MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

A COMPARATIVE STUDY ON THE HYPOGLYCEMIC ACTIVITY OF TURMERIC

(Curcuma longa) AND GARLIC (Tulbaghia violacea) ETHANOLIC

EXTRACT ON STREPTOZOTOCIN-INDUCED HYPERGLYCEMIC SPRAGUE-DEWLEY

RATS

________________________

A thesis presented to the Faculty

of the Department of Medical Technology

Medical Colleges of Northern Philippines

Tuguegarao City, Cagayan

________________________

In Partial Fulfillment

Of the Requirements for the Degree

BACHELOR OF SCIENCE IN MEDICAL TECHNOLOGY

________________________

RESEARCHERS:

Salvador, Karen Mae G.

Salvador, Shaira P.

Serrano, Adrian Louie T.

Tamot, Nemalyn A.

Torres, Ar- Kenneth A.

Torres, Divine D.
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

TABLE OF CONTENTS

Title Page 1

Table of Contents 2

Acknowledgment 4

Dedication 5

Abstract 6

Chapter 1:

THE PROBLEM AND REVIEW OF RELATED LITERATURE

Introduction 8

Review of Related Literature 8

Statement of the Problem 23

Hypotheses 24

Significance of the study 24

Conceptual Framework 26

Scope and Limitation 27

Definition of Terms 28

Chapter 2

Methodology 30

Subjects of the Study 32

Data Collection Procedure and Instrumentation 32

Data Analysis Procedure 37

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

Chapter 3

Presentation, Analysis, and

Interpretation of Data 39

Chapter 4

Summary of Findings 44

Conclusions 44

Recommendations 45

Bibliography 46

Appendix 49

Documentation 54
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

ACKNOWLEDGEMENT

We would like to express our dearest thanks and gratitude

to the persons who help us in making our research proposal
successful. This research was supported by the following
persons:

Mr. Enjay C. Javier, our thesis adviser, for his constructive

criticism, and sharing his ideas in the development of this
study.

Mr. Mark Angelo Binarao, our statistician, for helping us in our

statistical data and for his patience in teaching us on how to
interpret our data.

Dr. Abelardo Bas-Ong, for his guidance and persistent help

during the biological phase of our study.

Laboratory assistants, for their patience in giving us the

materials and chemicals needed in our study.

And above all, the almighty God, for His continuous

guidance.
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

DEDICATION

The researchers would like to dedicate this thesis to our

parents. Without their support, understanding, patience, ideas

and love the completion of this study would not be possible.

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

ABSTRACT

This study, entitled “A Comparative Study on the

Hypoglycemic Activity of Turmeric (Curcuma longa) and Garlic

(Tulbaghia violacea) Ethanolic Extract on Streptozotocin-

Induced Hyperglycemic Sprague-Dewley Rats was conducted at

Cagayan Valley Herbal Processing Plant for the biological assay.

For the authentication of plants, it was conducted at Department

of Agriculture and for the phytochemical screening; it was

conducted at DOST Industrial Technology Development Institute

Standards and Testing Division.

Based from the data obtained, Groups II, III and IV which

are the hyperglycemic groups received a dose of ethanolic

extract of turmeric and garlic respectively revealed a decrease

in their blood glucose level compared to their fasting blood

glucose. Both ethanolic extract of turmeric and garlic showed a

decrease in the blood glucose of the hyperglycemic rats after

30 and 120 minutes of treatment.

After comparing the effects of the ethanolic extract of

turmeric and garlic with the commercialized drug (pioglitazone),

all of them revealed a decrease in their blood glucose level

when compared to their fasting blood glucose level after three

hours of hyperglycemic induction. The hypoglycemic activity of

the turmeric and garlic extract was more effective compared to

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

the commercialized drug (pioglitazone) based from their

calculated means. Ethanolic extract of turmeric and garlic as

well as pioglitazone manifested their activity up to 120

minutes.

Thus, Turmeric (Curcuma longa) and Garlic (Tulbaghia

violacea) Ethanolic Extract was said to have a potential

hypoglycemic effect.
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

Chapter 1
THE PROBLEM AND REVIEW OF RELATED LITERATURE

Introduction

Diabetes mellitus (DM) is a chronic, metabolic disease of

the endocrine system characterized by hyperglycemia. High blood

glucose level may be a result of several factors such as

insufficient insulin production of pancreas, or resistance of

the body to the action of insulin. Diabetes due to absolute or

relative insulin deficiency or insulin resistance becomes a

widespread endocrine disorder that not only affects the glucose

homeostasis but also chronically alters lipids and protein

metabolism with increase in cellular oxidative stress. It is

estimated that there are 300 million people suffering from

diabetes in the world. (Aslan and Orhan, 2010).

The Philippines is one of the world’s emerging diabetes

hotspots. Ranked in the top 15 in the world for diabetes

prevalence, Philippines is home to more than 4 million people

diagnosed with the disease – and a worryingly large unknown

number who are unaware they have diabetes. The estimated

national prevalence of diabetes in the Philippines was 7.2% in

2008, which translates to approximately 6.377 million diabetics

in a national population of 88.57 million at that time. This

number is rapidly approximating the projected prevalence of 7.8%

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

in 2025 as predicted by the IDF. The estimated national

prevalence of pre-diabetes using the WHO criteria is 10.2% (IDF

Diabetes Atlas, 5th edition annual update, 2012)

In modern medicine, insulin and/or oral agents such as

sulfonylureas, biguanides and incretin mimetics are used for the

treatment of diabetes. On the other hand, traditional herbal

medicines are used all over the world by diabetic patients.

Researches on herbal medicines are encouraged to come up with

alternatives for treatment of diabetes since oral antidiabetic

agents have side effects in the long run (Samad et al., 2009).

Plants are considered to be the basis for deriving natural or

combined substance constituents that can be used against

diabetes (Aslan and Orhan, 2010). The antidiabetic effects of

some herbal medicine have been found to contain substances like

flavonoids, glycosides, terpenoids and prenolic compounds.

(Jarald et al., 2008; Rao et al., 2010). Investigations must be

carried out to identify the active chemical constituent(s)

responsible for the antidiabetic activity of the different

medicinal plants, and to understand their mechanism of action.

Management of diabetes without any side effect is still a

challenge to medical community. Therefore, it is prudent to look

for options in alternative medicine or herbal for diabetes as

well. Although, herbal medicines have long been used effectively

in treating disease in the Philippines and throughout the world,

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

yet the mechanism of most herbals used has not been defined

(Baily and Day, 2006).

