DEVELOPMENT OF TAGBAK (ALIPINIA ELEGANS) AND

TURMERIC (CURCUMA LONGA L.) RHIZOMES AS

AN IMMUNE BOOSTER CAPSULE

An Undergraduate Thesis Presented to the Faculty of

Nursing Department
Bicol University Polangui
Polangui, Albay

In partial Fulfillment of the Requirements for the Degree

Bachelor of Science in Nursing

Benitez, Johannah Rose C.

Bolalin, Camela D.
Dormetorio, Marjorie F.
Encinares, Alyssa Mae O.
Lisay, Maui Avril D.

2023
 Republic of the Philippines
BICOL UNIVERSITY POLANGUI
NURSING DEPARTMENT
Polangui, Albay

===============================================================

RECOMMENDATION FOR FINAL DEFENSE

This undergraduate thesis entitled, “Development of Tagbak (Alpinia elegans)

and Turmeric (Curcuma longa L.) as an Immune Booster Capsule”, prepared and

submitted by Johannah Rose C. Benitez, Camela D. Bolalin, Marjorie F. Dormetorio,

Alyssa Mae O. Encinares, and Maui Avril D. Lisay in partial fulfillment of the

requirements for the degree of Bachelor of Science in Nursing is hereby submitted to the

Thesis Committee for their consideration and approval to conduct the study.

CHRISTINE GENEVIEVE B. RIVERA, MAN

Thesis Adviser

ii
 Republic of the Philippines
BICOL UNIVERSITY POLANGUI
NURSING DEPARTMENT
Polangui, Albay

===============================================================

CERTIFICATION OF FINAL RATING

Thesis Title : DEVELOPMENT OF TAGBAK (Alpinia

Elegans) AND TURMERIC (Curcuma Longa L.)
RHIZOME AS AN IMMUNE BOOSTER
CAPSULE

Venue : Nursing Department

Date : September 4, 2023

Researchers : Johannah Rose C. Benitez

Camela D. Bolalin

Marjorie F. Dormetorio

Alyssa Mae O. Encinares

Maui Avril D. Lisay

Panel of Examiners Action Taken Signature

ARTEMIO JEROME S. RIVERA, JR., Ph.D. _______________ _____________

Chairperson

MA. BRENDA P. YAPCHIONGCO, MAN _______________ _____________

Member

MARVIN U. VELACRUZ, RN _______________ _____________

Member

iii
 Republic of the Philippines
BICOL UNIVERSITY POLANGUI
NURSING DEPARTMENT
Polangui, Albay

===============================================================

APPROVAL SHEET

This thesis entitled, “Development of Tagbak (Alpinia elegans) and Turmeric

(Curcuma longa L) Rhizome as an Immune Booster Capsule” prepared by Johannah
Rose C. Benitez, Camela D. Bolalin, Marjorie F. Dormetorio, Alyssa Mae O.
Encinares, and Maui Avril D. Lisay in partial fulfillment of the requirements for the
degree of Bachelor of Science in Nursing is hereby submitted to the Thesis Committee
for consideration and approval.

CHRISTINE GENEVIEVE B. RIVERA, MAN

Thesis Adviser

ARTEMIO JEROME S. RIVERA JR., Ph.D.

Chairperson

MA. BRENDA P. YAPCHIONGCO, MAN MARVIN U. VELACRUZ, RN

Member Member

Accepted and approved in partial fulfilment of the requirements for the degree of
Bachelor of Science in Nursing.

ARTEMIO JEROME S. RIVERA JR., Ph.D.

Department Chairperson

MARY JOY B. CATANGUI, Ed. D.

Dean

iv
 Republic of the Philippines
BICOL UNIVERSITY POLANGUI
NURSING DEPARTMENT
Polangui, Albay

===============================================================

CERTIFICATION OF THE GRAMMARIAN

This is to certify that the thesis entitled, “Development of Tagbak (Alpinia

elegans) and Turmeric (Curcuma longa L) Rhizome as an Immune Booster Capsule”

prepared by Johannah Rose C. Benitez, Camela D. Bolalin, Marjorie F. Dormetorio,

Alyssa Mae O. Encinares, and Maui Avril D. Lisay in partial fulfillment of the

requirement for the degree of Bachelor of Science in Nursing has been edited by the

undersigned.

ART V. MANGUBAT, MAEngEd

v
 ACKNOWLEDGEMENT

This study was made successful through the aid of significant persons who guided

and helped us, the researchers, throughout the completion of the study. We would like to

express our sincere gratitude to the following individuals for their invaluable support and

contributions to this research study:

First of all, we would like to thank the Almighty God for His enduring grace,

guidance and protection that He had bestowed upon us during the research process.

To Prof. Christine Genevieve B. Rivera, RM, MAN, our thesis adviser, we are

deeply thankful for your guidance, patience, and expertise throughout the entire research

process. Your insights and feedback were instrumental in shaping this study.

To Prof. Artemio Jerome S. Rivera Jr. Ph.D, our panel of evaluator chairperson

and to Prof. Ma. Brenda P. Yapchiongco, MAN and Prof. Marvin U. Velacruz, RN,

the members of the panel, who provided encouraging and constructive feedback and

valuable comments and suggestions in developing this research.

To Dr. Jose S. Sabater, veterinarian at his own veterinary clinic at Magurang,

Polangui, Albay for his expertise in collection of blood samples of the male albino mice as

well as for providing insights in care and handling of mice.

To Prof. Lester M. Narvaez, Ph.D., for his expertise in identifying and verifying

our plant sample Tagbak (Alpinia elegans).

To Sir. Edward Pescuela, our statistician for sharing his invaluable expertise in

the field of statistics that helped us understand the statistical methods and data analysis,

which significantly enhanced the quality and reliability of this study.

vi
 Deepest gratitude is also given to Bicol University College of Science for helping

the researchers to perform laboratory procedures for the data and findings needed in the

study.

To the researchers beloved parents Mr. and Mrs. Benitez, Mr. and Mrs. Bolalin,

Mr. and Mrs. Dormetorio, Mr. and Mrs. Encinares and Mr. and Mrs. Lisay, whose

unwavering support and encouragement have been instrumental in the successful

completion of this research study. From the start of this academic endeavor, you have been

our constant source of inspiration, guidance, and motivation. Your belief in our capabilities

and your willingness to provide both emotional and financial support have been invaluable

and propelled us forward.

To all of our friends, for being a source of encouragement and assistance

throughout the course of this research project. Your steadfast support during moments of

self-doubt and your enthusiastic applause at every achievement made this research journey

even more significant and rewarding.

Expressing gratitude is often underestimated, but it is a heartfelt way for the

researchers to convey their deep appreciation to all of you. Best wishes for your future

endeavors.

-The researchers

vii
 ABSTRACT
BENITEZ, JOHANNAH ROSE C., BOLALIN, CAMELA D., DORMETORIO,
MARJORIE F., ENCINARES, ALYSSA MAE O., LISAY, MAUI D.,
“Development of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.)
Rhizome as an Immune Booster Capsule” (Unpublished Undergraduate Thesis,
Bicol University Polangui, Polangui Albay, 2022-2023).

In response to the heightened susceptibility to illnesses caused by contemporary

lifestyle and climate changes, particularly among those facing financial constraints, the

researchers aim to provide an economically viable alternative for augmenting immunity.

The study investigated the phytochemical properties, evaluated the efficacy of Tagbak

(Alpinia elegans) and Turmeric (Curcumal longa L) rhizomes as an alternative immune

booster capsule, and determined the significant difference among the treatments and the

commercially available immune booster capsules. Findings revealed that Tagbak and

Turmeric rhizomes contained triterpenes, flavonoids, saponins, glycosides, and tannins,

except for alkaloids, curcumin, and vitamin C that are only present in Turmeric rhizomes.

Rhizomes were air-dried and pulverized, then the three-treatment concentration of 500 mg

combination was formulated based on the proportion: 60% Tagbak:40% Turmeric, 40%

Tagbak:60% Turmeric, and 50% Tagbak:50% Turmeric tested on Prednisone-induced,

malnourished male albino mice for 21 days. Then, evaluated for immune boosting activity

through three parameters: White Blood Cell count, Lymphocyte count, and

Immunoglobulin G level. Determined by the study, all treatments of different

concentrations were effective in regulating the immune system based on the statistical tool

Kruskal-Wallis H test. However, T3 - 50% Tagbak:50% Turmeric had the highest

increased rate and consistent level of 67.3% in WBC and 31.7% in Lymphocyte

parameters. Also, a result of 8.4 mg/ml in IgG parameter indicates an effective regulation
viii
of immune response in mice. The Mann-Whitney U test result proved that there is no

significant difference but with medium to large practical significance between the

formulated immune booster capsule and the commercially available ascorbic acid in terms

of WBC count (p=0.825) (r=0.48) and Lymphocyte count (p=0.383), (r=0.56). This reveals

that the two treatments are comparable and exhibit almost the same immune-boosting

effect.

Keywords: Capsule, Immune Booster, Rhizome, Tagbak, Turmeric

ix
 TABLE OF CONTENTS

PRELIMINARIES Page

Title Page i
Recommendation for Oral Defense ii
Certification for Final Rating iii
Approval Sheet iv
Certification of Grammarian v
Acknowledgement vi
Abstract viii
Table of Contents x
List of Tables xii
List of Figures xiv

Chapter 1 – THE PROBLEM

Introduction 1
Objectives of the Study 4
Hypothesis 5
Scope and Delimitation 5
Significance of the Study 6
References 8

Chapter 2 – REVIEW OF RELATED LITERATURE AND STUDIES

Related Literature 10
Related Studies 17
Synthesis of Reviewed Related Literature and Studies
And Gap to be Bridged by the Study 24
Theoretical Framework 26
Theoretical Paradigm 28
Conceptual Framework 29
Conceptual Paradigm 30
Definition of Terms 31
References 33

Chapter 3 – RESEARCH DESIGN

Research Method 38
Data Sources 38
Data Gathering Procedure 39
Statistical Tools 47

x
 References 49

Chapter 4 – DATA ANALYSIS AND INTERPRETATION

Phytochemical Composition of Tagbak (Alpinia elegans) and 50

Turmeric (Curcuma longa L.) Rhizome
Most Effective Tagbak (Alpinia elegans) and Turmeric (Curcuma
longa L.) Rhizome in Terms of White Blood Cell (WBC)
and Lymphocyte Count 56
Most Effective Tagbak (Alpinia elegans) and Turmeric (Curcuma
longa L.) Rhizome in Terms of Immunoglobulin G (IgG) 61
Significant Difference of the Most Effective Tagbak (Alpinia
elegans) and Turmeric (Curcuma longa L.) Concentration to the
Commercially Available Immune Booster 62
References 69

Chapter 5 – SUMMARY OF FINDINGS, CONCLUSIONS, AND

RECOMMENDATIONS
Summary of Findings 70
Conclusions 72
Recommendations 73

BIBLIOGRAPHY 75

APPENDICES

A. Letters 83
B. Certificates 89
C. Documentations 94
D. List of Tables 116
E. List of Figures 121

BIONOTE

xi
 LIST OF TABLES

Table Title Page

1 Phytochemical Composition of Tagbak (Alpinia elegans)

50
and Turmeric (Curcuma longa L.) rhizome.

2 Most Effective Formulation of Tagbak (Alpinia elegans)

and Turmeric (Curcuma longa L.) Rhizome in
Terms of White Blood Cell (WBC) and 57
Lymphocyte Count

3 Kruskal-Wallis H Test
59

4 Group Descriptive of Treatments (N, Mean, Standard

Deviation, Standard Error) in White Blood Cell
(WBC) and Lymphocyte Count 59

5 Most Effective Formulation of Tagbak (AIpinia elegans)

and Tumeric (Curcuma longa L.) Rhizome in Terms
of Immunoglobulin G (IgG) 61

6 Significant Difference of the Most Effective Tagbak

(Alpinia elegans) and Turmeric (Curcuma longa L.)
Concentration to the Commercially Available 63
Immune Booster (Ascorbic Acid)

7 Mann-Whitney U test for White Blood Cell (WBC) and

Lymphocyte Count [500 mg Tagbak (Alpinia
elegans) and Turmeric (Curcuma longa L.) Capsule 66
and 500 mg Ascorbic acid]

xii
8 Group Descriptive [500 mg Tagbak (Alpinia elegans and
Turmeric (Curcuma longa L.) and 500 mg Ascorbic
acid] in White Blood Cell (WBC) and Lymphocyte 66
Count

xiii
 LIST OF FIGURES

Figure Title Page

1 Theoretical Paradigm 28
2 Conceptual Paradigm 30
3 Tagbak (Alpinia elegans) and Turmeric
(Curcuma longa L.) as an Immune-
booster Capsule 41
4 Procedural Process of Administration of
Ascorbic Acid 44
5 Comparison of Mean White Blood Cell
counts of T3 and T4 after
Acclimatization, Post Prednisone
Induction, and Post Treatment Induction 64
6 Comparison of Mean Lymphocyte counts of
T3 and T4 after Acclimatization, Post
Prednisone Induction, and Post
Treatment Induction 65

xiv
 Chapter 1

THE PROBLEM

Introduction

Due to unexpected changes in the climate and lifestyle, specifically when a person

lacks sleep and exercise, has an imbalanced diet, or is exposed to viruses from social

interactions, they are more susceptible to developing certain diseases. As stated by Cheng

et al. (2020), the Philippines was located in the tropical climate region of Southeast Asia,

and its densely populated human and animal populations were thought to be a source of

novel viruses and global influenza epidemics. In relation to this, Alrehaili (2020)

emphasized that because of these emerging contagious respiratory diseases, people gained

awareness of the importance of taking natural immune boosters along with preventive

measures including correcting lifestyle habits, seasonal vaccinations, and other proper

practices to avoid the spread of viruses.

