Identification of dehydroepiandrosterone metabolites formed from human prostate homogenate using liquid chromatography–mass spectrometry and gas chromatography–mass spectrometry

The identification of the in vitro metabolites of dehydroepiandrosterone formed from human prostate homogenate was investigated by hyphenated techniques using the stable-isotope dilution method. A mixture of dehydroepiandrosterone and [2H4]dehydroepiandrosterone was incubated with hypertrophied human prostate tissue homogenate in the presence of NAD, NADH and NADPH. The metabolites were extracted with AcOEt–hexane, purified by solid-phase extraction, and then analyzed by LC–atmospheric pressure chemical ionization MS and/or GC–MS. Androst-5-ene-3β,17β-diol (major product), androst-4-ene-3,17-dione, testosterone, 5α-dihydrotestosterone, androsterone, and 7α-hydroxydehydroepiandrosterone were identified in comparison with authentic samples based on their chromatographic behavior and mass spectra.