Abstract
A gas chromatographic method is described using high-resolution open-tubular glass capillary columns for the simultaneous estimation of the following neutral steroids in urine: androsterone, etiocholanolone, dehydroepiandrosterone, 11-oxoandrosterone, 11-oxoetiocholanolone, pregnanolone, 11β-hydroxyandrosterone, 11β-hydroxyetiocholanolone, allo-pregnanediol. pregnanediol. 1 5-pregnenediol, pregnanetriol and cholesterol. The method involves an ether extraction of the urine after enzymatic hydrolysis and the gas chromatographic separation of the steroids as their trimethylsilyl ether derivatives on a 50 m × 0.25 mm I.D. OV-101 capillary column. Quantitation is effected by reference to the peaks derived from three mass internal standards added to the hydrolysed urine. Data are presented that show that the precision and sensitivity of the method are adequate for clinical purposes. Experiments using reference steroids demonstrated essentially quantitative recoveries and the specificity as determined by combined gas chromatography-mass spectrometry was found to be satisfactory for eleven of these neutral steroids. Preliminary studies using an auto-solid sampler and an automated gas chromatograph indicate that the method is readily amenable to routine application.
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