The Effect of Robusta Coffee (Coffea canephora) Fruit Extract Ethanol

on LDL Wistar Strain Male White Rat

Induced With High Fat Diet

Main Supervisor Co - supervisor

Dewi Ratih Handayani, dr., M.Kes Arinta Setyasari, dr., SpJP , FIHA
Oxa Putri Andini
4111191169
Faculty of Medicine
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CHAPTER I

INTRODUCTION
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Background

The effect of
What is inside Hypercholesterolemia and robusta coffee rind
Robusta coffee Antioxidant
robusta coffee skin hyperlipidemia in lowering LDL
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Identification of the Problems

Does the ethanol extract of robusta coffee rind at dose of

250 mg/KgBW/day, 500 mg/KgBW/day, 1000
mg/KgBW/day have an effect in lowering LDL levels in male
wistar strain rats induced by a high fat diet?

Identification of the Problems

What is the effective dose of ethanol extract of robusta

coffee rind at dose of 250 mg/KgBW/day, 500
mg/KgBW/day, 1000 mg/KgBW/day in reducing LDL levels
in male wistar strain rats induced by a high fat diet?
 Friday
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Purposes of the Research

General Purpose Special Purpose

1. To determine the effect of ethanol extract of robusta coffee

To determine the antihyperlipidemic
effect of robusta coffee rind on LDL
levels in wistar rats.
rind at a dose of 250 mg/KgBW/day, 500 mg/KgBW/day,
1000 mg/KgBW/day to reduce LDL levels in male Wistar
strain rats induced by a high-fat diet.
2. To determine the effect dose of ethanol extract of robusta
coffee rind at a dose of 250 mg/KgBW/day, 500
mg/KgBW/day, 1000 mg/KgBW/day in lowering LDL levels
in male wistar strain rats induced by a high-fat diet.
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Uses of the Research

Academic Benefits

To provide an understanding of the antihyperlipidemic effect

of robusta coffe rind on lowering LDL levels

Uses of the Research

Practical Benefits

To provide information to the public in utilizing robusta

coffee rind aa herbal medicine that give antihyperlipidemic
effects, to prevent cardiovascular disease.
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CHAPTER II

LITERATURE REVIEW
 Robusta Coffee

Robusta coffee rind is known to have phenolic compunds

that have an antioxidant role. These polyphenols are
known as active compunds that are widely contained at
coffee skin
 Antioxidant

Antioxidants are molecules that can neutralize the oxidation of

Reactive Oxygen Species in 2 ways, with Primary Defense and
Secondary Defense
 Reactive Oxygen Species

ROS refer to a group of metabolites derived from molecular

oxygen. ROS are produced as a result of the gradual reduction of
oxygen molecules and the univalent reduction of oxygen which
will produce highly reactive superoxide radicals
 Metabolism of Lipoprotein

Lipoprotein metabolism can be divided into 3 pathways, which are:

1. Exogenous Metabolic Pathway
2. Endogenous Metabolic Pathways
3. Reverse cholesterol transport pathway
 Lipoprotein
Lipoprotein Component of Lipids Source

Kilomikron Triacylglycerol Gut

VLDL Triacylglycerol Liver

LDL Cholesterol VLDL

HDL Phospholipids, Cholesterol Liver, gut

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Cholesterol LDL

LDL is a
lipoprotein that
Cholesterol is a has the highest
major constituent cholesterol
of plasma content and also
membranes and the lipoprotein
plasma that carries the
lipoproteins. most cholesterol
in the body
 Hyperlipidemia

Hyperlipidemia is a condition in which cholesterol and

triglyceride levels are elevated, or an increase in the
concentration of lipids or fats in the blood. In particular,
elevated LDL levels are one of the most common risk
factors and contribute to atherosclerosis.
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Pathway for plaque formation In hyperlipidemia

Monocytes and Appearance of fine Turns in to the

platelets stick to the Attach to the sit of cells in the intima beginning of plaque
Release of mediators
walls of blood endothelial damage and medial layers of appearance in
vessels blood vessels hyperlipidemia
 Extract Ethanol

Extraction is a process of separating the solubility of a substance with

the aim that the substance cannot be mixed with others. Usually there
are 2 solvent, Which are ethanol and methanol. Ethanol was chosen
because it’s relatively cheaper than the other one, easy to get, and more
polar
Framework
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Giving the ethanol extract of robusta coffee rind

in 250 mg/KgBW/day, 500 mg/KgBW/day, 1000
mg/KgBW/day has an effect in reducing LDL
levels in male Wistar strain rats induced by a
high-fat diet.

Hypothesis
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CHAPTER III

METHODS
 Research Design Research Subject

Laboratory Experimental Wistar strain male white rat

Analysis

Number of Samples

Upload
5 Experimental Groups (25 rats)
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Research Variables

Independent Variables Dependent Variable

the ethanol extract of robusta coffee LDL levels at plasma of male wistar
rind rats
 Operational Definition

1. The ethanol extract of robusta coffee rind

Thid robusta coffee rind extract will be extracted using an ethanol

2. Low Density Lipoprotein

LDL levels in wistar strain male white rats will be measured by using the CHOD –
PAP method (Cholesterol Oxidase – Peroxidase Aminoantypirin) on a photometric
system.

3. Wistar strain male white rat

Rats with increase cholesterol levels (hypercholesterolemia) which were rats with
increased in total cholesterol levels according to NCEP ATP III criteria, after treated with
a high fat diet and PTU
 Research Materials and Tools

1. MATERIALS : robusta coffee rind extract at a dose of 250 mg/kgBW/day, 500 mg/kgBW/day, and 1000
mg/kgBW/day, standard pellets for rats, high-fat diet, water for rats to drink, 0.01% propitiuracil and LDL
screening kit (Reagen for CHOD – PAP)

2. TOOLS : rat cage and place to eat for rats, water bottle, feeding tube 18G, sonde, scales, gloves,
measuring cups, refrigerator, scissors, labels, stationery, hematocrit pipe, and Autocheck® for checking
blood cholesterol.

The tools used to measure LDL are stopwatch, micropipette (size p1000, p2000, p20), test tube, LDL
examination reagent and spectrophotometer.
 Research Procedures

1. Making Ethanol Extract of Robusta Coffee Fruit Peel

2. Making a High-Fat Diet
3. Experimental Animal Treatment
4. Blood Drawing
5. Measurement of Rat Blood Cholesterol Levels
6. Measurement of LDL Levels
Research Flow
 Data Analysis

Normally test: Homogeneity Test:

One way anova test Post hock tukey
Shapiro - Wilk Lavene statistic
 Research Ethical Aspects

This study used Wistar Strain Male White Rats, so that the study was carried
out after obtaining an ethical license approved by the Research Ethics
Comission of the Faculty of Medicine, Jenderal Achmad Yani University
 Research Place and Time

Place:
- Experimental Animal Laboratiry of the Faculty of Medicine, Jenderal Achmad Yani
University
- Laboratory of Biochemistry and Biomolecular of the Faculty of Medicine, Jenderal
Achmad Yani University

Time;
April 2022 - February 2023
Research Schedule
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