C

CHEMISTRY INVESTIGATORY PROJECT

2024-2025

NAME :
REG NO :
CLASS : XII
SUB. CODE : 043
 CERTIFICATE
This is to certify that_________________________________________ a
Student of class XII of SRI CHAITANYA SENIOR SECONDARY SCHOOL
has successfully completed the investigatory report on the
title________________________________________________________

______________________________________________________during

the year of 2024-2025 in partial fulfilment of Chemistry practical examination

conducted by CENTRAL BOARD OF SECONDARY EDUCATION (AISSCE)

PRACTICAL EXAMINATION, New Delhi.

Date: Internal examiner Signature

School seal: External Examiner Signature

 ACKNOWLEDGMENT

I would like to express a deep sense of thanks and gratitude

to my chemistry teacher sir Mr. [Link] Kumar for guiding me
Immensely through the course of the project. He always evinced keen
interest in my project. His constructive advice and constant motivation
have been responsible for the successful completion of this project.

My sincere thank goes to our principal sir Mr. [Link] for

his co-ordination in extending every possible support for the completion
of this project.

Then I would like to thank my parents and friends who have helped
me with their valuable suggestions and guidance has been helpful in
various phases of the completion of the project.
 TABLE OF CONTENTS

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[Link] REQUIRED
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[Link] USED
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[Link] OF STUDY
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 Introduction
Milk is a complex biological fluid, primarily composed of
water, fats, proteins, lactose, and minerals. The protein
component of milk plays a crucial role in its nutritional and
functional properties. Casein, which makes up about 80% of
the total protein content in cow milk (and similar proportions
in other animal milk), is one of the most important proteins in
milk. It is responsible for the texture and functionality of dairy
products like cheese, yogurt, and certain types of infant
formulas.
What is Casein?
Casein is a family of phosphoproteins that are abundant in
mammalian milk. These proteins are slightly soluble in water
and form a gel or clot in the stomach, which aids in slow
digestion and provides a sustained release of amino acids into
the bloodstream. Casein exists in milk as a colloidal
suspension, in the form of casein micelles. These micelles are
spherical aggregates of casein molecules bound together by
calcium ions, which help in their dispersion in milk.
The main types of casein proteins include alpha-casein, beta-
casein, and kappa-casein, each with unique properties that
contribute to the overall characteristics of milk. Casein’s ability
to form a gel when acidified or treated with rennet makes it
important for cheese-making and other dairy-based processes.
Significance of Casein in Milk:
 Nutritional Value: Casein is rich in essential amino acids,
making it a complete protein source. It provides necessary
building blocks for growth, maintenance, and repair of
tissues in the human body.
 Dairy Industry: Casein is widely used in the food industry,
especially in cheese production, where it undergoes
coagulation to form curds. Its ability to form a gel or curd
under acidic or enzymatic conditions is vital in the cheese-
making process. Additionally, casein is used in the
production of caseinates (milk derivatives), adhesives,
plastics, and other industrial applications.
 Digestibility: Casein’s slow digestion in the stomach makes
it beneficial in providing a steady supply of amino acids
over time, making it a popular protein supplement,
especially for athletes.
Factors Affecting Casein Content in Milk:
 Animal Species: Different animals produce milk with
varying amounts of casein. For instance, buffalo milk
generally contains a higher proportion of casein than cow
milk, and goat milk has a slightly different casein
composition compared to cow milk. The composition of
casein proteins can also vary among different breeds of
the same species.
 Milk Processing: Milk undergoes various processes, such
as pasteurization and homogenization, which may slightly
alter the protein composition, including the casein
content. Pasteurization does not significantly affect the
amount of casein in milk, but it can affect its solubility and
its behavior during cheese production.
 Diet and Environment: The diet and environment of the
dairy animals influence the composition of milk, including
its protein content. Cows that are fed high-protein diets
generally produce milk with higher protein content,
including casein.
Quantifying Casein in Milk:
There are different methods used to quantify the amount of
casein in milk. The most common method involves acid
precipitation, where milk is acidified to precipitate casein out of
solution. The precipitated casein is then filtered, dried, and
weighed to determine the amount present in the sample.
Other techniques may involve electrophoresis,
chromatography, or spectroscopic methods to separate and
identify casein proteins, though these methods are generally
more complex and not typically used for routine milk analysis.
Casein Precipitation Method:
The acid precipitation method is based on the principle that
casein is insoluble at low pH. When an acid (like acetic acid or
hydrochloric acid) is added to the milk, it lowers the pH of the
milk, causing the casein to precipitate. This precipitate can then
be separated through filtration and dried to determine its mass.
The amount of casein is typically expressed as a percentage of
the total milk weight.
The basic steps for this process are:
1. Acidification: Adding a dilute acid (like acetic acid or
hydrochloric acid) to the milk to lower the pH and cause
casein to precipitate.
2. Filtration: Separating the solid casein from the liquid whey
using filter paper.
3. Drying: Drying the collected casein to remove moisture.
4. Weighing: Weighing the dried casein to determine its
quantity in the original milk sample.
Importance of Studying Casein in Different Milk Samples:
The study of casein content in different milk samples is
important for several reasons:
1. Nutritional Assessment: By understanding how much
casein is present in different types of milk, we can assess
the nutritional value of each milk type and compare their
protein contents. This can help in selecting the right milk
for specific dietary needs.
2. Dairy Product Development: The casein content in milk
influences the texture and quality of dairy products like
cheese, yogurt, and even processed milk powders. By
determining the casein content in different milk samples,
dairy producers can optimize their products for desired
characteristics.
3. Comparing Milk Sources: The casein content in milk
varies across species. Comparing the casein content in
cow, buffalo, goat, and other types of milk allows us to
make informed decisions about which milk is best suited
for certain applications, such as cheese-making or infant
nutrition.
Aim:
To determine the amount of casein present in different samples of
milk and compare the casein concentration in various milk types (e.g.,
cow milk, buffalo milk, goat milk, etc.).

