawback behind their pro-
motion. According to the Adverse Event Report, adulteration is not only due to the
planned herb but also due to the unplanned herbs. Herbal dealers or herbalists have
adopted some methods to create such high-quality adulteration which cannot be
determined without performing microscopic examination or chemical analysis. One
of the weaknesses behind the acceptance of herbal product formation is the lack of
quality control and standardization. It is quite difficult to establish the specific qual-
ity control standards due to the complex nature and innate unpredictability of chem-
ical constituents of herbal drugs. Adulteration of marketed herbal samples remains
the severe issue in domestic as well as in export markets due to the correct botanical
identification and confused nomenclature of traded herbal drugs. The morphologi-
cal and microscopic investigation and chemical profiling and DNA barcoding of
original as well as marketed samples are the multiple parameters to measure and
prevent adulteration. So it is a need of time to develop some sensitive, more effec-
tive, classical, as well as modern approaches for the authentication of herbal drugs
as adulteration is one of the weaknesses behind the acceptance of herbal product
formation. So our project is basically based upon the awareness of adulteration not
only to the scientific community but also to the local users of the herbal drugs not
only in Pakistan but also throughout the world where these herbal drugs are used as
medicines, and our research project is also based on the parameters by which we can
prevent this issue by using classical as well as modern scientific tools. The present
book is based on the ethnomedicinal surveys of many herbal markets of Pakistan,
collection of various samples from many herbal shops used for many ailments, and
then comparison of these herbal samples by the original herbal plants collected
Preface xi

from the field. Various techniques like taxonomic, phytochemical, organoleptic, and
DNA barcoding are used to determine adulterants in the marketed herbal samples.
We have many original case studies regarding this as this adulteration practice is
very common in many countries particularly in developing countries, which can
cause many harmful effects to human’s health and humanity also. So the present
book data is a significant contribution particularly in the herbal trade markets as this
highlights those particular herbal plants which are facing adulteration problem and
also estimates the extent of adulteration and substitution of the commonly used
herbal drugs in local as well as scientific community.

Lahore, Pakistan  Shabnum Shaheen


Acknowledgments

First of all, I bow my head before almighty Allah with humble thanks who has
enabled me to do this hard job. My all loves for Hazrat Muhammad (PBUH), a
torch of guidance for all mankind forever.
I deem it a real privilege and source of pleasure to express my profound and
cordial attitude to my mentor “Prof. Dr. Mir Ajab Khan” for his keen interest, sug-
gestions, guidance, encouragement, and support throughout the period of this
research work and book writing. I would like to thank him for his great understand-
ing and support during this whole period.
I offer my great regards to my coauthors for their sincere cooperation, hard work,
and throughout support. Special thanks to my parents, husband, and children for
their moral support and encouragement.
My appreciations are for my lab students and lab attendants for their hard work
and support.
Dr. Shabnum Shaheen

xiii
Contents

1 Adulteration A Global Issue��������������������������������������������������������������������    1


1.1 Adulteration A Global Issue��������������������������������������������������������������    1
1.1.1 Adulteration a Legal Term����������������������������������������������������    1
1.1.2 Federal Food, Drug and Cosmetics Act��������������������������������    3
1.2 Poisonous or Deleterious Substances������������������������������������������������    4
1.3 Filth and Foreign Matter of Adulteration������������������������������������������    4
1.4 Microbiological Contamination and Adulteration of Food��������������    6
1.5 Adulteration in Herbal Drugs������������������������������������������������������������    6
2 Types and Causes of Adulteration: Global Perspectives����������������������    9
2.1 Types of Adulteration and Food Items����������������������������������������������    9
2.1.1 Intentional/Deliberate Adulteration��������������������������������������    9
2.1.2 Incidental/Unknown Adulteration����������������������������������������   10
2.1.3 Metallic Contamination��������������������������������������������������������   10
2.2 Food Items and Adulterants��������������������������������������������������������������   10
2.3 Food Adulteration Categories ����������������������������������������������������������   12
2.3.1 Replacement��������������������������������������������������������������������������   12
2.3.2 Addition��������������������������������������������������������������������������������   13
2.3.3 Removal��������������������������������������������������������������������������������   13
2.4 Other Types of Food Adulteration����������������������������������������������������   13
2.5 Other Types of Common Adulteration����������������������������������������������   13
2.6 Reason for Food Adulteration����������������������������������������������������������   13
2.7 Precautions Against Food Adulteration��������������������������������������������   14
2.8 Law against Food Adulteration ��������������������������������������������������������   15
3 Why Study Herbal Plants?����������������������������������������������������������������������   17
3.1 Herbal Plants������������������������������������������������������������������������������������   17
3.2 Need to Study Herbal Plants ������������������������������������������������������������   19
3.3 Importance of Medicinal Plants in Healthcare System��������������������   21
3.4 Importance of Some Herbs with their Medicinal Values������������������   22
3.5 Adulteration in Medicinal Plants������������������������������������������������������   27
3.6 Reasons of Adulteration��������������������������������������������������������������������   28

