I take this opportunity to express my profound sense of gratitude and appreciation to all those who helped me throughout the

duration of this project.

First and foremost, I express my sincere gratitude to my project guide and chemistry teacher, Mrs. N.Menezes under whose able guidance I was able to take up and finish this project. I am indebted to her invaluable guidance rendered to me. I would also like to thank our

Im also thankful our chemistry lab assistant, Mr. R.K. Sharma, without who this project would not have been in its current shape.

I also take this opportunity to thank my parents and all my friends, who supported me throughout the duration of this project.

Milk is considered a complete food as it contains most of the nutrients required for a balanced diet. Milk is the lacteal secretion practically free from cholesterol, obtained by the complete milking of cows, buffaloes, goats etc. Its relative density varies from 1.018 to 1.045. Milk has a pH ranging from 6.4 to 6.8, making it slightly acidic. Throughout the world, there are more than 6 billion consumers of milk and milk products, the majority of them in developing countries. India is the world's largest producer and consumer of milk, yet neither exports nor imports milk. Milk is a key contributor to improving nutrition and food security particularly in developing countries. Improvements in livestock, dairy technology and milk quality may offer the most promise in reducing poverty and malnutrition in the world. Milk is a very complex substance and its constituents are influenced by various stages of treatment. The principal constituents of milk are proteins, fats, carbohydrates, vitamins and minerals. Substances like pigments, enzymes and phospholipids as well as gases and white blood corpuscles (leucocytes) are also present.

Proteins are an essential part of our diet. They are used for building tissues and strengthening the bones. Milk proteins consist of casein (80%) and whey (20%). These proteins are of a high biological value. Compared to egg, milk has smaller quantities of sulphur-containing amino acids.

Fat is a form of glycosides in emulsified form, and can be separated from milk by allowing it to stand for some time. After boiling, the fat rises to the surface and is separated by centrifugation. This is possible because the relative density of fats is lower than that of milk in solution. Carbohydrate is present in the form of the sugar lactose in milk. It is he only carbohydrate which mammals can synthesize. Its sweetness is relatively less, compared to cane sugar. Lactose is easily fermented by lactobacillis and is used to produce fermented milk that can be kept for longer periods than fresh milk.

Milk has got varying amounts of vitamin A, thiamine, riboflavin and nicotinic acid. It also contains vitamin B-12, which vegetarian foods lack. However milk is a poor source of vitamin C and vitamin E. Milk also has minerals like calcium, phosphorous, sodium and potassium, but it lacks iron. Milk is 85% water.

Milk casein combined with calcium, exists in milk in colloidal form and this is known as caseinogens. Casein is a phosphoprotein, meaning that phosphate groups are attached to hydroxyl groups of the same amino acids side chain. Fermentation of milk or addition of rennet leads to precipitation of insoluble calcium caseinate curds. There is a higher proportion of calcium and casein in animal milk and hence curds are harder to digest than human milk.

To extract casein and lactose from different samples of milk and thus compare their quantities in different samples.

Casein exists in milk as a calcium salt: calcium caseinate; present in the form of a micelle which has a negative charge. Thus, when an acid is added to the milk, the negative charge on the micelle is neutralized and casein gets precipitated. For example, 10% aqueous solution of acetic acid when added to milk, results in the precipitation of casein. Ca2 + Caseinate + 2CH2COOH(aq) Casein(s) + (CH3COOH)2Ca(aq)

Calcium caseinate has an isoelectric point of pH 4.6. The isoelectric point may be defined as the pH where no net migration of the protein under the influence of an external electric field takes place. At isoelectric pH, the solubility of a protein is minimum and thus it gets precipitated. The pH of milk is 6.6; therefore casein has a negative charge at this pH and is dissolved as a salt. But when acid is added, pH of the milk falls to 4.5-4.7 and hence casein gets precipitated.

500 ml beakers Filtration Flask Funnel Filter Paper Measuring Cylinders Glass Rod Dropper Weight Box Burner Tripod Stand Mortar Muslin Cloth

10% Acetic Acid (CH2COOH) Ethyl Alcohol (C2H5OH) 5% Sodium Hydroxide (NaOH) Rectified Spirit Ether Activated Charcoal

Isolation of Casein 1. Diluted 100ml of milk sample with 50 ml distilled water. 2. Warmed it to 40-50C and added 10% acetic solution drop wise until the precipitation was complete. Stirred the contents vigorously for 5-10 minutes and filtered the precipitated casein through a muslin cloth. 3. Preserved the filtrate for isolation of lactose. Purification of Casein 1. Washed the precipitated casein with water and ethanol. 2. Transferred the precipitate to a mortar, added ether and ground the mixture to remove fats. 3. Filtered and dissolved the precipitate in minimum quantity of NaOH solution. 4. Filtered off the insoluble material and the added ethanol. Dried it in a desiccator and recorded the yield.

Isolation of Lactose 1. Added NaOH solution to the filtrate left after the removal of casein to neutralize the acetic acid and warmed the contents to precipitate water soluble albumin. 2. Filtered off and reduced the volume of the filtrate by heating it in a steam bath.

3. Added a small quantity of ethanol and a little activated charcoal. Boiled the contents over the water bath to get a syrupy liquid, added more ethanol and kept it undisturbed for crystallization of lactose.

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1. Casein should be pressed by spatula during filtration. 2. Excess of acid should not be added, i.e. only the minimum required amount for precipitation should be added. 3. Milk should not be allowed to stand too long before use in examination. 4. Same volume of each sample should be used for the experiment.

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1. 2. 3. 4.

http://www.howstuffworks.com/dictionary/biology-terms/protein-info.htm http://en.wikipedia.org/wiki/Milk Together With Chemistry lab manual, 2012 edition NCERT Chemistry textbook, 2012 edition

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