Hypoglycemic Activity of Cacao ii

SAN PEDRO COLLEGE

12 de Guzman Street, Davao City
Philippines

APPROVAL SHEET

In partial fulfillment of the requirements for the degree of Bachelor of Science in

Pharmacy this THESIS entitled:

THE HYPOGLYCEMIC ACTIVITY OF THE CAPSULE

FORMULATION FROM THE FLAVONOID OF
CACAO (Theobroma cacao) HUSK EXTRACT

Have been prepared and submitted by JOCELYN B. BAQUIANO, MARRA LOREEN

L. HORDISTA, DARELLE D. LEONOR, CLAUDE DOMINIC R. NAVAREZ,
PATRICIA ROSE P. PADER, AMY ROSE A. POSADAS and YANICE GRACIELLE
S. VILLARUEL who are recommended for the ORAL EXAMINATION.

LOURDES G. BANGAYAN, MS Pharm

Adviser

APPROVED in partial fulfillment of the requirements for the degree of

Bachelor of Science in Pharmacy

By the Oral Examination Committee:

PROF. FATIMA MAY R. TESORO, MS Pharm PROF. JACQUELINE A. PADILLA, MS Pharm

Member Member

ACCEPTED in partial fulfillment of the requirements for the degree of

Bachelor of Science in Pharmacy

PROF. LEILANI A. BETONIO, MS Pharm

Dean

MARCH 2014
Date
 Hypoglycemic Activity of Cacao iii

ACKNOWLEDGEMENT

The researchers would like to express their deepest gratitude and heartfelt

appreciation to the following:

Our Parents and Family, for the continuous moral and financial support

throughout the duration of this research. They have taught good traits and values which

molded us to become responsible and dedicated researchers. Without them, we would not

have come to existence and everything would not have been possible.

The veterinarian, Dr. Criselda C. Lao, for her supervision on the bioassay of the

animals and for training us in handling experimental animals.

Our statistician, Prof. Carlo Comeling, for his assistance in the statistical

treatment of the gathered data.

Our adviser, Mrs. Lourdes Bangayan, for sharing her expertise and time, and for

assissting the researchers in conducting their study. Her assisstance is deemed valuable

by the researchers. Also, for the patience she had shown regarding the revisions of this

study. Lastly, for inculcating values to the researchers that gave them strength to

persevere and do well on this study.

The members of this panel, Prof. Jacqueline Padillaand Prof. Fatima Mae Tesoro,

for giving recommendations and constructive criticisms, and extending their time on

scrutinizing the study which helped the researchers improve their study.

Our school, San Pedro College, for including research on the curriculum and

giving the researchers quality education. The school has created professionals who are

globally competent and efficient.

 Hypoglycemic Activity of Cacao iv

And most especially, God, for giving the researchers knowledge, wisdom, good

health, and the heart to complete their requirements in the pursuit of this degree.
 Hypoglycemic Activity of Cacao v

Abstract
This study is designed to find out the hypoglycemic activity of the capsule formulation

from the flavonoid of Cacao (Theobroma cacao) husk extract. Prior to its assay,

qualitative and quantitative tests were done. The test animals used are 20 rabbits which

were divided into 5 groups with 4 rabbits each. Alloxan Monohydrate was used to induce

hyperglycemia to the rabbits. Metformin was used as the positive control and 1%

Carboxymethylcellulose as the negative control. The experimental group received 250

mg, 500 mg and 750 mg of the formulated capsule form the husk of Cacao. The drying,

phytochemical screening, formulation process and bioassay were conducted on the

research laboratory of San Pedro College. The researchers used One-way ANOVA and

Tukey HSD Test as the statistical tools in determining if there was significant difference

between the formulated capsules and positive control. Statistical results showed that there

was no significant difference on the hypoglycemic activity of the formulated capsule to

that of Metformin. It signified that the formulated capsule is comparable to Metformin in

reducing blood sugar level. It was the Cacao (250 mg) which is considered the most

effective dose since it has the highest percentage decrease mean among the three

formulated doses. Though the low dose (250 mg) of the formulated capsule exhibited a

higher percentage decrease mean than the positive control in reducing blood glucose

levels based on the comparison of the tabulated results, it had no significant difference to

that of Metformin (500 mg). Through quantitative and qualitative analysis, it was evident

that flavonoid is present in the formulation. The formulation passed the quality control

tests using the different parameters: Physical, Chemical, Toxicological and

Microbiological analysis. Stability tests confirmed that the formulation remained stable
 Hypoglycemic Activity of Cacao vi

on the different parameters: organoleptic analysis, pH, moisture determination and

chemical test. Based from the formula of Q10 method, the result showed that for 10 days

the formulated capsule remained stable and expected shelf-life of the product is 90 days

when stored at 25oC room temperature.

