Anion-exchange high-performance liquid chromatography assays of plasma lipoproteins and modified low-density lipoproteins using a ProtEx-DEAE column

Previously [Anal. Biochem., 232 (1995) 163-171], we reported a high-perform ance Liquid chromatography (HPLC) assay method for human plasma lipoprotein s using a diethylaminoethyl (DEAE)-glucomannan column, which is not commerc ially available. in this study, HPLC assay methods for Lipoproteins in plas ma samples of human and experimental animals, and modified low-density lipo proteins (LDLs) of rabbits have been developed using a commercially availab le anion-exchange ProtEx-DEAE column. For the assays of plasma lipoproteins , the method includes complete separation of high-density lipoproteins, LDL s and very low-density lipoproteins within 20 min using stepwise elution, a nd determination by post-column reaction with an enzymatic cholesterol reag ent as the total cholesterol (TC) level. Similarly, mild oxidative and arti ficially oxidised LDLs were separated into their subfractions using stepwis e elution, and determined based on the TC level. The methods using the DEAE -glucomannan and ProtEx-DEAE columns were cross-validated. There was an exc ellent correlation between the two methods. The obtained results reveal tha t the anion-exchange HPLC method using the ProtEx-DEAE column could be usef ul for the assays of plasma lipoproteins and modified LDLs. (C) 2001 Elsevi er Science B.V. All rights reserved.