QUANTITATION OF PLASMA-LIPIDS BY GAS-LIQUID-CHROMATOGRAPHY ON HIGH-TEMPERATURE POLARIZABLE CAPILLARY COLUMNS

We have previously demonstrated the potential usefulness of capillary columns coated with a high temperature polarizable phenylmethylsilicone liquid phase in plasma lipid profiling (Kuksis, A., J. J. Myher, and R Sandra. 1990. J Chromatogr. 500: 427-441). The present study reports improved operating conditions along with a practical application to the analysis of a series of human plasma samples in comparison to capillary gas-liquid chromatography on nonpolar columns. For this purpose the plasma lipids were dephosphorylated with phospholipase C and converted to the trimethylsilyl ethers. The molecular species of the plasma lipids were identified by comparing the relative retention times to reference standards. The species were quantitated using tridecanoylglycerol as internal standard. The recoveries of the lipid classes were determined by summing the molecular species within each carbon number and comparing the proportions of the carbon numbers obtained on polar and nonpolar columns. The relative recoveries varied with the lipid class and sample load and averaged as follows: FA C-16, 78%; FA C18, 78%; FC, 99%; tridecanoylglycerol (TD), 100% (by definition); CER 34:1, 92%; DG C34, 103%; DG C36, 98%; CE C-16, 73%; CE C18, 61%; TG C50, 93%; TG C52, 60%; TGC54, 32%. We conclude that high temperature polarizable capillary columns are suitable for qualitative and quantitative assessment of plasma lipids and provide more information per man-hour, instrument time, and unit cost than any other analytical method known.