 ACKNOWLEDGEMENT:

I would like to express my special thanks of

gratitude to my Chemistry teacher Mrs. Jagmeet
Chhabra for their able guidance and support in
completing this project. I would also like to
extend my gratitude to Principal Sir Mr. U.K Jha
for providing me the golden opportunity to do this
wonderful project.
During the accomplishment of the project I learnt
so many new things which benefited me and
helped me in gaining a lot of knowledge.
Lastly I would like to thank my parents and
friends who helped me in finalizing the project
within the limited time frame.

DARSHNA GUPTA

XII-A
 CERTIFICATE:
This is to certify that this “Chemistry
Investigatory Project” on the topic “Study the
quantity of casein in milk ” has been successfully
completed by DARSHNA GUPTA of class XII-A
under the guidance of MRS. JAGMEET CHHABRA
(subject teacher) for the academic year 2019-20
in particular fulfilment of the curriculum of
CENTRAL BOARD OF EDUCATION (CBSE).
All the works related to the project are done by
the candidate herself and the approach towards
the subject has been sincere and scientific.

Sign of Sign of Sign of

Principal: Teacher: External:
 INDEX:

S.NO CONTENT

1. AIM OF THE PROJECT

2. INTRODUCTION

3. MILK PROTEIN

4. CLASSIFICATION OF MILK PROTEIN

5. WHEY PROTEIN

6. CASEIN PROTEIN

7. EXPERIMENT

8. CONCLUSION

9. BIBLIOGRAPHY
 AIM OF THE PROJECT:

To study the quantity of Casein

present in milk
Introduction:
MILK:
Milk is a complete diet as it contains proteins, carbohydrates,
fats, minerals, vitamins and water. It is a multinutrient fluid
and is the primary source of nutrition for human. The average
composition of milk from different sources is given below:

SOURCE CARBO-
WATER MINERALS PROTEINS FATS
OF HYDRATES
(%) (%) (%) (%)
MILK (%)

Cow 87.1 0.7 3.4 3.9 4.9

Human 87.4 0.2 1.4 4.0 4.9

Goat 87 0.7 3.3 4.2 4.8

Sheep 82.6 0.9 5.5 6.5 4.5

Milk is an emulsion or colloid of butterfat globules within a

water-based fluid that contains dissolved carbohydrates and
protein aggregates with minerals. Because it is produced as a
food source for the young ones, all of its contents provide
benefits for growth. The principal requirements are (lipids,
lactose, and protein), biosynthesis of non-essential amino
acids supplied by proteins (essential amino acids and amino
groups), essential fatty acids, vitamins and inorganic
elements, and water.
Milk Protein:
There are several types of proteins in milk. The major
milk proteins are unique to milk (Not found in any other
tissue).
Milk contains 3.3% total protein. Milk proteins contain all
9 essential amino acids required by humans. Milk proteins
are synthesized in the mammary gland, but 60% of the
amino acids used to build the proteins are obtained from
the cow's diet. Total milk protein content and amino acid
composition varies with cow breed and individual animal
genetics.
Milk proteins are a source of nitrogen and amino acids
which are essential for the growth of the neonate and for
the maintenance of various bodily functions.
Milk proteins are used in variety of functional and
nutritional applications and some milk proteins also
possess biological activities.
Milk proteins possess functional properties that provide
desirable attributes to the final product and for this
reason have found numerous applications in
traditional dairy products and in other foods.
Classification of milk protein:
In the earlier years proteins were classified largely on the
basis of empirical fractionation procedures. In most cases
proteins were classified on the basis of their solubility and
their composition. Presently the proteins in milk are classified
on the basis of their fractionations and their behaviour during
electrophoresis, difference in their solubility in various
solutions, difference in their sedimentation rate etc.

The protein in the milk is classified into Casein and Whey

protein on the basis of their chemical composition and
physical properties. Milk protein consists of 80% casein and
20% whey protein. Insoluble milk proteins are called casein,
whereas soluble proteins are known as whey proteins. Both of
these groups of milk proteins are considered to be of excellent
quality, with a high proportion of essential amino acids and
good digestibility.
1. Whey Protein:
 Whey is a family of proteins, accounting for 20% of the
protein content in milk.

 Whey protein is a mixture of ß-lactoglobulin, α-

lactalbumin, bovine serum albumin and immunoglobins.

 Whey protein is considered a complete protein as it

contains all 9 essential amino acids but it is low in
lactose content.