xv
xvi Contents

3.6.1 Adulteration Caused due to the Similar Morphology ����������   28


3.6.2 Adulteration Caused due to Confusion in Vernacular Names������
  29
3.6.3 Lack of Elementary Knowledge about the Authentic Plant
Source ����������������������������������������������������������������������������������   30
3.7 Significance of Taxonomic Approaches in Resolving Adulteration Issue
of Medicinal Plants ��������������������������������������������������������������������������   30
3.7.1 Morphological Analysis��������������������������������������������������������   30
3.7.2 Microscopic Analysis������������������������������������������������������������   31
3.7.3 Organoleptic Analysis����������������������������������������������������������   31
3.8 Molecular Approaches in Medicinal Plants Identification����������������   31
4 History, Classification, Worldwide Distribution and Significance of
Herbal Plants��������������������������������������������������������������������������������������������   35
4.1 The History of Medicinal Plants ������������������������������������������������������   35
4.1.1 The Oral Tradition����������������������������������������������������������������   35
4.1.2 The First Written Texts ��������������������������������������������������������   36
4.1.3 Middle Ages��������������������������������������������������������������������������   37
4.1.4 The Contribution of the New World ������������������������������������   37
4.2 A Universal Legacy That Must Be Preserved ����������������������������������   39
4.3 Classification of Herbal Plants����������������������������������������������������������   39
4.3.1 On Basis of Their Growth Habit and Parts Utilized ������������   40
4.3.2 According to the Usage��������������������������������������������������������   41
4.3.3 According to Their Active Constituents��������������������������������   42
4.4 Distribution of Medicinal Plants by Families ����������������������������������   43
4.4.1 Medicinal Plants of the Compositae Family������������������������   43
4.4.2 Medicinal Plants of the Labiatae Family������������������������������   44
4.4.3 Medicinal Plants of the Umbelliferae Family����������������������   44
4.4.4 Medicinal Plants of the Leguminosae Family����������������������   45
4.4.5 Medicinal Plants of the Rosaceae Family����������������������������   45
4.4.6 Medicinal Plants of the Rutaceae������������������������������������������   45
4.4.7 Medicinal Plants of the Solanaceae Family��������������������������   45
4.4.8 Medicinal Plants of the Cruciferae Family ��������������������������   46
4.4.9 Medicinal Plants of the Liliaceae Family ����������������������������   46
4.4.10 Medicinal Plants of the Caryophyllaceae Family ����������������   47
4.4.11 Medicinal Plants of the Boraginaceae Family����������������������   47
4.4.12 Medicinal Plants of the Ranunculaceae Family��������������������   47
4.4.13 Medicinal Plants of the Papaveraceae Family����������������������   48
4.4.14 Medicinal Plants of the Malvaceae Family��������������������������   48
4.4.15 Medicinal Plants of the Cucurbitaceae Family ��������������������   48
4.4.16 Medicinal Plants of the Verbenaceae Family������������������������   48
4.4.17 Medicinal Plants of the Scrophulariaceae Family����������������   48
4.4.18 Medicinal Plants of the Phytolaccaceae Family ������������������   49
4.4.19 Medicinal Plants of the Euphorbiaceae Family��������������������   49
Contents xvii

5 Marketed Herbal Drugs: How Adulteration Affects����������������������������   51


5.1 Difference of Herbal and Conventional Drugs & Trends in Herbal
Medicine Use������������������������������������������������������������������������������������   51
5.1.1 Use of Whole Plants��������������������������������������������������������������   51
5.1.2 Herb Combining�������������������������������������������������������������������   51
5.1.3 Diagnosis������������������������������������������������������������������������������   52
5.2 Trends in Herbal Medicine Use��������������������������������������������������������   52
5.3 Market Value of Herbal Medicines ��������������������������������������������������   52
5.4 Indian Herbal Trade in World Scenario��������������������������������������������   53
5.5 World Wide Herbal Trade ����������������������������������������������������������������   53
5.6 Market Potential of Phytomedicines������������������������������������������������   54
5.7 Global Herbal Market ����������������������������������������������������������������������   54
6 Preventive Measures to Stop Adulteration in Marketed
Herbal Samples����������������������������������������������������������������������������������������   57
6.1 Trends and Challenges Related to Herbal Plants������������������������������   57
6.2 Constraints in Developing Traditional Medicines from Plants��������   57
6.3 The Role of the Private Sector����������������������������������������������������������   60
6.4 A Coherent Research Agenda Will Need to Address������������������������   60
6.4.1 Health������������������������������������������������������������������������������������   60
6.4.2 Biodiversity��������������������������������������������������������������������������   61
6.4.3 Social Dimension������������������������������������������������������������������   61
6.5 A Coherent National Programme on Medicinal Plants Should Include
the Following Institutions ����������������������������������������������������������������   61
6.5.1 Ministry of Health����������������������������������������������������������������   61
6.5.2 Ministry of Agriculture ��������������������������������������������������������   61
6.5.3 Universities ��������������������������������������������������������������������������   61
6.5.4 Pharmacy������������������������������������������������������������������������������   61
6.5.5 Public Health������������������������������������������������������������������������   62
6.5.6 Botany ����������������������������������������������������������������������������������   62
6.5.7 Taxonomy Identification ������������������������������������������������������   62
6.5.8 Clinical & Pharmaceutical Industry (Government and Private) ��
  62
6.5.9 Ministry of Trade������������������������������������������������������������������   62
6.6 A National Strategy on Medicinal Plants Should Highlight the
Following������������������������������������������������������������������������������������������   62
6.7 Priority Issues that Need Special Attention Include ������������������������   63
7 Medicinal Wealth of Pakistan ����������������������������������������������������������������   65
7.1 Medicinal Plants of Pakistan������������������������������������������������������������   65
8 Marketed Herbal Plants in Pakistan: Case Studies������������������������������   85
8.1 DNA Analysis ����������������������������������������������������������������������������������   96
8.1.1 Cinnamomum verum������������������������������������������������������������   97
8.1.2 Cinnamomum tamala������������������������������������������������������������ 102
8.1.3 Gymnema sylvestre�������������������������������������������������������������� 108
xviii Contents

8.1.4 Sphaeranthus indicus������������������������������������������������������������ 113


8.1.5 Artemisia maritima �������������������������������������������������������������� 118
8.1.6 Butea monosperma��������������������������������������������������������������� 123
8.1.7 Achillea millefolium ������������������������������������������������������������ 129
8.1.8 Morus nigra�������������������������������������������������������������������������� 138
9 List of Commonly Used Herbal Drugs throughout the World������������ 141
10 A Pictorial Guide of Marketed Herbal Plants: Comparison With the
Original Sample �������������������������������������������������������������������������������������� 171

Bibliography ���������������������������������������������������������������������������������������������������� 175


About the Authors

Shabnum Shaheen has completed her postdoctorate from Malaysia in Biological


Sciences. She is gold medal awardee in the International Conference on Botany and
also won Farogh-e-Taleem Gold Medal Award from Idara Farogh-e-Taleem and
Federal Minister Kashmir Affairs, Pakistan. She is the first scientist who introduced
EDS technique for grass phytoliths in epidermal leaf anatomy in Pakistan. Till yet,
she has three national patents which are submitted in the Intellectual Property
Organization, Government of Pakistan. She is a member of many national and inter-
national science societies/journals and also reviewer of many well-reputed journals.
She is the author of six books and published many research articles in national and
international journals.

