Abstract: A high performance liquid chromatography-electrospray ionization mass spectrometry (LC-ESI/MS) was developed to screen and determine trenbolone, tetrahydrogestrinone and gestrinone in human urine. The urine sample was enzymatically hydrolyzed with β-glucuronidase, then extracted with methyl tert-butyl ether. Chromatographic separation was performed on a Zorbax SB-C18 column (150 mm×2.1 mm, 5 μm) with ammonium formate buffer (pH 3.5) and acetonitrile as mobile phase. Using positive electrospray ionization mode, the effect of different parameters from electrospray chamber was investigated. The limits of detection based on signal noise ratio of 3 were between 1 and 5 ng. The method can be applied in screening and confirmation of the anabolic steroids in doping control.

Key words: anabolic steroids, doping detection , human urine, liquid chromatography-electrospray ionization mass spectrometry (LC-ESI/MS)