A scientific investigation of traditional herbal remedies

for diabetes may provide valuable leads for the development of

alternative drugs and strategies and also may guide the way to

let us perceive the variety and properties of herbal medicines

to be consumed. Alternatives are clearly needed for better

management of diabetes because of high cost and poor

availability of current therapies for many developing countries

(Bhattaram et al., 2002).

Based on the WHO recommendations, hypoglycemic agents of

plant origin used in traditional medicine are important (WHO

Expert Committee on Diabetes Mellitus, 2013). The attributed

anti hyperglycemic effects of these plants are due to their

ability to restore the function of pancreatic tissues by causing

an increase in insulin output or inhibit the pancreatic tissues

by causing an increase in insulin output or inhibit the

intestinal absorption of glucose or to the facilitation of

metabolites in insulin dependent processes (Neelesh Malviya,

Sanjay Jain and k; 2010).

The comparative study of anti-diabetic effects of turmeric

extract with that of Garlic (Tulbaghia violacea) has not been

documented. In the view of these reports, it seemed interesting

to search for more efficient, safe and cost effective single

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

anti-diabetic agent, which can also improve the profile in

diabetic patients. Therefore, it was considered worthwhile to

undertake this study to evaluate the hypoglycemic effect of

turmeric (Curcuma longa) and Wild Garlic (Tulbaghia violacea)

in hyperglycemic induced rats.

General and Specific objective:

To compare the hypoglycemic activity of Turmeric and Garlic

extract in STZ infected rat hyperglycemic.

Specific objective:

1. To compare the mean blood glucose of rats with and without

STZ injection.

2. To compare the effect of different dosages of turmeric and

garlic in STZ-induced hyperglycemic rats at 30 and 120 minutes

response time.

Diabetes

The review of related literature was based on different

studies, journals and with the help of internet. The information

started with the brief information about diabetes, followed by

hyperglycemic drug.

Diabetes is a defect in the body’s ability to convert

glucose (sugar) to energy. Glucose is the main source of fuel

for our body. When food is digested it is changed into fats,

protein, or carbohydrates. Foods that affect blood sugars are

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

called carbohydrates. Carbohydrates, when digested, change to

glucose. Examples of some carbohydrates are: bread, rice, pasta,

potatoes, corn, fruit, and milk products. Individuals with

diabetes should eat carbohydrates but must do so in moderation.

Glucose is then transferred to the blood and is used by the

cells for energy. In order for glucose to be transferred from

the blood into the cells, the hormone - insulin is needed.

Insulin is produced by the beta cells in the pancreas (the organ

that produces insulin). In individuals with diabetes, this

process is impaired. Diabetes develops when the pancreas fails

to produce sufficient quantities of insulin – Type 1 diabetes

or the insulin produced is defective and cannot move glucose

into the cells – Type 2 diabetes. Either insulin is not produced

in sufficient quantities or the insulin produced is defective

and cannot move the glucose into the cells

(www.diabeteswellness.net).

Diabetes is a global concern that cuts across geographical

boundaries regardless of race, sex, status and age. Diabetes and

its complications impose a heavy burden to the individual, his

family and society in general (World Health Organization 2016)

Diabetes is a chronic disease that occurs either when the

pancreas does not produce enough insulin or when the body cannot

effectively use the insulin, it produces (WHO, 2009). Diabetes

is a fast becoming a leading cause of morbidity, mortality and

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

disability across the world. Diabetes mellitus is a global

metabolic affecting essential biochemical activity in almost

every age group. (Gupta et al, 2008). It is a chronic metabolic

disorder with impaired glucose tolerance and high risk of

cardiovascular disease. Many oral synthetic anti-diabetic agents

have been developed. Hyperglycemia can be handled initially with

oral agents and insulin therapy, which sometimes require

achieving targeted glycemic leaves. However, this synthetic

agent produced some serious side effects and relatively

expensive for developing countries (Edoga C. O., Njoku O. O.,

Amadi E. N., Okeke J. J.; January 2013).

Diabetes has become an important chronic disease in the

world, and especially in developing countries. It comprises a

collection of heterogeneous diseases deferring in their

etiological, clinical and epidemiological characteristics but

have hyperglycemia and glucose tolerance in concurrence diabetes

affects quite a number of populations in the world about 200

million and is the emerging epidemic in Africa which threatens

to overwhelm the health care system (Maina J K., Kareru P G,

Gatebe E G, Ritich H, Githira P N, Njonge F, Kimani D and

Mutembei J K; 2014)

Clinical features of Diabetes Mellitus.

Normal blood sugar level in the body varies before and

after meals. The normal blood glucose is 4-7mmol/L before meals

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

and less than 10mmol/L after one and a half hour of meals. At

bed times the normal blood sugar level is about 7-10mmol/L. The

normal values of blood glucose level are 70-110mg/dL for fasting

(Mehta, Barehnholdz, Nyska, Ben-Shaul and Grossman, 2005).

Impairments caused by Diabetes Mellitus.

According to Tiwari, Pandey, Abidi and Rizvi (2013), the

causes of diabetes depend on the type of diabetes. Type I occurs

mainly due to Beta-cell destruction, mediated through either

immune mediated or idiopathic, whereas Type II diabetes occurs

mainly due to insulin resistance or with relative insulin

deficiency. Diabetes is also associated with lifestyle factors

and genetics. There are various types of other factors involved

in the development of diabetes such as chromosomal and

mitochondrial DNA mutation, lipoatrophic diabetes, congenital

rubella and cytomegalovirus infection.

Turmeric (Curcuma longa)

The researchers choose to use the pure extract because

normally, people who harvest and eat these plants would not

perform concentration of the extract.

Turmeric (Curcuma longa) is an herbaceous perennial plant

belonging to the Zingiberaceae family, more widely known as the

ginger family. It is native to south-east India, but can also

grow in other countries in Southeast Asia. The plant is about

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

1m high with elongated leaves and pink flowers, but it is the

rhizomes (roots) that are boiled, dried and then ground into a

yellow powder. This powder is commonly used as a spice in Asian

cooking, but it is also used to color cheese, butter and yogurt,

for example. The active component in turmeric is curcumin, which

may constitute 2 to 8% of the spice. The spice is utilized as a

traditional medicine in many countries, especially in Asia, and

numerous studies have shown that curcumin has antioxidant and

anti-inflammatory properties (Wickenberg, 2015)

Saxena Jyoti et.al (2012) have found that certain compounds

in turmeric such as (Carbohydrate, Proteins, Starch, Amino

acids, Steroid, Glycoside, Flavonoid, Alkaloid, Tannin and

Saponin) from the extracts of rhizomes of Curcuma longa that

work by increasing the amount of insulin in the blood by blocking

the liver’s inactivation of insulin, making more insulin

available to the body.

