Experiment No. 5.5: Object: To determine the chemical constituents of a given sample of milk.

Cows milk sample about 500ml, glass ware, different chemical solutions (e.g., Benedicts reagent, 1% acetic acid, NaOH, 1% CuSo4, ammonium molybdate, HNO3, potassium oxalate, H2So4), filter paper, and a burner.

Requirements:

Theory and principle: Milk is a type of the foods popularly used by people of all age groups, particularly children and elderly people. The reason is that it is easy to take, easy to digest and a good source of different important nutritional constituents. Nutritional status of the milk can be further increased by adding different supplements in it or by mixing different fruit juices or flavors in it. Different cereals and edibles are also made palatable by mixing them in milk. Children of all mammals including humans depend totally upon milk for their nutritional requirements for a certain period of their life. Apart from this nutritional importance, milk is a good source of substances like calcium and phosphate. Main sources of fresh milk are cows and buffalo but it is also obtained to some extent from sheep, goats and camels. Composition of human and cows milk is very similar in many aspects, but there are certain differences as well, like the amino acids in the human milk are ideal for absorption and utilization especially by the neonatal brain e.g., taurine. Also the main protein found in cows milk, Casein, forms a somewhat indigestible curd and has a high level of phenylalanine, tyrosine and methionine for which the infant has very little digestive enzymes. Taurine is not found in cows milk. During the period of increased energy and nutritional requirements like in the school going children, during adolescence when the growth rate is high, during and after illness, in females during pregnancy and lactation milk is the best supplement.

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Procedure: I. Formation of milk filtrate: a. Take a measuring cylinder and add the following chemicals: i. ii. iii. 10ml of milk. 10ml of 1% acetic acid. 20ml distilled water.

b. Shake the cylinder well and after few minutes filter it. The process of formation of milk filtrate is shown in the Fig. 5.5.1.

Fig. 5.5.1:

The bottle, funnel and filter paper are in position for the preparation of milk filtrate.

II. Reaction of milk filtrate with different chemicals: a. Test for the detection of carbohydrates: i. Take 5ml of Benedicts reagent in a test tube. ii. iii. iv. Add 1ml of the filtrate. Boil for a few minutes. Black red precipitate confirms the

presence of sugar. The result is shown in the Fig. 5.5.2.

Fig. 5.5.2: Appearance of black red precipitate on addition of Benedicts reagent and boiling represents the presence of glucose in the milk filtrate.

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b. Test for the detection of proteins: First method: i. ii. iii. Take 5.0ml of filtrate in a test tube. Heat the test tube over the burner. Coagulation of the filtrate confirms the presence of proteins.

Second method: i. ii. iii. iv. Take 4.0ml of the filtrate in a test tube. Add 3.0ml of NaOH. Add 1.0ml 1%CuSO4. Change of colour to purple or violet confirms the presence of proteins. c. Test for the detection of phosphate: i. ii. iii. iv. Take 2.0ml of the filtrate in a test tube. Add 3.0ml of ammonium molybdate. Add 1.0ml of HNO3. Appearance of yellow colour confirms the presence of phosphate. The result is shown in the Fig.5.5.3.

Fig. 5.5.3: Result of the test for the presence of phosphate in the milk filtrate.

d. Test for the detection of calcium: i. Take 2.0ml of the filtrate in a test tube. ii. Add 1.0ml 2% potassium oxalate. iii. Add 2.0ml . iv. Appearance of yellowish white

precipitate confirms the presence of calcium. The results is shown in the Fig. 5.5.4. 287

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Fig. 5.5.4: Yellowish white precipitate indicating the presence of phosphate in the milk filtrate.

e. Test for the detection of cholesterol: i. Take 3ml of the filtrate in a test tube. ii. Add few drops of

concentrated H2SO4. iii. Mix and allow the test to stand for some time. iv. Appearance of two layers i.e., upper red and lower yellow confirms the

presence of cholesterol.

f. Test for the detection of casein: i. ii. iii. iv. Take the milk residue in a test tube. Prepare its solution by mixing with 5.0ml of distilled water. Add 1%acetic acid drop by drop. Appearance of whitish yellow precipitate confirms the presence

Precautions: I. At least 500ml cows milk should be used for the experiment so that enough volume of the milk filtrate is formed. II. For experiment always prepare fresh milk filtrate. III. Dont throw away milk residue because it will be used for the detection of casein. 288

Department of Physiology, Faculty of Medicine, Umm Alqura University, Makkah, Kingdom of Saudi Arabia

Students report page

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Experiment No. 5.5 Object: To determine the chemical constituents of a given sample of cows milk. Observation and results: Write down the results of various tests in the space provided below in table 5.3.1.
Milk filtrate= 10ml of the filtrate + 10ml 1% acetic acid + 20ml distilled water

S. No.

Tests

Result
Write down the colour that must appear for the detection of a substance.

1. 2.

Carbohydrate: 1ml milk filtrate + 5ml benedicts reagent Protein:

a. 5ml filtrate + heating b. 4ml filtrate + 4ml Biuret reagent 3.

Phosphate: 2ml filtrate + 3ml ammonium molybdate + 1ml concentrated HNO3

4.

Calcium: 2ml filtrate + 1ml 2% potassium oxalate + 1ml concentrated HNO3

5.

Cholesterol: 3ml filtrate + few drops concentrated H2So4 + allow to stand for some time Casein: Milk residue + 5ml distilled water + 1% acetic acid

6.

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Answer the following questions: Q.1. Write down any two advantages of drinking milk:

a.

_____________________________________________________

b.

_____________________________________________________

Q.2. Name the chemical substance that is only present in the human milk and not present in cows milk: ________________________________________________________

Q.3. Write down any conditions in which the requirements for milk is increased:

a._____________________________________________________

b._____________________________________________________

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