The use of an immune booster is more valued by people nowadays than before

because of the existing contagious respiratory diseases such as common colds and

influenza that primarily affect and weaken the immune system. In a Rakuten Insight survey

from March 2022, respondents in the Philippines reported that 82% were taking dietary

supplements or nutraceuticals to boost and strengthen their immune systems. The increased

awareness of how immune boosters prevented oneself from acquiring these diseases

provided them with an understanding of the need to take immune boosters as part of their

primary prevention. According to Data Bridge Market Research (2021), in the forecasted
 2

period of 2021 to 2028, the market for immune system boosters is anticipated to develop

at a rate of 7.40%. The increasing prevalence of infectious and viral diseases globally has

enabled the growth of the market for immune booster supplements as demand continues to

rise.

Most people found it more efficient to use an immune booster made up of herbal

plants because of the idea that herbal medicine was "organic,” which they believe is more

effective because it does not have harmful effects on the body system than synthetic

medicines that are commercially available. According to Nisar et al. (2018), the majority

of people in rural areas had faith in herbal medicinal plants because they could treat any

disease without any lethal effects, and they also based their beliefs on their experiences

using herbal medicinal plants. In addition, people patronized the use of herbal medicine

because it was less expensive and less toxic than synthetic drugs. Traditionally, herbal

plants were widely and frequently used in most places, as they were easily accessible and

abundant locally. Due to the long-standing practice of utilizing herbal medicine, it is now

gaining attention locally as well as globally. The continuous transition to a more advanced

health and medical system allows herbal plants to be recognized. With the pharmacological

activities of these herbal plants, the advancement of technology would help formulate and

enhance these raw materials, which had the potential for greater good in terms of

prevention and treatment of mild illnesses.

Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) were both rhizomatous

herbaceous plants from the ginger family Zingiberaceae and were both native to South Asia

and widely distributed in the Philippines. Tagbak (Alpinia elegans) was mostly found in

wild forests, while Turmeric (Curcuma longa L.) was not found in the wild as it was
 3

cultivated indoors or in greenhouses; both conservation statuses were not threatened

because they are easy to grow. Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.)

were widely used as herbal remedies, and there was a lot of research and studies that

mentioned their properties. Tagbak (Alpinia elegans) had immune- stimulating effects;

anti-bacterial, anti-cancer, and antioxidant properties; anti-proliferative effects; and was a

potential treatment for osteoarthritis and inflammation. Turmeric (Curcuma longa L.) has

antiviral, antibacterial, and antimicrobial properties for immune boosters, as well as

antiprotozoal, anti-venom, anti-malarial, anti-inflammatory, anti- proliferative, anti-

angiogenic, anti-tumor, and anti-aging properties. Both were the best choices as immune

boosters, as the phytochemical composition of Tagbak (Alpinia elegans) rhizomes

contained constituents of triterpenes, flavonoids, saponins, glycosides, and tannins, while

Turmeric (Curcuma longa L.) contained constituents of saponins, flavonoids, tannins,

triterpenes, alkaloids, curcumin, and vitamin C

Considering the world's current situation with the rising incidence of viral

infections, many Filipinos were unable to protect themselves because they could not afford

dietary supplements and products because they were expensive. This prompted the

researchers to conduct a study about the development of Tagbak (Alpinia elegans) rhizome

and turmeric (Curcuma longa L.) capsules as an immune booster that aimed to improve the

immune system as a first line of prevention against acquiring the disease. The formulated

capsule from the raw materials, which were Tagbak (Alpinia elegans) and turmeric

(Curcuma longa L.), aspired to produce a more cost-efficient immune booster than

commercially available brands that still upheld safety and effectiveness. Making it
 4

affordable and accessible to all, especially immunocompromised individuals, and poor

groups of people.

Objectives of the Study

This study was intended to determine the potency of Tagbak (Alpinia elegans)

Rhizome and Turmeric (Curcuma longa L.) capsule as an immune booster, which

investigated:

1. To identify the phytochemical composition of Tagbak (Alpinia elegans) and

Turmeric (Curcuma longa L.) rhizome.

2. To determine the most effective formulation of Tagbak (Alpinia elegans) and

Turmeric (Curcuma longa L.) rhizome in terms of White Blood Cell (WBC) and

Lymphocyte count with a concentration of:

500 mg

a. 60% of Tagbak; 40% of Turmeric

b. 40% of Tagbak; 60% of Turmeric

c. 50% of Tagbak; 50% of Turmeric

3. The most effective formulation of Tagbak (AIpinia elegans) and Tumeric

(Curcuma longa L.) rhizome in terms of Immunoglobulin G (IgG).

4. To compare the significant difference between the most effective Tagbak (Alpinia

elegans) and Turmeric (Curcuma longa L.) rhizome formulation to commercially available

immune booster.
 5

Hypothesis

H0 (1): There is no significant difference between the three treatment concentrations of

Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) capsule.

H0 (2): Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) capsule had no

significant difference from the commercially available immune booster depending on the

blood serum composition of albino mice in terms of efficacy.

Scope and Delimitations

This experimental study focused on the use of Tagbak (Alpinia elegans) and

Turmeric (Curcuma longa L.) rhizome capsules and identified their phytochemical

composition as immune boosters. Tagbak (Alpinia elegans) and Turmeric (Curcuma longa

L.) rhizomes were the only raw materials used in the study. This study utilized fifteen (15)

male albino mice weighing 15 to 25 grams with an age of 12 weeks as subjects for testing

the efficacy of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) capsules as

immune boosters. The researchers conducted several blood tests using three parameters

that included White Blood Cell (WBC) count, Lymphocyte count and Immunoglobulin G

(IgG) to determine the effectiveness of the formulation.

The study was delimited to not use the stem, flower, roots, and fruits of both plants

since previous studies had already utilized those parts. Diseases such as pneumonia and

tuberculosis were not included in the study since the researchers focused only on boosting

the immune system.

 6

This research was conducted at Bicol University Polangui Campus during the first

semester of the academic year 2022-2023.

Significance of the study

The gathered findings in this study will benefit the following:

Immunocompromised Individuals. At the end of the experimentation, the

outcome in the form of a capsule will be beneficial to them. The immune booster capsule

can help strengthen their immune systems.

Community. This study can provide awareness and knowledge to the community

by providing them with information that Tagbak (Alpinia elegans) and Turmeric (Curcuma

longa L.) rhizomes can be used as an alternative immune booster. This study will lead the

townspeople to maximize the utilization of local resources, which will in turn lead to the

socio-economic improvement of the community.

National and Local Government. This research will enlighten national and local

governments on the medicinal properties of Tagbak (Alpinia elegans) and Turmeric

(Curcuma longa L.), allowing them to assist in the propagation of more Tagbak (Curcuma

longa L.) and Turmeric (Curcuma longa L.) plants.

Department of Health (DOH). The findings of the study will provide the

Department of Health information that Tagbak (Alpinia elegans) and Turmeric (Curcuma

longa L.) rhizomes have immune-boosting properties. They may recommend the

combination of these two as a much more cost-efficient and accessible alternative to

commercially available immune booster capsules.

 7

Department of Agriculture (DA). If the study proves that Tagbak (Alpinia

elegans) and Turmeric (Curcuma longa L.) rhizomes have immune-boosting properties, it

will promote the integration of research and development functions and enhance farmer

participation.

Philippine Institute of Traditional and Alternative Health Care (PITAHC).

This research will help the organization achieve its goal of developing and implementing

traditional and alternative healthcare in the country. It includes the recognition of

traditional and complementary medicine as legitimate medical fields.

Future Researchers. This study will serve as an input to future experimental

research and studies by further exploring the medicinal uses of Tagbak (Alpinia elegans)

and Turmeric (Curcuma longa L.).

 8

References

Alrehaili, A. (2020). Assessing the Understanding and Perception of the Significance of

Seasonal Immune Care among Taif Population during the Pilgrimage Seasons:

Emphasis on Influenza and Meningitis Vaccines.

https://pdfs.semanticscholar.org/1faf/73b60ed4d3d3132fecc8e3e0a74bdacc5626

Amalraj, A., Pius, A., & Gopi, S. (2017). Biological activities of curcuminoids, other

biomolecules from turmeric and their derivatives – A review. Science Direct.

https://www.sciencedirect.com/science/article/pii/S2225411016300293#bib2

Cheng, K.J.G., Rivera, A.S., Lam, H.Y., Ulitin, A.R., Nealon, J., Dizon, R., Wu, D.B.

(2020) Influenza-associated excess mortality in the Philippines.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7299398/

Immune system booster market – Global industry trends and forecast to 2028 | Data bridge

market research. (n.d.). Market Research Business Consulting and Strategy

Planning Firm | Data Bridge Market Research Private Ltd.

https://www.databridgemarketresearch.com/reports/global-immune-system-

booster-market

Nisar, B., Sultan, A., Rubab, S.L. (2018). Comparison of Medicinally Important Natural

Products versus Synthetic Drugs-A Short Commentary, 6.

https://www.iomcworld.com/open-access/comparison-of-medicinally-important-

natural-products-versus-syntheticdrugsa-short-commentary-2329-6836-1000308
 9

Philippines: Reasons for taking dietary supplements 2022. (2022, June 22). Statista.

https://www.statista.com/statistics/1183261/philippines-reasons-taking-dietary-

supplements/
 Chapter 2

REVIEW OF RELATED LITERATURE AND STUDIES

This chapter included gathered related literature and studies both from local and

foreign sources that helped the researchers in any aspect. Incorporated the synthesis,

conceptual and theoretical frameworks, conceptual and theoretical paradigm, the definition

of terms, and also the references for each gathered data.

Related Literature

Medicines were generally expensive and not accessible to individuals who were

part of the underprivileged group. It was hard for them to buy medicine in the pharmacy

because most of them are not capable due to lack of money and this results in the

progression of their illnesses and diseases. With this concern, the lawmakers recognized

the needs of the underprivileged groups and established the Republic Act No. 9502 or the

Universally Accessible Cheaper and Quality Medicines Act of 2008 which is an act

providing for cheaper and quality medicines. Additionally, as an alternative to expensive

medicines, underprivileged groups made use of herbal plants as their medicine. And these

were some of the herbal plants that are recognized by the Department of Health (DOH)

which includes Lagundi (Vitex negundo), Yerba Buena (Mentha villosa), Sambong

(Blumea balsamifera), Tsaang gubat (Ehretia microphylla), Niyog-niyogan (Quisqualis

indica L.), Bayabas (Psidium guajava L.), Akapulko (Senna alata), Ulasimang bato

(Peperomia pellucida), Bawang (Allium sativum), and Ampalaya (Momordica charantia).

Jointly, the Republic Act No. 8423 or the Traditional and Alternative Medicine Act
 11

(TAMA) of 1997 was responsible for the development of traditional and alternative health

care modalities such as herbal plants that were proven to be safe, effective, cost-efficient

and consistent with government standards on medical practice. Therefore, the researchers

came up with this study that aimed to help individuals achieve good health by having a

boosted immune system.

The Immune System and Conditions Related to the Immune System. Our body

consists of systems that function together and one of which is the immune system. The

immune system played a crucial role in defending the body against pathogens, toxic

chemicals, and cell alterations that made a person ill. The body's immune system fights

disease-causing microorganisms or pathogens such as bacteria, viruses, parasites, or fungi,

and expels them from the body. It also detected and destroyed harmful elements in the

environment; and to combat bodily modifications that cause disease such as cancer cells,

according to Informed Health (2020). Severe combined immunodeficiency (SCID), or an

immune deficiency that was present at birth and puts children at risk for infections from

bacteria, viruses, and fungi; temporary acquired immune deficiencies, like those caused by

illnesses like the flu virus, mono (mononucleosis), and measles; human immunodeficiency

viruses (HIV), and acquired immunodeficiency syndrome (AIDS) were common immune

system disorders that were mentioned in John Hopkins Medicine (2021).

Propagation of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.).

Tagbak (Alpinia elegans) required a larger area to cultivate because of its taller and larger

growth process, growing to 2 to 3 meters tall. It was commonly harvested from the wild in

the local forests of the Philippines. It had taken about 12 months to harvest. This was the

only estimated harvest of Tagbak (Alpinia elegans) rhizomes. Due to its location in wild
 12

forests, monitoring the number of months that this plant grows was not accurate. Most of

the harvested Tagbak (Alpinia elegans) were full grown and already bearing fruits (Mendez

& Acma, 2018). The process of propagation for the plant Tagbak (Alpinia elegans) was the

same as in the Turmeric (Curcuma longa L.). However, Turmeric (Curcuma longa L.) was

conventionally propagated through underground rhizomes, where rhizomes and roots were

cultivated at the end of its season. Storing the fresh rhizomes in a cool, dark and dry place

helped its sprouts to grow more. With that, it would be ready to be planted in the soil,

adding organic matter in the form of well-rotted manure, compost, or mulch throughout

the growing season, or fertilizing as necessary, could provide turmeric with the high levels

of nutrients it needs. Additionally, depending on the particular variety, it would take

between 6- and 9-months following planting for turmeric to be ready for harvest and

processing (Ryczkoski, 2021).

