Universities Research Journal 2011, Vol. 4, No.

Extraction of Casein and Determination of its Components from Three Kinds of Milk
Khine Zar Wynn Lae1 and Aye Aye Cho2

Abstract
In this research work, cow, goat and buffalo milk were collected and the protein and sugar tests were carried out which gave rise to positive for protein and trace amount of sugar. Moreover, casein protein was extracted from each sample. It was found that cow milk contains 45.36% of casein, goat milk contains 5.47% of casein and buffalo milk contains 14.07% of casein. According to these data, cow milk contains the largest amount of casein. The elemental analysis of casein compounds were determined by using Atomic Absorption Spectroscopy (AAS). From these determinations, all three milk samples contain the sufficient amount of minerals. In other words, the three samples contain adequate amount of nutrients in suitable proportion that maintain the health of an individual. Key words: casein protein, cow, goat, buffalo, Atomic Absorption Spectroscopy (AAS)

Introduction Casein (from Latin causes "cheese") is the predominant phosphoprotein that accounts for nearly 80% of proteins in cow milk and cheese. Casein protein is basically the most important protein that can be found in milk(Horne, 2002). An improved dietary supplement for mammals includes the essential dietary trace metals such as iron, copper and molybdenum. These metals are provided in a highly bioavailable state such as peptide and polypeptide chelates. This invention also includes a method for using natural proteinaceous starting materials for producing the peptide and polypeptide ligands for the chelates. The proteinaceous starting materials are obtained from a primary grown yeast, an isolated vegetable protein, dessicated tissues of animal origin and the like which, upon hydrolysis, yields the peptide and polypeptide ligands suitable for forming chelates with the iron, copper and molybdenum.

1. Assistant Lecturer, Dr, Department of Chemistry, University of Yangon 2. Professor and Head, Dr, Department of Chemistry, Myingyan Degree College

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Figure 1. Casein structure Casein protein is the majority of muscle repairing and rebuilding take place during sleep. For optimal recovery, the body needs a constant of nutrients, and most importantly, protein. When taken before bed, whey protein will be absorbed with about 45 minutes, leaving the body without a protein source for up to 8 hours. Micellar casein will provide the body with a protein source for up to 7 hours, greatly reducing muscle catabolism (Holt and Horne, 1996). The objective of the current study is to investigate the composition of casein protein in three kinds of milk. The essential analysis of casein compounds were determined by using Atomic Absorption Spectroscopy (AAS). Materials and Methods The cow, goat and buffalo milk samples were collected from Mandalay area. The protein tests such as Xanthoprotein test, Biuret test and Denaturation test, of three samples were carried out. Sugar tests were also determined. Caseins were isolated from the three kinds of milk samples which has pH of about 7. Casein will be separated as an insoluble precipitate of the milk to its isoelectric point (pH = 4.6). The fat that precipitates along with casein can be removed by dissolving it in alcohol. These precipitate milk products are protein. The identification will be achieved by performing a few important chemical test which are Biuret test, Ninhydrin test and Xanthoprotein test. The mineral contents of extracted casein protein from these three samples were determined by Atomic Absorption Spectroscopy (AAS).

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Results and Discussion The protein test for three milk samples were carried out. These data were tabulated in the following Table 1. Table 1. Results of protein tests No. 1. 2. 3. Protein Test Xanthoprotein Biuret Denaturation Observation Yellow Pink Denature Results Cow milk + + + Goat milk + + + Buffalo milk + + +

Based on the observation, these three milk samples gave positive test for protein. Therefore, these samples may contain casein protein. These three samples were tested the reducing sugar by using benedict solution. These were shown in the following Table 2. Table 2. Results of Sugar Test No. 1. 2. Sample Cow milk Goat milk Observation Green Green Results + +

3. Buffalo milk Green + The casein content of three milk samples were described in the following Table 3. Table 3. Casein protein from three milk samples No. 1. 2. 3. Sample Cow milk Goat milk Buffalo milk Casein protein (%) 45.36% 5.47% 14.07%

