Determination of sulfates of androsterone and epiandrosterone in human serum using isotope diluted liquid chromatography-electrospray ionization-mass spectrometry.
Determination of sulfates of androsterone and epiandrosterone in human serum using isotope diluted liquid chromatography-electrospray ionization-mass spectrometry.
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DOI：
10.1002/bmc.522
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发表时间：
2005-12
期刊：
影响因子：
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通讯作者：
K. Mitamura;Misako Setaka;K. Shimada;S. Honma;M. Namiki;Eitetsu Koh;A. Mizokami
中科院分区：
文献类型：
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作者：
K. Mitamura;Misako Setaka;K. Shimada;S. Honma;M. Namiki;Eitetsu Koh;A. Mizokami
建立了测定人血清中5α-雄激素、雄酮和表雄酮(A-S和表雄酮-S)硫酸盐的高效液相色谱-电喷雾电离-质谱法。用该方法对前列腺癌(PC)患者等男性的5α-还原酶活性标志物进行了检测。用乙腈去蛋白后，用固相萃取柱提取人血清中的雄激素硫酸盐和内标[7，7，16，16-2H4]脱氢表雄酮-S，并用正己烷洗脱。将萃取物重组后应用于选择的离子监测模式下的LC/ESI-MS系统。用10微L人血清进行检测，A-S在0.02-5微克/毫升，S在0.005-1.5微克/毫升范围内有效。该方法简便，不需要化学或酶促反应，不需要高效液相色谱分离纯化和衍生化反应，重现性和准确度令人满意。随着年龄的增长，这些雄激素硫酸盐水平呈下降趋势，健康男性和PC患者血清中A-S水平与其EPIA-S水平呈正相关。
A promising liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) for analysis of the sulfates of 5alpha-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 5alpha-reductase activity of males including patients with prostate cancer (PC). After deproteinization with acetonitrile, the androgen sulfates and the internal standard, [7,7,16,16-2H4]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/ESI-MS system operated in the selected ion monitoring mode. The method was validated over the range 0.02-5 microg/mL (A-S) and 0.005-1.5 microg/mL (EpiA-S) using 10 microL of human serum. The method was a concise procedure without chemical or enzymatic hydrolysis of the conjugates, purification by high-performance liquid chromatography and/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.