Although turmeric has shown remarkable effects on

diabetes and its associated complications in animal tests, no

clinical trials have yet been reported. A recent review revealed

that turmeric has beneficial effects on glycemic response in

animals, but the only clinical trial (including in vitro with

human cells) using turmeric/curcumin was on the treatment of

diabetic nephropathy, microangiopathy and retinopathy

(Wickenberg, 2015).
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

The (WHO) approves the use of plant, drugs for different

diseases, including diabetes mellitus. Therefore, studies with

plant extracts are useful in order to establish their efficacy,

mechanism of action and safety (Cromwell Kibiti, 2015 Oct)

Garlic (Tulbaghia violacea)

Garlic (Tulbaghia violacea) is an herbaceous plant and is

sometimes referred to as Wild Garlic or Society Garlic, it has

been used in folkloric medicine since early times to treat a

number of human diseases and ailments, including hypertension,

asthma, gastro-intestinal disorders, esophageal cancer, fevers

and colds, inflammation, tuberculosis and bacterial infections.

(Kogi Moodley, Kimane Joseph, Yougasphree Naidoo,

Shahidul Islam and Irene Mackraj; 2015). It belongs to

the Alliaceae family of herbs, similar to Garlic.

Tulbaghiaviolacea may contain a wide variety of compounds. Some

of these compounds include: 3 Cysteine derivates known as

Marasmin, Ethiin and Methiin, Kaempferol, From the leaves,

2,4,5,7-tetrathiaoctane-2,2-dioxide, 2,4,5,7-tetrathiaoctane,

and 2,4,5,7-tetrathiaoctane-4-oxide, Steroidal saponins (2011 -

2017 Examine.com Inc.)

Researchers have found that certain compounds in garlic

such as allicin, allyl propyl disulfide and S-allyl cysteine

sulfoxide work by increasing the amount of insulin in the blood

by blocking the liver’s inactivation of insulin, making more

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

insulin available to the body. (Priyanka Singh1, NidhiGauba

Dhawan2, Raaz K Maheshwari; 2014).

Despite the availability of medication for diabetes,

traditional remedies are desirable and are currently being

investigated. Wild Garlic, which is a common cooking spice, has

a long history as a folk remedy, has been reported to have

antidiabetic activity. The role of garlic as both an insulin

secretagogue and as an insulin sensitizer is reviewed. Evidence

suggests that garlic's anti-oxidative, anti-inflammatory, and

anti-glycative properties are responsible for garlic's role in

preventing diabetes progression and the development of diabetes-

related complications. Large-scale clinical studies with

diabetic patients are warranted to confirm the usefulness of

garlic in the treatment and prevention of diabetes (Cheng-Tzu

Liu, Lee-Yan Sheen, Chong-KueiLii; 2007).

Pioglitazone

Pioglitazone is an anti-diabetic drug (thiazolidinedione-

type, also called "glitazones") used along with a proper diet

and exercise program to control high blood sugar in patients

with type 2 diabetes. It works by helping to restore your body's

proper response to insulin, thereby lowering your blood sugar.

Controlling high blood sugar helps prevent kidney damage,

blindness, nerve problems, loss of limbs, and sexual function

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

problems. Proper control of diabetes may also lessen your risk

of a heart attack or stroke.

Pioglitazone treatment resulted in a decrease in

cardiomyocyte apoptosis as revealed by a decrease in cardiac

caspase-3, lactate dehydrogenase (LDH) levels and DNA

fragmentation, and an increase in Na+K+ATPase (sodium-potassium

pump) levels in diabetic rats. Cardiac histology of diabetic

control rats showed dense focal fatty infiltration in the

myocardial cells whereas normal architecture with regular

morphology and well preserved cytoplasm was observed with

pioglitazone treatment. Pioglitazone treatment significantly

reduced the heart rate, mean arterial blood pressure, body mass

index (BMI) and levels of serum glucose, leptin, insulin, HOMA-

IR, total cholesterol (TC) and triglycerides (TGs),

apolipoprotein-B glycosylated hemoglobin (HbA1c) levels and

atherogenic index, and increased the levels of serum high

density lipoprotein cholesterol (HDL-C) and cardiac antioxidant

enzymes. Pioglitazone, a member of the PPAR-gamma agonist drug

family, has been demonstrated to improve both metabolic and

vascular insulin resistance when applied to patients with Type

II diabetes mellitus. The drug is well tolerated with fluid

retention and weight gain being the most frequently described

side effects. The observed effects (e.g., improvements in

glucose and lipid metabolism, improvements of endothelial

function and microcirculation, reduction of surrogate markers

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

of atherosclerosis and inflammation and an improvement in

hypertension) have made pioglitazone one of the frequently

prescribed antidiabetic drugs in the US and Europe. Several

trials have shown its potency to reduce carotid intima-media

thickness, and outcome studies with pioglitazone have shown its

potential to delay the progression of Type II diabetes and

atherosclerosis and even reduce cardiovascular mortality. The

purpose of this review is to provide an overview about recently

published clinical results with pioglitazone. They underline the

value of this drug when used alone or in combination with other

antidiabetic drugs for a successful management of Type II

diabetes mellitus (Pfützner A1, Weber MM, Forst T. 2011)

Streptozotocin

Streptozotocin is a glucosamine-nitrosourea compound

derived from streptomycesachromogenes that is clinically as

chemotherapeutic agent in the treatment of pancreatic B-cell

carcinoma. Streptozotocin damages pancreatic B-cells, resulting

to hypoinsulinemia and hyperglycemia. It is naturally occurring

chemical that is particularly toxic to the insulin-producing

beta cells of the pancreas in mammals. It is used in medicines

for treating certain cancers of the Islets of Langerhans and

used in medical research to produce in animal model for

hyperglycemia in a large dose as well as type II diabetes or

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

type 1 diabetes with multiple low doses. (Elazu, Sonia

Chukwuma,and Udeme Nelson Essien; 2013).

Streptozotocin (STZ)-induced diabetes mellitus (DM) offers

a very cost effective and expeditious technique that can be used

in most strains of rodents, opening the field of DM research to

an array of genotypic and phenotypic options that would

otherwise be inaccessible. Despite widespread use of STZ in

small animal models, the data available concerning drug

preparation, dosing and administration, time to onset and

severity of DM, and any resulting morbidity and mortality are

often limited and inconsistent. Because of this, investigators

inexperienced with STZ-induced diabetes may find it difficult

to precisely design new studies with this potentially toxic

chemical and account for the severity of DM it is capable of

inducing (MC Deeds, JM Anderson, AS Armstrong, DA Gastineau, HJ

Hiddinga,A Jahangir, NL Eberhardt, and YC Kudva; 2014 Feb 7)

STZ is a broad-spectrum antibiotic that is toxic to the

insulin producing β cells of pancreatic islets. It is currently

used clinically for the treatment of metastatic islet cell

carcinoma of the pancreas and has been used investigational in

a wide variety of large and small animal species. The method of

STZ action in β cell depletion has been studied extensively over

the years. It is generally assumed that STZ is taken up via the

cell membrane GLUT2 glucose transporter and causes DNA

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

alkylation and eventual β cell death, although streptozotocin’s

actions as a protein alkylating agent and nitric oxide donor may

contribute to its cytotoxicity. Because STZ enters the cell via

GLUT2, the toxic action is not specific to β cells and can cause

damage to other tissues including the liver and kidney.