Rhizome. Rhizome was the most commonly used part of the plant as herbal

medicine, especially those plant species from the Zingiberaceae family. Tagbak (Alpinia

elegans) and Turmeric (Curcuma longa L.) were both from the Zingiberaceae family and

were known for their pharmacological properties. Tagbak (Alpinia elegans) rhizome was

primarily used by the community through decoction to relieve their ailments (Dalisay, et

al., 2018). Turmeric (Curcuma longa L.) rhizome was where the most active medicinal

ingredients were found, which was the foundation for several clinical studies on the health

benefits of turmeric (Brittany, 2022). In this study, rhizomes of Tagbak (Alpinia elegans)

and Turmeric (Curcuma longa L.) were used as the main ingredients in the formulation of

the capsule.
 13

Tagbak (Alpinia elegans) as Herbal Remedy. Tagbak (Alpinia elegans) came

from the plant family called Zingiberaceae and were widely distributed in the tropics of

Southeast Asia. It could also be found along streams at low or medium altitudes (Stuart,

2022). Researches and studies found out that plant Tagbak (Alpinia elegans) contains

antioxidant properties (Naive, et al., 2019); anti-cancer and anti-proliferative (Lintao and

Medina, 2021); and a treatment for osteoarthritis or anti-inflammatory (Tumaneng and

Castillo, 2021). With this, the researchers will utilize the rhizomes of the Tagbak (Alpinia

elegans) as a raw material and turn it into a capsule to be used to strengthen the immune

system.

Turmeric (Curcuma longa L.) as Herbal Remedy. Turmeric (Curcuma longa L.)

was a plant from the ginger family Zingiberaceae, which was native to Southeast Asia and

grown commercially in that region. In the country of India, Turmeric (Curcuma longa L.)

was traditionally used for disorders of the skin, upper respiratory tract, joints, and digestive

system according to the National Institutes of Health (2022). The plant Turmeric (Curcuma

longa L.) was said to be filled with the substance called curcumin, which was a powerful

antioxidant that helped the body's repair and keep inflammation away (The Medical City,

2021). Additionally, it also contains properties such as antioxidant, anti-protozoal, anti-

microbial, anti-venom, anti-microbial, anti-malarial, anti-inflammatory, anti-proliferative,

anti-angiogenic, anti-tumor, and anti-aging. The researchers came up with the decision to

combine Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) to develop a more

effective way to help increase the immune system.

Use of Vitamin C as an Immune Booster. A person having lowered defenses

against infection increases a person's vulnerability to certain illnesses. It was mentioned by

 14

the Cleveland Clinic (2022), that one of the most effective immune system boosters was

vitamin C. According to Carr and Maggini (2017), vitamin C supports different cellular

processes of the innate and adaptive immune systems which aids in immune protection.

The use of vitamin C was further enumerated by Raman (2020), first, vitamin C helps in

production of Lymphocytes and phagocytes, two types of white blood cells that aid in

defending the body against infection. Second, vitamin C both improved the efficiency with

which these white blood cells perform as well as defending them against injury from

potentially hazardous substances like free radicals. Thirdly, vitamin C was crucial to the

skin's defense mechanism. It was actively carried to the skin, where it can function as an

antioxidant and support the skin's protective barriers. Vitamin C supplements were said to

be the most effective immune booster because it happens to be a free radical scavenger.

Free radicals were simply molecules that attacked the body and lowered its immunity.

Different Tests to Determine Immune Booster. An important part of assessing

the immune system was laboratory testing. One common laboratory test was the conduct

of blood tests. Blood tests could assess the quantities of blood cells and immune system

cells as well as to ascertain whether an individual has normal levels of the infection-

fighting proteins known as immunoglobulins. Blood cell counts that fall outside of the

normal range could be a sign of an immune system problem as stated by the Immune

Deficiency Foundation (2022). In this study, the researchers decided to use blood tests in

determining if the immune system of the male albino mice had changed after the intake of

the combination of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) immune

booster capsules.
 15

Immunoglobulin G (IgG) and White Blood Cell (WBC) Test. Immunoglobulin

makes antibodies in order to fight with bacteria, viruses, and allergens in the body that

lower the immune system. Immunoglobulin G (IgG) was a specific and common type of

antibody that could be found in blood and other body fluids that protects against bacterial

and viral infections. An immunoglobulin G (IgG) test helped indicate immunodeficiencies.

It could be suspected that a person had immunodeficiency if there was also a presence of

infections. Symptoms such as diarrhea, fever, skin rash, and weight loss might indicate

infection and low immunity. Low levels of immunoglobulin means that the immune system

was weak and had an infection caused by bacteria and viruses. High levels of

immunoglobulin means that there was an allergic reaction and autoimmune disorders

(WebMD, 2021). On the other hand, the White Blood Cell Count monitors the quantity of

white blood cells in the blood, which aids the body in warding off infections and other

disorders, according to the National Library of Medicine (2021). For instance, when a

person has fallen ill, the body produces more white blood cells to combat the bacteria,

viruses, or other foreign substances that were causing the sickness; this increases the white

blood cell count. On the other side, several cancers and viral infections like human

immunodeficiency viruses (HIV), and acquired immunodeficiency syndrome (AIDS)

could make the body produce fewer white blood cells than was necessary, decreasing the

white blood count. Therefore, white blood cells were also a crucial component of the

body’s immunological response.

Male Albino Mice. The use of laboratory mice in most experimental research had

become an inclination both in local and international testing. Laboratory mice played a big

role in the creation of new medications, therapies, and preventative measures in addition
 16

to assisting researchers in deciphering human genomes. According to Charles River

Laboratories (2022), some reasons why laboratory mice were widely used in research

includes the following, first, lab mice could be inbred through brother or sister mating to

yield genetically uniform strains for more accurate and reproducible experiments; second,

lab mice could be genetically engineered to model virtually any human disease or

condition; third, lab mice had an accelerated lifespan one mouse year equals approximately

30 human years, making it faster to study a life cycle and age-related conditions; fourth,

they were well characterized and widely utilized by the biomedical research community;

and lastly, they were small, quick to reproduce, and relatively easy to handle and transport.

That was why, in this experimental study, the researchers made the decision to utilize

laboratory mice as subjects for testing for each formulation in order to gather accurate

results by the end of the experimentation period that would help in the success of the study.

Prednisone as Immunosuppressant. Prednisone is a glucocorticoid medication

which is effective and commonly used as anti-inflammatory and immunosuppressive

agents. Steroid drugs, such as prednisone, work by lowering the activity of the immune

system. The immune system is the body’s defense system. Steroids work by slowing the

body’s response to disease or injury. Prednisone can help lower certain immune-related

symptoms, including inflammation and swelling, because prednisone inhibits the immune

system, it decreases the ability to fight infection and contributes to slow wound healing

(National Kidney Foundation, 2023). In this study, the researchers used Prednisone as a

causative agent because it caters to what the researchers need and that was to lower the

immune system of the male albino mice.

 17

In this experimental study, a capsule was formulated from the rhizome of Tagbak

(Alpinia elegans) and Turmeric (Curcuma longa L.) that aimed to boost the immune

system. With a strong immune system this could help reduce the likelihood of developing

other health problems during this time because diseases that were caused by a weak

immune system could contribute to vulnerability to certain diseases such as common colds

and influenza. Both Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) rhizomes

underwent phytochemical analysis to ascertain its immune boosting property. These

ingredients went through preparations and processes including washing, chopping, air

drying, grinding, sifting until it became a powder. Formulation of Tagbak (Alpinia elegans)

and Turmeric (Curcuma longa L.) rhizome was then tested to male albino mice assessing

its effectiveness by significant results on the set biomarkers involving White Blood Cell

(WBC) and Lymphocyte count as well as Immunoglobulin G (IgG).

Related Studies

Improving the body's immune system were identified in the means of practicing a

healthy living such as engaging in regular physical exercises, eating a healthy balanced

diet, reducing stress, and avoiding vices. In addition, improving the body's immune system

was also through immune boosters, which were popularly known as consuming nutritional

supplements including vitamins, antioxidants, probiotics, and foods such as fruits and

vegetables. These immune boosters were simply recognized by lay persons as

supplementary materials that can boost their immune system (Macedo, et. al., 2019).

In the study of Jiang, et al. (2021) entitled “Potential Application of Plant-Based

Functional Foods in the Development of Immune Boosters” stated that plant-based

 18

functional foods contain an active ingredient that could help strengthen immunity. These

active ingredients were bioactive secondary metabolites including the saponins,

polysaccharides, flavonoids, and alkaloids. It was further discussed in the study that

saponins extracted in plants had demonstrated anti-tumor and immune-enhancing

regulatory properties. Flavonoids that were commonly found in plants can boost the

cytotoxic T cells, and the destructive activities of the NK cells, which results in the release

of cytokines and would improve the immune organ index, as well as improve the

functionality of the immune system. Polysaccharides also provided promising immune

regulating effects which was enhanced by its activation of antigen-specific immune

systems. Moreover, polysaccharides possess anti-tumor and anti-inflammatory activities.

Alkaloids was known for its role in enhancing immunity by managing the proliferation of

thymic and splenic lymphocytes as well as secretions of cytokines.

The research of Lintao and Medina, (2021), entitled “Screening for Anticancer

Activity of Leaf Ethanolic Extract of Alpinia elegans (“tagbak”) on Human Cancer Cell

Lines'', concluded in the results that in the phytochemical analysis of Tagbak (Alpinia

elegans) leaf, it showed a multitude of phytochemical traces of alkaloids, flavonoids,

saponins, glycosides and tannins, abundant amounts of triterpenes, and moderate number

of sterols, in the crude ethanolic extraction of the Tagbak (Alpinia elegans) leaf.

Furthermore, it was concluded in this research study that Tagbak (Alpinia elegans) leaf

extract can exhibit anti-proliferative activity which claim for its anticancer activity.

Additionally, a study about the scavenging effects of Tagbak (Alpinia elegans)

leaves conducted by Naive et al. (2019) entitled “Free radical scavenging effects of the

Philippine endemic medicinal plant Alpinia elegans (Zingiberaceae), concluded that the
 19

leaves of Tagbak (Alpinia elegans) revealed a presence of secondary metabolites including

the heavy composition of flavonoids, steroids, and tannins; and moderate amount of

saponins, alkaloids, and cyanogenic glycosides, but an absent anthraquinone. It was also

addressed in this study that the leaves of Tagbak (Alpinia elegans) have the potential to be

an antioxidant because of its phytochemical compounds that had scavenging effects. This

study compared the ethanolic extracts of Tagbak (Alpinia elegans) leaves to L-ascorbic

acid as a benchmark for the scavenging action. However, the result showed that Alpinia

elegans ethanolic extract was twenty times lower compared to the L-standard ascorbic acid

in terms of its antioxidant activity. The process used in this study to differentiate the two

materials was through inhibition of DPPH radical with different concentrations.

Along with this, another study was conducted that researched about the potential of

Tagbak (Alpinia elegans) leaf as a prevention for cartilage degradation. The study was

conducted by Tumaneng and Castillo, (2021) entitled “Immunoregulatory and

Chondroprotective Activity of Philippine Endemic Plant Alpinia elegans (C. Presl) K.

Schum Leaf Extracts against Osteoarthritis”, it was also included in the study the utilization

of crude ethanolic extract and sub-extracts of Alpinia elegans leaves. Hydroxyl radical and

nitric oxide scavenging assays, as well as hydrogen peroxide, were subject in-vitro testing

of the leaves of Tagbak (Alpinia elegans) in rats induced with proteoglycan loss in

cartilages. It was shown in the results that the ethyl acetate and crude ethanolic extract of

Tagbak (Alpinia elegans) leaf could be a therapeutic alternative for the development of

osteoarthritis treatment. In addition, with the undergone different assessment and analysis

of the Tagbak (Alpinia elegans) leaf extracts, this study also found out that it contained an

accepted compound particularly the flavonoids and polyphenols which were good
 20

regulators of antioxidants and cytokines that were important in immune system

maintenance.

The study of Dalisay et al. (2018) entitled “Taxonomic Studies and Ethnomedicinal

uses of Zingiberaceae in the Mountain Ranges of Northern Antique, Philippines”, asserted

that in northern Antique there were numerous species of Zingiberaceae including the

Tagbak (Alpinia elegans), and were traditionally utilized by Antiqueños to alleviate and

cure certain diseases. It was said in the study that the rhizome of the Tagbak (Alpinia

elegans) was the part used and prepared through decoction. According to the Antiqueños

they used the Tagbak (Alpinia elegans) as a cure for headache, migraine, stomachache,

postpartum - relapse (binat), and a cough reliever by drinking the boiled rhizome extracts.

