Chinese Journal of Chromatography

Chinese Journal of Chromatography

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Determination of endogenous steroids in urine by liquid chromatography-tandem mass spectrometry

WANG Mengye1,2, XIANG Ping2, YAN Hui1,2, SHEN Baohua2, SHEN Min2

  

  1. 1.Department of Forensic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China;
    2.Institute of Forensic Science, Ministry of Justice, PRC, Shanghai 200063, China
  • Received:2007-08-13 Revised:2007-11-09 Online:2008-01-30 Published:1984-09-25
  • Contact: Min Shen

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Abstract:

A method was developed for the determination of endogenous steroids in urine using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with methyltestosterone as internal standard. After enzymatic hydrolysis by β-glucuronidase and liquid-liquid extraction, the urine sample was chromatographed on a Cosmosil C18 column with a mixture of methanol and ammonium acetate-formic acid (68:32, v/v) as mobile phase, then detected using MS/MS system with electrospray ionization (ESI) in multi-reaction monitoring (MRM) mode. The detection limits ranged from 0.01 ng/mL to 10 ng/mL. The recoveries ranged from 96.7% to 106.5%, and the intra- and inter-day precisions (measured as relative standard deviations) were less than 7% and 11%, respectively. With simple and fast sample preparation, the method was sensitive and specific for simultaneous determination of these 5 kinds of endogenous steroids in urine. The method has been successfully applied in pharmacokinetic study and is thus a potential alternative for gas chromatography-mass spectrometry (GC-MS) based procedures in routine analysis of endogenous steroids such as DHEA in human urine.

Key words: urine , dehydroepiandrosterone (DHEA), doping agents, endogenous steroids, liquid chromatography-tandem mass spectrometry (LC-MS/MS)