CHAPTER I

INTRODUCTION

This chapter covers the background of the study, statement of the problem,

significance of the study, and scope and delimitation.

Background of the Study

We all know that everyone can have diabetes and everyone can have it at some

point of their life. Diabetes is a disease or condition in which causes your blood

glucose, or blood sugar levels to become high. When you have diabetes it affects the

body’s ability to produce or use insulin. There are two common types of diabetes, its

either your body does not make insulin (type 1 diabetes) or your body does not make or

use insulin well (type 2 diabetes). Without enough insulin, the glucose will stays in your

blood, and if the glucose stays in our blood for too long, it can lead to potential

complications such as heart disease, stroke, kidney damage, and nerve damage. And

we all know that these complication that results in diabetes are some of the common

cause of death, in short diabetes should be treated as soon as possible so it may not

lead to some of those complications or something even worse from that.

Bitter melon is linked to lowering the body’s blood sugar. This is because the

bitter melon has properties that act like insulin, which helps bring glucose into the cells

for energy. The consumption of bitter melon can help your cells utilize glucose and

move it to your liver, muscles, and fat. ( Natalie Silver, 2018)

Bitter gourd (Momordica Charantia) is a popular vegetable which is traditionally

known to have health beneficial effects not only, but mainly on diabetes. Bitter gourd
could be a cheap possibility to help the poor in these and other countries to control their

blood glucose levels. This review describes anti-diabetic effects of bitter gourd reported

in the literature and discusses what still needs to be clarified for developing an

evidence-based and safe use of the bitter gourd for diabetes. Analyses of bioactive

compounds have shown that bitter gourd is rich in nutrients and phytochemicals of

which some have anti-diabetic effects. Juices, powders, extracts, and isolated

compounds have been tested in vitro and in vivo. Bitter gourd increases insulin

secretion of the pancreas, decreases intestinal glucose uptake, and increases uptake

and utilization of glucose in peripheral tissues. (Habicht SD et al., 2014)

Statement of the Problem

This study aims to determine the effectiveness of Bitter Melon Seeds in lowering

the blood sugar level of the diabetic animal specifically the rabbit (Oryctolagus

Cuniculus). Specifically it will sought answer to the following questions:

1. What are the chemical compositions of Bitter Melon (Momordica Charantia)

Seeds based on the phytochemical analysis?

2. What are the of effects of Bitter Melon (Momordica Charantia) Seeds on the

blood sugar level of the diabetic rabbit?

3. Is there any significant difference between the initial and final blood test results of

the diabetic rabbit or Oryctolagus Cuniculus?

Significance of the Study

The main purpose of this study is to help diabetic individual in lowering their

blood sugar level. The researchers ensure that Bitter Melon (Momordica Charantia)

Seed will be effective on lowering the blood sugar level. If the result of the study was

proven that has a significant effect on lowering the blood sugar level of the rabbit, this

might produce also a significant effect to the following:

Community

Each individual of the community can be able to use available plants in their

community such as bitter melon seed as an alternative medicine for the people whose

suffering from diabetes

Future Researchers

The result of the study can also be use as a guide to help other researchers for

their other studies. This work therefore, is commit to authenticate the effectiveness of

bitter melon seed on lowering the blood sugar level.

Medical Field

It can also contribute to the knowledge of the medical field in the area of biology

and pharmacology in which it can help the next generations in terms of herbal

medicines.
Diabetic Individual

Specially, it can also help diabetic people because they can be inform that bitter

melon seed can be use as an alternative medicine to lower or reduce their blood sugar

level.

Scope and Delimitations

This study will be conduct to determine the in vitro effect of Bitter Melon

(Momordica Charantia) Seeds on lowering the blood sugar level through the use of

rabbit as a specimen. The age and the variety of samples are not included in the study.

This study will be conduct at Cainta, Rizal for the preparation of the bitter melon

seeds, while the phytochemical analysis will be conduct at registered institute -

Department of Science and Technology, testing division at Taguig, Metro Manila.

This study is limited only on the study of rabbits as specimen sample. The age

and variety of the sample were taken into considerations. The rabbit that will be use is

going to be brought only at Cainta, Rizal.

 CHAPTER II

REVIEW OF RELATED LITERATURE AND STUDIES

This chapter includes the review of related literature and studies which are found

significant in the study.

Bitter Melon (Momordica Charantia)

Bitter Melon is used as a traditional medicinal plant for the treatment of various

diseases in Bangladesh as well as other developing countries like Brazil, China,

Colombia, Cuba, Ghana, Haiti, India Mexico, Malaya, Nicaragua, Panama, and Peru.

The most common traditional use of the plant is to treat diabetes in different

countries around the globe. It is also used for the treatment of various other pathological

conditions such as dysmenorrhea, eczema, emmenagogue, galactagogue, gout,

jaundice, kidney (stone), leprosy, leucorrhoea, piles, pneumonia, psoriasis, rheumatism,

and scabies.