The research study of Barbosa and Minguillan (2020), entitled “Antioxidant

activity and total phenolic content of fresh and cured rhizomes of Turmeric Curcuma longa

L. and Etlingera philippinensis”, studied the total antioxidant activity of fresh and cured

Turmeric (Curcuma longa L.) rhizome using the phosphomolybdenum method. Upon

using the method, it was shown in their result that the total antioxidant activity of Turmeric

(Curcuma longa L.) rhizome was higher when it was prepared cured rather than fresh. The

study concluded that the total antioxidant activity of the Turmeric (Curcuma longa L.)

rhizome depends on the process of preparation.

The research study of Chanda and Ramachandra (2019), entitled “Phytochemical

and Pharmacological Importance of Turmeric (Curcuma longa L.)”, applied the method of

drying and powdering using the turmeric rhizome. To ascertain the saponin and flavonoid

content of turmeric rhizome, Zn and foam tests were used to analyze these phytochemical
 21

compositions. The researchers of this study concluded that saponin and flavonoids made

up the majority of turmeric. Moreover, it was asserted in this study that turmeric has anti-

inflammatory and antioxidant properties due to these phytochemical components, which

are comparable to those of Vitamin C and E.

In connection with this, the study of Jyotirmayee and Mahalik (2022) entitled “A

review on selected pharmacological activities of Curcuma longa”, asserted that the

curcumin component of turmeric was considered as the most potent known antioxidant. It

had a fat-soluble extract that has an equivalent antioxidant activity to Vitamins C and E.

Curcumin assists in preventing and reducing the risk of heart disease, high blood pressure,

macular degeneration, and high cholesterol. It possessed antioxidant, immunomodulatory,

and anti-inflammatory effects suggesting that these special plant materials were potential

therapeutic agents for treating oxidative, immune dysregulation, and inflammatory

disorders. Furthermore, it was also included in this reviewed study the application of

turmeric to immunosuppressed albino mice to test the efficacy of curcumin in the immune

system. The mice were sensitized with SRBCs acquired from sheeps, which were given

orally with varied dosage of curcumin. The experiment utilized the ethanolic extract of

turmeric, and tested in albino mice for its immunomodulatory activity, which had a good

response right away after administration. Due to this, it was concluded that curcumin had

immune-enhancing and anti-inflammatory effects.

Additionally, the study of Permatananda et al. (2021) entitled “Phytochemical and

antioxidant capacity test on turmeric extract (Curcuma longa) traditionally processed in

Bali'' revealed that in the Quantitative phytochemical tests that the plant extracts included

the presence of 67.38% starch, 3.42% protein, 2709.39 mg/100 gr flavonoids, tannins
 22

291.64 mg/100 gr, phenol 1584.04 mg/100 gr, and vitamin C 0.06 mg/100gr. Qualitatively,

turmeric extract contained triterpenes, alkaloids, and saponins, but did not contain steroids.

The antioxidant capacity of turmeric extract was 70.9 mg/L GAEAC. Turmeric extract is

a traditional medicine made from nature that is most commonly used by almost everyone

and has the potential to be developed as an antibacterial, antioxidant, anti-inflammatory,

or other benefits that still need further investigation.

Ascorbic acid was employed as a positive control in this study to show the

difference in effectiveness between the Tagbak (Alpinia elegans) and Turmeric (Curcuma

longa L.) immune booster capsule. The right delivery of the preparations and the ascorbic

acid to the subject’s male albino mice must be determined in order to produce an accurate

result of the effectiveness of the materials. Therefore, gaining additional information

regarding the ascorbic acid oxidation process was essential.

With reference to the study of Njus et al. (2020), entitled “Ascorbic Acid: The

Chemistry Underlying Its Antioxidant Properties”, stated that 35% of Vitamin C ascorbic

acid added in tap drinking water was oxidized into dehydroascorbic acid which was a form

of oxidized ascorbic acid, within 15 minutes. And after a 3-hour incubation period at room

temperature, 93% of added 2mM (Millimolar) ascorbic acid has been oxidized. However,

in commercially sold domestic bottled water samples, it revealed that ascorbic acid did not

result in rapid oxidation of Vitamin C.

The research study of Biancatelli et al. (2020) entitled “Quercetin and Vitamin C:

An Experimental, Synergistic Therapy for the Prevention and Treatment of SARS-CoV-2

Related Disease (COVID-19)”, asserted that different murine models were able to survive
 23

fatal infections because of Vitamin C. (1) The mice infected with Venezuelan encephalitis

virus had lessened its potential mortality and lowered the concentration of virus in its body

because of ideal Vitamin C administration of 50 mg/kg. (2) The mice which were unable

to synthesize the Vitamin C were the ones infected with influenza. (3) The mice which did

not get any supplemental Vitamin C concluded for higher lung pathology score. (4) The

mice with a virus-induced pneumonia and supplemented with Vitamin C, results in

decreased capillary-alveolar structural damage. Included also in this study was the ideal

number of Vitamin C administration in those herpes zoster patients. 5 g/day twice/week

given intravenously was the required amount of administration. While in influenza-

associated pneumonia patients, a 300 mg/day of Vitamin C supplementation was said to

reduce the hospital stays of these patients. However, the average and ideal oral

administration of Vitamin C in adult humans is 70 to 150 mg daily.

Another study was conducted entitled “Combination of Vitamin C and Curcumin

Safeguards Against Methotrexate-Induced Acute Liver Injury in Mice by Synergistic

Antioxidant Effects” studied by Khudhair et al. (2022). In this research study, the duration

of the experiment was 10 days, and the intervention drugs were given from 1 to 10 days.

Different sample groups of mice were experimented using different administrations of

Methotrexate, and Curcumin in combination with Vitamin C, wherein oral administration

using gavage feeding was also included. This study concluded that Vitamin C and

Curcumin together had a synergistic hepatoprotective effect that was superior to

pretreatment with Curcumin alone but appeared to be comparable to 200 mg/kg/day of

Vitamin C. Therefore, a combination of Vitamin C and Curcumin or a high dose of Vitamin

C might be beneficial against hepatotoxicity brought on by Methotrexate.

 24

Synthesis of Reviewed Literature and Studies and

Gap Bridged by the Study

The potential application of plant-based functional foods in the development of

immune boosters was studied by Jiang (2021), and it revealed that plants that contain active

ingredients such as saponins, polysaccharides, flavonoids, and alkaloids can strengthen a

person's immune system. Research by Lintao and Medina (2021), showed a multitude of

phytochemical traces in the leaf extract of Tagbak (Alpinia elegans) that exhibited anti-

proliferative activity. Additionally, in the study conducted by Naive et al. (2019), the

antioxidant property of Tagbak (Alpinia elegans) leaf extract. Another study by Tumaneng

and Castillo (2021), utilized Tagbak (Alpinia elegans) leaf extract that showed its potential

as an alternative for osteoarthritis treatment and a good antioxidant. According to Dalisay

et al. (2018), Tagbak (Alpinia elegans) rhizomes were traditionally used by the Antiqueños

through decoction to alleviate and cure their headache, migraine, stomachache, postpartum

relapse, and cough. The use of Tagbak (Alpinia elegans) leaf extract was used by several

researchers and there was also a study that showed the traditional utilization of Tagbak

(Alpinia elegans) rhizomes through decoction which cures certain diseases.

Studies about the plant, Turmeric (Curcuma longa L.) revealed that it contains

several properties that particularly could contribute to improving the health of humans. The

research study by Chanda and Ramachandra (2019), entitled “Phytochemical and

Pharmacological Importance of Turmeric (Curcuma longa L.) concluded that turmeric

contains saponins and flavonoids by using Zn and foam test which determines these

phytochemical components. Therefore, it was asserted in this study that turmeric had anti-

inflammatory and antioxidant properties. Supported by the study conducted by

 25

Jyotirmayee and Mahalik (2022) entitled “A review on selected pharmacological activities

of Curcuma longa”, emphasized that the principal constituent of turmeric, curcumin was

considered as the most potent known antioxidant. Additionally, it was mentioned that

turmeric and its curcumin components, the water and fat-soluble extracts had an equivalent

antioxidant activity to vitamins C and E. The curcumin of turmeric also assists in

preventing and reducing the risk of heart disease, high blood pressure, macular

degeneration, and high cholesterol, including its anti-inflammatory and antioxidant

activity. It was shown that both Tagbak (Alpinia elegans) and Turmeric (Curcuma longa

L.) had the potential properties to be an immune booster supported by the different

literature and studies mentioned above. In addition, there was no literature and studies that

explored the combination of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.).

The synopsis of all collected related literature and studies allowed the researchers

as well as the readers to have an in-depth comprehension of this study. In the Philippines,

the practice of traditional medicine is widely used. These herbal plants were being used as

an alternative to treat illnesses and diseases. Through the presented literature and studies,

the abovementioned plants had a potential in the development of immune boosters.

Immune boosters are products that claim to be able to contribute to strengthening the

immune system of individuals. In relation to this study, the researchers used endemic plants

which have healing properties. Specifically, the rhizomes of Tagbak (Alpinia elegans) and

Turmeric (Curcuma longa L.) both have notable properties recognized by past researchers.

Therefore, in this study, the researchers focused on the immune booster capacity of

the Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.). Considering its essential
 26

the researchers developed a capsule from the rhizome of Tagbak (Alpinia elegans) and

Turmeric (Curcuma longa L.) as an immune booster.

Theoretical Framework

This research utilized the Health Promotion Model of Nola J. Pender published in

1982. This theory noted that every individual had unique personal characteristics and

experiences that would eventually affect their decisions and actions. It aimed to incorporate

health-promoting behaviors in people that would result in improved health, enhanced

functional ability, and better quality of life at all stages. Health promotion, as defined in

the Health Promotion Model, is an approach to wellness in which an individual is motivated

by a desire to improve well-being and actualize full potential in terms of health (Gonzalo,

2021). This theory assisted the researchers in knowing the factors that affect an individual's

health-promotion behavior, particularly the three personal factors: first, personal biological

factors including age, gender, body mass index, pubertal status, aerobic capacity, strength,

agility or balance; second, personal psychological factors such as self-esteem, self-

motivation, personal competence, perceived health status, and definition of health; third,

personal socio-cultural factors including race, ethnicity, acculturation, education, and

socio-economic status. It also aided the researchers in understanding the major

determinants of health behaviors as a foundation for behavioral counseling to promote

well-being and healthy lifestyles.

Denham Harman's Mitochondrial Free Radical Theory, published in 1955, was also

relevant to this research. According to Meinger and Yeager (2019), free radicals react with

molecules in cell membranes, specifically unsaturated lipid cell membranes such as

 27

mitochondria, lysosomes, and nuclear membranes. It took over the membrane's receptor

sites, preventing interaction with other substances that would normally use this site. As a

result, these cell membranes were no longer as efficient, resulting in increased

permeability. When the body loses or gains too much fluid, internal homeostasis is

disrupted, and cell death may result.

Harman (1956) was the first to propose that administering chemicals that prevent

free radical propagation would increase life span or delay the aging process. Animal studies

revealed that antioxidant administration did increase the average length of life, possibly

due to the delayed appearance of diseases that would eventually kill the animals studied. It

appears that taking antioxidants delays the onset of diseases such as cardiovascular disease

and cancer, two of the leading causes of death. Antioxidants appear to influence the decline

of the immune system as well as degenerative neurologic diseases, both of which have an

impact on morbidity and mortality (Meinger & Yeager, 2019).

The paradigm adapted two theories related to the raw materials that were used in

the development of an immune booster capsule, Tagbak (Alpinia elegans) and Turmeric

(Curcuma longa L.). According to the Mitochondrial Free Radical Theory, free radicals

contribute to aging and disease development, whereas antioxidants improve health by

increasing lifespan and delaying the aging process. The raw materials used in this study

have antioxidant properties. The Health Promotion Model reflects the factors that influence

an individual's behavior and assists researchers in incorporating health-promoting

behaviors into the community, resulting in improved health and quality of life.
 28

Figure 1. Theoretical Paradigm

 29

Conceptual Framework

This section describes the conceptual framework that the researchers used to

address the current demands theoretically and practically. The researchers utilized the

Input-Output-Process model to relate the study and generate results. The presentation was

based on the objectives of the study.

As shown in the paradigm, the input involved the related literature, studies, readings

as well as narratives related to Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.).

Also included the phytochemical analysis of Tagbak (Alpinia elegans) and Turmeric

(Curcuma longa L.) which involved Alkaloids, Steroids, Anthraquinones, Flavonoids,

Saponins, and Tannins. For the process, it consisted of data gathering, data analysis, and

data interpretation. Data gathering was comprised of the preparation of subjects for the

experimentation, particularly the male albino mice; harvesting of the Tagbak (Alpinia

elegans) and Turmeric (Curcuma longa L.); washing and chopping, air-drying of rhizomes,

grinding; formulation of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.)

capsule; induction of causative agent; blood test; monitoring of the vital signs specifically

Temperature, Respiratory rate, Heart rate, and behaviors of the mice. The data analysis and

interpretation consisted of results from the phytochemical analysis, blood tests, vital signs,

and behaviors. Lastly for the output, the researchers focused on the development of Tagbak

(Alpinia elegans) rhizome and Turmeric (Curcuma longa L.) capsule as an immune

booster.
 30

Figure 2. Conceptual Paradigm

 31

Definition of Terms

In this part, terms used in this experimental study were defined according to its

conceptual and operational use.

