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Determination of sulfates of androsterone and epiandrosterone in human serum using isotope diluted liquid chromatography–electrospray ionization–mass spectrometry
Author(s) -
Mitamura Kuniko,
Setaka Misako,
Shimada Kazutake,
Honma Seijiro,
Namiki Mikio,
Koh Eitetsu,
Mizokami Atsushi
Publication year - 2005
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.522
Subject(s) - chemistry , chromatography , electrospray ionization , androsterone , derivatization , mass spectrometry , solid phase extraction , extraction (chemistry) , enzymatic hydrolysis , hydrolysis , steroid , biochemistry , hormone
A promising liquid chromatography–electrospray ionization–mass spectrometry (LC–ESI‐MS) for analysis of the sulfates of 5 α ‐androgen, androsterone and epiandrosterone (A‐S and EpiA‐S) in human serum was developed. The method was used to assess one of the markers of 5 α ‐reductase activity of males including patients with prostate cancer (PC). After deproteinization with acetonitrile, the androgen sulfates and the internal standard, [7,7,16,16‐ 2 H 4 ]dehydroepiandrosterone‐S, were extracted from human serum using a solid‐phase extraction cartridge and washed with hexane. The extract was reconstituted and applied to the LC[sol ]ESI‐MS system operated in the selected ion monitoring mode. The method was validated over the range 0.02–5 µg[sol ]mL (A‐S) and 0.005–1.5 µg[sol ]mL (EpiA‐S) using 10 µL of human serum. The method was a concise procedure without chemical or enzymatic hydrolysis of the conjugates, purification by high‐performance liquid chromatography and[sol ]or derivatization, and proved to be satisfactory in its reproducibility and accuracy. The levels of these androgen sulfates tended to decrease during aging, and the A‐S levels in the sera obtained from both healthy males and patients with PC were correlated with their EpiA‐S levels. Copyright © 2005 John Wiley & Sons, Ltd.