Sehrish Ramzan is a PhD scholar in Biological Sciences and a plant scientist. Her
PhD work is focused on the adulteration of herbal drugs. Till yet, she has published
ten research articles in national and international journals and is member of many
societies. Her research interests include biodiversity, plant systematics, adultera-
tion, and medicinal plants.

Farah Khan did her postdoctorate from Ohio State University, Columbus, USA
(2011). She is professor/chairperson at the Department of Botany, LCWU, Lahore.
She is a member of many research journal editorial boards. So far, she has super-
vised 62 research theses including 2 PhDs and has published 52 research papers in
impact factor, HEC, and UGC recognized international and national journals and 34
abstracts/proceedings in different international and national conferences and semi-
nars. Furthermore, she has on her record 31 poster (research) presentations and 4
monographs published by international publishers. She has won many awards
including a gold medal by IFT and PK.

Mushtaq Ahmad is plant biologist in the field of biodiversity. He has authored


387 research publications (to date) in top international journals and more than 3000
citations, with high IF and H and I indices. He has also published 20 international
books circulated internationally. He is awardee of various national and international

xix
xx About the Authors

awards (TWAS, CAS-PIFI, and USEA). Moreover, he is an editorial board member


of many well-reputed ISI journals and book series. He has supervised 13 PhD and
65 MPhil research scholars in applied disciplines of natural plant products, plant
biodiversity, wild edible plants, medicinal plants, energy, and environment.
List of Figures

Fig. 1.1 Food adulteration.


(Source: https://foodpoisoningsymptom.blogspot.com/2015)���������   2
Fig. 1.2 The way how to spot adulteration.
(Source: https://fusion.werindia.com/social-buzz/simple-
ways-check-food-adulteration)���������������������������������������������������������   2
Fig. 1.3 Is your food safe today.
(Source: https://foodpoisoningsymptom.blogspot.com/2015)���������   4
Fig. 1.4 Microbiological Contamination.
(Source: https://foodpoisoningsymptom.blogspot.com/2015)���������   7
Fig. 1.5 Herbal drugs adulteration. (Source: https://ghscientific.com/
safety-herbal-medicine/)�������������������������������������������������������������������   7
Fig. 2.1 Intentional Adulteration. (Source: https://www.ien.com/food-
beverage/news/21065550/intentional-adulteration-inspections-
to-begin-next-march)������������������������������������������������������������������������ 10
Fig. 2.2 Incidental Adulteration.
(Source: https://www.foodsafetymagazine.com/fsm/cache/file/
F1E98A73-63B0-40C7-B35780984BEC4637.jpg)������������������������� 11
Fig. 2.3 Metallic adulteration. (Source: https://www.slideshare.net/
AroobaAnwar/food-safety-case-study)�������������������������������������������� 11
Fig. 2.4 Other food adulteration.
(Source: https://www.foodfraudadvisors.com/facts-stories-
news-advice/)������������������������������������������������������������������������������������ 14
Fig. 3.1 Why study Herbal plants. (Source: https://www.pinterest.com/
pin/478437160385965282)�������������������������������������������������������������� 19
Fig. 3.2 Need to study herbal plants. (Source: https://study.com/articles/
Herbology_How_Can_I_Become_a_Herbalist.html)���������������������� 20
Fig. 3.3 Medicinal value of herbal plants.
(Source: https://www.organicfacts.net/health-benefits/herbs-
and-spices)���������������������������������������������������������������������������������������� 24
Fig. 3.4 Reasons for adulteration. (Source: https://ars.els-cdn.com/
content/image/1-s2.0-S2225411017300020-gr1.jpg)����������������������� 28

xxi
xxii List of Figures

Fig. 3.5 Standard reasons of adulteration.


(Source: https://ars.els-cdn.com/content/image/1-s2.0-
S2225411017300020-gr2.jpg)���������������������������������������������������������� 29
Fig. 4.1 History of medicinal plants.
(Source: https://en.wikipedia.org/wiki/Medicinal_plants)��������������� 36
Fig. 4.2 written map of medicinal plants. (Source: http://www.openculture.
com/2018/04/the-illustrated-medicinal-plant-map)�������������������������� 37
Fig. 4.3 History of medicinal plants in Middle ages. (Source: Jenny
and Nancy 1992)������������������������������������������������������������������������������� 38
Fig. 4.4 New world contribution towards herbal plants.
(Source: http://wocmap2019.org/)���������������������������������������������������� 39
Fig. 5.1 Marketed herbal medicine preparation
(Source: Djordjevic 2017)���������������������������������������������������������������� 55
Fig. 6.1 Challenges to herbal plants��������������������������������������������������������������� 58
Fig. 8.1 (a) Phylogenetic analysis of matK sequences of marketed
samples of Cinnamomum verum. Other sequences were
retrieved from the databases based on BLAST analysis.
Neighbour joining algorithm was used for phylogenetic tree
construction in Mega 6 software tool. (b) Phylogenetic analysis
of nrITS sequences of marketed samples of Cinnamomum verum.
Other sequences were retrieved from the databases based
on BLAST analysis. Neighbour joining algorithm was used for
phylogenetic tree construction in Mega 6 software tool. (c)
Phylogenetic analysis of rbcL sequences of marketed samples
of Cinnamomum verum. Other sequences were retrieved from
the databases based on BLAST analysis. Neighbour joining
algorithm was used for phylogenetic tree construction in Mega 6
software tool. (d) Phylogenetic analysis of psbA-trnH sequences
of marketed samples of Cinnamomum verum. Other sequences
were retrieved from the databases based on BLAST analysis.
Neighbour joining algorithm was used for phylogenetic tree
construction in Mega 6 software tool. (e) PCR amplification of
Cinnamomum verum fresh sample and market sample with matK
primers (Panel A). Lane A, B, C, D showing amplified fragment
from market sample (1,5,10 and 12). Lane Fr showing fresh
sample and Lane NC is Negative Control. Lane L is showing
1 kb Ladder (Thermo Fisher Scientific, USA). With nrITS primers
(Panel B). Lane A and B showing from market samples (1, 10).
Lane NC for Negative Control and Lane L is showing 1 kb
Ladder (Thermo Fisher Scientific, USA). With rbcL primers
(Panel C). Lane A, B, C and D showing amplified PCR band
from market samples (1,3,10,11 and 12 respectively). Lane Fr
showing fresh sample fragment and Lane L is showing 1 kb
List of Figures xxiii

