Abstract
A method has been developed for the microassay of serum cholesteryl ester fatty acid (CEFA) patterns by densitometry of the charred spots produced after chromatography on silica gel impregnated glass paper. Two important features of the procedure are “humidity conditioning” of the gel paper prior to and during chromatography, and the application of sulfuric acid to the chromatogram by a quasi-chromatographic technique instead of by spraying. The method has been extended to the analysis of not only serum extracts but also microliter volumes of serum by a directspotting technique. The fatty acid composition of the ester groups separated from human serum by the gel-paper chromatography has been established by silicic acid column chromatography and gas-liquid chromatography, as well as by the use of pure reference standards, both nonradioactive and “tagged” with cholesterol-4-C 14. The reproducibility of the method was demonstrated both with lipid extracts of human and rat serum and with “direct-spotting” assay of the same sera. Satisfactory recoveries were obtained of pure cholesteryl esters added to lipid extracts of serum.
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