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A new technique for serum lipoprotein quantitation by high performance liquid chromatography (HPLC) was evaluated by comparing the analytical data with the electrophoretic method using agarose gel film. The elution pattern of human serum (5μl) monitored by cholesterol gave three clear peaks according to particle-size differentiation: peak 1, chylomicrons+very low density lipoproteins (VLDL); peak 2, low density lipoproteins (LDL); peak 3, high density lipoproteins (HDL). As for a group (n=92) of normolipidemic men and women, cholesterol levels in these three fractions were determined by the HPLC method (X), and they were compared with those by the electrophoretic method (Y). A good correlation was obtained for peak 2 to β lipoprotein (r=0.961, Y=0.990X+3.43) and for peak 3 to α lipoprotein (r=0.911, Y=0.945X+7.21). Cholesterol level of peak 1 (chylomicrons +VLDL) was 5.3±4.0mg/dl, but that of pre-β liporrotein could not be measured by the electrophoretic method. Precision of determination (coefficient of variation, CV%) by the HPLC method was less than 1% both for peak 2 (LDL) and peak 3 (HDL). The HPLC method was found to have better precision and resolution for lipoprotein quantitation than the electrophoretic method.

Comparison of Serum Lipoprotein Quantitation between High Performance Liquid Chromatography and Agarose Gel Electrophoresis