Separation and Quantification of Bovine Milk Proteins by Reversed-Phase High-Performance Liquid Chromatography

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Abstract
Current analytical methods for milk proteins lack the capacity to simultaneously separate and quantify the six major bovine milk proteins and their genetic variants. A method is described that simultaneously separates and quantifies the six major bovine milk proteins. The separation is based on reversed-phase partitioning of the six major milk proteins and several genetic variants of κ-casein, β-casein, and β-lactoglobulin. The described method has for each of the six milk proteins a linear quantitative response, precision (coefficient of variation below 5.1% within days of analysis, and below 7.1% between days of analysis), resolution (over 2.5 between proteins), peak efficiency (theoretical plate numbers between 8 000 and 50 000), and a sample treatment without filtration steps that together with the analysis takes 2 h. The composition of protein from milk of each of 234 cows was determined using the current method, and the results were similar to reference values for milk proteins. Keywords: Casein; whey; milk protein; reversed-phase high-performance liquid chromatography; quantification