Stable isotope dilution analysis of human urinary metabolites of 17α-methyltestosterone |
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Authors: | Yoshihiko Shinohara Koichiro Isurugi Takao Hashimoto |
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Institution: | a School of Pharmacy, Tokyo University of Pharmacy and Life Science, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392, Japan;b Kuramoto Memorial Hospital, 696-1 Toyotomi, Funabashi, Chiba 274, Japan |
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Abstract: | A method based on gas chromatography–mass spectrometry–selected-ion monitoring was developed to measure the main metabolites of 17α-methyltestosterone, 17α-methyl-5α-androstan-3α,17β-diol and 17α-methyl-5β-androstan-3α,17β-diol, in human urine. 17α-Methyl-2H3]-5α-androstan-3α,17β-diol and 17α-methyl-2H3]-5β-androstan-3α,17β-diol were used as internal standards. The methods involved purification using a Sep-Pak C18 cartridge, hydrolysis by β-glucuronidase from Ampullaria and derivatization with N-methyl-N-trimethylsilyl-trifluoroacetamide/dithioerythriol/ammonium iodide. Quantitation was achieved by selected-ion monitoring of the characteristic fragment ions ((M+H)−2×TMSOH]+) of the di-TMS derivatives on the chemical ionization mode. The method provides a specific, sensitive and reliable technique to determine the urine levels of 17α-methyl-5α-androstan-3α,17β-diol and 17α-methyl-5β-androstan-3α,17β-diol, and can be applied to pharmacokinetic studies of 17α-methyltestosterone. |
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Keywords: | Steroids 17α -Methyltestosterone |
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