Ladder (Thermo Fisher Scientific, USA). With TrnH-PsbA


primers (Panel D). Lane A showing amplified PCR band from
market samples (11). Lane Fr showing fresh sample fragment,
Lane NC for Negative Control and Lane L is showing 1 kb
Ladder (Thermo Fisher Scientific, USA)����������������������������������������� 98
Fig. 8.2 (a) Phylogenetic analysis of matK sequences of marketed
samples of Cinnamomum tamala. Other sequences were retrieved
from the databases based on BLAST analysis. Neighbour joining
algorithm was used for phylogenetic tree construction in Mega 6
software tool. (b) Phylogenetic analysis of nrITS sequences of
marketed samples of Cinnamomum tamala. Other sequences
were retrieved from the databases based on BLAST analysis.
Neighbour joining algorithm was used for phylogenetic tree
construction in Mega 6 software tool. (c) Phylogenetic analysis
of rbcL sequences of marketed samples of Cinnamomum tamala.
Other sequences were retrieved from the databases based
on BLAST analysis. Neighbour joining algorithm was used for
phylogenetic tree construction in Mega 6 software tool. (d)
Phylogenetic analysis of psbA-trnH sequences of marketed
samples of Cinnamomum tamala. Other sequences were retrieved
from the databases based on BLAST analysis. Neighbour joining
algorithm was used for phylogenetic tree construction in Mega 6
software tool. (e) PCR amplification of Cinnamomum tamala
fresh sample and market sample with matK primers (Panel A).
Lane A, B, C, D, E, F, G, H and I showing amplified fragment
from market sample (2, 3, 5, 6, 8, 9, 10 and 12). Lane Fr showing
fresh sample and Lane NC is Negative Control. Lane L is showing
1 kb Ladder (Thermo Fisher Scientific, USA). With nrITS primers
(Panel B). Lane A, B, C, D, showing from market samples (1, 10).
Lane NC for Negative Control and Lane L is showing 1 kb Ladder
(Thermo Fisher Scientific, USA). With rbcL primers (Panel C).
Lane A, B, C and D showing amplified PCR band from market
samples (1,3,10,11 and 12 respectively). Lane Fr showing fresh
sample fragment and Lane L is showing 1 kb Ladder (Thermo
Fisher Scientific, USA). With TrnH-PsbA primers (Panel D).
Lane A showing amplified PCR band from market samples (11).
Lane Fr showing fresh sample fragment, Lane NC for Negative
Control and Lane L is showing 1 kb Ladder (Thermo Fisher
Scientific, USA)�������������������������������������������������������������������������������� 103
Fig. 8.3 (a) Phylogenetic analysis of matK sequences of fresh and
marketed samples of Gymnema sylvestre. Other sequences were
retrieved from the databases based on BLAST analysis.
Neighbour joining algorithm was used for phylogenetic tree
construction in Mega 6 software tool. (b) Phylogenetic analysis
of nrITS sequences of fresh and marketed samples of Gymnema
xxiv List of Figures

sylvestre. Other sequences were retrieved from the databases


based on BLAST analysis. Neighbour joining algorithm was
used for phylogenetic tree construction in Mega 6 software tool.
(c) Phylogenetic analysis of rbcL sequences of fresh and marketed
samples of Gymnema sylvestre. Other sequences were retrieved
from the databases based on BLAST analysis. Neighbour joining
algorithm was used for phylogenetic tree construction in Mega 6
software tool. (d) Phylogenetic analysis of psbA-trnH sequences
|of fresh and marketed samples of Gymnema sylvestre. Other
sequences were retrieved from the databases based on BLAST
analysis. Neighbour joining algorithm was used for phylogenetic
tree construction in Mega 6 software tool. (e) PCR amplification
of Gymnema sylvestre fresh sample and market sample with matK
primers (Panel A). Lane A, B, C and D showing amplified
fragment from market sample (2, 4, 9 and 11). Lane Fr showing
fresh sample and Lane NC is Negative Control. Lane L is
showing 1 kb Ladder (Thermo Fisher Scientific, USA). With
nrITS primers (Panel B). Lane A and B showing from market |
samples (1and 6). Lane Fr showing fresh sample and NC for
Negative Control. Lane L is showing 1 kb Ladder (Thermo
Fisher Scientific, USA). With rbcL primers (Panel C). Lane
A, B, C, D, E and F showing amplified PCR Fig. 8.3a(continued)
band from market samples (5,6,7,8,9 and 11 respectively).
Lane Fr showing fresh sample fragment, lane NC for negative
control and Lane L is showing 1 kb Ladder (Thermo Fisher
Scientific, USA). With TrnH-PsbA primers (Panel D).Lane
showing amplified PCR band from market samples. Lane Fr
showing fresh sample fragment, Lane NC for Negative Control
and Lane L is showing 1 kb Ladder (Thermo Fisher Scientific,
USA)������������������������������������������������������������������������������������������������� 109
Fig. 8.4 (a) Phylogenetic analysis of matK sequences of marketed
samples of Sphaeranthus indicus. Other sequences were retrieved
from the databases based on BLAST analysis. Neighbour joining
algorithm was used for phylogenetic tree construction in Mega
6 software tool. (b) Phylogenetic analysis of nrITS sequences
of fresh and marketed samples of Sphaeranthus indicus. Other
sequences were retrieved from the databases based on BLAST
analysis. Neighbour joining algorithm was used for phylogenetic
tree construction in Mega 6 software tool. (c) Phylogenetic
analysis of rbcL sequences of fresh and marketed samples of
Sphaeranthus indicus. Other sequences were retrieved from the
databases based on BLAST analysis. Neighbour joining
algorithm was used for phylogenetic tree construction in Mega 6
software tool. (d) Phylogenetic analysis of psbA-trnH sequences
of fresh and marketed samples of Sphaeranthus indicus. Other
List of Figures xxv

