B.

DETERMINATION OF CASEIN FROM MILK WITH AN ACID (VINEGAR)

INTRODUCTION Milk is the most nutritionally complete food found in nature. All kinds of milk, human or animal, contain vitamins (principally thiamine VitB1, riboflavin VitB2, pantothenic acid VitB5, and vitamins A, B12, D), minerals (calcium, potassium, sodium, phosphorus, and trace metals), proteins(mostly casein), carbohydrates (principally lactose), and lipids (fats). Whole milk is an oil-in-water emulsion, containing its 3.9% fat dispersed as micronized globules. The fat emulsion is stabilized by complex phospholipids and proteins that are adsorbed on the surfaces of the globules. Because the fat in milk is so finely dispersed, it is digested more easily than fat from any other source. The globules are lighter than water, thus coalesce on standing and eventually rise to the surface of the milk as cream. Vitamins A and D are fatsoluble substances and are thus concentrated in the cream. The fats in milk are primarily triglycerides, which are esters of saturated and unsaturated carboxylic acids with glycerol, a trialcohol. OBJECTIVE To determine a casein from milk with an acid (vinegar).

MATERIALS / APPARATUS Soymilk Pasteurized whole milk Vinegar pH meter pH buffer 4 and 7

Beaker , 250 ml

PROCEDURE 1. The pH meter with pH buffer 4 and 7 is calibrated 2. The empty beaker is weighed and the weight is recorded 3. The weight of 120 ml of milk in the beaker is weighed and recorded. 4. The pH of the milk is measured 5. The beaker with the milk is placed on a hot plate. The plate is turned off and the beaker is removed 6. 11ml of vinegar is added to the warm milk and stirred for 2 minutes, and then the milk is allowed to sit for 5 minutes. The casein is precipitated into heavy white curds. The remaining liquid was the whey. 7. The pH of the liquid portion is measured again 8. A piece (2-3 layers) of cheesecloth is cut out to cover the top and 2 inches down the sides of a beaker. 9. The cheesecloth over the top of the beaker is fasten using the rubber band 10. The curdled milk is poured into the beaker, the curds are collected in the cheesecloth and the vinegar and whey is allowed to drain off into the bottom of the beaker. 11. The cheesecloth is gathered up with the casein and rinsed in cool water by dipping into another beaker containing water. 12. The casein is squeezed until almost dried and the cheesecloth is spreader out to let the casein dry for 5 minutes 13. The precipitate is weighed. e ed e

RESULTS Determination of Casein in Whole Milk and Soymilk Whole Milk pH before added acid and before heated 3. 6.82, 24. pH after added acid and after heated 1. 4.88 , 33 C 3. 6.99, C 1. 6.90 Soymilk

1. 4.70, 37.8 C 2. 4.56, 32.0 C 3. 4.44 , 36.7 C 1. 125.29 2. 123.21 3. 124.36 1. 1.9290 2. 16.5392 3. 3.8957 Creamy White Colour Soft

2. 4.94 , 26.3 C 3. 4.92 , 30.3 C

Weight of milk

1. 120.18 2. 121.97 3. 121.21

Weight of curd or casein

1. 16.6298 2. 18.7700 3. 17.0757

Colour of curd or casein Texture of curd or casein

Creamy Yellow Colour Harder

DISCUSSION For this experiment, we have learned how to determine casein from milk with an acid (vinegar). We use whole milk and soy milk as our sample for this experiment. Milk composed of many protein .Casein and whey protein is the main groups. Proteins are polymers of amino acids. While for casein, Casein was first separated from milk by adding acid to milk, thus establishing its existence as a distinct protein. The whey proteins were separated into globulin and albumin fractions. It was subsequently shown that casein is made up of a number of fractions and is therefore heterogeneous. The whey proteins are also made up of a number of distinct proteins. For the determination of casein from milk with an acid, the pH of whole fat milk before added acid is 6.85and after added acid (+) heated is 4.91. While for soy milk is 6.98 and 4.57. But there is a difference in weight of casein for whole fat milk and soy milk .Soy milk has lower quantity of casein which is 16.5392 compare to whole fat milk which is 17.4918. For soy milk, there have error for trial one and three. The value we get is 1.9290 and 3.8957. This result is error because time of drying does not enough and the temperature use is too low. The colour also are different where by whole fat milk give a creamy yellow colour and soymilk give a creamy white colour .The texture of casein for soymilk is soft compare to whole fat milk which is harder . As the general, Casein is easily separated from milk, either by acid precipitation or by adding rennin. Casein is dispersed in milk in the form of micelles. The micelles are stabilized by c en en re e -

ydr p b c bu -casein contains a hydrophilic portion known as the

glycomacropeptide and it is this that stabilizes the micelles when the pH of milk is changed, the acidic or basic groups of the proteins will be neutralized.

At the pH at which the positive charge on a protein equals exactly the negative charge, the net total charge of the protein is zero. This pH is called the isoelectric point of the protein (pH 4.6 for casein). If an acid is added to milk, or if acid-producing bacteria are allowed to grow in milk, the pH falls. As the pH falls the charge on casein falls and it precipitates. Hence milk curdles as it sours, or the casein precipitates more completely at low pH.

CONCLUSION The objective of this experiment is to determine the pH of milk and to determine casein from milk with an acid (vinegar). The pH of milk changes over time. When increasing the period of the day, the milk goes sour, it becomes more acidic and the pH gets lower. For Casein, it is easily separated from milk, either by acid precipitation or by adding rennin. We are also able to observe the texture of the casein after added acid and heated. The objective was accepted.

QUESTION 1. What causes the reduction in the pH of milk? The reduction in the pH of milk is primarily caused by bacteria present in milk or its surrounding. As the bacteria grow and multiply in the milk, it will contaminate the milk and eventually change the odor and flavor of the milk making it tastes sour. That is when the pH of milk will start to decrease.

2. Compare the amount of acid casein precipitate from the low fat with the amount of soy protein coagulated from the soymilk? How do your result compare with the nutrition facts label for each product?

3. List 3 factors that influence the precipitation of casein in milk. Temperature, acidity, and calcium content of the milk.

4. W

p r nce

fc en n

e f d industry?

Casein is used in the chemical industry and as a glazing additive in paper manufacturing.

REFERENCE 1) http://biology.unm.edu/ccouncil/Biology_124/Summaries/Macromol.html 2) http://www.accessexcellence.org/AE/AEPC/IFT/unit_three.php 3) http://dspace.library.uu.nl/handle/1874/765 4) https://www.uoguelph.ca/foodscience/cheese-making-technology/section-c-milk/rawmilk-quality/milk-growth-medium 5) http://www.picotech.com/experiments/ph_of_milk/results.html