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New Potential Biomarker For Methasterone Misuse In Human Urine By Liquid Chromatography Quadrupole Time Of Flight Mass Spectrometry

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES(2016)

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摘要
In this study, methasterone urinary metabolic profiles were investigated by liquid chromatography quadrupole time of flight mass spectrometry (LC-QTOF-MS) in full scan and targeted MS/MS modes with accurate mass measurement. A healthy male volunteer was asked to take the drug and liquid-liquid extraction was employed to process urine samples. Chromatographic peaks for potential metabolites were hunted out with the theoretical [M - H](-) as a target ion in a full scan experiment and actual deprotonated ions were studied in targeted MS/MS experiment. Fifteen metabolites including two new sulfates (S1 and S2), three glucuronide conjugates (G2, G6 and G7), and three free metabolites (M2, M4 and M6) were detected for methasterone. Three metabolites involving G4, G5 and M5 were obtained for the first time in human urine samples. Owing to the absence of helpful fragments to elucidate the steroid ring structure of methasterone phase II metabolites, gas chromatography mass spectrometry (GC-MS) was employed to obtain structural information of the trimethylsilylated phase I metabolite released after enzymatic hydrolysis and the potential structure was inferred using a combined MS method. Metabolite detection times were also analyzed and G2 (18-nor-17 beta-hydroxymethyl-2 alpha, 17 alpha-dimethyl-androst-13-en-3 alpha-ol-xi-O-glucuronide) was thought to be new potential biomarker for methasterone misuse which can be detected up to 10 days.
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关键词
methasterone,new biomarker,human urine,doping control,liquid chromatography time-of-flight tandem mass spectrometry
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