Rapid separation and quantification of major caseins and whey proteins of bovine milk by capillary electrophoresis.
A rapid capillary zone electrophoresis (CZE) method was established for separating and quantifying major casein and whey proteins in milk. Optimum sample preparation and electrophoretic conditions in a coated capillary maintained at 40 degrees C allowed accurate and reproducible quantification of milk proteins in a single analysis. Sample and run buffer allowed caseins to be maintained in solution by using a combination of urea and a nonionic detergent in phosphate buffer at pH 2.5. Quantitative CZE protein data were derived by calculating percentages and concentrations (mg/mL) of alpha-casein, beta-casein, alpha-lactalbumin, and beta-lactoglobulin. Calibration curves followed linear relationships with highly significant (p < 0.1) correlation coefficients. Relative standard deviations of less than 0.82 (%) for migration times and 2.18 (%) for percent protein indicated that the technique was reproducible. Electrophoretic protein profiles of fresh bovine milk and rehydrated dry milk showed marked quantitative differences in whey protein concentrations. Whey protein represented 12.37 +/- 0.07% beta-lactoglobulin and 3.05 +/- 0.08% alpha-lactalbumin of total protein in typical fresh milk, while only 1.90 +/- 0.16% beta-lactoglobulin and 0.86 +/- 0.04% alpha-lactalbumin of total protein were detected in a commercial rehydrated milk powder. By quantifying these differences, the established technique may allow the detection of substitution of fresh milk with rehydrated milk powder. The accuracy and reproducibility of the technique permitted the quantitation of individual protein concentrations in milk samples, which agreed with ranges reported in the literature. CZE may be well suited for routine use by dairies and regulatory agencies, since it allows the determination of milk proteins in less than 60 min.
Vallejo-Cordoba
B
B
Centro de Investigación en Alimentación y Desarrollo, A.C. (CIAD), Hermosillo, Sonora, Mexico.
D000818:Animals; D002364:Caseins / Q000302:isolation & purification*; D002417:Cattle; D019075:Electrophoresis, Capillary / Q000379:methods*; D008892:Milk; D008894:Milk Proteins / Q000302:isolation & purification*; D000067816:Whey Proteins
D002364:Caseins; D008894:Milk Proteins; D000067816:Whey Proteins