OJAS  Vol.3 No.3 , July 2013
Determination of caprine serum albumin in milk using bromocresol green dye
Abstract: The objective of this study was to develop and validate a colorimetric assay to measure caprine serum albumin (CSA) concentrations in milk. The principle of the say was based on the binding of albumin to bromocresol green (BCG) dye at pH 4.0. The absorption of the dye-albumin complex was determined spectrophotometrically at 640 nm. A linear relationship existed between CSA concentrations (0.01 to 8.0 mg/ml) and absorbance values. Intra-assay and interassay coefficients of variation for the proposed assay were 2.6% and 12% respectively. Recovery from milk samples spiked with albumins was 89.15% ± 7.6%. The developed assay was validated using 126 milk samples collected from lactating Saanen goats at various stages of lactation`. Caprine serum albumin concentrations in milk ranged between 0.31 and 0.52 mg/ml. The concentrations of CSA were significantly higher (p < 0.05) at the beginning of lactation, declined rapidly and remained relatively stable in mature milk. Caprine serum albumin correlated positively (p < 0.05) with milk somatic cell counts, total solids, protein, fat and whey and correlated negatively (p < 0.05) with milk lactose and casein concentrations. It was concluded that the proposed BCG dye-binding method is easy to perform, rapid, sensitive, reliable and can be used to determine CSA concentrations in milk collected from goats under different physiological conditions.
Cite this paper: Olaniyan, B. , Zhao, X. and Mustafa, A. (2013) Determination of caprine serum albumin in milk using bromocresol green dye. Open Journal of Animal Sciences, 3, 261-267. doi: 10.4236/ojas.2013.33039.

[1]   Lieske, B., Jantz, A. and Finke, B. (2005) An improved analytical approach for the determination of bovine serum albumin in milk. Lait, 85, 237-248. doi:10.1051/lait:2005018

[2]   Leitner, G., Merin, U. and Silanikove, N. (2004) Changes in milk composition as affected by subclinical mastitis in goats. Journal of Dairy Science, 87, 1719-1726. doi:10.3168/jds.S0022-0302(04)73325-1

[3]   Keay, G. and Doxey, D.L. (1983) Serum albumin values from healthy cattle, sheep and horses determined by the immediate bromocresol green reaction and by agarose gel electrophoresis. Veterinary Research Science, 1, 58-60.

[4]   Affonso, A. and Lasky, F.D. (1985) Bromocresol purple dye-binding method for the estimation of serum albumin adapted to the SMA 12/60. Clinical Biochemistry, 18, 285-289. doi:10.1016/S0009-9120(85)80033-3

[5]   Akram, M.E. (2005) Extractive spectrophotometric methods for the determination of oxomemazine hydrochloride in bulk and pharmaceutical formulations using bromocresol green, bromocresol purple and bromocresol blue. Archiv der Pharmazie—Chemistry in Life Sciences, 338, 190-197.

[6]   Barber, B.J. and Stanhope, V.L. (1992) Bromocresol green assay is nonspecific for rat plasma albumin. American Journal of Physiology, 262, 292-302.

[7]   Guzman, A.A., Perez, M.D., Marquez, R.M. and Garza, J.R. (1986) Colorimetric determination of bovine serum albumin in milk using bromocresol green dye. Journal of Dairy Science, 69.

[8]   Bouchard, L.S., Blais, C., Desrosiers, X., Zhao, X. and Lacasse, P. (1999) Nitric oxide production during endotoxin-induced mastitis in the cow. Journal of Dairy Science, 82, 2574-2581. doi:10.3168/jds.S0022-0302(99)75512-8

[9]   Levieux, D., Morgan, F., Geneix, N., Masle, I. and Bouvier, F. (2002) Caprine immunoglobulin G, β-lactoglobulin, α-lactalbumin and serum albumin in colostrum and milk during the early post partum period. Journal of Dairy Research, 69, 391-399. doi:10.1017/S0022029902005575

[10]   Gustafsson, J.E.C. (1976) Improved specificity of serum albumin determination and estimation of acute phase reactants by use of the BCG reaction. Clinical Chemistry, 22, 616-622.

[11]   Association of Official Analytical Chemists (1999) Official methods of analysis. 16th Edition, Gaithersburg, 2000.

[12]   SAS/STAT (1989) Statistical software and user’s guide. Version 6. 4th Edition, SAS Institute, Inc., Cary.