 The whey proteins exist as individual units dissolved in

the water phase of milk and functions of many whey
proteins are not clearly defined, and they may not have a
specific function in milk but may be an artifact of milk
synthesis.

STRUCTURE-
APPLICATIONS-

Benefits~
i. Whey proteins have been associated with
many beneficial health effects, such as decreased blood
pressure, improved mood during periods of stress,
weight loss and low cholesterol level.

ii. Whey protein is excellent for growing and maintaining

muscles. As a result, it’s a popular supplement among
athletes and bodybuilders. It is commonly marketed as
a dietary supplement, typically sold in powdered form
for mixing into beverages.

iii. Whey protein could improve the immune response in

children with asthma. One small study involving 11
children, published in the International Journal of Food
Science and Nutrition, found that children with asthma
who were supplemented with 10 gram whey protein
twice daily for 1 month had an improved immune
response.

Possible Dangers~

Some people who are allergic to milk may be specifically

allergic to whey. In moderate doses, whey protein does not
typically cause any adverse events. However, consuming
very high doses can cause: stomach pains, cramps, nausea,
headache, fatigue etc.

Consistent high doses of whey protein may also cause acne.

From a nutritional point of view, whey protein is very unusual
and does not have a natural equivalent.
2. Casein Protein:
 The primary group of milk proteins are the caseins which
forms the majority — or 80% — of proteins in milk. There
are 3 or 4 caseins in the milk of most species; the different
caseins are distinct molecules but are similar in structure.
All other proteins found in milk are grouped together
under the name of whey proteins.

 Casein is composed of phosphoprotein and occurs in milk

as calcium salt (calcium caseinate) in the form of micelle.
Several similar proteins form a multi-molecular, granular
structure called casein micelle. In addition to casein
molecules, the casein micelle contains water and salts
(mainly calcium and phosphorous). The casein micelles
consist of subunits of the different caseins (α-s1, α-s2 and
ß) held together by calcium phosphate bridges on the
inside. These micelles are spherical and are 0.04 to 0.3
µm in diameter, much smaller than fat globules. The
casein micelles are porous structures that allow the water
phase to move freely in and out of the micelle.

 The micelle has a negative charge. So when an acid is

added to the milk, the negative charges are neutralized.

Calcium caseinate + acetic acid → casein(s) + calcium acetate(aq)

 Casein is the most predominant phosphoprotein found in

milk and cheese. When coagulated with rennet, casein is
sometimes called Para casein. British terminology, on the
other hand, uses the term caseinogen for the uncoagulated
protein and casein for coagulated protein. As it exists in
milk, it is a salt of calcium.

 Casein is a complete protein meaning that it contains all

of the essential amino acids, which the body cannot
manufacture on its own. Caseins have an appropriate
 amino acid composition that is important for growth and
development of the nursing young. This high quality
protein in cow milk is one of the key reasons why milk is
such an important human food.

 Caseins are highly digestible in the intestine and are a

high quality source of amino acids. Most whey proteins
are relatively less digestible in the intestine, although all
of them are digested to some degree. When substantial
whey protein is not digested fully in the intestine, some
of the intact protein may stimulate a localized intestinal
or a systemic immune response. This is sometimes
referred to as milk protein allergy and is most often
thought to be caused by ß-lactoglobulin. Milk protein
allergy is only one type of food protein allergy.

 Some enzymes are associated with casein micelles, too.

The micellar structure of casein in milk is an important
part of the mode of digestion of milk in the stomach and
intestine, the basis for many of the milk products
industries (such as the cheese industry), and the basis for
our ability to easily separate some proteins and other
components from cow milk.

 Centrifugation of the skim milk in an ultracentrifuge

(usually about 50,000 x g or greater) results in pelleting
of the casein and in a supernatant called whey (also
sometimes called the serum phase of milk) which
contains the water, lactose and soluble non-casein
proteins. Once casein is removed, then by definition every
other protein left in the milk preparation is a whey
protein.

 Casein has isoelectric pH of about 4.7 and can be easily

separated around this isoelectric pH. Casein molecules
can also be separated from the whey by precipitation of
the casein with acid (similar to what happens in the
 stomach when milk is consumed) or by disrupting the
micellar structure by partial hydrolysis of the protein
molecules with a proteolytic enzyme. In the stomach of
the young of many species is an enzyme
called rennin which specifically hydrolyses the part of the
casein micelle resulting in formation of a curd.