sequences were retrieved from the databases based on BLAST


analysis. Neighbour joining algorithm was used for phylogenetic
tree construction in Mega 6 software tool. (e) PCR amplification
of Sphaeranthus indicus fresh sample and market sample with
matK primers (Panel A). Lane A showing amplified fragment
from market sample (8). Lane Fr showing fresh sample and Lane
NC is Negative Control. Lane L is showing 1 kb Ladder (Thermo
Fisher Scientific, USA). With nrITS primers (Panel B). Lane
A, B, C, D and E showing from market samples (2,5,6,7 and 8).
Lane NC for Negative Control and Lane L is showing 1 kb
Ladder (Thermo Fisher Scientific, USA). With rbcL primers
(Panel C). Lane A, B, C, D, E, F and G showing amplified PCR
band from market samples (2,6,7,8,9,10 and12 respectively).
Lane Fr showing fresh sample fragment and Lane L is showing
1 kb Ladder (Thermo Fisher Scientific, USA). With TrnH-PsbA
primers (Panel D). Lane A, B and C showing amplified PCR
band from market samples (6,7 and 8 respectively). Lane Fr
showing fresh sample fragment, Lane NC for Negative Control
and Lane L is showing 1 kb Ladder (Thermo Fisher Scientific,
USA)������������������������������������������������������������������������������������������������� 114
Fig. 8.5 (a) Phylogenetic analysis of matK sequences of fresh and
marketed samples of Artemisia maritima. Other sequences were
retrieved from the databases based on BLAST analysis.
Neighbour joining algorithm was used for phylogenetic tree
construction in Mega 6 software tool (b) Phylogenetic analysis
of nrITS sequences of fresh and marketed samples of Artemisia
maritima. Other sequences were retrieved from the databases
based on BLAST analysis. Neighbour joining algorithm was
used for phylogenetic tree construction in Mega 6 software tool.
(c) Phylogenetic analysis of rbcL sequences of fresh and marketed
samples of Artemisia maritima. Other sequences were retrieved
from the databases based on BLAST analysis. Neighbour joining
algorithm was used for phylogenetic tree construction in Mega
6 software tool. (d) Phylogenetic analysis of psbA-trnH
sequences of marketed samples of Artemisia maritima. Other
sequences were retrieved from the databases based on BLAST
analysis. Neighbour joining algorithm was used for phylogenetic
tree construction in Mega 6 software tool. (e) PCR amplification
of Artemisia maritima fresh sample and market sample with matK
primers (Panel A). Lane A,B,C and D showing amplified fragment
from market sample (7,8,9 and 10). Lane NC is Negative Control
and lane L is showing 1 kb Ladder (Thermo Fisher Scientific,
USA). With nrITS primers (Panel B). Lane A, B, C, D, E and F
showing from market samples (4,5,7,8,9 and 11). Fr showing
fresh sample fragment and Lane NC for Negative Control. Lane
xxvi List of Figures

L is showing 1 kb Ladder (Thermo Fisher Scientific, USA).


With rbcL primers (Panel C). Lane A, B, C, D, E and F showing
amplified PCR band from market samples (6,7,9,10,11 and12
respectively). Lane NC for Negative Control and lane L is
showing 1 kb Ladder (Thermo Fisher Scientific, USA). With
TrnH-PsbA primers (Panel D). Lane A, B, C and D showing
amplified PCR band from market samples (5,7, 9 and 11
respectively). Lane Fr showing fresh sample fragment, Lane
NC for Negative Control and Lane L is showing 1 kb Ladder
(Thermo Fisher Scientific, USA)������������������������������������������������������ 119
Fig. 8.6 (a) Phylogenetic analysis of matK sequences of fresh and
marketed samples of Butea monosperma. Other sequences were
retrieved from the databases based on BLAST analysis. Neighbour
joining algorithm was used for phylogenetic tree construction in
Mega 6 software tool. (b) Phylogenetic analysis of m sequences
of fresh and marketed samples of Butea monosperma. Other
sequences were retrieved from the databases based on BLAST
analysis. Neighbour joining algorithm was used for phylogenetic
tree construction in Mega 6 software tool. (c) Phylogenetic
analysis of rbcL sequences of fresh and marketed samples of
Butea monosperma. Other sequences were retrieved from the
databases based on BLAST analysis. Neighbour joining
algorithm was used for phylogenetic tree construction in Mega 6
software tool. (d) Phylogenetic analysis of TrnH-psbA sequences
of fresh and marketed samples of Butea monosperma. Other
sequences were retrieved from the databases based on BLAST
analysis. Neighbour joining algorithm was used for phylogenetic
tree construction in Mega 6 software tool. (e) PCR amplification
of Butea monosperma fresh sample and herbalist provided sample
with matK primers (Panel A). Lane A showing amplified fragment
from market sample (herbalist provided). Lane Fr showing fresh
sample fragment and lane L is showing 1 kb Ladder (Thermo
Fisher Scientific, USA). With rbcL primers (Panel C).
Lane A showing amplified PCR band from market samples
(herbalists provided sample). Lane Fr for fresh sample
fragment and lane L is showing 1 kb Ladder (Thermo Fisher
Scientific, USA). With TrnH-PsbA primers (Panel D).
Lane A showing amplified PCR band from market samples
(herbalists provided sample). Lane Fr showing
fresh sample fragment, Lane NC������������������������������������������������������ 124
Fig. 8.7 (a) Phylogenetic analysis of matK sequences of fresh
and marketed samples of Achillea millefolium. Other sequences
were retrieved from the databases based on BLAST analysis.
Neighbour joining algorithm was used for phylogenetic tree
construction in Mega 6 software tool. (b) Phylogenetic analysis of
List of Figures xxvii