[13]   Macasek, F., Gerhart, P., Malovikova, A. and Celkova, A. (1996) Speciation of native cations and added radionuclides in raw bovine milk. Journal of Radioanalytical Nuclear Chemistry, 208, 163-182. doi:10.1007/BF02039758

[14]   Perez, M.D., Puyol, P., Ena, J.M. and Calvo, M. (1993) Comparison of the ability to bind lipids of beta-lactoglobulin and serum albumin of milk from ruminant and non ruminant species. Journal of Dairy Research, 60, 55-63. doi:10.1017/S0022029900027345

[15]   Walstra, P. and Jenness, R. (1984) Dairy chemistry and physics. Wiley, New York.

[16]   Shuster, D.E. and Harmon, R.J. (1990) Enzyme immunoassay of bovine lactoferrin and serum albumin in acid precipitated and ultracentrifugal wheys. Journal of Dairy Science, 73, 3104-3111. doi:10.3168/jds.S0022-0302(90)78998-9

[17]   Webster, D. (1977) The immediate reaction between bromocresol green and serum as a measure of albumin content. Clinical Chemistry, 23, 663-665.

[18]   Sordillo, L.M., Oliver, S.P., Duby, R.T. and Rufner, R. (1984) Effects of colchine on milk yield, composition and cellular differentiation during caprine lactogenesis. International Journal of Biochemistry, 16, 1135-1141.

[19]   Belewu, M.A. and Adewusi, B.C. (2001) Distribution of protein fractions in the milk of West African dwarf goat. Journal of Food Technology, 6, 8-10.

[20]   Leitner, G., Chaffer, M., Shamay, A., Shapiro, F., Merin, U., Ezra, E., Saran, A. and Silanikove, N. (2004) Changes in milk composition as affected by subclinical mastitis in sheep. Journal of Dairy Science, 87, 46-52. doi:10.3168/jds.S0022-0302(04)73140-9

[21]   De Wit, J.N. (1998) Nutritional and functional characteristics of whey proteins in food product. Journal of Dairy Science, 81, 597-608. doi:10.3168/jds.S0022-0302(98)75613-9

[22]   De Cremoux, R. (1995) Relations entre les numerations cellulaires du lait et les infections mammaires chez la chevre (Relations between milk somatic cell counts and intramammary infections in goats). Vet. Doctor Thesis. Paul Sabatier-Toulouser University, Toulouser.

[23]   Anderson, K.L., Kindahl, H., Smith, A.R. and Gustafsson, B.K. (1985) Arachidonic acid metabolism in milk of cows during acure coliform mastitis. American Journal of Veterinary Research, 46, 1573-1577.

[24]   Klei, L., Yun, J., Sapru, A., Lynch, J., Barbano, D., Sears, P. and Galton, D. (1998) Effects of milk somatic cell counts on cottage cheese yield and quality. Journal Dairy Science, 81, 1205-1213. doi:10.3168/jds.S0022-0302(98)75680-2

[25]   Nguyen, D.D. and Neville, M.C. (1998) Tight junction regulation in the mammarygland. Journal of Mammary Gland Biology and Neoplasia, 3, 233-273. doi:10.1023/A:1018707309361

[26]   Haenlein, G.F.W. (2002) Relationship of somatic cell counts in goat milk to mastitis and productivity. Small Ruminant Research, 45, 165-178. doi:10.1016/S0921-4488(02)00097-4

[27]   Byeng, R.M., Grant, T. and Steve, P.H. (2007) Effect of subclinical intramammary infection on somatic cell counts and chemical composition of goats’ milk. Journal of Dairy Research, 74, 204-210. doi:10.1017/S0022029906002378

[28]   Fernandes, A.M., Oliveira, C.A.F. and Tavolaro, P. (2004) Relationship between somatic cell counts and composition of milk from individual Holstein cows. Arquivos do Instituto Biológico, 71, 163-166.

[29]   Rayna-Ljutovac, K., Pirisi, A., De Cremoux, R. and Gonzalo, C. (2007) Somatic cells of goat and sheep milk: Analytical, sanitary, productive and technological aspects. Small Ruminant Research, 68, 126-144. doi:10.1016/j.smallrumres.2006.09.012

[30]   Nagamoto, H., Miyaoka, T., Shimizu, T., Katayama, H. and Takahashi, H. (1996) Quantitative comparison between serum components and somatic cells in bovine quarter milk. Journal of Veterinary Medical Sciences, 58, 1121-1123. doi:10.1292/jvms.58.11_1121

[31]   Politis I., Barbano D.M. and Gorewit R.C. (1992) Distribution of plasminogen and plasmin in fractions of bovine milk. Journal of Dairy Science, 75, 1402-1410. doi:10.3168/jds.S0022-0302(92)77893-X