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Clinical Chemistry 36: 519-521, 1990;
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Clinical Chemistry, Vol 36, 519-521, Copyright © 1990 by American Association for Clinical Chemistry

Electron-capture gas chromatography as a sensitive method for measuring subnanogram amounts of cholesterol in saliva and urine

HA Schwertner, ER Johnson and TE Lane
Internal Medicine Branch, USAF School of Aerospace Medicine, Brooks Air Force Base, TX 78235-5301.

This is a sensitive method, suitable for measuring subanogram amounts of cholesterol. Cholesterol and the internal standard, epicoprostanol (5-beta-cholestan-3-alpha-ol), are derivatized with pentafluorobenzoyl chloride and detected by electron-capture gas chromatography. The pentafluorobenzoyl esters of cholesterol and the internal standard are easily formed and possess excellent chromatographic and electron- capturing properties. The lower limit of detection of the method, approximately 100 pg injected, is about 500-fold as sensitive as chromatographic methods involving flame-ionization detection. Within- day and between-day coefficients of variation were 4.2% and 8.2%, respectively, for determinations of a urinary cholesterol concentration of 570 micrograms/L (1.47 mumol/L). Such sensitivity permits analysis for cholesterol in (e.g.) physiological fluids, tissue samples, and cell cultures that contain very low concentrations of cholesterol.




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