Abstract
The determination of α-tocopherol, free cholesterol, esterified cholesterols and triacylglycerols in human plasma and in fractions containing individual lipoproteins was achieved by reversed-phase high-performance liquid chromatography (HPLC). The lipoprotein fractions, such as chylomicron, VLDL, LDL, HDL 2 and HDL 3, were collected by ultracentrifugation of human plasma. The chromatographic separation was accomplished with a column packed with Hitachi Gel 3057, which is a spherical octadecylsilica of particle size 3 μm. The mobile phase was acetonitrile-2-propanol (75:25, v/v), and the elute was monitored with ultraviolet absorption and fluorescence detectors connected in series. Qualitative analysis of the main chromatographic peaks collected during the HPLC of a plasma sample was done with the use of field-adsorption mass spectrometry. The determination analysis of α-tocopherol, free cholesterol and esterified cholesterols was affected with a single chromatographic run with n-hexane extracts of plasma or lipoprotein fraction. The separation and determination of these fat-soluble components required as little as 5 μl of plasma or lipoprotein fraction.
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