nrITS sequences of marketed samples of Achillea millefolium.


Other sequences were retrieved from the databases based
on BLAST analysis. Neighbour joining algorithm was used for
phylogenetic tree construction in Mega 6 software tool. (c)
Phylogenetic analysis of rbcL sequences of marketed
samples of Achillea millefolium. Other sequences were
retrieved from the databases based on BLAST analysis.
Neighbour joining algorithm was used for phylogenetic
tree construction in Mega 6 software tool. (d) Phylogenetic
analysis of TrnH-psbA sequences of marketed samples
of Achillea millefolium. Other sequences were retrieved
from the databases based on BLAST analysis. Neighbour
joining algorithm was used for phylogenetic tree construction
in Mega 6 software tool. (e) PCR amplification of Achillea
millifolium L fresh sample and market sample with matK primers
(Panel A). Lane A, B showing amplified fragment from market
samples (1 and 2 respectively). Lane Fr showing fresh sample and
Lane NC is Negative Control. Lane L is showing 1 kb Ladder
(Thermo Fisher Scientific, USA). With nrITS primers (Panel B).
Lane A, B, C, D and E showing from market samples (1, 7, 8, 9
and 11 respectively). Lane NC for Negative Control and Lane
L is showing 1 kb Ladder (Thermo Fisher Scientific, USA).
With rbcL primers (Panel C). Lane A, B, C, D, E, F and G showing
amplified PCR band from market samples (3,5,6,8,9,10 and 11
respectively). Lane Fr showing fresh sample fragment, Lane NC
for Negative Control and Lane L is showing 1 kb Ladder (Thermo
Fisher Scientific, USA). With TrnH-PsbA primers (Panel D).
Lane A and B showing amplified PCR band from market samples
(9 and 10 respectively). Lane Fr showing fresh sample fragment,
Lane NC for Negative Control and Lane L is showing
1 kb Ladder (Thermo Fisher Scientific, USA)��������������������������������� 130
Fig. 8.8 (a) Phylogenetic analysis of matK sequences of marketed
samples of Morus nigra. Other sequences were retrieved from
the databases based on BLAST analysis. Neighbour joining
algorithm was used for phylogenetic tree construction in Mega
6 software tool. (b) Phylogenetic analysis of nrITS sequences of
marketed samples of Morus nigra. Other sequences were retrieved
from the databases based on BLAST analysis. Neighbour joining
algorithm was used for phylogenetic tree construction in Mega 6
software tool. (c) Phylogenetic analysis of rbcL sequences of
marketed samples of Morus nigra. Other sequences were retrieved
from the databases based on BLAST analysis. Neighbour joining
algorithm was used for phylogenetic tree construction in Mega 6
software tool. (d) Phylogenetic analysis of TrnH-psbA sequences
of marketed samples of Morus nigra. Other sequences were
xxviii List of Figures

retrieved from the databases based on BLAST analysis. Neighbour


joining algorithm was used for phylogenetic tree construction in
Mega 6 software tool. (e) PCR amplification of Morus nigra fresh
sample and market sample with matK primers (Panel A). Lane
A,B,C and D showing amplified fragment from market sample
(2,9,10 and 11). Lane NC is Negative Control and lane L is
showing 1 kb Ladder (Thermo Fisher Scientific, USA). With
nrITS primers (Panel B). Lane A, B, C and D showing from
market samples (6,10,11 and 12). Fr showing fresh sample
fragment and Lane NC for Negative Control. Lane L is showing
1 kb Ladder (Thermo Fisher Scientific, USA). With rbcL primers
(Panel C). Lane A, B, C, D and E showing amplified PCR band
from market samples (5, 6,10,11 and12 respectively). Lane Fr
showing fresh sample fragment, Lane NC for Negative Control
and lane L is showing 1 kb Ladder (Thermo Fisher Scientific,
USA). With TrnH-PsbA primers (Panel D). Lane A, B and C
showing amplified PCR band from market samples (6, 10 and
12 respectively). Lane Fr showing fresh sample fragment, Lane
NC for Negative Control and Lane L is showing 1 kb Ladder
(Thermo Fisher Scientific, USA)������������������������������������������������������ 134
List of Tables

