Steroid spectrum in human urine as revealed by gas chromatography. II. Quantitative analysis of C19--C21O2-3 steroids

The amounts of C19-C21O2-3 steroids excreted in the urine was determined in 35 individuals representing a normal healthy fraction of the adult population. The metabolites were determined after acid hydrolysis and simultaneous toluene extraction, in the form of acetyl derivatives, under isothermic conditions, on two columns. For the quantitative determination of the components and for ensuring reliability, two internal standards were applied. The preparatory steps, beginning with extraction from acidic media, were controlled by the addition to the urine of 4 beta, 5 beta-epoxycholestanone as a "marker" internal standard, while the "measurement" standard applied in gas chromatography was 5 alpha-cholestane. Quantitative measurements were carried out on a SE-30 column, androsterone and etiocholanolone were separated on a CHDMS column; the reliability of the determinations was checked by statistical analysis. The physiological values of some 8-9 of the most important steroid metabolites excreted in the urine, as averaged from the values measured for 23 women and 12 men, are discussed and compared with the data in the literature.