Quantitative Characterization of Bovine Serum Albumin, a-Lactalbumin and b-Lactoglobulin in Commercial Whey Sample by RP-LC Goutam Mukhopadhyay 1,& , Jasmina Khanam 2 , Arunabha Nanda 2 , Arindam Basu Sarkar 3 1 B.C.D.A College of Pharmacy and Technology, 78 Jessore Road (S) Hridaypur, Barasat, Kolkata 700127, India; E-Mail: goutam_bst@yahoo.com 2 Department of Pharmaceutical Technology, Jadavpur University, Kolkata 700032, India 3 School of Pharmacy, The University of Findlay, 1000 North Main Street, Findlay, OH 45840, USA Received: 6 August 2009 / Revised: 12 January 2010 / Accepted: 22 January 2010 Abstract A simple, sensitive and precise reverse phase liquid chromatographic method has been developed and validated for quantification of bovine serum albumin (BSA), a-lactalbumin (a-La) and b–lactoglobulin (b-Lg) that are removed from whey waste by foam fractionation method. The data is reproducible over a wide concentration range. This optimized method allowed analysis of BSA, a-La, b-Lg in a mixture within 5 min and could be applied to the analysis of a variety of commercial and laboratory whey products within a short time. Keywords Column liquid chromatography Commercial waste whey Bovine serum albumin a-Lactalbumin, b-lactoglobulin Introduction Whey and whey products have been used successfully in the food industry for the past 30 years. Cost effectiveness and quality improvement are key drivers in using whey products. The nutritional value of whey products is also an impor- tant reason why an increasing number of food manufacturers worldwide include whey products in their formulations. The principal whey proteins are b–lactoglob- ulin (b-Lg, MW 18.3 kDa), a–lactalbu- min (a-La, MW 14.1 kDa) and bovine serum albumin (BSA, MW 69.3 kDa). Continuous effort is persisting in order to improve cost efficiency and quality of whey products. Since variation of com- position in whey from natural sources is observed, it is important to develop a reliable and fast analytical method to analyze proteins in source and products. There is already an abundance of litera- ture concerning whey protein analysis. Methods include gel electrophoresis [1, 2], chromatography and electropho- resis [3], capillary electrophoresis [4–6] and immunochemical detection [7]. By far the most used method for whey protein analysis is liquid chromatography. LC has become one of the main techniques in the dairy industry as it combines versa- tility, short analysis time and high reso- lution with increasingly sophisticated automation and superior column perfor- mance. There are many methods described for ion-exchange, gel perme- ation, hydrophobic interaction and reversed-phase (RP) LC [3–9], but not a single method is suitable enough to quantify all the whey proteins in a single injection. Here in this paper, our main objective was to develop an easy, suitable and time saving method by reverse phase liquid chromatography and apply this method to quantify the amount of major whey proteins like BSA, a-lactalbumin and b-lactoglobulin after recovering proteins from commercial whey using the foam fractionation method [11, 12]. Pure proteins were used as standard. Method development was aimed at accurate quantitation, maximum resolution, and reliable validation by LC with UV detection. DOI: 10.1365/s10337-010-1501-5 Ó 2010 Vieweg+Teubner | GWV Fachverlage GmbH Full Short Communication