Quantitation of lipids in major lipoprotein classes

The content of lipid in each lipoprotein class can be obtained from the peak area of the elution pattern monitored by absorbance (A) at 550 nm or 500 nm using the enzyme reagent. The directly measured A should be corrected for the dilution of the effluent with the enzyme solution in order to obtain the absolute lipid concentration in the effluent. The peak area (P ) which is proportional to the absolute lipid concentration in the effluent is independent of the flow rate of eluent (Fjjj) or that of the enzyme solution (F ) as presented in Eq. (10). 

The relation of the peak area (P ) of the elution pattern to lipid concentration in a loaded sample was examined using the standard lipoprotein (LDL) sample prepared by ultracentrifugation for three lipids, and a linear relationship was obtained in the following concentration ranges 5 - 54 pg for TC (Fig. 15), 17 - 51 pg for TG and 3 - 85 pg for PL. 

Column G4000SW+G3000SW. Sample serum from normal female (A) and hyperlipidemia type IV (B). Fraction 1, chylomicrons+VLOL 2, LDL 3, HDL2 4, HOL3 5, VHDL. Other HPLC conditions as in Fig. 14. 

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