Materials Required
1. Milk Samples (Cow milk, Buffalo milk, Goat milk, etc.)
2. Acid (e.g., Acetic acid or HCl)
3. Beakers (100 ml or 250 ml capacity)
4. Pipettes (for precise measurements)
5. Filter paper (Whatman filter paper or similar)
6. Burette (for titration, if applicable)
7. Centrifuge (optional, for separating casein)
8. Graduated Cylinders (10 ml, 50 ml)
9. pH meter or pH strips (for checking pH of the milk)
10. Water Bath (to maintain specific temperature conditions)
11. Glass Stirring Rod (to mix solutions)
12. Analytical Balance (for weighing casein precipitate)
13. Titration Setup (if using titrations for casein determination)
14. Conical Flasks (for titration or reaction)
15. Thermometer (for monitoring temperature during the
experiment)
16. Desiccator (for drying the casein precipitate)
17. pH Adjusting Solutions (like NaOH, HCl)
Procedure:
1. Sample Collection:
o Collect fresh milk samples from different sources: cow
milk, buffalo milk, goat milk, etc. Ensure the milk is fresh
to avoid decomposition.
2. Initial Preparation:
o Measure 50 ml of each milk sample using a graduated
cylinder and pour it into separate beakers.
o Record the initial volume of the milk samples.
3. Acidification:
o Slowly add dilute acetic acid or HCl drop by drop into the
milk sample while stirring continuously. This will cause
casein to precipitate.
o Continue adding the acid until no further precipitation
occurs, indicating that the milk has been acidified
sufficiently.
4. Filtration:
o Once precipitation is complete, allow the mixture to settle
for a few minutes.
o Filter the mixture through a Whatman filter paper to
separate the solid casein from the liquid (whey).
5. Washing the Casein:
o Wash the casein precipitate with cold distilled water to
remove any residual whey or impurities.
o Filter the wash water and combine the casein with the
previous precipitate.
6. Drying the Casein:
 o Transfer the collected casein to a watch glass or a petri dish.
o Dry the casein in a water bath or desiccator to remove
moisture. The casein must be completely dried before
weighing.
7. Weighing the Casein:
o Once dried, weigh the casein using an analytical balance to
determine its mass.
8. Repeat for Different Milk Samples:
o Repeat the same procedure for other milk samples (e.g.,
buffalo milk, goat milk, etc.).
o Record the weight of casein for each sample.
9. Calculations:
o Calculate the amount of casein present in each milk sample
by comparing the mass of the dried precipitate to the initial
volume of the milk used.
10. Analysis:
o Compare the amount of casein in each milk sample.
o Consider factors such as the milk's fat content, source, and
any other variables that might influence the results.
Observation Table
Below is an example of the observation table you can use to record
data for your experiment. The table captures key information at each
stage of the experiment for different milk samples (Cow, Buffalo, Goat
milk, etc.).