Capsule. According to the U.S. National Cancer Institute (2021), a capsule is a type

of drug packaging that is typically cylindrical in shape with rounded ends. In this study,

out of the best formulation of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.)

the immune booster was produced in the form of a capsule.

Immune booster. According to Harvard Health (2021), products that claim to be

able to support the immune system. In this study, the researchers decided to use blood tests

to determine whether the immune system of the male albino mice had changed after the

intake of the formulation of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.)

immune booster capsules.

Immunoglobulin G Test. According to Nemours Kids health (2020), it measures

the level of types of antibodies in the blood. The immune system makes antibodies to

protect the body from bacteria, viruses, and allergens. In this study, an Immunoglobulin G

(IgG) test was one of the parameters that determined the efficacy of the best formulation

of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) immune booster capsule.

Lymphocyte Count. According to Cleveland Clinic, it was measured as part of the

complete blood count and expressed as a percentage. Lymphocytes are a type of white

blood cell. In this study, lymphocyte count was one of the parameters that indicated the

effectiveness of the formulated immune booster capsule.

 32

Rhizomes. According to Encyclopedia Britannica, a rhizome is a horizontal

underground plant stem capable of producing the shoot and root systems of a new plant. In

this study, the rhizomes of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.)

were the specific parts that were used as the main ingredients in the formulation of the

capsule.

Tagbak (Alpinia elegans). According to the Philippine Medicinal Herb and

Philippine Alternative Medicine (2022), a stout herbaceous plant, growing 2 to 4 meters in

height with stout rootstocks. The stems have a swollen base and are leafy throughout.

Leaves were leathery, spreading, or reflexed. In this study, Tagbak (Alpinia elegans) was

one of the raw materials used to formulate an immune booster capsule.

Turmeric (Curcuma longa L.). According to the National Center for

Complementary and Integrative Health, a type of herb belonging to the ginger family,

widely grown in the southern and south western tropical Asia regions. In this study,

Turmeric (Curcuma longa L.) was one of the raw materials used to formulate an immune

booster capsule.

White Blood Cell (WBC) Count. According to MedlinePlus, it monitors the

quantity of white blood cells in the blood, which aids the body in warding off infections

and other disorders. In this study, white blood cell count was one of the parameters that

indicated the effectiveness of the formulated immune booster capsule.

 33

References

Barbosa, G. B. & Minguillan, J.M.O. (2021). Antioxidant activity and total phenolic

content of fresh and cured rhizomes of Curcuma longa and Etlingera

philippinensis. http://www.ifrj.upm.edu.my

Biancantelli, R.M.L.C., Berill, M., Catravas, J.D., Marik, P.E. (2020). Quercetin and

Vitamin C: An Experimental, Synergistic Therapy for the Prevention and

Treatment of SARS-CoV-2 Related Disease (COVID-19), 11.

https://doi.org/10.3389/fimmu.2020.01451

Blood Test: Immunoglobulins (IgA, IgG, IgM). (2020). KidsHealth.

https://kidshealth.org/en/parents/test-immunoglobulins.html

Brittany, C. (2022). Turmeric Rhizome or Root. HerbaZest.

https://www.herbazest.com/herbs/turmeric/turmeric-rhizome-or-root

Can supplements help boost your immune system? (2020, January 1). Harvard Health.

https://www.health.harvard.edu/staying-healthy/can-supplements-help-boost-

your-immune-system

Capsule. (2022). openMD. https://openmd.com/define/capsule

Chanda, S., & Ramachandra, T.V. (2019). Phytochemical and Pharmacological

Importance of Turmeric (Curcuma longa): A Review, 9.

Dalisay, J.A.G.P., Bangcaya, P.S., Naive, M.A.K., (2018). Taxonomic Studies and

Ethnomedicinal uses of Zingiberaceae in the Mountain Ranges of Northern

Antique, Philippines.

https://www.researchgate.net/publication/327387794_Taxonomic_Studies_and_
 34

Ethnomedicinal_uses_of_Zingiberaceae_in_the_Mountain_Ranges_of_Norther

n_Antique_Philippines

Disorders of the immune system. (2021). Johns Hopkins Medicine, based in Baltimore,

Maryland. https://www.hopkinsmedicine.org/health/conditions-and-

diseases/disorders-of-the-immune-system

Gonzalo, A. (2021). Nola Pender: health promotion model. Nurseslabs.

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Herbal medicine. (2021). Johns Hopkins Medicine, based in Baltimore, Maryland.

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medicine

Herbal medicine plants approved by the DOH. (2017). RNpedia.

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notes/herbal-medicine-plants-approved-doh/

How does the immune system work? - InformedHealth.org - NCBI bookshelf. (2020).

National Center for Biotechnology Information.

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Jiang, L., Zhang, G., Li, Y., Li, Y., Shi, G., Li, M. (2021). Potential Application of Plant-

Based Functional Foods in the Development of Immune Boosters, 12.

www.frontiersin.org

Jyotirmayee, B. & Mahalik, G. (2022). A review on selected pharmacological activities of

Curcuma longa L.25. https://doi.org/10.1080/10942912.2022.2082464

Khudair, D.H., Al-Gareeb, A., Al-kuraishy, H.M., El-Kadem, A.Y., Elekhnawy, E., Negm,

W.A., Saber, S., Cavalu, S., Tirla, A., Alotaibi, S.S., Batiha, G.S. (2022).
 35

Combination of Vitamin C and Curcumin Safeguards Against Methotrexate-

Induced Acute Liver Injury in Mice by Synergistic Antioxidant Effects.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9047671/

National Kidney Foundation. (2023). What You Need To Know About Prednisone.

https://www.kidney.org/content/what-you-need-know-about-prednisone

Lintao, R. & Medina, P.M. (2021). Screening for Anticancer Activity of Leaf Ethanolic

Extract of Alpinia elegans (“tagbak”) on Human Cancer Cell Lines, 22.

Lymphocytes: Function, definition, levels & ranges. (n.d.). Cleveland Clinic.

https://my.clevelandclinic.org/health/body/23342-lymphocytes

Macedo, A., Faria, A., Ghezzi, P. (2019). Boosting the Immunity System, From Science to

Myth: Analysis of Infosphere With Google.

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Meiner, S., & Yeager, J. (2019). Gerontologic nursing (6th ed.). Elsevier.

Mendez, N., & Acma, F.M. (2018). In vitro Studies on Pollen Viability, Pollen

Germination and Pollen Tube Growth of Hornstedtia conoidea Ridl. – a

Philippine Endemic Ginger Species.

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en_Viability_Pollen_Germination_and_Pollen_Tube_Growth_of_Hornstedtia_c

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 36

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children

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 37

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 Chapter 3

RESEARCH DESIGN

This chapter presented the different steps and statistical procedures made in

completing the study. These procedures were not just limited to the kind of research utilized

but also included the sources, data gathering tools and procedures followed in analyzing

the data collected.

Research Method

This study used an experimental design that focused on the effectiveness of the

combination of the rhizomes of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa

L.) to formulate an immune booster capsule. The researchers conducted a qualitative

phytochemical analysis of Tagbak (Alpinia elegans) rhizomes to further and accurately

determine a generous amount of certain phytochemicals containing medicinal uses that

could boost immunity. The researchers then identified which formulation of the Tagbak

(Alpinia elegans) and Turmeric (Curcuma longa L.) combinations was the most effective

in a 500 mg dosage: T1 - 300 mg tagbak, 200 mg turmeric; T2 - 200 mg tagbak, 300 mg

turmeric; T3 - 250 mg tagbak, 250 mg turmeric. The treatments were tested on male albino

mice determining their effectiveness as an immune booster.

Sources of Data

This research used both primary and secondary data sources. During the

experiment, primary data were collected through careful observation of mice behavior,
 39

including increased vigilance, hypoactivity, and suppressed food consumption, as

well as the monitoring of vital signs, specifically temperature and heart rate. Additionally,

phytochemical analysis of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.)

rhizomes and blood test parameters, such as White Blood Cell (WBC) count, Lymphocyte

count, and Immunoglobulin G (IgG), were conducted.

The researchers collected secondary data from sources such as books, related

unpublished undergraduate studies, journals, dictionaries, search engine libraries, and other

relevant publications.

Data Gathering Procedure

To authorize the conduct of the study, the researchers secured a letter to ask

permission from the Dean, Assistant Dean, Head of the Nursing Department and Thesis

Adviser to allow the researchers to investigate in a laboratory test outside the school,

purchase of the male albino mice, and blood serum test in Health Quarters.

Before starting the laboratory testing for the phytochemical analysis, the

researchers harvested 15 kilograms of mature rhizomes of Tagbak (Alpinia elegans) and

Turmeric (Curcuma longa L.) early in the morning at Pintor, Polangui, Albay as the

primary source of raw materials. Then, the researchers verified the Tagbak (Alpinia

elegans) and Turmeric (Curcuma longa L.) rhizomes from the Bicol University College of

Agriculture and Forestry (BUCAF). After the plant verification and validation, Tagbak

(Alpinia elegans) and Turmeric (Curcuma longa L.) underwent manual separation of

rhizomes from the plants. Then washed until all unnecessary dirt was thoroughly removed,

and chopped each rhizome into thin pieces. The chopped rhizomes were hung in a warm,
 40

humid area for air drying. For this to prevent exposure to direct sunlight and to not reduce

the phytochemical properties present in the rhizomes. After 2-3 weeks of air drying, the

desired color and texture were achieved. The rhizomes were grinded using a blender, and

pulverized to the desired outcome, weighing 500 mg then placed into the transparent

gelatin capsule. The study used five (5) treatments with three replicates: negative control

(-) which is a placebo effect; T1 - 300 mg Tagbak, 200 mg Turmeric; T2 - 200 mg Tagbak,

300 mg Turmeric; T3 - 250 mg Tagbak, 250 mg Turmeric; positive control (+) which is a

commercially available immune-booster capsule in 500 mg. After formulation, the

treatments were given once daily (8:00 am) for three to four (3-4) consecutive days to the

stressed, Prednisone induced, and malnourished male albino mice to test the effectiveness

of the formulated Tagbak and Turmeric capsule as an immune-booster. The researchers

conducted a baseline White Blood Cell (WBC) count and Lymphocyte count tests, vital

signs (heart rate and temperature) and behavior monitoring of mice after acclimatization

period to establish a reference point for comparison. After induction of

immunosuppressant, the researchers then again, conducted a WBC count and Lymphocyte

count tests, and monitored the mice’s VS and behavior to ensure that the mice were

immunosuppressed. Then, the researchers conducted a final WBC count and Lymphocyte

count tests, as well as VS and behavior monitoring after administering the treatments to

evaluate the effectiveness of the product in terms of boosting the immunity.

Immunoglobulin G (IgG) parameter was also tested after determining the most effective

treatment concentration. Intensive observation and documentation were done to evaluate

the findings and differentiate the results.

 41

Verification of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.)

In this study, Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) rhizomes

were utilized as raw materials. For the verification and validation of the chosen variety of

Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.), the researchers went to Bicol

University College of Agriculture and Forestry (BUCAF). The said institution verified and

validated the raw materials as Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.)

belonging to the family of ginger Zingiberaceae.

Figure 3. Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) as an Immune-

booster Capsule
 42

Ethanolic Extraction of Tagbak (Alpinia elegans) rhizome

An initial and vital step in producing a quality research output is the preparation of

medicinal plants for experimentation. It entails extracting and determining the quality and

quantity of bioactive elements before proceeding with the desired biological testing.

(Abubakar & Haque, 2020).

The researchers went to Bicol University College of Science (BUCS) Laboratory

to conduct the ethanolic extraction of the Tagbak (Alpinia elegans) and Turmeric

(Curcuma longa L.) rhizomes. Prior to extraction, the Tagbak (Alpinia elegans) rhizomes

were cut into pieces and then soaked in 95% ethanol for 24 hours. During this soaking

period, the soluble components of the extract were separated by the ethanol. After that, the

researchers removed and filtered out all solid materials; only the solution of ethanol and

the extract remained. To separate the ethanol and the extract, the researchers utilized the

rotary evaporator which uses the vacuum to reduce the boiling point of ethanol, resulting

in faster evaporation. The thereby produced extract has been completely free of ethanol

and was used for phytochemical analysis.

The Process of Formulation of Tagbak (Alpinia elegans) and Turmeric (Curcuma

longa L.) Capsule

The Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) rhizomes are the

primary sources and raw materials which serve as the main ingredient in the formulation

of immune booster capsules. The rhizomes were freshly harvested at 7:00 am in Pintor,

Polangui, Albay. Then, manually separated from the other parts of the plants and washed

with running water until all the soil was thoroughly removed and drained. Next, the
 43

researchers chopped each rhizome into thin pieces and were hung in a warm, humid area

for air drying using a net and basket. To preserve their phytochemical properties, the

rhizomes were air-dried in a shaded area for 2-3 weeks, with daily monitoring and

repositioning to ensure even drying. After achieving the desired color and texture, the

rhizomes were ground into a powder using a blender and weighed precisely at 500 mg.

This powdered mixture was then carefully filled into transparent gelatin capsules to create

the immune booster capsules.