 It dissolves slightly in water, extensively in alkalis or

strong acids. Individual molecules of casein alone are not
very soluble in the aqueous environment of milk.
However, the casein micelle granules are maintained as a
colloidal suspension in milk. If the micellar structure is
disturbed, the micelles may come apart and the casein
may come out of solution, forming the gelatinous material
of the curd. This is part of the basis for formation of all
non-fluid milk products like cheese.

 Casein micelles are stable but dynamic structures that do

not settle out of solution. They can be heated to boiling or
cooled, and they can be dried and reconstituted without
adverse effects. ß-casein, along with some calcium
phosphate, will migrate in and out of the micelle with
changes in temperature, but this does not affect the
nutritional properties of the protein and minerals.

 The caseins are stable to heat treatment. Typical high

temperature short time (HTST) pasteurization
conditions will not affect the functional and nutritional
properties of casein proteins. High temperature
treatments can cause interactions between casein and
whey proteins that affect the functional but not the
nutritional properties. For example, at high temperatures,
ß-lactoglobulin can form a layer over the casein micelle
that prevents curd formation in cheese.
STRUCTURE-
APPLICATIONS-
Benefits~
i. In addition to being
consumed in milk, casein in
used in the manufacturing of
various types of adhesives,
binders, protective coatings,
plastics (such as for knife
handles and knitting
needles), fabrics, food
additives and many other
products. It is used in paints
for fast drying water-soluble medium. Casein based glues
are formulated from the mixture of casein, water,
hydrated lime and sodium hydroxide and are popular for
woodworking, including for aircraft.

ii. It is commonly used by

bodybuilders and athletes as
a slow-digestive source of
amino acids as opposed to
the fast-digesting whey
protein because the body can
slowly digest casein protein,
which means muscle tissues
have a longer opportunity to
use the amino acids for
repair work and growth. Another reason for its use by the
bodybuilders is that Casein helps to build new muscular
tissues and promote lean muscle growth. It’s usually
known as an “anti-catabolic” protein.
iii. Casein is frequently found in
otherwise non-dairy cheese
substitutes to improve the
consistency especially when
melted. Around the world,
there are more than six
billion consumers of milk
and milk products. Over 750
million people live in dairy
farming households. Several
foods, creamers, and toppings all contain a variety of
caseinate. Sodium caseinate acts as a greater food
additive for stabilizing processed foods, however
companies could opt to use calcium caseinate to increase
calcium content and decrease sodium levels in their
products.

Caseinate Presence and Function in Different Products

Product Caseinate % Function

Meat 2–20 Texture and nutrition

Cheese Matrix formation, fat, and water

3–28
binding

Ice Cream 1–7 Texture and stabilizer

Whipped toppings
2–11 Fat stabilization

Pasta 2–18 Texture, nutrition, and taste

Baked goods 1–15 Water binding

Possible dangers~

i. Casein can cause minor side effects in people with a

dairy allergy. Due to casein being free of lactose, a lot
more people are able to consume it than they are other
dairy protein sources. But you can still be allergic to it if
you have a dairy allergy. The side effects of this can be
nausea, bloating, headaches, and similar minor
complaints, but nothing too serious.

ii. Casein has been found with low levels of

contamination. One issue that has surfaced recently is
contamination, a recent study by the clean label project
found that many protein powders had levels of lead,
cadmium, and BPA within them. Now there should be
some context given here, while the levels were higher
than expected, they were still well within safe levels.
Secondly, milk and egg based proteins (including casein)
were found to be much safer than vegan and plant based
protein powders.

iii. Casein may lead to a higher risk of mortality in a higher

protein diet. There is a lot of debate at the moment on
whether high protein diets are safe or not. The worry
about kidney problems occurring has finally been laid to
rest (protein will not affect healthy kidneys at all) but
some studies indicate that lower protein diets lead to a
lower risk of mortality in under 65(s). That same study
also found that people who were over 65 actually
benefited from higher protein, possibly due to the
increased requirement of Leucine in the elderly.
Experiment:
Aim- To study quantity of casein in different samples
of milk.