Table 2.1 Some common food articles, their adulterants and their
effects on humans����������������������������������������������������������������������������� 12
Table 3.1 Most Commonly used DNA Markers in Plants Identification��������� 32
Table 3.2 Disadvantages of Most Commonly used DNA Markers
in Plants Identification���������������������������������������������������������������������� 32
Table 6.1 Standardization tools to overcome constraints in herbal drug
preparations�������������������������������������������������������������������������������������� 59
Table 7.1 Some medicinal plant sources and traditional uses in Pakistan������� 66
Table 7.2 Medicinal wealth of Pakistan����������������������������������������������������������� 70
Table 7.3 Medicinal plants of the Miandam Area (Pakistan) with their
medicinal properties, and biological, ecological and
chorological characteristics�������������������������������������������������������������� 74
Table 8.1a Cinnamomum verum Presl. vs Canella winterana (L.) Gaertn.������� 86
Table 8.2a Cinnamomum tamala (Buch.-Ham.) T. Nees & Eberm. vs
Cinnamomum obtusifolium (Roxb.) Nees���������������������������������������� 87
Table 8.3a Gymnema sylvestre (Retz.) R.Br. ex Sm. vs
Gymnema lactiferum (L.) R.Br. ex Schult���������������������������������������� 88
Table 8.4a Sphaeranthus indicus Linn. vs Sphaeranthus africanus L.�������������� 89
Table 8.5a Artemisia maritima Linn.vs Artemisia absinthium L.���������������������� 90
Table 8.6a Butea monosperma (Lam.) Taub. vs Averrhoa carambola L.���������� 91
Table 8.7a Achillea millefolium L. vs Adhatoda vasica Nees���������������������������� 93
Table 8.8a Morus nigra L. vs Morus alba L.����������������������������������������������������� 95
Table 9.1 List of commonly used herbal drugs throughout the world����������� 142

xxix
List of Plates

Plate 10.1 Cinnamomum verum Presl. (a) Light green leaves with
3 mid ribs. Canella winterana (L.) Gaertn (b) Dark green
leaves with single mid ribs������������������������������������������������������������ 171
Plate 10.2 Cinnamomum tamala (Buch.-Ham.) T.Nees & Eberm.
(a) Alternate leaves. Cinnamomum obtusifolium (Roxb.)
Nees. (b) Oblong to acute leaves �������������������������������������������������� 171
Plate 10.3 Gymnema sylvestre (Retz.) R.Br. ex Sm. (a) ovate leaves.
Gymnema lactiferum (L.) R.Br. ex Schult. (b) ovate but
pubescent leaves���������������������������������������������������������������������������� 172
Plate 10.4 Sphaeranthus indicus Linn. (a) Small toothed leaves.
Sphaeranthus africanus L. (b) large pubescent leaves������������������ 172
Plate 10.5 Artemisia maritima Linn. (a) dull green leaves.
Artemisia absinthium L. (b) bright green leaves���������������������������� 172
Plate 10.6 Butea monosperma (Lam.) Taub. (a) Orangish red flower.
Averrhoa carambola L. (b) Purplish pink flower�������������������������� 173
Plate 10.7 Achillea millefolium L. (a) Narrow pointed leaves.
Adhatoda vasica Nees. (b) long broad leaves�������������������������������� 173
Plate 10.8 Morus nigra L. (a) Leaves of Morus nigra. Morus alba L.
(b) Leaves of Morus alba �������������������������������������������������������������� 173

xxxi
Chapter 1
Adulteration A Global Issue

1.1 Adulteration A Global Issue

1.1.1 Adulteration a Legal Term

Adulteration is a legal term meaning that a food or herbal product fails to meet legal
standards. Adulteration is basically found in two major fundamentals of life.
1. Adulteration in food items
2. Adulteration in herbal drugs
These substances may be other available food items or non-food items. Among
meat and meat products some of the items used to adulterate are water or ice, car-
casses, or carcasses of animals other than the animal meant to be consumed
(Gaur 2008) (Fig. 1.1).
In recent years, adulteration has become an increasing problem globally in the
food industry and for consumers. Since food adulteration is a huge concern in all
parts of the globe, there is voluminous literature on various aspects of food adul-
teration including its detection. In fact, methodological procedures of food adul-
teration detection have been reviewed extensively (Druml et al. 2015). Adulteration
and fraudulent labeling of a foodstuff’s origin costs the global food industry an
estimated $10 to $15 billion per year. Adulteration usually refers to mixing other
matter of an inferior and sometimes harmful quality with food or drink intended to
be sold. As a result of adulteration, food or drink becomes impure and unfit for
human consumption (Fig. 1.2).

© Springer Nature Switzerland AG 2019 1


S. Shaheen et al., Adulteration in Herbal Drugs: A Burning Issue,
https://doi.org/10.1007/978-3-030-28034-5_1
2 1 Adulteration A Global Issue

Fig. 1.1 Food adulteration. (Source: https://foodpoisoningsymptom.blogspot.com/2015)

Fig. 1.2 The way how to spot adulteration. (Source: https://fusion.werindia.com/social-buzz/


