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Analysis of whey proteins by high performace liquid chromatogrpahy

Authors: R.J. Pearce


High performance liquid chromatography (HPLC) has been applied to the analysis of whey proteins. Using a short alkyl chain (C6) reverse phase column with an acid saline / acetonitrile gradient the major whey proteins bovine serum albumin, α-lactalbumin and β-lactoglobulin have been completely resolved in a 30 min. analysis. In addition the A and B generic variants of β-lactoglobulin have been completely resolved to better than 70% in the same analysis. Reproducibly checks of peak retention times and peak areas yielded standard deviations of better than 1% and 3% respectively about mean values (n=5). Analysis of purified whey proteins by HPLC demonstrated the presence of levels of impurity not readily detected by electrophoresis. This technique has been applied to the analysis of whole Cheddar cheese whey and when protein concentrates (WPC) derived from Cheddar, hydrochloric acid and sulphuric acid casein wheys. Significant differences could be detected in the protein elution profiles of the whey proteins in the various WPC. Whey protein analysis by this HPLC method is sensitive enough to detect subtle changes in the whey protein which have occurred as a result of processing.

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