[Determination of endogenous steroids in urine by liquid chromatography-tandem mass spectromretry].
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 beta-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.
Wang
Mengye
M
Department of Forensic Medicine, Shanghaii Medical College, Fudan University, Shanghai 200032, China. 0158047@fudan.edu.cn
Xiang
Ping
P
Yan
Hui
H
Shen
Baohua
B
Shen
Min
M
D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D003687:Dehydroepiandrosterone / Q000008:administration & dosage / Q000652:urine; D004300:Doping in Sports; D005260:Female; D006801:Humans; D057230:Limit of Detection; D008297:Male; D015203:Reproducibility of Results; D013256:Steroids / Q000008:administration & dosage / Q000652:urine*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D016482:Urinalysis / Q000379:methods*
D013256:Steroids; D003687:Dehydroepiandrosterone