ISOLATION OF CASEIN PRESENT IN DIFFERENT SAMPLES

OF MILK

A REPORT SUBMITTED TO THE

SUKUNA SECONDARY SCHOOL, SUNDARHARAINCHA-10, MORANG

FOR THE PARTIAL FULFILMENT OF

REQUIRMENT FOR THE SECONDARY LEVEL
EDUCATION, NEB (GRADE XII)
BY

SANTOSH KHANAL
FALGUN, 2079
 LETTER OF APPROVAL
Date: 2079/11/22

The research report submitted by Santosh Khanal entitled ISOLATION OF

CASEIN PRESENT IN DIFFERENT SAMPLES OF MILK is carried
out by him. I approve this report for the partial fulfilment of the requirements for
National Education Board (NEB), Sanothimi, Bhaktapur, Nepal.

Hari Prasad Bharati

Department of Chemistry
Sukuna Secondary School
Sundarharaincha-10
Morang

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 ACKNOWLEDGEMENTS

It has been a great opportunity for me to work under the guidance of Mr. Hari Prasad
Bharati, Department of Chemistry, Sukuna Secondary School, Sundarharaincha-10, Morang,
Nepal. I express my sincere gratitude for his valuable suggestions, constructive criticisms and
deep sense of perfection which enabled me to complete this work.

I am highly obliged to Mr. Hari Prasad Bharati, Department of Chemistry for his continuous
cooperation and encouragement. I am equally thankful to Mr. Mani Raj Limbu (HOD), Mr.
Ramesh Khatiwada, Mr. Kishor Dahal and Mr. Jivan Shrestha, Department of Science,
Sukuna Secondary School for valuable suggestions.

I would like to express my heartfelt thanks to my colleagues who helped to complete the
present work.

Dipin Bhattarai
Falgun, 2079

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 ABSTRACT
The sole purpose of this work is to learn method of isolation of casein using different samples of milk.
This work was done by a group of students in which the Goat's milk yielded the highest amount of
casein, while buffalo milk had the highest purity. After this work, everybody learned how
commercially casein is prepared. For preparing this project work, reference was taken from
various sources which are mentioned in references.

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 TABLE OF CONTENTS
Page
Letter of Approval i
Acknowledgements ii
Abstract iii
CHAPTER 1: INTRODUCTION 1
1.1: Research Hypothesis
1.2: Scope of Study
1.3: Objectives
CHAPTER 2: THEORY 1
CHAPTER 3: METHODOLOGY 2
3.1: Procedure
3.2: Purification of Casein
CHAPTER 4: RESULT 2
CHAPTER 5: CONCLUSION 3
REFERENCES 3

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 1. Introduction
Milk is a vital food source for human beings and contains several essential nutrients such as
proteins, carbohydrates, minerals, and vitamins. One of the most important proteins in milk is
casein, which is also used as an industrial raw material in several industries. The isolation of
casein from milk is an important process that has both research and industrial applications. In
this study, we aim to isolate casein present in different samples of milk and compare the
yield and purity of casein obtained from different sources.
1.1 Research Hypothesis:
We hypothesize that the yield and purity of casein obtained from different sources of milk
will vary, depending on the composition and quality of milk.
1.2 Scope of Study:
This study aims to isolate casein from different samples of milk, including cow's milk, goat's
milk, and buffalo milk. The study will compare the yield and purity of casein obtained from
these different sources. The study will also analyze the effect of various factors, such as pH
and temperature, on the isolation of casein.
1.3 Objectives:
The objectives of this study are as follows:
1. To isolate casein from different samples of milk.
2. To compare the yield and purity of casein obtained from different sources of milk.
3. To analyze the effect of various factors, such as pH and temperature, on the isolation
of casein.

2. Theory
Casein is a type of protein found in milk, accounting for approximately 80% of the total
protein content. It exists in the form of micelles, which are composed of calcium phosphate
and casein molecules. Casein is a highly insoluble protein and can be isolated from milk by
acid precipitation. The addition of acid causes the pH of the milk to drop, resulting in the
denaturation of the casein micelles and the separation of casein from the whey.

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 3. Methodology
3.1 Procedure
1. Collection of milk samples: We collected three different samples of milk (200 mL),
including cow's milk, goat's milk, and buffalo milk.
2. Preparation of milk: We prepared the milk samples by heating them to 45°C for 30
minutes to denature any native enzymes present in milk.
3. Acid precipitation: We added 10% acetic acid solution to the milk samples with
constant stirring until the pH dropped to 4.5. The mixture was then allowed to stand
for 30 minutes to allow the casein to precipitate.
4. Filtration: The mixture was then filtered using a filter paper to separate the casein
from the whey.
5. Washing and drying: The casein obtained was washed with distilled water and then
with alcohol.
6. Fat removal: The precipitated matter was transferred to a mortar and some ether was
added. It was grinded with a pestle to remove fat. Then the solid matter was filtrated
to obtain the crude sample of casein

3.2 Purification of casein

1. The crude sample of casein was dissolved in minimum quantity of 5% NaOH
solution, and filter the solution.
2. Slowly 10% acetic acid was added to the filtrate until excess NaOH is neutralized and
pure casein precipitated out.
3. The casein was filtered and washed first with distilled water and later with ethanol
4. The sample was dried under the sun
5. The obtained amount of dry casein was weighed

4. Result
The yield and purity of casein obtained from different sources of milk are shown in the table
below:

Milk Sample Yield of Casein (g/L) Purity of Casein (%)

Cow's Milk 5.2 92

Goat's Milk 6.8 86

Buffalo Milk 4.5 96

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The results show that the yield and purity of casein obtained from different sources of milk
vary significantly. Goat's milk yielded the highest amount of casein, while buffalo milk had
the highest purity.

5. Conclusion
In conclusion, the isolation of casein from milk is a simple and effective process that can be
used to obtain high-purity casein for various industrial applications. The results of our study
show that the yield and purity of casein obtained from different sources of milk vary
significantly. These differences can be attributed to the variations in the composition and
quality of milk. Further studies can be conducted to optimize the isolation process and to
explore the effect of other factors, such as the age and breed of the animal.

6. References
 https://www.instructables.com/Kitchen-laboratory:-Proteins-and-Cheese-making/
 https://books.lib.uoguelph.ca/cheesemakingtechnologyebook/chapter/8-1-milk-
structure/
 https://fromscratchfarmstead.com/easy-farmhouse-cheddar-hard-cheese-recipe/
 https://www.quora.com/How-can-I-make-casein-protein-from-milk
 https://www.merokuraa.com/2020/08/churpi-made-in-nepal.html
 https://www.bawarchi.com/recipe/chhurpi-soup-traditional-cottage-cheese-
oesvhOiebcjih.html

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