Experiment No. 4 Isolation and Characterization of Protein From Milk/Soymilk

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Experiment No.

4
Isolation and Characterization of Protein from Milk/Soymilk
I. Introduction
Soy milk or soya milk is a plant milk produced by soaking dry soybeans and
grinding with water. A traditional stable of asian cuisine that is caused by emulsion
of oil, water and pectin.
Milk is produced by mammary glands of mammals. It is the primary source of
nutrition for young mammals due to its digestible characteristic. Milk contains
calcium that is needed for the growth and development of bones and teeth.

II. Objectives
At the end of the experiment, the students should be able to:

Isolate casein from milk


Coagulate protein from soymilk using magnesium sulfate
Characterize protein from milk/soymilk

III. Materials
50 g. milk

1.6 g. magnesium sulfate

100 ml. water


Glacial acetic acid

Mayonaisse

HNO3
NaOH

Egg
Gelatin

95 % ethanol

Copper Sulfate Solution

125 ml soymilk

Cheese

III. Procedure:
A. Milk
1. Measure the mass of 250 ml beaker then add 50 g. of milk and dilute it
with 100 ml. water.
2. Prepare a water bath by placing 200 ml of water in a beaker. Heat the
water bath at 40
degrees celcius.
3. Slowly add 10 drops of glacial acetic acid to the milk while stirring
4. Filter the mixture to another beaker. Scrape the solid into a beaker. Add 25
ml of 95 % ethanol from the curd.
5. Stir the mixture for 5 minutes. Decant the liquid into another beaker

6. Weigh the solid and determine the percentage yield.


B. Soymilk
1. Weigh an empty beaker and record its weight. Then add cup of soymilk
in the beaker.
2. Heat the beaker with soymilk and let it boil.
3. Add 1.6 g. of magnesium sulfate to the hot soymilk and stir. Wait until the
curds are floating.
4. Filter the mixture into another beaker by pouring it through cheesecloth.
5. Scrape the solid into a beaker. Add 25 ml of 95 % ethanol from a curd.
6. Stir the mixture for about 5 minutes. Then decant the solid into another
beaker.
7. Weigh the solid and determine the % yield.

C. Biuret Test
1. Add an equal amount of NaOH to a solution of the food, mix carefully. .
2. Add a few drops of 1% CuSO4, do not shake the mixture.
3. a PURPLE/MAUVE COLOR is a positive result: protein is present.
4. Repeat steps 1-3 to the other samples
D. Xanthroproteic Test
1. Place 2 ml of matter that will be tested to the test tube, add with 1 ml of
strong HNO3 to the test tube.
2. If the sample turns to Yellow; then the protein is present.
IV. Guide Questions:
1.) Explain why the temperature in Part A should not exceed with 40 degrees
celcius?
2.) What is important of adding acetic acid and ethanol in isolating protein
from milk?
3. ) Compare the amount of acid casein precipitated from the whole milk with
the amount of
soy protein coagulated from the soymilk?
4.) What is the purpose of biuret test and xanthoproteic acid?
V. Conclusions.

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