Comparison of dairy phospholipid preparative extraction protocols in combination with analysis by high performance liquid chromatography coupled to a charged aerosol detector

Different fat extraction methodologies, Röse Gottlieb (RG), Folch with CaCl2 (F1) and Folch without CaCl2 (F2) were evaluated alongside a high performance liquid chromatography (HPLC)-based analytical protocol involving a charged aerosol detector (CAD) to optimise the separation and quantification of dairy phospholipids (PLs). Total PL recovery by F2 proved to be 1.8- and 2.5-fold greater than RG and F1, respectively. Alteration of the HPLC elution program maximised peak separation and reduced co-elution of individual PLs detected by CAD. Two-fold reduction in retention time variability was achieved, <0.15 min, compared with the original method. Improved PL recovery was reflected in the higher PL values obtained from different dairy streams using the optimised protocol, 35.32 ± 0.01% and 46.09 ± 0.01% total PL, for buttermilk and butter serum respectively. Phosphatidylinositol and phosphatidylserine, in particular, demonstrated increases of between 2% and 7%, of total PL, compared with previous studies.