 Hypoglycemic Activity of Cacao vii

TABLE OF CONTENTS

Page

TITLE PAGE ...................................................................................................................... i

APPROVAL SHEET ......................................................................................................... ii
ACKNOWLEDGEMENT ................................................................................................ iii
ABSTRACT ....................................................................................................................... v
LIST OF TABLES ............................................................................................................ ix
LIST OF FIGURES ............................................................................................................x

CHAPTER I INTRODUCTION
Review of Literature ................................................................ 3
Theoretical Framework .......................................................... 16
Conceptual Framework .......................................................... 18
Statement of the Problem ....................................................... 19
Null Hypothesis ..................................................................... 20

CHAPTER II METHOD
Design ..................................................................................... 21
Study Flow .............................................................................. 22
Setting ..................................................................................... 23
Participants ..............................................................................23
Measures ................................................................................. 23
Procedure ................................................................................ 24
Data Analysis .......................................................................... 37

CHAPTER III RESULTS .......................................................................................... 38

CHAPTER IV DISCUSSION ....................................................................................50

CHAPTER V SUMMARY, CONCLUSION AND RECOMMENDATION ...........54

REFERENCES .............................................................................................................58

APPENDICES

Appendix A Plant Authentication..................................................................... 63

Appendix B Phytochemical Screening............................................................. 64
Appendix C Capsule Formulation....................................................................68
Appendix D Quality Control Tests ................................................................. 70
Appendix E Microbiological Analysis Result..................................................77
Appendix F Toxicological Analysis Result......................................................78
Appendix G Animal Certification.....................................................................81
Appendix H Veterinarian Certification.............................................................82
 Hypoglycemic Activity of Cacao viii

Appendix I Bioassay........................................................................................83
Appendix J Raw Data of the Blood Glucose Reduction in an 8 Hour
Interval..........................................................................................85
Appendix K One-way ANOVA.........................................................................86
Appendix L Descriptives (Computation)..........................................................87
Appendix M Post Hoc (Tukey HSD Test).........................................................89
Appendix N Homogenous Subsets (Tukey HSD Test).....................................92
Appendix O Percentage Decrease Mean...........................................................94
Appendix P Bar Graphs of the Percentage Reduction in an 8 Hour Interval
(All Groups) ...............................................................................95
Appendix Q Bar Graphs of the Percentage Reduction Mean
among the 3 Doses of the Formulated Capsule............................97
Appendix R Bar Graph of the Percentage Reduction Mean between
the Most Effective Dose (Cacao 250 mg) and Metformin............98
Appendix S Moisture Content Determination..................................................99
Appendix T Disintegration.............................................................................100
Appendix U Timeline......................................................................................101

CURRICULUM VITAE .................................................................................................103

 Hypoglycemic Activity of Cacao ix

LIST OF TABLES

Tables Page

1 Group of Plant Constituent Distributed in the Extracts for Spotting....................31

2 Developing Solvent Systems for TLC of Solution A, B, C..................................31

3 List of Plant Constituents, their Visualizing Agents, and Positive Results...........31

4 Test for the Presence of Flavonoids......................................................................38

5 Determination of the Pesence of Ethyl Alcohol....................................................39

6 Determination of the Presence of Flavonoid using TLC.......................................39

7 Capsule Formulation .............................................................................................40

8 Testing the Significant Difference in an 8-hour Interval Between Groups using

One-Way ANOVA.................................................................................................41

9 Comparative Table of Difference in the Hypoglycemic Activity of Metformin

Between the Experimental and Negative Control Group......................................42

10 Percentage Decrease Mean among Doses of the Formulated Capsule..................43

11 Percentage Decrease Mean Between Most Effective Dose (Cacao 250mg) &
Metformin (500mg)...............................................................................................44

11 Organoleptic and pH Analysis...............................................................................45

12 Moisture Content Determination...........................................................................46

13 Flavonoid Content Determination using HPLC.....................................................46

14 Microbial Analysis Results....................................................................................48

15 Toxicological Analysis Results.............................................................................48

 Hypoglycemic Activity of Cacao x

LIST OF FIGURES

Figure Page

1 Conceptual Framework .............................................................................18

2 Schematic Diagram of the Processes Involved in the Hypoglycemic

Activity of the Capsule Formulation from the Flavonoid of the Cacao
(Theobroma cacao) Husks ....................................................................... 22

3 Percentage Decrease Mean among Doses of the Formulated Capsule .....43

4 Percentage Decrease Mean Between Most Effective Dose (Cacao 250mg)

& Metformin (500 mg) ............................................................................ 44

5 HPLC Sample Result ................................................................................47