Theory- Milk contains 3-4% casein suspended in

water in the colloidal form. It is precipitated in a
weakly acidic medium. The name of casein is related to
the family of phosphoproteins. These proteins are
commonly found in the mammalian milk.
This study deals with the precipitation of casein from
the various milk samples such as cow milk, goat milk,
buffalo milk and also the samples that are available in
the market. The technique of precipitation of casein is
used to predict the protein content in the milk samples.
When dried, it is a white, amorphous powder without
taste and odour.
The quantity, quality and fat-content from the various
milk samples differ with the type of particular
mammals and their fodder. The composition of milk
varies with according to the animals from which it
comes, providing the correct growth rate and
development for the young of that species. The primary
function of protein in living cells is to promote growth
and maintenance.
Requirements-
Apparatus and Chemicals
 250 ml beakers
 Funnel
 Glass rod
 Chemical balances
 Test tubes
 Filtration flask
 Bunsen burner
 Different samples of milk
 1% acetic acid solution
 Saturated ammonium sulphate solution
Procedure-
1. Wash the beaker (250 ml) with the distilled water
and dry it.
2. Take 20ml of sample cow milk in 250ml beaker and
find its weight.
3. Add 20ml of saturated solution of ammonium
sulphate slowly with stirring. Fat and casein will
separate out as precipitate.
4.Filter the above solution and transfer the
precipitate in another beaker.
5. Treat the above precipitate with 30ml distilled
water. Casein dissolves forming milky solution
whereas fat remains as such.
6.Warm the above contents of the beaker to 40 - 45°C
on a low flame. Now, add 1% acetic acid solution
drop wise with stirring when casein gets
precipitated.
7. Filter the precipitated casein and wash with
distilled water and dry it.
8.Find the weight of dry precipitate.
9.Repeat the whole experiment with goat’s milk and
buffalo’s milk.
Observations-
Volume of milk taken in each case = 20 ml
Weight of milk taken = W₁ g
Weight of Casein isolated = W₂ g
Percentage of casein = Weight of Casein x 100
Weight of milk

YIELD OF CASEIN
S.NO SAMPLES
(grams)
1. Cow milk 7.8 g

2. Buffalo milk 4.0 g

3. Goat milk 6.4 g

4. Milk A 6.8 g

5. Milk B 5.5 g
Result-
According to our analysis of various samples of milk,
we conclude that:
Cow milk contains 5% casein.
Goat milk contains 3.2% casein.
Buffalo milk contains 4.2% casein.
According to the research findings, cow milk contains
the largest amount of casein protein. Thus, the cow
milk is suitable for the best muscle growth and basic
body building achievements. It was found that goat
milk contains the small amount of casein protein.
Although the mineral content of goat's milk and cow's
milk is generally similar, goat's milk contains more
calcium, potassium, iron, magnesium and sodium. All
milk has lots of casein but there are different types of
casein and for someone who has casein sensitivity, goat
milk may provide an alternative to which they do not
react.
According to above results, we conclude that Cow’s milk
is most beneficial for human beings.
Precautions-
 Use only fresh milk
 Handle apparatus and chemicals carefully.
 Add ammonium sulphate solution very slowly.
 Stir milk while adding chemicals.
 Use the same amount of each sample during
the experiment.
 Do not disturb milk after adding ammonium
sulphate solution and wait some time for fat and
casein to precipitate out.
 Take the amount readings carefully with digital
weighing machine only.
Conclusion:
This study clearly indicated that the amount of casein
precipitated from the cow milk was higher than that of
the other milk samples. The quantitative analysis of
casein precipitated from the various milk samples
provide the ample scope to the cottage cheese
manufacture.
According to the research findings, cow milk contains
the largest amount of casein protein. Thus, the cow
milk is suitable for the best muscle growth and basic
body building achievements. It was found that goat
milk contains the small amount of casein protein.
Although the mineral content of goat's milk and cow's
milk is generally similar, goat's milk contains more
calcium, potassium, iron, magnesium and sodium. All
milk has lots of casein but there are different types of
casein and for someone who has casein sensitivity, goat
milk may provide an alternative to which they do not
react.
 BIBLIOGRAPHY:
 https://healthtrends.com/casein/
 https://en.wikipedia.org/wiki/Milk
 https://www.uoguelph.ca/foodscience/book-
page/milk-proteins
 https://www.slideshare.net/dineshpol/amount-of-
casein-in-milk
 https://www.seminarsonly.com/Engineering-
Projects/Chemistry/amount-of-casein.php
 https://www.sciencedirect.com/topics/agricultural-
and-biological-sciences/milk-proteins
 http://ansci.illinois.edu/static/ansc438/Milkcomps
ynth/milkcomp_protein.html