Albino Rats

From formulating new cancer drugs to testing dietary

supplements, mice and rats play a critical role in developing

new medical wonders. In fact, 95 percent of all lab animals are

mice and rats, according to the Foundation for Biomedical

Research (FBR).

Scientists and researchers rely on mice and rats for

several reasons. One is convenience: rodents are small, easily

housed and maintained, and adapt well to new surroundings. They

also reproduce quickly and have a short lifespan of two to three

years, so several generations of mice can be observed in a

relatively short period of time. Mice and rats are also

relatively inexpensive and can be bought in large quantities

from commercial producers that breed rodents specifically for

research. The rodents are also generally mild-tempered and

docile, making them easy for researchers to handle, although

some types of mice and rats can be more difficult to restrain

than others (Remy Melina | November 16, 2010)

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

Most of the mice and rats used in medical trials are inbred

so that, other than sex differences, they are almost identical

genetically. This helps make the results of medical trials more

uniform, according to the National Human Genome Research

Institute.

The researchers utilized Albino rats as experimental animal

since they are easy to handle, response quickly. Furthermore,

it is the most common experimental object used in researches.

Statement of the Problem

The study aims to compare the hypoglycemic effect of the

Turmeric (Curuma longa) and Garlic (Tulbaghiaviolacea) ethanolic

extract in streptozotocin-induced hyperglycemic albino rats.

The study aims to answer the following questions:

1. What is the blood glucose level of the male albino rats before

and after the induction of streptozotocin?

2. Is there a significant difference in the blood glucose level

of the albino rats before and after the induction of

streptozotocin?

3. What is the glucose level of the streptozotocin-induced

albino rats in the negative, positive and experimental groups

after 30 and 120 minutes of treatment?

4. Is there a significant difference in the blood glucose level

of the streptozotocin-induced albino rats in the negative,

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

positive and experimental groups after 30 and 120 minutes of

treatment?

Hypotheses

This study is pursued on the basis of these hypotheses:

Ho: There is no significant difference in the glucose level of

albino rats before and after induction of Streptozotocin.

Ho: There is no significant difference in the glucose level of

albino rats after 30 and 120 minutes of treatment in different

groups.

Significance of the Study

The study is expected to provide valuable information on

how to utilized Turmeric (Curcuma Longa) and Garlic (Tulbaghia

violacea) extract properly to its maximum and results could be

most particularly useful or can be used in a number of ways to

the following groups or agencies:

Diabetic Patients: This study provides information in

relation to the medical use of turmeric (Curcuma Longa) and

Garlic (Tulbaghia violacea) that provides possible alternative

medicine to expensive drugs for diabetes that are available in

the market.

Community and Public Consumers: The study will benefit the

community and public consumers by providing cost-effective

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

alternative, inexpensive, and accessible to place in the

country.

Health Care Professions: The study serves as base-line

information for the concern profession that will guide

researchers in the future.

Government: This research will offer some experimental

message to the Department of Health (DOH) and the Department of

Science and Technology (DOST) by providing them information

about the potential use of Turmeric (Curcuma Longa) and Garlic

(Tulbaghia violacea) to decrease the prevalence of diabetes and

for promotion of health and wellness of the Filipinos.

Future Researchers: This study can serve as a guide and

basis to undertake future studies along with this.

 MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

Conceptual Framework

Collection of Turmeric and Garlic

Preparation of Turmeric and Garlic

Extract

Ethanolic Extract from Turmeric and Garlic Ethanolic Extract of Turmeric and Garlic

Selected Phytochemical Screening Biological Assay: Hypoglycemic

Effect
a. Alkaloids
b. Saponin
Stage I:
c. Steroid
Hyperglycemic Induction
d. Tannins

Group I: Normal control - Given isotonic saline

(10ml/kg/day).
Group II: Diabetic rats - Given Pioglitazone
Stage II: (6mg/kg/day)
Turmeric and Garlic Extract Group III: Diabetic rats - Given Turmeric extract
(300mg/kg/day).
Group IV: Diabetic rats - Given Garlic extract
Blood glucose levels at the after 30 minutes and 120 (300mg/kg/day).
minutes.
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

The study looked at the specificity of Turmeric (Curcuma longa)

and Garlic (Tulbaghia violacea) ethanolic extract along with

pioglitazone, the positive control of the blood glucose level

of the streptozotocin-induced male albino rats.

Scope and Limitations

Twenty (20) male albino rats will be obtained in Zone 5

Malvar, Santiago City, Isabela. The rats weigh 100-175 grams,

2-3 months old and acclimatize for one week in order to adapt

to the research environment. The rats will be assessed by a

veterinarian in good condition by the Philippine Institute of

Traditional and Alternative Health Care (PITAHC) at Carig,

Tuguegarao City, Cagayan.

The Turmeric (Curcuma longa) and Garlic (Tulbaghia

violacea) will be used in the study. The Turmeric will be

collected at City of Ilagan, Isabela while the Wild Garlic will

be collected at Tagga, Tuguegarao City and will be verified by

the Department of Agriculture for the assurance of the variety.

The preparation of turmeric (Curcuma longa) and Garlic

(Tulbaghia violacea) will be done through ethanolic extraction

and to be done at St. Paul University Philippines.

Blood samples will be obtained by pricking the top of the

rat's tail with a sterile lancet, wiping the first drop of blood,

and then placing a drop of blood on the glucose strip to be read

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

by a glucometer. The values will be recorded and taken as the

baseline blood glucose values in mg/dL.

Pioglitazone, which is a standard drug for diabetes, will

be administered to compare its effect to that of the extract.

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

DEFINITION OF TERMS

The following are the definition used by the researchers

in doing this study.

 Active Constituent

- secondary metabolite present in Turmeric (Curcuma longa)

and Garlic (Tulbaghia violacea) possessing hypoglycemic

activity.

 Blood Glucose

- this is the parameter that will be ran for chemical

bioassay analysis.

