Simultaneous determination of androstenediol 3-sulfate and dehydroepiandrosterone sulfate in human serum using isotope diluted liquid chromatography–electrospray ionization-mass spectrometry

A simple method for simultaneous determination of androstenediol 3-sulfate (Adiol-3S) and dehydroepiandrosterone sulfate (DHEA-S) in human serum using isotope diluted liquid chromatography–electrospray ionization-ion trap-mass spectrometry (LC–ESI-ion trap-MS) was developed. After addition of deuterated internal standards ([2H5]Adiol-3S and [2H4]DHEA-S), human serum (100 μl) was deproteinized with acetonitrile and then applied to a solid-phase extraction cartridge, Oasis HLB. The obtained steroid sulfates fraction was washed with hexane and then analyzed by LC–ESI-MS operated in the negative ion mode. The quantification ranges of Adiol-3S and DHEA-S were 10–400 ng/ml and 0.05–8 μg/ml, respectively. The method does not require the chemical or enzymatic hydrolysis of the conjugates and purification with high-performance liquid chromatography, and shows satisfactory reproducibility and accuracy. The concentrations of these sulfates in the sera of healthy male volunteers (n=14) were 19.2–245.3 mg/ml (Adiol-3S) and 0.175–5.16 μg/ml (DHEA-S), and those of patients with prostate cancer (n=19) were 15.3–182.7 ng/ml (Adiol-3S; four samples, not detectable) and 0.110–2.421 μg/ml (DHEA-S).