Group 4
Group members
Sofiya kulkarni(53)
Rishabh Kataria (49)
Sreehari Kariat (46)
Karishma Ravichandran (47)
Leena Karunakaran (48)
Shoban Kathiresan (50)
Sumedh Malandkar (60)
Pankaj Kokate (51)
Sowmya Kolanjiappa (52)
Aishwarya Kumbhar (54)
Siddheshwar Laddha (55)
Nganthoi Laishram (56)
Dilip Lonkar (57)
Pranav Mahendra (58)
Soni Mahendran (59)
POTENTIAL LIPID LOWERING EFFECT OF MALUNGGAY (Moringa oleifera) SEED SAPONIN FRACTION IN
HIGH CHOLESTEROL FED ALBINO MICE
CHAPTER 1 - introduction
Background of the Study
Objectives
Significance of the Study
Scope & Limitation
Definition of Terms
List of Acronyms
 Background of the study
Obesity is a serious and increasing health problem consisting of an
excessive growth of adipose tissue.
Because of the adverse effects associated with this available lipid-
lowering agent, there is a growing interest in herbal remedies, aiming
to find well-tolerated naturally-effective drugs.
Moringa oleifera seed oil possess hypo-cholesterolemic activity &
hypolipidemic effects as it is found to lower serum cholesterol,
triglyceride, VLDL & LDL cholesterol levels with an increase in the HDL
cholesterol levels..
Moringa Oleifera seeds contain saponins in them. Saponins are natural
high-molecular-weight glycosides of triterpene or steroids. They are
anticholesterolemic due to the formation of a complex with cholesterol in
gastrointestinal tract thus preventing absorption
Till date saponins from M. oleifera have not been reported hence this
study was an attempt to isolate saponin component from the pods and to
see it's anti- hyperlipidemic effects . This investigation was conducted to
evaluate the effects of saponins from Moringa oleifera on the serum lipid
profile of hyperlipidemic albino mice.
GENERAL OBJECTIVES

To determine the effects of Moringa oleifera seed saponin fraction

on hyperlipidemic mice.
SPECIFIC OBJECTIVES
1.To determine the mean weight &BMI of albino mice on day 0,14 and 28 days of
treatment and high cholesterol diet on the following groups
A. Negative control (Saline)
B. Positive control (Atorvastatin)
C. 50 mg/kg of M.oleifera seed saponin suspended in ______
2.To determine the mean LDL level on day 0,14 and 28 days of the treatment group.
3.To determine if there is significant difference in the mean pre & post treatment LDLof
the treatment groups.
4.To determine the saponin content of Malunggay methanolic seed extract using HPLC
analysis.
SIGNIFICANCE
oLipids are a constitutive and important part of our diet, but hyperlipidemia
which encompasses elevation in cholesterol and HDL is also one of the major
reasons for adult morbidity, mortality and decrease in the quality of life due
to diseases like hypertension, atherosclerosis, Myocardial Infarction and
Coronary Heart Disease.

oMoringa is one of the easily available plants, which is often used in regular
households. Hence, if the hypolipidemic effects of Moringa Olifera are
proven, they can be used as prophylactic means to prevent all disorders and
problems that can be caused by elevated lipid levels in the blood
SCOPE AND LIMITATIONS
• Our study has some limitations within which our findings need to be
interpreted carefully. Some limitations of our study should be notated.
• First due to constraint time we have to finish our study within the time period
of 2 to 3 months.
• The study will be conducted on the limited expenses which will be convenient
to the students doing research.
• The study is only limited to the Swiss albino mice and we will be using limited
amount of mice for our study
THEORETICAL FRAMEWORK
Saponins are naturally-occurring surface-active glycosides . Their structure
consists of a sugar moiety linked to a hydrophobic aglycone called sapogenin.
The restoration of the insulin response, the increase of plasma insulin levels and
the induction of the hypoglycemic action of saponins seems to be due to different
mechanisms of action, such as the release of insulin from the pancreas.