Preparation of Ascorbic acid

Ascorbic acid is a Food and Drug Administration (FDA) approved drug that can be

bought with prescription or over-the-counter drugs that is helpful in boosting the immune

system.

Five hundred (500) mg of ascorbic acid were administered to three (3) male albino

mice which are three replicates that were intended for positive control. The ascorbic acid

was diluted to 1 cc of water. Then, the researchers administered the diluted ascorbic acid

through gavage feeding into the male albino mice for accurate treatment.

To determine the significant difference between the commercially available immune

booster to Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) capsule, the data

was obtained from the different parameters including White Blood Cell (WBC) count and

Lymphocyte count as well as the observation and documentation of the behavior and vital

signs of the albino mice after administering the ascorbic acid.

 44

Figure 4. Procedural Process of Administration of Ascorbic Acid

Rearing of Male Albino Mice and Induction of Immunosuppressant

The researchers utilized fifteen (15) male albino mice. Mice were used in the

experiment due to their similarities to humans in terms of anatomical, physiological, and

genetics and also many symptoms of human conditions can be replicated in mice. The mice

were all of the same age of 12 weeks and the same weight that ranges from 15 - 25 grams.
 45

The researchers purchased the male albino mice from MOTS Animal House in Brgy.

Malusak, Santa Rosa, Laguna, and provided with the Certificate of Animal Welfare

Registration. Then, the male albino mice were transported to Polangui, Albay where the

researchers reside and were placed in a clean and safe container provided with enough

space, food, water and air. The fifteen (15) male albino mice underwent a week of

acclimatization to adapt to their new environment. We had five (5) cages for mice and each

cage had three (3) mice.

Prior to the experimentation, the researchers underwent a 1-day seminar and

training for ethical use of laboratory animals in research with the Bicol University Animal

Care and Use Committee (BU ACUC), which tackled the BU ACUC protocol review

process and the animal care and use requirements. The researchers also underwent training

regarding the proper animal handling and restraint, routes of drug administration, methods

of blood collection, and the euthanasia and tissue or organ collection.

After the acclimatization period, the researchers conducted a White Blood Cell

(WBC) count and Lymphocyte count tests, vital signs (heart rate and temperature) and

behavior monitoring (hypoactivity, increased vigilance, suppressed food consumption) of

mice that served as the baseline data. Then, a day after, the researchers induced an

immunosuppressant agent which is Prednisone, a glucocorticoids medication that

suppresses the immune system. To ensure that the mice were immunosuppressed, the

researchers also exposed mice to stress and malnutrition by manipulating their environment

as well as limiting their access to food and water. According to Coutinho & Chapman

(2011), exogenous glucocorticoids or persistent stress promote thymic atrophy, which

suppresses rodent immune systems. They also claim that restricting calories can lower
 46

humoral reactions and T-cell suppressor activity. Protein-calorie deficiency can lower

antibody affinity for antigens. To confirm its immunosuppressant effect, the researchers

conducted a White Blood Cell (WBC) count and Lymphocyte count tests after three (3)

days of immunosuppressant induction and monitored the vital signs and behavior of the

mice.

Experimental Procedure

For the experimentation proper, the researchers utilized fifteen (15) male albino

mice that were grouped into five (5) treatments, with three (3) mice per treatment. Each

mouse was labeled as (-) control A, (-) control B, and (-) control C for negative control

treatment; T1A, T1B, and T1C for treatment 1; T2A, T2B, and T2C for treatment 2; T3A,

T3B, and T3C for treatment 3; and (+) control A, (+) control B, and (+) control C for

positive control treatment.

To test for the immune boosting effect of the formulated immune booster capsules,

each mice beside negative control were induced with 500 mg capsule diluted in a 1 ml of

distilled water with different treatments: T1 - 300 mg Tagbak, 200 mg Turmeric; T2 - 200

mg Tagbak, 300 mg Turmeric; T3 - 250 mg Tagbak, 250 mg Turmeric and a positive

control treatment - commercially available ascorbic acid. The treatments were

administered once daily (8:00 am) after meals for 3-4 days to the stressed, Prednisone-

induced and malnourished male albino mice through gavage feeding with a volume of

0.125-0.25 ml in 25 g mice depending on every mouse’s weight to test the effectiveness of

Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) immune booster capsules. A

final WBC count, lymphocyte count, and Immunoglobulin Test were done after the next

3-4 days of treatment administration. The researchers also monitored and documented for
 47

any possible changes in albino mice's behaviors (hypoactivity, increased vigilance,

suppressed food consumption) and vital signs obtaining the temperature, and heart rate.

Intensive observation and documentation were done to evaluate the findings and

differentiate the results.

Statistical Treatment

Kruskal-Wallis H Test was used by the researchers to determine the most ideal

dosage of the combined Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.)

rhizome. Kruskal-Wallis H Test also called the “one-way ANOVA (Analysis of Variance)

on ranks” is a test that can be used to determine if there were statistically significant

differences between two or more groups of an independent variable on a continuous or

ordinal dependent variable (Laerd Statistics, 2018). This statistical tool was utilized to

determine if there is a significant difference between the three conditions using alpha =

0.05 using the different concentrations.

Kruskal-Wallis H Test formula:

Where,

H = denotation test statistic for Kruskal-Wallis Test

k = the number of comparison groups

N = the total sample size

 48

nj = the sample size in the jth group

Rj = the sum of the ranks in the jth group

After determining the most effective concentration, the researchers utilized the

Mann-Whitney U Test to evaluate the significant difference between the most effective

concentration of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) from the

commercially available immune booster in the market. The Mann-Whitney U Test is used

to compare differences between two independent groups when the dependent variable is

either ordinal or continuous, but not normally distributed (Laerd Statistics, 2018).

Mann-Whitney U Test formula:

Where,

U = denotation test statistic for Mann Whitney U Test

n1 and n2 = the size of the two samples

R1 = sum of the ranks for group 1

R2 = sum of the ranks for group 2

 49

References

Administration Of Drugs and Experimental Compounds in Mice and Rats (IACUC) | Office

of Research. (n.d.). © 2023 Boston University.

https://www.bu.edu/research/forms-policies/administration-of-drugs-and-

experimental-compounds-in-mice-and-rats-iacuc/

Bevans, R. (2022, November 14). An Introduction to t Tests | Definitions, Formula and

Examples. Scribbr. https://www.scribbr.com/statistics/t-test/

Coutinho, A., & Chapman, K. (2011). The anti-inflammatory and immunosuppressive

effects of glucocorticoids, recent developments and mechanistic insights.

Molecular and Cellular Endocrinology, 335(1), 2–13.

https://doi.org/10.1016/j.mce.2010.04.005

Kruskal-Wallis H Test in SPSS Statistics | Procedure, Output and Interpretation of the

Output Using a Relevant Example. (n.d.). https://statistics.laerd.com/spss-

tutorials/kruskal-wallis-h-test-using-spss-statistics.php

Mann Whitney U test (Wilcoxon rank sum test). (n.d.).

https://sphweb.bumc.bu.edu/otlt/mph-

modules/bs/bs704_nonparametric/bs704_nonparametric4.html
 Chapter 4

DATA ANALYSIS AND INTERPRETATION

This chapter presented the discussion, analysis, interpretation, and narratives of the

data collected from the actual experiment. It also included the tables, figures, and graphical

representations regarding the result of the experimentation. The study on the development

of Tagbak (Alpinia elegans) rhizome and Turmeric (Curcuma longa L.) capsule as an

immune booster was accomplished with the following results.

Phytochemical Composition of Tagbak (Alpinia elegans) and Turmeric (Curcuma

longa L.) Rhizome

The phytochemicals of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.)

rhizome as an immune booster capsule were recognized and investigated. The analysis of

phytochemical composition in rhizomes is essential in identifying and verifying the

primary compound present within plant extracts through established protocols. This

assessment was also significant in terms of pharmacological benefits and potential health

risks.

Table 1. Phytochemical Composition of Tagbak (Alpinia elegans) and Turmeric (Curcuma

longa L.) Rhizome

Tagbak (Alpinia elegans) Turmeric (Curcuma longa L.)

Test Parameters Results Test Parameters Results

Sterols - Curcumin +

Triterpenes + Triterpenes +
 51

Flavonoids + Flavonoids +

Alkaloids - Alkaloids +

Saponins + Saponins +

Glycosides + Vitamin C +

Tannins + Tannins +

Table 1 shows the summary of the phytochemical composition of Tagbak (Alpinia

elegans) and Turmeric (Curcuma longa L.) rhizomes. The result of the analysis conducted

in the DOST Laboratory Taguig shows that Tagbak (Alpinia elegans) rhizomes contained

triterpenes, flavonoids, saponins, glycosides, and tannins. Sterols and alkaloids were not

present. A positive (+) result means the presence of constituents, while a negative (-) result

means the absence of constituents. The ethanolic extracts of Tagbak (Alpinia elegans)

rhizome were used to test the presence and absence of constituents using different specific

methods. The essentiality of the evaluation of the phytochemical composition of Tagbak

(Alpinia elegans) was also emphasized in several studies including the study of the

anticancer activity of leaf ethanolic extract of Tagbak (Alpinia elegans) on human cancer

cell lines (Lintao & Medina, 2021). Also, the study about the scavenging effects of Tagbak

(Alpinia elegans) leaves (Naive et al., 2019)

For the analysis of Turmeric (Curcuma longa L.), the researchers did not conduct a

phytochemical testing for turmeric (Curcuma longa L.) rhizome due to several studies that

already conducted research about its phytochemical composition. Saponins, flavonoids,

tannins, triterpenes, alkaloids, curcumin, and vitamin C are the phytochemicals present in

the rhizome of Turmeric (Curcuma longa L.) according to the studies about the
 52

phytochemical and pharmacological importance of turmeric and its antioxidant capacity in

the studies of Chanda and Ramachanda (2019), Permanatanda et al. (2021), and

Jyotirmayee and Mahalik (2022).

The following phytochemical constituents below were present in the Tagbak

(Alpinia elegans) and Turmeric (Curcuma longa L.) rhizomes, and each were provided

with further analysis including the test method used, description, and its benefits in the

immune system.

Triterpenes

Triterpenes belongs to saponin’s compound group. Triterpenes and saponins almost

have the same properties. Triterpenes were revealed to be a broad chemical group of active

plants used against disorders that impair the immune system. Triterpenes are anti-

inflammatory, antimicrobial, antiviral, antitumoral, and immunomodulator compounds

found in plants.

Liebermann-Burchard Test was the method used in testing the presence of

triterpenes. 1 ml of sulphuric acid was added to 10 mg of rhizome ethanolic extract. The

presence of pinkish color in the solution confirms the presence of triterpenes.

Flavonoids

Flavonoids are the most common type of polyphenols. Polyphenols are plant

compounds that can act as antioxidants which help in neutralizing harmful free radicals

that could damage human cells and increase the risk of heart disease, diabetes, and cancer.

Flavonoids present their effects through altering membrane permeability and inhibiting

membrane-bound enzymes such as ATPase and phospholipase A2 which means that its
 53

mechanism of action includes inhibition of oxidative stress and inflammatory diseases. It

was reported that flavonoids have anti-inflammatory, antioxidant, cytotoxic, antispasmodic

and antimicrobial properties (Pacaña & Galarpe, 2017). Flavonoids are also known as

immunoadjuvants, which boost immune responses to vaccines without eliciting specific

antigen effects particularly in viral infections (Sotto et al., 2020).

Shinoda Test was the method used in testing the presence of flavonoid content. The

Tagbak (Alpinia elegans) rhizome extract was obtained by using 95% ethanol and extracted

through rotary evaporation. Warmed and filtered, added with a few drops of magnesium

(Mg) concentrated Hydrochloric acid (HCl) in the filtrate. Then, observe for the appearance

of red color which confirms the presence of flavonoids.

Saponins

Saponins are glycoside compounds found naturally in plants. Saponins in plants

were reported to have antioxidant properties due to their free radical scavenging abilities.

It can boost the cell-mediated immune system and increase antibody production. Saponins

contain anti-inflammatory, immune-stimulating, antibacterial, expectorant, antioxidant,

and anticoagulant effects (Naïve et al., 2019). Saponins are also known as

immunostimulants because they serve to increase the body's innate immunological

defenses, allowing the body's homeostasis to be restored and maintained (Sotto et al.,

2020).

The Froth Test was the method used to test the presence of saponins in the Tagbak

(Alpinia elegans) rhizome. Included in this test method was the foam test which used 1 ml

solution of the Tagbak (Alpinia elegans) rhizome ethanolic extract, diluted in 5 ml distilled
 54

water and shaken in a graduated cylinder for about 15 minutes. The development of white

foam confirms the presence of saponins.

Glycosides

Glycosides are also natural compounds in plants that are considered as secondary

metabolites likewise with other phytochemical constituents found in the Tagbak (Alpinia

elegans) rhizome. Similarly, to saponins, glycosides are renowned for their

immunostimulant effects, which aids in the activation of innate and adaptive immunity.

Fehling’s Test was used to test the presence of glycosides in the Tagbak (Alpinia

elegans) rhizome. This procedure includes reduction of sugar in glycosides present in the

rhizome’s ethanolic extract. 1 mg of ethanolic extract was dissolved in 4 ml of distilled

water and kept in a tube soaked in water. Few drops of blue solution, which is Fehling's

solution, were added and the development of red precipitate in the solution confirms the

presence of glycosides.