simple-ways-check-food-adulteration)
1.1 Adulteration A Global Issue 3

1.1.2 Federal Food, Drug and Cosmetics Act

The Federal Food, Drug, and Cosmetic (FD&C) Act provides that food is “adulter-
ated” if it meets any one of the following criteria (https://www.law.cornell.edu/
uscode/text/21/342):
• it bears or contains any “poisonous or deleterious substance” which may render
it injurious to health;
• it bears or contains any added poisonous or added deleterious substance (other
than a pesticide residue, food additive, color additive, or new animal drug, which
are covered by separate provisions) that is unsafe;
• its container is composed, in whole or in part, of any poisonous or deleterious
substance which may render the contents injurious to health;
• or it bears or contains a pesticide chemical residue that is unsafe. (Note: The
United States Environmental Protection Agency (EPA) establishes tolerances for
pesticide residues in foods, which are enforced by the FDA).
Food also meets the definition of adulteration if:
• it is, or it bears or contains, an unsafe food additive;
• it is, or it bears or contains, an unsafe new animal drug;
• it is, or it bears or contains, an unsafe colour additive;
• it consists, in whole or in part, of “any filthy, putrid, or decomposed substance”
or is otherwise unfit for food;
• or it has been prepared, packed, or held under unsanitary conditions (insect,
rodent, or bird infestation) whereby it may have become contaminated with filth
or rendered injurious to health.
Further, food is considered adulterated if:
• it has been irradiated and the irradiation processing was not done in conformity
with a regulation permitting irradiation of the food in question (the FDA has
approved irradiation of a number of foods, including refrigerated or frozen
uncooked meat, fresh or frozen uncooked poultry, and seeds for sprouting
(Adullah and Rehbein 2014).
• it contains a dietary ingredient that presents a significant or unreasonable risk of
illness or injury under the conditions of use recommended in labeling (for exam-
ple, foods or dietary supplements containing aristolochic acids, which have been
linked to kidney failure, have been banned (21 USC §331, Federal Food, Drug,
and Cosmetic Act (Grossman 2008);
• a valuable constituent has been omitted in whole or in part or replaced with
another substance; damage or inferiority has been concealed in any manner; or a
substance has been added to increase the product’s bulk or weight, reduce its
quality or strength, or make it appear of greater value than it is (this is “economic
adulteration”);
• or it is offered for import into the United States and is a food that has previously
been refused admission, unless the person reoffering the food establishes that it
is in compliance with U.S. law (Fig. 1.3).
Another random document with
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civilised world, and surprise can hardly be felt when we remember
that a sentence of mutilation was carried out in England little more
than 300 years ago. Camden’s Annals for the year 1581 contain
an account of the mutilation of one Stubbs, for publishing an
attack upon Queen Elizabeth’s proposed marriage with the Duke
of Alençon. The historian was an eyewitness of the scene, which
has been utilised by Sir Walter Scott in the Fortunes of Nigel,
chap. xiii.
[38]A Moorish beehive is made from the bark of the cork-tree.
In the summer months, when the sap rises, a vertical incision
about four feet long is made through the cork to the inner bark,
and the part to be removed, having been cut above and below, is
hammered with a heavy mallet. The cork is separated from the
stem of the tree, and being elastic, is taken off entire. Two circular
pieces of cork are inserted in the orifices at each end and
fastened with wooden pegs. The bees close with wax the cracks
which may appear. The hive is warm, and keeps out both wet and
sun.
[39]Mr. Reade was Consul, Mr. Green Private Secretary. The
latter, as Sir William Kirby Green, succeeded Sir John Hay as
Minister to the Court of Morocco in 1886.
[40]Jebel Kebír, now known as ‘The Hill.’
[41]These were troops from the seat of war not yet disbanded.
The Sultan evidently desired to impress Mr. Hay with the strength
of his army.
[42]The duties on the export of wheat and barley were never
added to those noted above, in spite of Sir John’s constant and
unceasing endeavours.
[43]In allusion to the manner in which, in ancient times, Jews
and Christians in Morocco were put to death. The victims were
suspended by large iron hooks through the flesh of their backs;
one of these hooks was still to be seen on a gate of the city of
Marákesh in 1846; or a spit was run through their bodies, and
they remained transfixed till death put an end to their tortures.
[44]The late Sultan Sid Mohammed, the descendant of Sultan
Mulai Ahmed, was a good mathematician, and also very clever as
a mechanist. He mended and cleaned his own watches. When I
presented H.M. with a breech-loading gun, and at his request
took it and the lock to pieces, I bungled in putting them together.
H.M., taking the gun from me, at once re-adjusted it.—J. H. D. H.
[45]On the site now occupied by the chief mosque.
[46]Pauper, or holy man.
[47]A delicate paste, partaking of the nature of Italian paste,
but round in form, the best being no larger than dust shot.
[48]Ovis musimon.
[49]Cedrus Atlantica and Callitris quadrivalvis.
[50]Yet, according to Marmol, it may be inferred that by this
pass the ‘Almoravides’ entered Western Barbary from Numidia.
[51]On this, as on all his other Missions, the members of Sir
John Hay’s family and his ‘private friends’ were his personal
guests, the ‘officials’ travelled at the expense of Government.
[52]Sultan Mulai Hassan.
[53]A white but much sunburnt Moorish servant of Sir J. H. D.
H.
[54]Though this permission was then granted, the laying of the
cable was delayed until 1886-87.
[55]His son, then Consul at Mogador.
[56]The ‘arum arisarum,’ called ‘yerna’ by the Moors, is used
by the inhabitants of Western Barbary as an article of food in
times of great scarcity, though it is held by them to be poisonous
without careful preparation. The tubers when collected are cut up
in small pieces, which they wash in many waters and then steam,
as they do their ‘siksu,’ after which they pound them into meal, of
which they make cakes, mixed if possible with a little ‘dra’ (millet)
meal. They also make this arum meal into a kind of porridge. This
food appears to contain few nourishing qualities, and those who
are reduced to live on it suffer much in health.
[57]Journal of Society for Psychical Research, March, 1891, p.
40.
[58]Mashallah.
[59]The loan referred to was that raised in England in 1862 to
enable the Sultan to pay the Spanish war indemnity. See chapter
xv. p. 218.
[60]Then Spanish Minister in Morocco.
[61]Sheríf of Wazan.
[62]Though Ordega acknowledged that the dead Moor had
received two hundred lashes.
[63]Moorish Minister for Foreign Affairs.
[64]German Minister.
[65]‘Mellah,’ the Jewish quarter in all Moorish towns.
[66]Baba, father.
[67]A large covert a short distance off.
[68]Evil Genius.
[69]Moors have a superstition that in hunting the lion the man
who first reports having seen the ‘S’ba’ (lion), and mentions the
word, will be the first victim.
Transcriber's note:

pg x Added comma after: flourished in the environs of Tangier


pg 87 Changed: the Luxis of the ancients to: Lixus
pg 103 Paragraphs starting with "On March 28" and the following one formatted as
being from the Editors' perspective
pg 180 Paragraph starting with "Ten days later" formatted as not belonging to the
surrounding quoted correspondence
pg 270 Changed: admit the cortége to: cortège
pg 344 Changed: formidable porportions to: proportions
pg 356 Changed: she decares she detests to: declares
Minor changes in punctuation and quotation mark placement have been done
silently.
Other spelling errors or inconsistencies have been left unchanged.
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