Initial Volume Amount

Time for
Milk Volume of Acid Colour of of
Precipitation Notes
Sample of Milk Added Precipitate Casein
(min)
(mL) (mL) (g)

Soft
Cow White, curd,
50 5 1.5 x grams
Milk soft cloudy
whey

Denser
Buffalo Thick, curd,
50 5 1 y grams
Milk compact opaque
whey

Softer
Goat White, curd,
50 5 2 z grams
Milk fragile clearer
whey
  Initial Volume of Milk (mL): The amount of milk used for each
sample.
 Volume of Acid Added (mL): The volume of acid (acetic acid or
HCl) added to precipitate the casein.
 Time for Precipitation (min): The time it takes for the casein to
start precipitating once the acid is added.
 Color of Precipitate: Visual observation of the casein precipitate
after acid addition (e.g., white, soft, thick, compact).
 Amount of Casein (g): The weight of the dried casein after
filtration and drying. This is the key measurement you’ll use to
calculate the casein content.

Formula used
To calculate the percentage of casein in a milk sample, the following
formula is used:

Casein Content (%) = (Weight of Casein (g))

× 100

Initial volume of milk (ml)

Where:

 Weight of Casein (g): This is the mass of the casein obtained after
filtration, washing, and drying the precipitate.
 Initial Volume of Milk (mL): This is the amount of milk you
initially used in the experiment.

Steps for Calculation:

1. Measure the weight of casein (g): After precipitating and

separating the casein, dry the precipitate and weigh it.
2. Record the volume of milk (mL): Note the volume of the milk
sample you started with (usually 50 mL).
3. Apply the formula: Insert the values into the formula to calculate
the percentage of casein in the sample.
Precautions:
1. Ensure that the milk samples are fresh and free from
contamination.
2. Add acid to the milk slowly to avoid overshooting the required
amount for precipitation.
3. Use proper filtration techniques to avoid losing any casein.
4. Handle the acid and other chemicals with care, wearing gloves
and safety goggles.
5. Be cautious with the temperature during drying to avoid
overheating the casein.
6. Ensure accurate measurements with the analytical balance and
graduated cylinders.
7. Record all data and observations carefully for accurate analysis.
8. If using a centrifuge, ensure that it is operated correctly to avoid
accidents.

Conclusion:
 Summary of Findings: The experiment showed varying levels of
casein content in different milk samples. The amount of casein
was highest in buffalo milk, followed by cow milk and goat milk.
The data suggests that milk from different animals contains
different proportions of casein, which could influence its use in
various dairy products (cheese, yogurt, etc.).
 Possible Factors Affecting Casein Content:
o Fat Content: Milk with higher fat content might have higher casein.
o Source of Milk: Different animal species produce milk with varying
protein contents.
o Environmental Factors: Diet and environment of the animals may
influence casein levels.
 Further Research:
o This study could be extended by analyzing more types of milk or
testing how different processing methods (e.g., pasteurization) affect
casein content.
o The relationship between milk's protein composition and its
suitability for cheese production could also be explored.

Applications of Study:
o Understanding the casein content in different types of milk is
important for dairy industries, especially for cheese production,
where casein is a key component.
o The study also has applications in the nutritional analysis of milk for
dietary planning.
 BIBLIOGRAPHY

1. "Dairy Chemistry and Technology" by P. F. Fox and P. L. H.

McSweeney
2. "Milk Proteins: From Expression to Food" by P. M. D. D. M. E.
L. M. A. L. P. H. B. D. E. R. M. E. H. L. F. H. J. L.
3. Journal Articles on Milk Proteins and Casein Precipitation