They are anticholesterolemic due to the formation of a complex with

cholesterol in gastrointestinal tract thus preventing absorption
CONCEPTUAL FRAMEWORK

Moringa Oleifera Lipid lowering

Saponin fraction effect
Independent Dependent
variable variable
Chapter 2 - METHODOLOGY
Research Design
Research Locale
Data Gathering Instruments
Procedures
Ethical Considerations
Dummy Tables
Budget
Research design
The design of the Research is Experimental as we will conduct an experiment to
test the hypolipidemic effects of Moringa oleifera.
Research locale
1. The Moringa oleifera seed SAPONIN will be be extracted on 8th floor Research
Center, College Of Medicine Building ,Davao Medical School Foundation College,
Medical School Drive, Bajada, Davao City.

2. The HPLC will be done on 8th Floor Research Centre, College Of Medicine Building ,
Davao Medical School Foundation College,Medical School Drive, Bajada, Davao City.

3. The LDL testing will be done in Dmsf Hospital, Medical School Drive, Bajada, Davao
City.
DATA GATHERING INSTRUMENTS
1. Measurment of the LDL- The mice will be tested for their serum LDL
cholesterol levels for the baseline by using a Mouse LDL-Cholesterol Assay kit
by Crystal Chem.

2. Determination & quantification of active components of Malunggay seeds

using Phytochemical screening & HPLC
SAMPLING TECHNIQUE
Fifteen mice will be randomly divided GROUP DOSE [mg]

into three groups of five animals each

naming A to C (negative control, one
group treated with saponins and A Positive ( Atorvastin)
another one treated with atorvastatin –
positive control) and are given free
access to a high cholesterol diet B Negative Control
(HCD).

C 50
 PROCEDURE
1.The mice will be tested for their serum LDL cholesterol levels for the
baseline by using a Mouse LDL-Cholesterol Assay kit by Crystal Chem. The kit
uses a 3-microliter sample volume. The kit has a high sensitivity and can
measure from 4.5 to 24 mg/dL.
The pre test will be performed on all the groups of mice. The mice will be
subjected to fasting for approximately 10 hours. The test will be done early in
the morning at 8 am.
2. After the test, the Positive Control Group will be administered the drug
atoravastin, an anti cholesterol drug. The negative control group will not be
given any medications to control Serum LDL cholesterol levels. The
Experimental Group will be administered the Moringa seed extract equivalent
to a Saponin concentration of 50mg/kg. The drug and the Moringa extract will
be administered orally to the mice, everyday in the morning for a total period
of four weeks.
3. Following the administration of the dosage, the serum LDL
Cholesterol levels will be checked after every two weeks to see if there
is a significant reduction in the levels of serum LDL Cholesterol levels.
The post test will be done using the same Mouse LDL - Cholesterol Assay
Kit by Crystal Chem. The sample used will be 3 microliter of serum. The
test will be done for all the groups at the same time on the same day,
after a ten hour fasting.
4. The results of each test will be recorded and tabulated. Any increase
or decrease in serum LDL cholesterol levels will be duly noted
 PROCEDURE
Moringa oleifera seeds

Phytochemical Screening

HPLC

Extraction of Saponins

Categorise of the mice in 3 groups from A to C

C
Positive control Negative control
50 mg
 Total Saponin test in UIC
Methanolic sample preparation Rotavap
Solvent preparation

Rotavap

Freeze Drug

HPLC

Fraction Collector

Rotavap

Perform on Mice
ETHICAL CONSIDERATION
All the mice which was used in the study which remained hyperlipedemic after
the study will be justified accordingly on how much stress and pain was incurred
during the study and be treated appropriately. If there is unrelieved pain or
distress which necessitates the killing of the mice, euthanasia will be performed
by a skilled veterinarian.
RESULTS
• TABLES (EXAMPLE)
Group Post test LDL value

Positive Control
Mice:
1.
2.
3.
Negative Control
Mice:
4.
5.
6.
Experimental group
Mice:
7.
8.
9.
THANK YOU