Separation of Whey Proteins by Reversed-Phase High-Performance Liquid Chromatography

초록

A reversed-phase high-performance liquid chromatography was successfully developed for the simultaneous and rapid separation of the main proteins of a-lactalbumin and b-lactoglobulin contained in whey. This method consisted of a linear gradient of the two mobile phases of 0.1% trifluoroacetic acid in water and 0.1% trifluoroacetic acid in acetonitrile. The total run time for this separation was approximately 30min, and a-lactalbumin was eluted and followed by b-lactoglobulin. The injection volume was fixed at 20ml and the flow rate was 1ml/min. The optimum mobile phase composition and gradient conditions to separate a-lactalbumin and b-lactoglobulin (A+B) was experimentally obtained at the particle sizes of 15㎛ with the pore size of 300Å on the linear-gradient mode