 Blood Glucose Level

- blood glucose that will be obtained after 30 and 120

minutes.

 Experimental Group

- this includes the rats which will be treated with

Turmeric (Curcuma longa) and Garlic (Tulbaghia violacea)

extracts.

 Extraction

- a way to separate a desired substance when it is mixed

with others. The mixture is brought into contact with a

solvent in which the substance of interest is soluble,

but the other substances present are insoluble

(Chemicool, 2014)
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

 Fasting Blood Sugar

- blood sugar that will be obtained after 18 hours of

fasting.

 Glucometer

- device used in measuring the blood glucose level of

the mice.

 Hyperglycemia

- increased blood glucose level of the rats.

 Hypoglycemia

- decreased blood glucose of the rats as measured.

 Steptozotocin

- chemical used to induce hyperglycemia.

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

Chapter 2
METHODOLOGY

This chapter outlines the research methodology of this

study, particularly the research design and the subject of the

study, data gathering, procedures and data analysis.

Acclimatization

The study was done to compare the effect of TURMERIC and

GARLIC extract on the blood glucose levels of streptozotocin

rendered diabetic rats. Twenty (20) 2 months old male rats will

be used following randomized block design with four treatments

and five blocks. Initial blood glucose levels will be obtained

after 18 hours of fasting without food except water. Injection

with streptozotocin (75mg/dl) will be allowed a duration of 3

days with oral glucose tolerance test (2g/kg) confirms the

diabetic status of each animal in 30 and 120 minutes. Rats with

close difference in glucose levels will be arranged in 5 blocks

and 4 treatments groups. Following 30 and 60 minutes of oral

intake of treatments, the blood glucose will be drawn and

analyzed with RCBD ANOVA. For this purpose, 20 healthy Sprague-

dewley rats will be selected and grouped as follows:

Subjects of the Study

The rats will be acclimatized in the PITAHC-DOH

experimental animal house before carrying out experimental work

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

in a well-ventilated animal house under natural photoperiod

conditions for a period of 1 week prior to experimentation. All

rats will receive basal diet and water will be provided ad

libitum during the entire period of the study. Rats will be

monitored closely every day and will be weighed every week.

(Sabry M. El- Bahr, et al. 2014)

Group I: Normal control Given isotonic saline (10ml/kg/day).

Group II: Diabetic rats Given Pioglitazone (6mg/kg/day)

Group III: Diabetic rats Given turmeric extract (300mg/kg/day).

Group IV: Diabetic rats Given garlic extract (300mg/kg/day).

Data Collection Procedures and Instrumentation

1. Procurement of rats

The rats will be obtained from the Santiago City, Isabela

and will be assessed by a veterinarian in good condition.

2. Acclimatization period

The rats will be acclimatized in the PITAHC-DOH

experimental animal house before carrying out experimental work

in a well-ventilated animal house under natural photoperiod

conditions for a period of 1 week prior to experimentation. All

rats will receive basal diet and water will be provided ad

libitum during the entire period of the study. Rats will be

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

monitored closely every day and will be weighed every week.

(Sabry M. El- Bahr, et al. 2014)

3. Collection of Turmeric (Curcuma longa) and Wild Garlic

(Tulbaghia violacea)

The Turmeric (Curcuma longa) that will be used in the study

will be collected at City of Ilagan, Isabela while the Wild

Garlic (Tulbaghia violacea) that will be used in the study will

be collected at Tagga, Tuguegarao City. Collection will be done

in the morning so that water dews will be dried by solar heat

before gathering and after which leaves will be freed from any

adhering materials on the surface of the rhizomes using tap

water followed by a sterile distilled water and will be subjected

to air dry under a shed at room temperature. It will be cut and

crushed to small pieces and will be placed on a plastic container

for the extraction process (Kimberly, Acoba et al. 2010)

4. Plant Authentication

Turmeric (Curcuma longa) and Wild Garlic (Tulbaghia

violacea) is to be authenticated at the Department of

Agriculture at San Gabriel, Tuguegarao City for the assurance

of the variety.

5. Preparation of turmeric and garlic ethanolic extract

The prepared rhizomes will be cut into small pieces. The

cut rhizomes will be soaked in 95% ethanol, 1000 mL for each 200
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

grams of the cut rhizomes. These mixtures will be kept in a

tightly closed amber glass jar for a period of 5 days at room

temperature and will be shaken or stirred several times daily.

After 5 days, the mixture will be filtered and expressed

mechanically using the muslin cloth. The volume of extracted

liquid will be measured and subjected to rotatory evaporation

for 5-6 hours to remove the solvent ethanol. The remaining liquid

will be measured for volume and percentage yield. It will be

keep in an amber glass jar at 2-8C until further use.

%yield= volume of the extract after rotatory evaporation x 100

volume of the extract before rotatory evaporation

6. Standard Drug (Pioglitazone)

Pioglitazone is an anti-diabetic drug (thiazolidinedione-type,

also called "glitazones") used along with a proper diet and

exercise program to control high blood sugar in patients with

type 2 diabetes. It works by helping to restore your body's

proper response to insulin, thereby lowering your blood sugar.

Single dose of 6 mg/kg body weight once a day orally

on streptozotocin-induced diabetic rats to restore its body's

proper response to insulin, thereby lowering the blood sugar.

(JeevangiSantoshkumar et al. 2013)

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

7. Streptozotocin Preparation

Streptozotocin (60mg/kg b.w.) will be dissolved in cold

0.01 M citrate buffer (pH 4.5). STZ injections will be given

intraperitoneally and the doses will be determined according to

the body weight of animals. (Tank et al. 1989).

8.Bioassay

8.1 Determination of Baseline Glucose Levels

Before the administration of the treatments in the

test animals, initial glucose levels will be determined. The

animals will be fasted for at least 18 hours with free access

to water and blood glucose levels will measured. Blood samples

will be obtained by pricking the top of the rat's tail with a

sterile lancet, wiping the first drop of blood, and then placing

a drop of blood on the glucose strip to be read by a glucometer.

The values will be recorded and taken as the baseline blood

glucose values in mg/dL (M. Abuzo et al. 2015).

8.2 Hyperglycemic induction

After taking the baseline glucose values, Diabetes will be

induced by administering intraperitoneal injection of a freshly

prepared solution of streptozotocin (60mg/kg b.w) in 0.1 M cold

citrate buffer to the fasted rats (Tank et al.1989). and to be

tested using oral glucose tolerance test to confirm the diabetic

status of each animal in 30 and 120 minutes.