Tannins

Tannins are another type of polyphenols found in plants. Similarly, to flavonoids,

tannins are also considered as immunoadjuvants. Other than its immunological effects,

tannins also possessed antioxidant and anti-inflammatory properties (Tong et al., 2021).

Ferric Chloride Test was the test done to observe the presence of tannins in Tagbak

(Alpinia elegans) rhizomes. The ethanolic extract was warmed and filtered. 5 ml of its

filtrate was added with 1 ml of Ferric Chloride solution. Development of dark green to

black color indicates the presence of tannins.

 55

Alkaloids

Alkaloids derived from plants are known to contain large amount of antibacterial

properties, and are vital to be used as analgesics and narcotics. Alkaloid in turmeric plants

indicates that in treating headache caused by hypertension, cold, chronic catarrh, and

migraine, turmeric could be used.

Curcumin

Curcumin is a polyphenol of hydrophobic nature with very low water-solubility,

low stability, higher metabolism and very low absorption rate. Curcumin has good

therapeutic and preventive potential against several major human conditions such as

cardiovascular, inflammation suppression, antimicrobial, obesity, tumorigenesis, chronic

tiredness, antidepressant & neurological function, anxiety, muscle & bone loss, and

neuropathic pain (Mughal MH, 2019).

Vitamin C

Vitamin C acts as a natural antioxidant and potent chemopreventive agent in

Turmeric (Curcuma longa L.). Since vitamin C plays a role in numerous immune system

functions, boosting immunity is one of the main reasons individuals take vitamin C

supplements. Vitamin C aids in the promotion of the creation of lymphocytes and

phagocytes, two types of white blood cells that aid in the body's defense against infection.
 56

Most Effective Formulation of Tagbak (Alpinia elegans) and Turmeric (Curcuma

longa L.) Rhizome in Terms of White Blood Cell (WBC) and Lymphocyte Count

To test for the effectiveness of the Tagbak (Alpinia elegans) and Turmeric

(Curcuma longa L.) capsule as an immune booster, the researchers prepared three different

treatments: T1 - 300 mg of Tagbak (Alpinia elegans) and 200 mg of Turmeric (Curcuma

longa L.), T2 - 200 mg of Tagbak (Alpinia elegans) and 300 mg of Turmeric (Curcuma

longa L.), T3 - 250 mg of Tagbak (Alpinia elegans) and 250 mg of Turmeric (Curcuma

longa L.).

To prepare the solution, each formulation was diluted to 1 cc of sterile water and

was induced using gavage feeding. Approximately only about 0.125-0.25 cc/ml were

induced in the test subjects. Daily monitoring of behavior, vital signs and weighing was

done and food was given to evaluate the effectiveness of the product. After 3-4 days of

post-induction, which was day 7, the researchers again collected blood samples with the

help of a licensed veterinarian to test for WBC and Lymphocyte count and results revealed

that there was a change in these two parameters.

Immune-system activity induces changes in animal behavior. These changes are

mediated by proinflammatory cytokines, and the administration of cytokines produces the

same profile of behavior change as do infection and inflammation.

The researchers delved into the impact of the treatments as well as the

immunosuppressant on the vital signs and behavior of the experimental subjects. We

closely monitor key indicators such as heart rate and temperature to gain insights into their

physiological responses. Additionally, we observe changes in their behavior, which include

 57

heightened vigilance, hypoactivity, and suppressed food consumption when the

immunosuppressant was administered. These behavioral changes returned to normal after

the treatments were administered.

Throughout the experimentation process, the researchers observed that the vital

sign levels consistently remained within the normal range. Notably, we observed a slight

decrease in body temperature, measuring between 36.0°C and 36.2°C during the induction

of immunosuppressant and remained until the administration of treatments, compared to

the reference range of 36.5°C and 38.0°C. Additionally, the heart rates of the mice fell

within the range of 320 bpm to 424 bpm, aligning with the reference range of 310 to 840

bpm. Therefore, these vital signs and behavioral results conclude that the treatment

concentrations helped in regulating the immune system of the subjects which was observed

through vital signs and behavioral changes.

Table 2. Most Effective Formulation of Tagbak (Alpinia elegans) and Turmeric (Curcuma

longa L.) Rhizome in Terms of White Blood Cell (WBC) and Lymphocyte Count

Treatment T1 (60:40) T2 (40:60) T3 (50:50)

White Blood Cell (WBC) Count (x103/μL) MEAN

Baseline Data 12 9.5 9.3

Post Prednisone Induction (Day 3) 5.27 7.27 4.73

Treatment Induction (Day 7) 12.0 13.7 14.5

Lymphocyte Count (%) MEAN

Baseline Data 65.6 55.5 59.2

Post Prednisone Induction (Day 3) 48.2 46.1 41.3

 58

Treatment Induction (Day 7) 58.1 50.1 60.5

Scale Descriptor/ Level

8.1- 21.5 x103/μL White Blood Cell Count Normal range

30-70% Lymphocytes Normal range

Table 2 represents the summary of the results in the different parameters throughout

the treatment period. The parameters are White Blood Cell (WBC) and Lymphocyte Count.

White Blood Cell (WBC) Count in x103/μL and Lymphocyte Count in % was obtained by

a registered medical technician. The results revealed that T3 - 50% Tagbak: 50% Turmeric

has gained the highest mean in White Blood Cell (WBC) and Lymphocyte Count post

treatment induction and lowest mean post prednisone induction, which suggest that it was

the most effective concentration in comparison to T1 and T2 .

In this study, Kruskal-Wallis H Test was utilized to determine the significant

difference between the different treatment concentrations and parameters.

H(0); μ(60:40) = μ(40:60) = μ(50:50)

H(1); not all μ’s are equal

α = 0.05
 59

Table 3. Kruskal-Wallis H Test

Kruskal-Wallis

H-value df P-value

White Blood Cell 0.36 2 0.837

Lymphocyte 2.49 2 0.288

Statistically, the result revealed that there is no significant difference in terms of

WBC and Lymphocyte count regulation between the three concentrations of Tagbak

(Alpinia elegans) and Turmeric (Curcuma longa L.) capsule. In the WBC parameter, the

H(2) = 0.36 which is less than the critical value X 2crit = 5.99, p = 0.837 which is > 0.05.

Similarly in the Lymphocyte parameter, the H(2) = 2.49 which is less than the critical value

X2crit = 5.99, p = 0.288 which is > 0.05. Therefore, the researchers accepted the null

hypothesis that all three treatment concentrations are equal. However, there was still a

normal difference present in the different treatment concentrations.

In relation to Kruskal-Wallis statistical result above, a group descriptive of results

was utilized to determine the normal difference between the different treatment

concentrations.

Table 4. Group Descriptive of Treatments (N, Mean, Standard Deviation, Standard Error)

in White Blood Cell (WBC) and Lymphocyte Count

Table presented in the next page.

 60

Treatment N Mean SD SE

White Blood Cell (WBC) Count

T1 (60:40) 3 12.0 2.94 1.70

T2 (40:60) 3 13.7 6.91 3.99

T3 (50:50) 3 14.5 5.92 3.42

Lymphocyte Count

T1 (60:40) 3 58.1 12.25 7.07

T2 (40:60) 3 50.1 7.66 4.42

T3 (50:50) 3 60.5 8.91 5.14

Table 4 revealed that in WBC count parameter, T1 mice replicates treated with T1

(60 Tagbak:40 Turmeric) concentration have a Mean of M=12.0 and a Standard Deviation

of SD=2.94. T2 mice replicates are treated with T2 (40 Tagbak:60 Turmeric) concentration

(M=13.7, SD=6.91). And T3 mice replicates are treated with T3 (50 Tagbak:50 Turmeric)

concentration (M=14.5, SD=5.92). The table indicated that T3 - 50 Tagbak:50 Turmeric

concentration scored a higher increased rate and consistent level than the other treatments.

In lymphocyte count parameter, T1 mice replicates treated with T1 (60 Tagbak:40

Turmeric) concentration have a Mean of M=58.1 and a Standard Deviation of SD=12.25.

T2 mice replicates are treated with T2 (40 Tagbak:60 Turmeric) concentration (M=50.1,

SD=7.66). And T3 mice replicates are treated with T3 (50 Tagbak:50 Turmeric)

concentration (M=60.5, SD=8.91). The table indicated that T3 - 50 Tagbak:50 Turmeric

concentration scored a higher increased rate and consistent level than the other treatments.

These findings concluded that there is a difference but not significant as it is a normal
 61

difference between the three treatment concentrations in terms of WBC and lymphocyte

count regulation.

Most Effective Formulation of Tagbak (AIpinia elegans) and Tumeric (Curcuma

longa L.) Rhizome in Terms of Immunoglobulin G (IgG)

The third objective was to test the most effective formulation of Tagbak and

Turmeric treatment concentration in terms of Immunoglobulin G (IgG). In connection to

objective two, the most effective treatment concentration in terms of White Blood Cell

(WBC) and Lymphocyte Count was the T3 - 50% Tagbak: 50% Turmeric. Therefore, the

researchers conducted an Immunoglobulin G (IgG) test for this treatment concentration to

validate its immune boosting effect via serum test, which analyzes the presence or level of

antibodies present in the blood.

Table 5. Most Effective Formulation of Tagbak (AIpinia elegans) and Tumeric (Curcuma

longa L.) Rhizome in Terms of Immunoglobulin G (IgG)

Immunoglobulin G (IgG)

Normal Range Treatment 3

7 – 10 mg/ml 8.4 mg/ml

The researchers opted to utilize Immunoglobulin G (IgG) as a benchmark for

evaluating the effectiveness of the developed immune-boosting capsule in comparison to

other types of immunoglobulins. This decision was based on the fact that mouse IgG

constitutes approximately 75% of serum immunoglobulins as stated by Fortis Life Sciences

in 2023. This approach ensures that the measurement of IgG levels in mice provides a more
 62

precise estimation of the quantity of antibodies present in mice treated with the most potent

combination of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) capsules. IgG

plays a critical role in binding to a variety of pathogens, including bacteria, viruses, and

fungi. This binding mechanism serves to protect cells and prevent the body from

succumbing to infections.

Table 5 revealed the result of the IgG test of the most effective treatment which was

T3 with a concentration of 50% Tagbak:50% Turmeric. The researchers only used the mice

replicate in T3 as it was the most effective treatment among other treatments. It showed

that T3 - 50% Tagbak: 50% Turmeric IgG result was in a normal range which is 8.4 mg/ml.

Therefore, T3 concentration was effective in regulating the IgG levels in mice and its

immune response.

Significant Difference of the Most Effective Tagbak (Alpinia elegans) and Turmeric

(Curcuma longa L.) Concentration to the Commercially Available Immune Booster

(Ascorbic Acid)

The fourth objective of the study was to determine the significant difference of the

500 mg (50% Tagbak:50% Turmeric) Immune Booster capsule which was the most

effective treatment of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) Immune

Booster capsule to the commercially available 500 mg ascorbic acid. The Tagbak (Alpinia

elegans) and Turmeric (Curcuma longa L.) with 500 mg capsule and Ascorbic acid 500

mg capsule was administered to the male Albino mice through oral gavage feeding once a

day for 3-4 days at 8:00 o'clock in the morning.

 63

The parameters used are White Blood Cell (WBC) Count and Lymphocytes. The

researchers obtained every 3 days starting at baseline and carefully watched out for any

untoward signs that affect the health of the Albino mice. The effectiveness of the

formulated capsule was evaluated by determining the significant differences using Mann-

Whitney.

Table 6. Significant Difference of the Most Effective Tagbak (Alpinia elegans) and

Turmeric (Curcuma longa L.) Concentration to the Commercially Available Immune

Booster (Ascorbic Acid)

Treatments Treatment 3 (50:50) Treatment 4 (Commercially

available Ascorbic Acid)
White Blood Cell Count in x103/μL
Replicates A B C Mean A B C Mean
Baseline Data 8.7 10.4 8.0 9.0 14.3 10.7 8.0 11
Post Prednisone
1.8 7.6 4.8 4.7 7.4 10.6 9.8 9.3
Induction (Day 3)
Treatment Induction
12.0 10.3 21.3 14.5 12.6 12.0 13.0 12.5
(Day 7)
Lymphocytes Count in %
Replicates A B C Mean A B C Mean
Baseline Data 70.6 56.5 50.6 59.20 67.5 70.2 45.6 61.10
Post Prednisone 41.9 36.3 45.6 41.30 44.7 39.6 59.5 47.90
Induction (Day 3)
Treatment Induction 59.3 52.2 69.9 60.50 69.8 43.3 50.5 54.50
(Day 7)
 64

Scale Descriptor/ Level

8.1- 21.5 x103/μL White Blood Cell Count Normal range

30-70% Lymphocytes Normal range

Table 6 represents the result of the WBC count as well as the Lymphocytes count

of Treatment 3 which was the most effective concentration of Tagbak (Alpinia elegans)

and Turmeric (Curcuma longa L.) Immune Booster capsule and Treatment 4 which was

the commercially available Ascorbic Acid. It also shows the corresponding mean of each

result after every blood test of each treatment.

