Following the oral administration of one capsule of “1-Androsterone”, corresponding to 115 mg of 3-hydroxy-5-androst-1-en-17-one (3), to one healthy male volunteer the combined unconjugated and glucuronidated metabolite fractions were analyzed as per-TMS derivatives as used in routine sports doping control. The parent compound (3) was detectable in the urines for about two days with the highest concentration detected in the 0–3 h urine. Its 3-isomer (4) was identified by comparison with the unpurified reference obtained by reduction of 1-androstenedione with LS-Selectride. It was found to be the metabolite detectable for the longest time. The highest concentration was found in the 3–7.5 h urine, while its presence could still be confirmed up to seven days (ions used: m/z 432, 275,and 169) according to the regulations applicable in human sports doping control [15]. Suspicious results in screening analysis were possible for two more days. Applying these criteria, 5-androst- 1-ene-3,17-dione (m/z 415, 430, and 194) and 1-testosterone (m/z 194, 432, and 417) were detectable for six and almost four days, respectively. The 5-androst-1-ene-3,17-diols (5) and (6) were unambiguously identified for about one (3-) and three days (3-) due to their elution in a region of relatively high background in the chromatograms. An optimized sample pretreatment may eliminate this in case of confirmation of a positive screening result. Positive screening results were obtained for about six (1), eight (2), four (5), and three days (6). Additionally, two more metabolites were detected in the early post-administration urines (HO-Dione1 and HO-Dione2). Based on their mass spectra (Fig. 7), structures of androst-1-ene-3,17-diones hydroxylated most likely at position C-18 (OH-Dion1) and C-19 (OH-Dion2) are proposed due to the loss of 103 u (TMS-O-CH2 •) from the molecule resulting in m/z 415.