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

8.3 Determination of glucose levels after hyperglycemic

induction

After 72 hours, fasting blood glucose levels will be

monitored. Animals having blood glucose values above 200 mg/dl

on the third day after streptozotocin administration will be

considered as diabetic rats (Tank et al. 1989). Then the

treatment will start on the third day after streptozotocin

administration and it will be considered as the first day of

treatment.

8.4 Turmeric and Garlic Extract Induction

8.4.1 Group I – Control:

 After 72 hours, the fasting blood sugar will

be determined.

8.4.2 Group II - Hyperglycemic rats receiving

pioglitazone:

 This group will receive a dose of the standard

drug solution based on their weight.

8.4.3 Group III - Hyperglycemic rats receiving

Ethanolic Turmeric Extract:

 This group will receive a dose of ethanolic

turmeric extract based on their weight.

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

8.4.4 Group IV - Hyperglycemic rats receiving

Ethanolic Garlic Extract:

 This group will receive a dose of ethanolic

turmeric extract based on their weight.

8.5 Blood Glucose Level Testing

After a single dose administration of Turmeric and Garlic

extract, blood samples will be collected and estimated for blood

glucose levels after 30 and 120 minutes by using glucometer.

9. Selected Phytochemical Screening

Crude Extraction:
200 gm Turmeric and Garlic will be soaked in 500 ml of

Ethanol solvents in the cold for 3 days with occasional shaking.

The solvent from the total extract will be filtered and then

dried under shade and it will be used for the qualitative

analysis of secondary metabolites.

Identification Test:
The individual extract will be subjected to the qualitative
phytochemical screening for the presence of some chemical
constituents. Phytochemical test will be carried out adopting
standards procedure (Trease et.al 1983, Kokate et.al 1997, Hegde
et.al 2010).

 Alkaloids: A quantity (3 ml) of concentrated extract

will be taken into a test tube and 1 ml HCl will be added
to the mixture heated gently for 20 min cooled and
filter, the filtrate will be used for following test.
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

 Saponin: 5 ml extract will be mixed with 20 ml of

distilled water then agitated in graduated cylinder for
15 min, formation of foam indicates Saponin.
 Sterols: 1ml extract will be dissolved in 10 ml of
chloroform & equal volume of concentrated H2SO4 acid will
be added from the side of test tube. The upper layer
turns red and H2SO4 layer showed yellow with green
fluorescence. This indicates the presence of steroid.
 Tannin: 4ml extract will be treated with 4 ml FeCl3
formation of green color indicates that presence of
condensed tannin.
 Flavonoid: Alkaline reagent test: Extract will be
treated with 10 % NaOH solution; formation of intense
yellow color indicates presence of Flavonoid.

Data Analysis

The significant difference between the baseline values will

be calculated with One-way ANOVA. Paired t-test will be used to

compare the effect of treatments before and after induction. All

values will be expressed as mean with their Standard Deviation

(SD) P values <0.05 will be considered as significant.

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

CHAPTER 3
PRESENTATION, ANALYSIS AND INTERPRETATION OF DATA

A paired-samples t-test was conducted to compare blood

glucose levels in the untreated fasted and streptozotocin

treated validated glucose tolerance diabetic rats (Table 1).

There was a significant difference in the levels for the

untreated and fasted rats (M=84.90, SD=11.77) and with

streptozotocin GTT validated levels (M=251.60, SD=144.91)

(Appendix table 1), t (20), 5.2398, p = 0.0001 (table 1). These

results suggest that streptozotocin is a potent hyperglycemic

activity. The results suggest that when rats were given a dose

of Intra-peritoneal streptozocin, the amount of un-metabolized

blood glucose levels increases, hence the significant rise in

the glucose levels of the test animals.

Table 1. Paired t test analysis of the mean difference of

blood glucose levels of rats obtained with and without
streptozotocin injection

Paired difference
Mean Std 95% confidence t Df{decrease Sig
error interval of the of freedom t-
Mean difference tailed
lower upper

Pair 166.7 31.814 100.11 233.29 5.2398 19 0.0001

initial
and
with
stz
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

A one way RCBD ANOVA was conducted to determine the effect

of various treatments 30 and 120-minute response time on the

blood glucose of rats rendered hyperglycemic with streptozotocin

validated with glucose tolerance test. The blood glucose levels

of experimental groups are shown in Table 2.

Table 2. Blood levels of rats due to treatments.

Response Experimental Groups F P
time to
treatments Isotonic Pioglitazone Turmeric Garlic
(minutes)
(10ml/kg/day) (6mg/kg/day) (300mg/kg/day) (300mg/kg/day)

30 339.8a 143.6b 185.2b 193.8b 4.42* 0.025

(127.5) (196) (154.4) (145.8)

120 339.6a 129.4b 162.6b 197.7b 6.6** 0.006

(127.5) (26.7) (108.4) (104.7)

Note *= p ≤ 0.05** = p ≤ 0.01. Standard deviations appear in parentheses below means.

Means with differing subscripts within rows are significantly different at the p ≤
0.05 based on TUKEY HONEST SIGNIFICANT DIFFERENCE post hoc paired comparisons.

The treatments were as follows: 5 rats was assigned in

the isotonic dosage group had an average glucose level of 339.8

mg/dl (SD = 127.5); the 5 rats in the pioglitazone dosage group

had an average level of 143.6mg/dl (SD = 196), the 5 rats in

the turmeric dosage group had a mean of 185.2mg/dl (SD = 154.4),

and the last 5 rats in the garlic dosage group had a mean of

193.8mg/dl (SD=145.8) The effect of treatments, therefore, was

significant, F(3,12) = 4.42, p=.025 while the effect of blocks

was not, F (4,12)=2.67. Post hoc analyses using the Tukey HSD

(honest significant difference) post hoc criterion for

significance indicated that the average levels of blood glucose

were significantly higher in the isotonic dosage group (M =

339.6, SD = 127.5) than in the other three groups combined (M =

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

143.6, SD = 196), (M = 185.3, SD = 154.4), (M = 193.8, SD =

145.8).

The effect of the treatments on the blood glucose of test

rats after 120 minutes was analyzed as before showing the effect

of treatments are significantly different, F (3, 12) =6.6,

p=0.0069, while the effect of blocks is not, F (4, 12) =3.125.

Post hoc analyses using the Tukey HSD (honest significant

difference) post hoc criterion for significance indicated that

the average levels of blood glucose were significantly higher

in the isotonic dosage group (M = 339.6, SD = 127.5) than in the

other three groups combined (M = 129.4, SD = 26.7), (M = 162.6,

SD = 108.4), (M = 197.2, SD = 104.7).