Recent scientific exploration on this plant elucidated potential biological effect on

both animal and clinical studies. Apart from its potential antibacterial [36] and antiviral

activities, bitter melon extracts are also effective against cancer and were found to be

effective for the treatment of ulcer, malaria, pain and inflammation, psoriasis,

dyslipidaemia, and hypertension. Bitter Melon also contains biologically active chemical

compounds such as glycosides, saponins, alkaloids, fixed oils, triterpenes, proteins, and

steroids. Several other biologically active chemical constituents have so far been

isolated from different parts of the plant, including the leaves, fruit pulp, and seeds. (Md

Ashraful Alam et al., 2015)

Blood Sugar (Glucose)

Blood glucose (also called blood sugar) is an essential measure of your health.

Too much glucose in the blood is the common factor between all types of diabetes.

Whenever the glucose (sugar) level in one's blood rises high temporarily, this condition

is known as hyperglycemia. The opposite condition, low blood sugar, is

called hypoglycemia. Glucose comes from most foods, and the body uses other

chemicals to create glucose in the liver and muscles. The blood carries glucose (blood

sugar) to all the cells in the body. To carry glucose into the cells as an energy supply,

cells need help from insulin. Insulin is a hormone made by the pancreas, an organ near

the stomach. The pancreas releases insulin into the blood, based upon the

blood sugar level. Insulin helps move glucose from digested food into cells. Sometimes,

the body stops making insulin (as in type 1 diabetes), or the insulin does not work

properly (as in type 2 diabetes). In diabetic patients, glucose does not enter the cells

sufficiently, thus staying in the blood and creating high blood sugar levels. (Ferry,

Robert Jr., 2017)

Rabbit (Oryctolagus Cuniculus)

All varieties of domestic rabbits used in modern biomedical research were

developed from the European rabbit, Oryctolagus Cuniculus. The European rabbit

possesses a mixture of features that make it an appealing model for in vivo research: an

intermediate body size, tractable disposition, ability to breed readily in captivity, and a

short generation time. Given its obvious utility as a research subject, the European

rabbit is richly represented in the scientific literature from the Age of Enlightenment

onward. When breed standards improved and production facilities moved from the
backyard to commercial rabbi tries, rabbits were increasingly accepted as research

models. The obvious physical and behavioural differences between European rabbits

and other leporids ultimately led to the recognition of Oryctolagus as a separate genus,

with O. Cuniculus as the only member species. The role of the European rabbit as a

game species, agricultural commodity, research model, invasive pest, endangered

species, and companion animal has increased.(Naff and Craig, 2012)

Definition of Terms

Alkaloids- any of a class of nitrogenous organic compounds of plant origin which have

pronounced physiological actions on humans. They include many drugs (morphine,

quinine) and poisons (atropine, strychnine).

Bitter Melon- also known as bitter gourd or Momordica Charantia, is a tropical fruit-like

gourd said to offer a variety of benefits. Bitter melon may be consumed as a food, as a

juice known as karela juice, or as a tea. Bitter melon contains compounds thought to aid

in the treatment of conditions like diabetes.

Blood Sugar (Glucose)- The blood sugar level, blood sugar concentration, or blood

glucose level is the concentration of glucose present in the blood of humans and other

animals. Glucose is a simple sugar and approximately 4 grams of glucose are present

in the blood of a 70-kilogram human approximately.

Diabetic- an individual having or suffering from diabetes.

Glycosides- a compound formed from a simple sugar and another compound by

replacement of a hydroxyl group in the sugar molecule. Many drugs and poisons

derived from plants are glycosides.

Hyperglycemia- an excess of glucose in the bloodstream, often associated with

diabetes mellitus.

Hypoglycemia- is deficiency of glucose in the bloodstream.

In vitro- In vitro studies are performed with microorganisms, cells, or biological

molecules outside their normal biological context. Colloquially called "test-tube

experiments", these studies in biology and its sub disciplines are traditionally done in lab

ware such as test tubes, flasks, Petri dishes, and microliter plates.

In vivo- Studies that are in vivo (Latin for "within the living"; often not italicized in

English) are those in which the effects of various biological entities are tested on whole,

living organisms or cells, usually animals, including humans, and plants, as opposed to

a tissue extract or dead organism.

Phytochemicals- are any of various biologically active compounds found in plants.

Rabbit (Oryctolagus Cuniculus)- The European rabbit or coney is a species of rabbit

native to south-western Europe and to northwest Africa. It has been widely introduced

elsewhere, often with devastating effects on local biodiversity.

Saponins- Saponins are a class of chemical compounds found in particular abundance

in various plant species. More specifically, they are amphipathic glycosides grouped

phenomenologically by the soap-like foam

Steroids- any of a large class of organic compounds with a characteristic molecular

structure containing four rings of carbon atoms (three six-membered and one five). They

include many hormones, alkaloids, and vitamins.

Triterpenes- any of a group of terpenes found in plant gums and resins, having

unsaturated molecules based on a unit with the formula C30H48.

