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-performance liquid chromatography (HPLC) assay method for human plasma lipoproteins using a diethylaminoethyl (DEAE)-glucomannan column, which is not commercially available. In this study, HPLC assay methods for lipoproteins in plasma samples of human and experimental animals, and modified low-density lipoproteins (LDLs) of rabbits have been developed using a commercially available anion-exchange ProtEx-DEAE column. For the assays of plasma lipoproteins, the method includes complete separation of high-density lipoproteins, LDLs and very low-density lipoproteins within 20 min using stepwise elution, and determination by post-column reaction with an enzymatic cholesterol reagent as the total cholesterol (TC) level. Similarly, mild oxidative and artificially oxidised LDLs were separated into their subfractions using stepwise elution, and determined based on the TC level. The methods using the DEAE-glucomannan and ProtEx-DEAE columns were cross-validated. There was an excellent correlation between the two methods. The obtained results reveal that the anion-exchange HPLC method using the ProtEx-DEAE column could be useful for the assays of plasma lipoproteins and modified LDLs.
Faculty of Pharmaceutical Sciences, Mukogawa Women's University, Nishinomiya, Hyogo, Japan. haginaka@mwu.mukogawa-u.ac.jp
J
J
Haginaka
Y
Y
Yamaguchi
M
M
Kunitomo
D000818:Animals; D002138:Calibration; D015243:Cholesterol, VLDL / Q000097:blood; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D006949:Hyperlipidemias / Q000097:blood; D008074:Lipoproteins / Q000097:blood*; D008075:Lipoproteins, HDL / Q000097:blood; D008077:Lipoproteins, LDL / Q000097:blood*; D008351:Mannans; D051379:Mice; D010084:Oxidation-Reduction; D011817:Rabbits; D051381:Rats; D015203:Reproducibility of Results
D015243:Cholesterol, VLDL; D008074:Lipoproteins; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D008351:Mannans; C022901:(1-6)-alpha-glucomannan