These results suggest that medication with our test extract

of turmeric and garlic really do have an effect on levels of

blood glucose in streptozotocin hyperglycemic rats comparable

with pioglitazone. Specifically, our results suggest that when

test rats were medicated the extract 30 and 120 minutes response

time, the amount of blood glucose remained lower with turmeric

and garlic extract intake than when they are given isotonic

solution alone.

Discussion

Turmeric and garlic are separate plant species carrying

different natural chemicals which could alter or potentiate the

effect of an agonist drug. In our study, diabetes was induced

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

with agonist chemical in streptozotocin administered i.p.

(intraperitoneal). A review of the diabetogenic mechanism of the

chemical (Szkudelski T.(2001)) shows that damage to B cells by

necrosis is brought about by the entrance of streptozotocin in

B cells via the glucose transport (GLUT2) causing alkylation to

DNA thereby activating the poly ADP-ribosylation which depletes

the NAD+ and ATP. This enhance ATP dephosphorylation provides a

substrate for xanthine oxidase to form superoxide radicals,

hydrogen peroxide, and hydroxyl radicals. Nitric oxide was

another chemical in the equation which was observed to inhibit

aconitase and putting this activity all together participates

in the DNA damage. The significantly lower amount of glucose

obtained from test rats medicated separately with the test

plants extract lead us to assume that such damage to the B cell

by necrosis could have been lessened by reaction of the extract

chemicals during the metabolic processes in-vivo. The

antioxidants activity of the plant extract could have lessened

the formation of hydrogen peroxide and other free radicals

resulting from streptozotocin injection which is why we believe

that the B cells could have regenerated awhile thus some amount

of insulin could have been produced in response to the oral

glucose intake.
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

CHAPTER 4
FINDINGS, CONCLUSION AND RECOMMENDATIONS

Summary

The study was done to compare the effect of TURMERIC and

GARLIC extract on the blood glucose levels of streptozotocin

rendered diabetic rats. Twenty (20) 2 months old male rats were

used following randomized block design. Initial blood glucose

levels were obtained after 18 hours of fasting without food

except water. Injection with streptozotocin (75mg/dl) was

allowed a duration of 3 days with oral glucose tolerance test

(2g/kg) confirms the diabetic status of each animal in 30 and

120 minutes. Rats with close difference in glucose levels were

arranged in 5 blocks and 4 treatments groups. Following 30 and

120 minutes of oral intake of treatments, the blood glucose were

drawn and analyzed with RCBD ANOVA. Results shows a prior blood

glucose of rats before and after administration of

streptozotocin were significantly increased. Whereas, blood

glucose levels with intake of the plant extract are

significantly lower than those given isotonic solution alone.

Conclusion

We therefore conclude that rats rendered diabetic with

streptozotocin injection and validated with glucose tolerance

test would tolerate the amount of glucose in experimental group

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

hence rats in the turmeric and garlic dosage group had the lower

amount of glucose compared with the isotonic dosage group. We

also conclude that hence pioglitazone did not differ in the mean

amount of glucose after treatment with the plant extract given

separately that these plants had the same effect. We cannot

conclude however as to what mechanism they were brought in to

cause this effect.

Recommendation

Based on the foregoing results, the two plant extract

having shown significantly lowering effect in streptozotocin

diabetic rats is recommended due to the hypoglycemic effect at

par with pioglitazone, a synthetic oral hypoglycemic drug with

caution.
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

BIBLIOGRAPHY

Jarald E, Joshi SB, Jain DC (2008). Diabetes and herbal

medicines.Iranian J. Pharmacol .Ther. 7(1):97-106.

WHO Expert (2013). Committee on Diabetes Mellitus. Technical

Report Series 874.Geneva and World Health Organization.

DuduÖzkum, Ömrüm Akı and Hale ZerrinToklu. (2013). Herbal

medicine use among diabetes mellitus patients in Northern

Cyprus. Near East University, Nicoisa, Northern Cyprus (TRNC).

Maina J K., Kareru P G, Gatebe E G, Ritich H, Githira P N, Njonge

F, Kimani D and Mutembei J K. (2014). Hypoglycemic effects of

selected herbal drug formulations from the Kenyan market.

Nairobi, Kenya.

Samad A, Shams MS, Ullah Z, Wais M, Nazish I, Sultana Y, Aqil

M. (2009). Status of herbal medicines in the treatment of

diabetes: a review. Curr. Diabetes Rev. 5(2):102-111.

NeeleshMalviya, Sanjay Jain and SapnaMalviya. (2010).

Antidiabetic potential of medicinal plants.Madhya Pradesh,

India.

Edoga C. O., Njoku O. O., Amadi E. N., Okeke J. J. (2013). Blood

Sugar Lowering Effect of MoringaOlifeira Lam in Albino

Rats.Nsukka, Enugu State, Nigeria

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

Maina J K., Kareru P G, Gatebe E G, Ritich H, Githira P N, Njonge

F, Kimani D and Mutembei J K. (2014). Hypoglycemic effects of

selected herbal drug formulations from the Kenyan

market.Nairobi, Kenya.Krup V, Prakash LH, Harini A (2013)

Pharmacological Activities of Turmeric (Curcuma longa linn): A

Review. J HomeopAyurv Med 2:133. doi: 10.4172/2167-1206.1000133

Hoda W. El Gawly, Mona K. Tawfik, Maha F. Rashwan,Ali S. Baruzaig

(2009) The effect of pioglitazone on the liver of

streptozotocin-induced diabetic albino Wistar rats Eur Rev Med

Pharmacol Sci.

Eleazu, C. O., Eleazu, K. C., Chukwuma, S., &Essien, U. N.

(2013). Review of the mechanism of cell death resulting from

streptozotocin challenge in experimental animals, its practical

use and potential risk to humans. Journal of Diabetes and

Metabolic Disorders, 12, 60.http://doi.org/10.1186/2251-6581-

12-60

MC Deeds, JM Anderson, AS Armstrong, DA Gastineau, HJ

Hiddinga,A Jahangir, NL Eberhardt, and YC Kudva (2014) Single

Dose Streptozotocin Induced Diabetes: Considerations for Study

Design in Islet Transplantation Models Lab Anim; 45(3): 131–140.

Remy Melina (2010) Albino mice http://www.livescience.com

SachinArora, Shreesh Kumar Ojha and DivyaVohora. (2009).

Characterization of Streptozotocin Induced Diabetes Mellitus in

Swiss Albino Mice.Global Journal of Pharmacology, 3 (2): 81-84.

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

M. Abuzoet. al (2015). Anti-diabetic activity of Turmeric leaf

ethanolic extract (calophylluminophyllum) on alloxan- induced

diabetic spraguedawleyrats.Pharmacy Research Journal.St. Paul

University Philippines. School of Health Sciences. Pharmacy

Department

Edoga C. O., Njoku O. O., Amadi E. N., Okeke J. J. (2013). Blood

Sugar Lowering Effect of MoringaOlifeira Lam in Albino

Rats.Nsukka, Enugu State, Nigeria.