 CHAPTER III

METHODOLOGY

This chapter presents the methods to be used in the study, research locale and

research methodology.

This study will determine the effectiveness of Bitter Melon (Momordica Charantia)

seeds on lowering the blood sugar level of the rabbit or the Oryctolagus Cuniculus.

Methods of Research

The researcher used experimental method of research because it involves

control and manipulation of data or variables, where it can establish a cause-effect. It is

for the researcher to determine if the Bitter Melon (Momordica Charantia) Seeds have

an effect on lowering the blood sugar level of the variables that are being manipulated.

Materials Used

Bitter Melon (Momordica Charantia) seeds was the main material that is used in

the conduction of the study, where in the rabbit is the respondent used. Mortar and

Pestle and a frying pan were used in the process due to the grinding and drying of the

seeds. Gloves together with the lab gown were also used on the conduction of the

study. In addition, 6 syringes with 21 gauge needle and 2 Grey-top tube (Potassium

Oxalate/Sodium Fluoride) were provided in the initial and final extraction of blood

sample and also for the injection of the extract. Moreover, a cage with a temperature set

on about 15-21°C and humidity at 50% were provided together with the food and water

that is needed for the living of the rabbit.

Research Locale

The experimental method of the study was done under strict laboratory

conditions. The extraction and supervision of the specimen were conducted at Buenmar

Ave. Greenland Subd. Brgy. San Juan Cainta, Rizal. The collection of the Bitter Melon

Seed was also done at Greenland Subd Brgy. San Juan Cainta, Rizal. The blood

extraction of the rabbit were conducted at Rowenas Garden Ortigas Ext, Dolores,

Taytay, 1920 Rizal in Nano Veterinary Clinic while the testing of the blood samples

was under the supervision of Femar, Room 219 -221, street, Barangay, M. L. Quezon

Ave, Antipolo, 1870 Rizal.

Research Methodology

The experimental study has five major phases: I. Phytochemical Analyses of

Bitter Melon (Momordica Charantia) seeds II. Initial blood testing in rabbit III.

Preparation of the Bitter Melon Seeds VI. Testing of the Extract on the rabbit V. Final

blood testing of the rabbit.

Phase I. Phytochemical Analyses

Test for Sterols and Triterpenes: Liebermann-Burchard Test

Evaporate about 5 ml of the chloroform solution of each sample to a small

volume, about 1ml of acetic anhydride was added followed by 2 ml of sulphuric acid

poured carefully on the wall of the test tube to form a lower layer. A reddish brown

colour appeared indicates the presence of triterpenes while when there is a grass-

green coloration were observe it indicates the presence of the sterols.

Test for Flavonoids: Magnesium Turning Test

The stock solution (1 mL) was taken in a test tube and added few drop of dilute

NaOH solution. An intense yellow colour was appeared in the test tube. It became red

when on addition of a few drop of dilute acid that indicated the presence of flavonoids.

Test for Alkaloids: Mayer’s test

It is precipitated from neutral or slightly acidic solution by Mayer's reagent

(potassiomercuric iodide solution). A cream coloured yellowish precipitate indicates the

presence of the alkaloids.

Test for Saponins: Froth Test

Three millilitres (3 mL) of the aqueous solution of the extract were mixed with 10

mL of distilled water in a test-tube. The test-tube was stoppered and shaken vigorously

for about 5 min, it was allowed to stand for 30 min and observed for honeycomb froth,

which was indicative of the presence of saponins.

Test for Glycosides: Fehling’s Test

Two millilitres (2 mL) of the aqueous solution of the extract in a test tube was

added into 5 mL mixture of equal volumes of Fehling’s solutions I and II and boiled in a

water bath for about 2 min. The brick-red precipitate was indicative of the presence of

reducing sugars.
Test for Tannins: Ferric Chloride Test

Two millilitres (2 mL) of the aqueous solution of the extract were added to a few

drops of 10% Ferric chloride solution (light yellow). The occurrence of blackish blue

colour showed the presence of Gallic tannins and a green-blackish colour indicated

presence of catechol tannins.

Phase II. Initial Blood Testing of the Rabbit

Preparing the rabbit for getting the initial Collecting the blood sample through the
blood sample back of the neck of the rabbit

Phase III. Preparation of the Bitter Melons seeds

Prepare and Gather all the materials Prepare also the frying pan for the drying
needed especially for the sanitation of the seeds
Phase III. Preparation of the Bitter Melons seeds Phase III. Preparation of the

Bitter Melons seeds

Phase VI: Testing of the Extract on the rabbit

Injecting of the extract on the rabbit

through the back of the neck

Phase V: Final Blood Testing of the Rabbit

Collecting the blood sample through the
back of the neck
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In Vitro Effect of Bitter Melon (Momordica Charantia)

Seeds on Lowering the Blood Sugar Level of

Diabetic Rabbit (Oryctolagus Cuniculus)

Karl Darwin A. Legaspi
Proponent
Mrs. Ofelia D. Astapan
 Teacher
G9- Narra
Gr. and Sec.