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50.00% 45.00% casein protein content (%) 40.00% 35.00% 30.00% 25.00% 20.00% 15.00% 10.00% 5.00% 0.00% Cow milk Goat milk milk samples Buffalo milk

Figure 2. Casein protein from three milk samples The elemental analyses of these three casein proteins were shown in the following Table 4. Table 4. Elemental analyses of casein proteins from three milk samples Results No. 1. 2. 3. 4. 5. 6. 7. Protein Test Calcium (Ca) Iron (Fe) Potassium (K) Magnesium (Mg) Manganese (Mn) Sodium (Na) Zinc (Zn) Cow milk (ppm) 507 140 348 189 12 219 58 Goat milk (ppm) 662 148 1879 330 10 767 50 Buffalo milk (ppm) 523 117 621 165 16 395 31

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Cow milk (ppm) Goat milk (ppm) Buffalo milk (ppm) Measuring value (ppm) 2000 1600 1200 800 400 0

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Figure 3. Elemental analyses of casein proteins from three milk samples Calcium, magnesium, sodium and potassium are macrominerals for human that are required in amounts greater than 100 mg per day. Iron, zinc and manganese are microminerals for trace elements that are required in amounts less than 100 per day. According to these results, all three samples contain the sufficient amount of minerals. In other words, the three samples contain adequate amount of basic nutrients in suitable proportion that maintain the health of an individuals. Casein and lactose are found in the milk of every mammal, although in varying ratios. According to these data, cow milk contains the largest amount of casein protein. Thus, the cow milk is suitable for the best muscle growth and basic body building achievement. Goat milk contains the small amount of the casein protein content. Goat milk protein forms a softer curd which makes the protein more easily and rapidly digestible. Theoretically, this more rapid transit through the stomach could be an advantage to infants and children who regurgitate cow's milk easily. Goat's milk may also have advantages when it comes to allergies. Goat's milk contains only trace amounts of an allergenic casein protein, alpha-S1, found in cow's milk. Goat's milk casein is more similar to human milk, yet cow's milk and goat's milk contain similar levels of the other allergenic protein, beta lactoglobulin. Some mothers are certain that their child tolerates goat's milk better than cow's milk.

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So people who have an unusual milk allergy, or who are merely lactose may find goat milk more digestible other than cow's milk (Kurek et al., 1992 and Millward et al., 2008). Conclusion From this research work, it was found that three samples of milk contain casein protein. Moreover, the casein contents were observed 45.36%, 5.47% and 14.07% in cow milk, goat milk and buffalo milk respectively. According to the research findings, cow milk contains the largest amount of casein protein. Thus, the cow milk is suitable for the best muscle growth and basic body building achievements. It was found that goat milk contains the small amount of casein protein. Although the mineral content of goat's milk and cow's milk is generally similar, goat's milk contains more calcium, potassium, iron, magnesium and sodium. All milk has lots of casein but there are different types of casein and for someone who has casein sensitivity, goat milk may provide an alternative to which they do not react. Acknowledgements
The authors would like to thank Rector, Dr Tin Tun, University of Yangon and Professor and Head, Dr Nilar, Department of Chemistry, University of Yangon, for their valuable advice and permission.

References
Holt C., and Horne D.S., (1996), "The hairy casein micelle: evalution of the concept and its implication for dairy technology". Neth. Milk Dairy J.50: 85-111. Horne D.S., (2002), "Casein structure, self-assembly and gelation, current opinion in colloid and interface", Sci, 7: 456-461. Kurek M., Przybilla B., Hermann K. and Ring J., (1992), "A naturally occurring opioid peptide from cow's milk, beta-casomorphine-7, is a direct histamine releaser in man", 97. Millward C., Ferriter M., Calver S., and Connell-Jones G., (2008), "Gluten and casein-free diets for autistic spectrum disorder", Cochrane Database of systematic Review, Issue 2, Art. No. CD003498. DOI; 10.1002.