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Steroid spectrum in human urine as revealed by gas chromatography. I. Qualitative analysis of C19--C2102-3 steroids.

C19C21O2 steroids of urine can be analyzed under isothermal conditions by gas chromatography after a pretreatment consisting of mild acid hydrolysis, simultaneous extraction with toluene (or, in the case of of C21O3 steroids, enzymatic hydrolysis followed by ether extraction), a short purification step and the formation of derivative. Separation of acetyl derivatives could be achieved by three columns: 3%SE-30,1.3%NGS and 3%QF-1. These were needed because the separation of 3alpha-hydroxy-5alpha-androstan-17-one from 3alpha-hydroxy-5beta-androstan-17-one and that of 3alpha-20alpha-dihydroxy-5beta-pregnane from 3alpha, 20alpha-dihydroxy-5alpha-pregnane could be accomplished only in the NGS fluid phase and QF-1 stationary phase, respectively. At the same time, this method gives information on the secretion of anabolic androgenic steroids and on the secretion and metabolism of progesterone and 17-hydroxy-progesterone in the human body. The application of a retention-index system specified for the steroid skeleton and substituents offers the possibility to correlate retention times to molecular structure, which facilitates the identification of problematic metabolites. Steroid spectra as shown by gas chromatography seem to be a promising tool in the early diagnosis of pregnancy on the basis of the increase in the excretion of 3alpha-20alpha-dihydroxy-5beta-pregnane.