서강대학교

초록/요약

5-androsten-3,16α-diol-17-one-acetate is a prohormone. A prohormone is a committed intra-glandular precursor of a hormone, usually exhibit minimal hormonal effects by itself. It is known to increase muscle growth and strength, so bodybuilders often use it. Prohormone has fewer side effect than anabolic steroids, but still cause the gynecomastia, virilism, skin diseases, headache and fatigue. Urinary 5-androsten-3,16α-diol-17-one-acetate and its metabolites following oral administration of 5-androsten-3,16α-diol-17-one-acetate to volunteer were isolated from urine by way of PAD-1 adsorption, enzymatic hydrolysis with β-glucuronidase from E-coli and liquid/liquid extraction with methyl tert-butyl ether at pH 7. The extracted 5-androsten-3,16α-diol-17-one-acetate and its metabolite were compared to blank urine, spiked urine and dosed urine sample by LC/ MS. 5-androsten-3,16α-diol-17-one-acetate was not detected in dosed urine but M1-5-androsten-3,16α-diol-17-one , M2-5-androsten-3β, 16α , 17β-triol and M3-5-abdrosten-3β, 16α, 17α-triol were found in dosed human urine and also identified as endogenous metabolites. For confirmation, these were derivatized with MSTFA/NH4I/DTE (500:2:1 (w/w/w)) at 70 ℃ for 30 min and detected by GC/MS. The structure of M1 was estimated that acetic group at 16-carbon position was replaced with hydroxyl group and the structures of M2 and M3 were estimated that ketone group at 17-carbon position was reduced to hydroxyl group. Comparing chromatogram of retention time and mass spectrum with each of the reference material, it was confirmed that M1 was 5-androsten-3,16α-diol-17-one, M2 was 5-androsten-3β, 16α, 17β-triol and M3 was 5-abdrosten-3β, 16α, 17α-triol.. It was confirmed that M1-5-androsten-3,16α-diol-17-one, M2-5-androsten-3β, 16α, 17β-triol and M3-5-abdrosten-3β, 16α, 17α-triol formed not sulfuric acid conjugates but glucuronic acid conjugates. Also, the endogenous metabolites of 5-androsten-3,16α-diol-17-one-acetate in urine samples showed the same excretion tendency in five urine samples which increased and decreased as time goes on. To determine whether taking the 5-androsten-3,16α-diol-17-one-acetate is a prohormone. A prohormone is a committed intra-glandular precursor of a hormone, usually exhibit minimal hormonal effects by itself. It is known to increase muscle growth and strength, so bodybuilders often use it. Prohormone has fewer side effect than anabolic steroids, but still cause the gynecomastia, virilism, skin diseases, headache and fatigue. Urinary 5-androsten-3,16α-diol-17-one-acetate and its metabolites following oral administration of 5-androsten-3,16α-diol-17-one-acetate to volunteer were isolated from urine by way of PAD-1 adsorption, enzymatic hydrolysis with β-glucuronidase from E-coli and liquid/liquid extraction with methyl tert-butyl ether at pH 7. The extracted 5-androsten-3,16α-diol-17-one-acetate and its metabolite were compared to blank urine, spiked urine and dosed urine sample by LC/TOF/MS. 5-androsten-3,16α-diol-17-one-acetate was not detected in dosed urine but M1-5-androsten-3,16α-diol-17-one , M2-5-androsten-3β, 16α , 17β-triol and M3-5-abdrosten-3β, 16α, 17α-triol were found in dosed human urine and also identified as endogenous metabolites. For confirmation, these were derivatized with MSTFA/NH4I/DTE (500:2:1 (w/w/w)) at 70 ℃ for 30 min and detected by GC/MSD. The structure of M1 was estimated that acetic group at 16-carbon position was replaced with hydroxyl group and the structures of M2 and M3 were estimated that ketone group at 17-carbon position was reduced to hydroxyl group. Comparing chromatogram of retention time and mass spectrum with each of the reference material, it was confirmed that M1 was 5-androsten-3,16α-diol-17-one, M2 was 5-androsten-3β, 16α, 17β-triol and M3 was 5-abdrosten-3β, 16α, 17α-triol. Average max amount and total excretion amount of M1-5-androsten-3β, 16α-diol-17-one , M2-5-androsten-3β, 16α, 17β-triol and M3-5-androsten-3β, 16α, 17α-triol in experimental groups were certainly larger than them in control group. In addition, the most accurate method for determining administration of drugs could be inspected after 7~10 hr from suspected time of administrating drugs. From the above results, M1-5-androsten-3β, 16α-diol-17-one can be used as a bioindicator which could predicted the administration of 5-androsten-3,16α-diol-17-one-acetate.