To show further the significant differences between the most effective treatment

concentration of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) to the

commercially available immune booster capsule, a Figure presentation was also presented.

Figure 5. Comparison of Mean White Blood Cell counts of T3 and T4 after

Acclimatization, Post Prednisone Induction, and Post Treatment Induction

 65

Figure 6. Comparison of Mean Lymphocyte counts of T3 and T4 after Acclimatization,

Post Prednisone Induction, and Post Treatment Induction

Figures 5 and 6 show the comparison of T3 (50% Tagbak:50% Turmeric) and T4

(commercially available Ascorbic Acid). The average WBC and Lymphocyte count was

obtained from the Mean of each replication of treatment 3 and treatment 4. After the

acclimatization period, the mean baseline WBC of Treatment 4 is M=11.0, compared to

M=9.0 in Treatment 3. Meanwhile, the Mean Lymphocyte count of Treatment 4 is

M=61.10%, compared to M=59.20% in Treatment 3. Prednisone induction results in a

decrease in WBC count of a mean of M=9.3 in Treatment 4 and M=4.7 in Treatment 3. At

the same time, the average Lymphocyte count in Treatment 4 is M=47.90%, while

M=41.30% in Treatment 3. The increase in WBC count in Treatment 4 was evident after

Ascorbic Acid induction with an average of M=12.5, but the most effective concentration

of Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.), which is Treatment 3,
 66

obtained a higher result of M=14.5. Conversely, the Mean Lymphocyte count of T4 is

M=54.50% and M=60.50% in Treatment 3 after treatment induction.

It was shown that both Treatment 4 and Treatment 3 are effective when it comes to

immune-boosting activity. It also appears to be the most effective concentration of Tagbak

(Alpinia elegans) and Turmeric (Curcuma longa L.) was slightly more effective compared

to the commercially available Immune Booster capsule when it comes to boosting

immunity based on each treatment's average. To test this hypothesis, the Mann-Whitney U

test was used as the statistical tool to analyze if there is a significant difference between T4

(commercially available Ascorbic acid) and T3 (the most effective concentration of Tagbak

(Alpinia elegans) and Turmeric (Curcuma longa L.), and it will be revealed on the

following pages.

Table 7. Mann-Whitney U test for White Blood Cell (WBC) and Lymphocyte Count [500

mg Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) Capsule and 500 mg

Ascorbic acid]

Stat z-value p-value Effect size

White Blood Mann-Whitney 3.50 -0.06 0.825 Pearson’s 0.48

Cell (WBC) U r

Lymphocytes 2.00 8.80 0.383 0.56

Table 8. Group Descriptive [500 mg Tagbak (Alpinia elegans and Turmeric (Curcuma

longa L.) and 500 mg Ascorbic acid] in White Blood Cell (WBC) and Lymphocyte Count

Table presented in the next page.

 67

White Blood Cell (WBC) Count

Group N Mean Median SD SE

500 mg Tagbak (Alpinia elegans) and 3 14.5 12 5.92 3.42

Turmeric (Curcuma longa L.) capsule

in 50:50 concentration

Commercially available Ascorbic Acid 3 11 12.6 0.50 0.29

Lymphocyte Count

500 mg Tagbak (Alpinia elegans) and 3 60.5 59.3 8.91 5.14

Turmeric (Curcuma longa L.) capsule

in 50:50 concentration

Commercially available Ascorbic Acid 3 54.5 50.5 13.7 7.91

Table 7 and 8 shows the results of the Mann-Whitney U test and group descriptive

presentation regarding the two parameters, which is the White Blood Cell (WBC) and

Lymphocyte Count. In WBC count parameter, the results revealed that the Male Albino

mice that were treated with 500 mg (50% Tagbak:50% Turmeric) Immune Booster capsule

exhibited an increase in White Blood Cell (WBC) Count (M=14.5, SD=5.92) relative to

Male Albino mice that were treated with 500 mg Commercially available Ascorbic acid

(M=11, SD=0.50), u(4)=3.50, p=0.825. The effect size is medium (r=0.48). These findings

suggest that the 500 mg Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.)

Immune Booster capsule has more practical significance when it comes to raising or

stabilizing WBC levels after being immunosuppressed than the commercially available

Immune Booster capsule.

 68

In Lymphocyte count parameter, the male albino mice treated with 500 mg (50%

Tagbak:50% Turmeric) Immune Booster capsule had an increase in lymphocytes (M=60.5,

SD=8.91) compared to the male albino mice treated with 500 mg Ascorbic acid (M=54.5,

SD=13.7), u(4)=2.00, p=0.383. The effect size is large (Pearson's r=0.56). These findings

imply that the 500 mg Tagbak (Alpinia elegans) and Turmeric (Curcuma longa L.) Immune

Booster capsule has more practical significance than the commercially available Immune

Booster capsule when it comes to boosting or maintaining lymphocyte levels after

immunosuppression.
 69

References

Fortis Life Sciences. (2023). What is Mouse IgG?

https://www.fortislife.com/products/articles/what-is-mouse-igg/What-is-Mouse-

IgG#:~:text=Mouse_IgG_is_make_up,_involved_in_autoimmune_condition_dev

elopment

Naive, M.A.K., Dalisay, J.A.G.P., Maglangit, E.P.T., Cafe, G.C., Nuňeza, O.M. (2019).

Free radical scavenging effects of the Philippine endemic medicinal plant Alpinia

elegans (Zingiberaceae)

https://www.researchgate.net/publication/337971762_Free_radical_scavenging_ef

fects_of_the_Philippine_endemic_medicinal_plant_Alpinia_elegans_Zingiberace

ae

Pacaňa, J.S. & Galarpe, V.R.K.R. (2017). Antibacterial property of Atuna racemosa Rafin,

Chrysobalanaceae shell and kernel extracts (aqueous, methanol, ethyl acetate, and

decoction).

https://www.researchgate.net/publication/319165494_Antibacterial_property_of_

Atuna_racemosa_Rafin_Chrysobalanaceae_shell_and_kernel_extracts_Aqueous_

Methanol_Ethyl_Acetate_and_Decoction

Sotto, A., Vitalone, A., Giacomo, S. (2020). Plant-Derived Nutraceuticals and Immune

System Modulation: An Evidence-Based Overview.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7563161/

Tong, Z., He, W., Fan, X., Guo, A. (2021). Biological Function of Plant Tannin and Its

Application in Animal Health. Vol. 8.

https://www.frontiersin.org/articles/10.3389/fvets.2021.803657/full
 Chapter 5

SUMMARY OF FINDINGS, CONCLUSIONS, AND RECOMMENDATIONS

This chapter presents the summary of the study and its findings. The conclusions

and recommendations are also included.

Summary of Findings

This study developed an immune booster capsule product using Tagbak (Alpinia

elegans) and Turmeric (Curcuma longa L.) and evaluated its efficacy as an immune booster

that helps in regulating the WBC and lymphocyte levels, as well as the Immunoglobulin G

(IgG). It answered specific questions including its phytochemical properties and evaluated

the effectiveness of the immune booster capsule in three different treatment concentrations

in terms of Complete Blood Count (CBC) and Immunoglobulin G (IgG) parameters.

Lastly, it determined the significant difference between the formulated immune booster

capsule and the commercially available immune booster.

After a detailed analysis and interpretation of the collected data, the following findings

were obtained.

The phytochemical properties of Tagbak (Alpinia elegans) and Turmeric (Curcuma

longa L.) rhizomes which were the raw materials used for immune booster capsule,

revealed that Tagbak (Alpinia elegans) rhizomes contained triterpenes, flavonoids,

saponins, glycosides, and tannins. Turmeric (Curcuma longa L.) rhizomes also contained

saponins, flavonoids, tannins, triterpenes, alkaloids, curcumin, and vitamin C. All of these

phytochemical properties were reported to be beneficial in regulating the immune system

function.
 71

As determined by the study, all treatments of different Tagbak (Alpinia elegans) and

Turmeric (Curcuma longa L.) concentrations were effective in regulating the WBC and

Lymphocyte levels involving the immune system. However, T3 - 50 Tagbak:50 Turmeric

(250 mg Tagbak: 250 mg Turmeric) concentration has the highest increased rate and

consistent level of 67.3% from M=4.73 x103/μL post prednisone induction to treatment

induction with M=14.5 x103/μL in WBC count for 7 days. In comparison to T2 using 40

Tagbak:60 Turmeric (200 mg Tagbak: 300 mg Turmeric) concentration which resulted in

a 46.93% increased rate from M=7.27 x103/μL to M=13.7 x103/μL. T1 using 60

Tagbak:40 Turmeric (300 mg Tagbak: 200 mg Turmeric) concentration resulted in a 56.1%

increased rate from M=5.27 x103/μL to M=12.0 x103/μL. Similarly, to lymphocyte count

changes, T3 resulted in a higher increased rate of 31.7% from M=41.3% post prednisone

induction to M=60.5% after treatment for 7 days. T2 with 7.9% increased rate from

M=46.1% to M=50.1% and T1 from M=48.2% to M=58.1% increased rate. In IgG

(Immunoglobulin G) result, T3 - 50 Tagbak:50 Turmeric (250 mg Tagbak: 250 mg

Turmeric) concentration as the most effective treatment has a normal level of IgG level

resulting in 8.4 mg/ml which indicates that T3 was effective in regulating the immune

response in mice.

Based on the statistical treatment, T3 - 50 Tagbak:50 Turmeric (250 mg Tagbak:

250 mg Turmeric) concentration has the highest increased rate among the three treatments

garnering a weighted mean of M=14.5 x103/μL in WBC count and M=60.5% in

Lymphocyte count, in comparison to T4 - commercially available Ascorbic acid with a

result of M=12.5 x103/μL in WBC count and M=54.5% in Lymphocyte count. The data

reveals that the WBC p-value of 0.825 and Lymphocyte p-value of 0.383 is greater than
 72

the significance level of 0.05. Therefore, there is enough evidence to say that there is no

significant difference between the Tagbak (Alpinia elegans) and Turmeric (Curcuma longa

L.) Immune booster capsule and the commercially available immune booster in terms of

efficacy, statistically. However, with the effect size of 0.48 for WBC count and 0.56 for

Lymphocyte levels, the study reveals that the 500 mg Tagbak (Alpinia elegans) and

Turmeric (Curcuma longa L.) Immune Booster capsule has a medium to large practical

significance than the commercially available Immune Booster capsule.

Conclusions

Based on the aforementioned findings, these were the conclusions drawn by the

researchers:

1. Objective one tackled the presence of phytochemical properties of the rhizomes of

Tagbak (Alpinia elegans). After undergoing phytochemical analysis, it revealed

that the plant particularly its rhizomes contained the following phytochemical

properties which includes triterpenes, flavonoids, saponins, glycosides, and

tannins. The researchers did not conduct a phytochemical testing for turmeric

(Curcuma longa L.) rhizome due to several studies that already conducted research

about its phytochemical composition. Saponins, flavonoids, tannins, triterpenes,

alkaloids, curcumin, and vitamin C are the phytochemicals present in the rhizome

of turmeric (Curcuma longa L.) according to the studies about the phytochemical

and pharmacological importance of turmeric and its antioxidant capacity in the

studies of Chanda and Ramachanda (2019), Permanatanda et al. (2021), and

Jyotirmayee and Mahalik (2022).

 73

2. The second objective of the study included the determining of the most effective

formulation in terms of Immunoglobulin G (IgG), White Blood Cell (WBC) and

Lymphocyte Count, which resulted to T3 - 50 Tagbak:50 Turmeric (250 mg Tagbak:

250 mg Turmeric) concentration as the one to have the higher increased rate and

consistent level among other treatment concentrations.

3. The researchers accepted the null hypothesis that there is no significant difference.

Statistical treatment showed a WBC p-value of 0.825 and Lymphocyte p-value of

0.383. This reveals that the two treatments are comparable and exhibited almost the

same immune boosting effect before the researchers conducted the experiment.

Hence, there is enough evidence to say that Tagbak (Alpinia elegans) and

Turmeric (Curcuma longa L.) capsule has no significant difference from the

commercially available immune booster in terms of immune boosting efficacy.

Nevertheless, a medium effect size indicates a practical significance of the research

outcome.

Recommendations

The researchers had seen the potential of the combined raw materials Tagbak

(Alpinia elegans) and Turmeric (Curcuma longa L.) in boosting the immune system. And

in order to further strengthen the study, the researchers recommend the following:

1. Further testing of different concentrations with increased dose. This would help

future researchers to compare the effectiveness between various treatment

formulations.
 74

2. Further study using other methods of drying: dehydrator drying, sun drying and

microwave drying.

3. The researchers recommend finding a bigger size & weight for the subject since it

has an effect on the blood that can be extracted out of it.

4. Further study to extend the days to more than 3 days of daily administration of the

treatment to achieve more significant changes in parameter results.

5. Further study to increase the frequency of CBC and Immunoglobulin G (IgG)

testing in days to prove a more significant result in the changes in the subject’s

immune system.

6. Undergo several tests including toxicity tests for human consumption of immune

booster capsule products.

7. Incorporate neutrophil levels as parameters to strengthen the effectiveness of the

treatment concentration outcomes within the immune system.

 75

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