Hoda W. El Gawly, Mona K. Tawfik, Maha F. Rashwan,Ali S. Baruzaig

(2009) The effect of pioglitazone on the liver of

streptozotocin-induced diabetic albino Wistar rats Eur Rev Med

Pharmacol Sci.
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

APPENDIX

Appendix Table 1. Initial and Stz-induced blood glucose

levels of rats confirmed with glucose tolerance test
BEFORE AFTER
Initial Blood STZ induced GTT (3 After GTT
Glucose days) 72 hours
1 84 g/dL 0.3 mL 446 g/dL
2 84 g/dL 0.3 mL 142 g/dL
3 80 g/dL 0.4 mL 281 g/dL
4 93 g/dL 0.4 mL 412 g/dL
5 113 g/dL 0.4 mL 417 g/dL
6 81 g/dL 0.4 mL 141 g/dL
7 95 g/dL 0.4 mL 171 g/dL
8 97 g/dL 0.4 mL 107 g/dL
9 87 g/dL 0.4 mL 163 g/dL
10 67 g/dL 0.4 mL 294 g/dL
11 81 g/dL 0.4 mL 100 g/dL
12 80 g/dL 0.3 mL 541 g/dL
13 80 g/dL 0.3 mL 390 g/dL
14 60 g/dL 0.3 mL 201 g/dL
15 105 g/dL 0.4 mL 447 g/dL
16 80 g/dL 0.4 mL 117 g/dL
17 86 g/dL 0.4 mL 310 g/dL
18 84 g/dL 0.3 mL 166 g/dL
19 78 g/dL 0.3 mL 101 g/dL
20 83 g/dL 0.3 mL 85 g/dL
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

Appendix table 2. Descriptive statistics of the pre and post

blood glucose levels of rats.

Group Initial Final

Mean 84.90 251.60
SD 11.77 144.91
SEM 2.63 32.40
N 20 20

Paired t test results:

P value and statistical significance:
The two-tailed P value is less than 0.0001
By conventional criteria, this difference is considered to
be extremely statistically significant.

Confidence interval:
The mean of Group One minus Group Two equals -166.70
95% confidence interval of this difference: From -233.29 to
-100.11

Intermediate values used in calculations:

t = 5.2398
df = 19
standard error of difference = 31.814

Appendix Table 3. RCBD. The mean Blood glucose of rats due to

treatments 30 minutes thereafter.
Blocks
Treatments B1 B2 B3 B4 B5 Treatment

Total Mean

Isotonic 446 142 281 412 417 1698 339.6

Pioglitazone 140 147 118 116 198 719 143.8

Turmeric 117 122 130 414 143 926 185.2

Garlic 113 116 110 275 355 969 193.8

D
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY
Block total 816 527 639 1217 1113 4312

Block mean 204 132 160 304 278 215.6

Appendix Table 4. Data Summary after 30 minutes of treatment.

Appendix Table 5. RCBD ANOVA of the blood glucose levels 30 minutes

after treatment.

Source of SS Df MS F P
variation
Treatments 109653.2 3 36551.0667 4.42 0.025923
(between 5%=3.49
groups)
Blocks 88273.8 4 22068.45 2.67ns
5%=3.26
Error 99165.8 12 8263.8167
Totals 297092.8 19

Tukey HSD Test

HSD [.05] = 170.85; HSD [.01] = 223.71
M1 vs M2 P<.05
M1 vs M3 nonsignificant
M1 vs M4 nonsignificant
M2 vs M3 nonsignificant
M2 vs M4 nonsignificant
M3 vs M4 nonsignificant
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

Appendix Table 6. Blood glucose after 2 hours of treatment.

treatments B1 B2 B3 B4 B5 Treatment
Total Mean
Isotonic 446 142 281 412 417 1698 339.6
A
Pioglitazone 133 125 106 110 173 647 129.4
B
Turmeric 104 109 118 356 128 813 162.6
C
Garlic 140 113 112 294 327 986 197.2
D
Block total 823 489 617 1172 1045 4144
Block mean 206 122 154 293 261 207

Appendix Table 7. Data summary glucose levels 2 hours after

treatment.
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

Appendix Table 8. ANOVA of the blood glucose levels 2 hours after

treatment.

Source of SS Df MS F P
variation
(sum of (mean (Fisher’s
(decrease
Square) square) test)
of
freedom)

Treatments 128358.8 3 42786.2667 6.6 0.006961

(between 5%=3.49
groups)
Blocks 81006.2 4 20,251.55 3.125ns
5%=3.26
Error 77762.2 12 6480.1833
Totals 287127.2 19

Tukey HSD Test

HSD [.05] = 151.3; HSD [.01] = 198.1
M1 vs M2 P<.01
M1 vs M3 P<.05
M1 vs M4 nonsignificant
M2 vs M3 nonsignificant
M2 vs M4 nonsignificant
M3 vs M4 nonsignificant
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

DOCUMENTATION

ACCLIMATIZATION

COLLECTION OF TURMERIC AND GARLIC

MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

CURRICULUM VITAE

NAME: AR-KENNETH A. TORRES

AGE: 21
DATE OF BIRTH: February 19, 1998
HOME ADDRESS: #9 Diversion Road, Ugac Norte, Tuguegarao City
CIVIL STATUS: Single
RELIGION: Roman Catholic
CITIZENSHIP: Filipino
PARENTS: Armando P. Torres, Jr.
Janette A. Torres
SIBLINGS: Jan-Kenneth A. Torres
Mary Andrea A. Torres
Ma. Lovella A. Torres
SCHOOLS ATTENTED:
ELEMENTARY: Baggao East Central Elementary School
SECONDARY: Cagayan National High School
TERTIARY: Medical Colleges of Northern Philippines
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

NAME:
AGE:
DATE OF BIRTH:
HOME ADDRESS:
CIVIL STATUS:
RELIGION:
CITIZENSHIP:
PARENTS:
SIBLINGS:
SCHOOLS ATTENTED:
ELEMENTARY:
SECONDARY:
TERTIARY:
MEDICAL COLLEGES OF NORTHERN PHILIPPINES COLLEGE OF MEDICAL TECHNOLOGY

NAME:
AGE:
DATE OF BIRTH:
HOME ADDRESS:
CIVIL STATUS:
RELIGION:
CITIZENSHIP:
PARENTS:
SIBLINGS:
SCHOOLS ATTENTED:
ELEMENTARY:
SECONDARY:
TERTIARY: