SIR C. R.

REDDY COLLEGE FOR WOMEN

(Affiliated to Adikavi Nannaya University, Rajamahendravaram)

ELURU, W.G.Dist.,(A.P)

CERTIFICATE

This is to certify that this is a bonafide record of the project work entitled
“Determination of Blood Grouping and Determination of Casein in Milk” done by
_______________________________________________________________ bearing
Regd. No: ______________________________, a student of III B.Sc., VI Semester in the
Department of Chemistry, Sir C. R. R. College for Women, Eluru, during the academic year
2021 -2022.

Project Supervisor Head of the Department Examiner

 ACKNOWLEDGEMENT

We feel highly grateful to Smt. P.Sailaja, Principal, Sir C.R.R College for Women,
Eluru, for being a motivating factor throughout the course of the project.

We are thankful to Smt B. Tulasi Koteswari, Head of the Department, Department of

Chemistry, Sir C R Reddy College for Women, Eluru for providing necessary facilities during
our work

We take this opportunity to express my deep sense of gratitude to our project guide,
Smt P. Ramya Krishna, for her keen interest and valuable guidance right from the inception
of the project till the complete execution of the same. We also thank other staff for their help.

I am using this opportunity to express my gratitude to all he staff who supported me

throughout the course of this project. I am thankful for their aspiring guidance, invaluably
constructive criticism and friendly advice during the project work.

We will be failing in our duty if we do not acknowledge the continued encouragement

and support we have had from all our family members, without whose help this project would
not have been completed.
 DECLARATION

I hereby declare that the work reported in this project work entitled
“Determination of Blood Grouping and Determination of Casein in Milk” is
entirely an original work carried out by me independently in the Department of
Chemistry , Sir C R Reddy College for Women, Eluru under the supervision of Smt P.
Ramya Krishna. I further declare that this dissertation has not been submitted by us
anywhere else for the award of any other degree.

Regd. No:

Place:
 INDEX

S.No Date Name of the Experiment Page No

1 Determination of Blood Group

Blood Sample - 1 1

Blood Sample - 2 3

Blood Sample - 3 5

2 Determination of Casein in Milk

Milk Sample - 1 7

Milk Sample - 2 9

Milk Sample - 3 11
 DETERMINATION OF BLOOD GROUP

BLOOD SAMPLE-1

AIM:
To determine the blood group of an individual by slide agglutination method.

PRINCIPLE :
The reaction behind the blood group determination is agglutination reaction that
results in clump Formation.

When particular antigen and its specific antibody are combined ,the antibody of
antiserum causes

The cellular antigens to add have to one and another to form clumps. The antibodies
that causes agglutination are Called “agglutinins” and particular antigens are called
agglutinogens. RBC agglutinating reactions are called “Haemoagglutination” reactions.

REQUIREMENTS:

1. Blood typing sera-anti A, anti B, anti D

2. 70%of ethanol
3. Microscopic slide
4. Applicator stick etc.,

Page | 1
PROCEDURE:
1. A clean glass slide was taken and marked out at 3 points as A,B and D.
2. The middle finger tip was wiped with the help of cotton , soaked in 70%of
ethanol and allowed to dry.
3. The disinfected area of fingertip was sprinkled with a sterile needle and squeezed
the finger to ooze the blood out.
4. A drop of blood was placed at each point marked on the slide.
5. Each 1 drop of anti A, anti B, anti D sera was added to this points A, B and D
respectively.
6. The blood drop and antiserum was mixed well at each point using separate
applicator sticks and allowed to agglutinate.

OBSERVATION:
When the antisera solutions were mixed with blood drops, agglutination was observed
with anti A and anti D.

DISCUSSION:
The human ABO blood group system was given by Landsteiner.
They are A,B,AB and O based on type of antigen present on the RBC.

Blood Group Type of Antigen Type of Anti Body

A A B
B B A
AB A,B No AB’s
O No AB’s A,B

Anti D solution indicates the presence of Rh factor.

RESULT:

The blood group of the individual was found to be

Page | 2
 DETERMINATION OF BLOOD GROUP
BLOOD SAMPLE-2

AIM:

To determine the blood group of an individual by slide agglutination method.

PRINCIPLE :

The reaction behind the blood group determination is agglutination reaction that
results in clump Formation.

When particular antigen and its specific antibody are combined ,the antibody of
antiserum causes

The cellular antigens to add have to one and another to form clumps. The antibodies
that causes agglutination are Called “agglutinins” and particular antigens are called
agglutinogens. RBC agglutinating reactions are called “Haemoagglutination”
reactions.

REQUIREMENTS:

1. Blood typing sera-anti A, anti B, anti D

2. 70%of ethanol
3. Microscopic slide
4. Applicator stick etc.,

Page | 3
PROCEDURE:
1. A clean glass slide was taken and marked out at 3 points as A, B and D.
2. The middle finger tip was wiped with the help of cotton , soaked in 70%of
ethanol and allowed to dry.
3. The disinfected area of fingertip was sprinkled with a sterile needle and squeezed
the finger to ooze the blood out.
4. A drop of blood was placed at each point marked on the slide.
5. Each 1 drop of anti A, anti B, anti D sera was added to this points A,B and D
respectively.
6. The blood drop and antiserum was mixed well at each point using separate
applicator sticks and allowed to agglutinate.

OBSERVATION:
When the antisera solutions were mixed with blood drops, agglutination was observed
with anti A and anti D.

DISCUSSION:
The human ABO blood group system was given by Landsteiner. They are A,B,AB
and O based on type of antigen present on the RBC.

Blood Group Type of Antigen Type of Anti Body

A A B
B B A
AB A,B No AB’s
O No AB’s A,B

Anti D solution indicates the presence of Rh factor.

RESULT:

The blood group of the individual was found to be

Page | 4
 DETERMINATION OF BLOOD GROUP

BLOOD SAMPLE-3

AIM:

To determine the blood group of an individual by slide agglutination method.

PRINCIPLE:

The reaction behind the blood group determination is agglutination reaction that results
in clump Formation.

When particular antigen and its specific antibody are combined ,the antibody of
antiserum causes

The cellular antigens to add have to one and another to form clumps. The antibodies
that causes agglutination are Called “agglutinins” and particular antigens are called
agglutinogens. RBC agglutinating reactions are called “Haemoagglutination” reactions.

REQUIREMENTS:

1. .Blood typing sera-anti A, anti B, anti D

2. 70%of ethanol

3. Microscopic slide

4. Applicator stick etc.,

Page | 5
PROCEDURE:

1. A clean glass slide was taken and marked out at 3 points as A,B and D.
2. The middle finger tip was wiped with the help of cotton , soaked in 70%of
ethanol and allowed to dry.
3. The disinfected area of finger tip was sprinkled with a sterile needle and
squeezed the finger to ooze the blood out.
4. A drop of blood was placed at each point marked on the slide.
5. Each 1 drop of anti A, anti B, anti D sera was added to this points A,B and D
respectively.
6. The blood drop and antiserum was mixed well at each point using separate
applicator sticks and allowed to agglutinate.

OBSERVATION:

When the antisera solutions were mixed with blood drops, agglutination was observed
with anti A and anti D.

DISCUSSION:

The human ABO blood group system was given by Landsteiner. They are A,B,AB
and O based on type of antigen present on the RBC.

Blood Group Type of Antigen Type of Anti Body

A A B
B B A
AB A,B No AB’s
O No AB’s A,B

Anti D solution indicates the presence of Rh factor.

RESULT:

The blood group of the individual was found to be

Page | 6
 DETERMINATION OF CASEIN IN MILK

MILK SAMPLE-1

AIM:

To study quantity of casein in buffalo’s milk.

THEORY:

Milk contains 3 to 4% casein suspended in the colloidal form. It is precipitated in a

weakly acidic medium

APPARATUS REQUIRED:

a. Funnel,
b. Funnel Stand
c. Glass Rod
d. Filter Paper
e. Weight Box
f. Test Tubes
g. Pestles
h. Mortar

CHEMICALS REQUIRED:

a. Sample of buffalo’s milk

b. Saturated ammonium sulphate solution
c. 1% acetic acid solution.

Page | 7
PROCEDURE:

1. Wash the beaker (250ml) with the distilled water and dry it.

2. Take 20ml of buffalo’s milk in 250ml beaker and find its weight.

3. Add 20ml of saturate 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 40-45oc 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 cow’s milk, goat’s milk, sheep’s milk.

PRECAUTIONS:

1. Handle apparatus and chemicals carefully.

2. Add ammonium sulphate solution very slowly
3. Stir milk while adding chemicals.
4. Do not disturb milk after adding ammonium sulphate solution and wait some
time for fat and casein to precipitate out.
5. Take the amount readings carefully with digital weighing machine only.

RESULT:

The percentage of casein present in buffalo’s milk is

Page | 8
 DETERMINATION OF CASEIN IN MILK

MILK SAMPLE-2

AIM:

To study quantity of casein in cow’s milk.

THEORY:

Milk contains 3 to 4% casein suspended in the colloidal form. It is precipitated in a

weakly acidic medium

APPARATUS REQUIRED:

❖ Funnel,
❖ Funnel Stand
❖ Glass Rod
❖ Filter Paper
❖ Weight Box
❖ Test Tubes
❖ Pestles
❖ Mortar

CHEMICALS REQUIRED:

➢ Sample of cow’s milk

➢ Saturated ammonium sulphate solution

➢ 1% acetic acid solution.

Page | 9
PROCEDURE:

a) Wash the beaker (250ml) with the distilled water and dry it.

b) Take 20ml of cow’s milk in 250ml beaker and find its weight.

c) Add 20ml of saturate solution of ammonium sulphate slowly with stirring. Fat
and casein will separate out as precipitate.

d) Filter the above solution and transfer the precipitate in another beaker.

e) Treat the above precipitate with 30ml distilled water. Casein dissolves forming
milky solution whereas fat remains as such.

f) Warm the above contents of the beaker to 40-45oc on a low flame. Now add
1%acetic acid solution drop wise with Stirring when casein gets precipitated.

g) Filter the precipitated casein and wash with distilled water and dry it.

h) Find the weight of dry precipitate.

i) Repeat the whole experiment with cow’s milk, goat’s milk, sheep’s milk.

PRECAUTIONS :

1. Handle apparatus and chemicals carefully.

2. Add ammonium sulphate solution very slowly.

3. Stir milk while adding chemicals.

4. Do not disturb milk after adding ammonium sulphate solution and wait some
time for fat and casein to precipitate out.

5. Take the amount readings carefully with digital weighing machine only.

RESULT:

The percentage of casein present in cow’s milk is

Page | 10
 DETERMINATION OF CASEIN IN MILK

MILK SAMPLE-3

AIM:

To study quantity of casein in goat’s milk.

THEORY:

Milk contains 3 to 4% casein suspended in the colloidal form. It is precipitated in a

weakly acidic medium

APPARATUS REQUIRED:

❖ Funnel,
❖ Funnel Stand
❖ Glass Rod
❖ Filter Paper
❖ Weight Box
❖ Test Tubes
❖ Pestles
❖ Mortar

CHEMICALS REQUIRED:

➢ Sample of goat’s milk

➢ Saturated ammonium sulphate solution

➢ 1% acetic acid solution.

Page | 11
PROCEDURE:

1. Wash the beaker (250ml) with the distilled water and dry it.

2. Take 20ml of goat’s milk in 250ml beaker and find its weight.

3. Add 20ml of saturate 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 40-45oc 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 cow’s milk, goat’s milk, sheep’s milk.

PRECAUTIONS:

1. Handle apparatus and chemicals carefully.

2. Add ammonium sulphate solution very slowly.

3. Stir milk while adding chemicals.

4. Do not disturb milk after adding ammonium sulphate solution and wait some
time for fat and casein to precipitate out.

5. Take the amount readings carefully with digital weighing machine only.

RESULT:

The percentage of casein present in goat’s milk is

Page | 12