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Profiling of urinary steroids by gas chromatography-mass spectrometry detection and confirmation of androstenedione administration using isotope ratio mass spectrometry
Authors:Wang Jingzhu  Wu Moutian  Liu Xin  Xu Youxuan
Institution:National Anti-Doping Laboratory, China Anti-Doping Agency, Beijing 100029, China
Abstract:Androstenedione (4-androstene-3,17-dione) is banned by the World Anti-Doping Agency (WADA) as an endogenous steroid. The official method to confirm androstenedione abuse is isotope ratio mass spectrometry (IRMS). According to the guidance published by WADA, atypical steroid profiles are required to trigger IRMS analysis. However, in some situations, steroid profile parameters are not effective enough to suspect the misuse of endogenous steroids. The aim of this study was to investigate the atypical steroid profile induced by androstenedione administration and the detection of androstenedione doping using IRMS. Ingestion of androstenedione resulted in changes in urinary steroid profile, including increased concentrations of androsterone (An), etiocholanolone (Etio), 5α-androstane-3α,17β-diol (5α-diol), and 5β-androstane-3α,17β-diol (5β-diol) in all of the subjects. Nevertheless, the testosterone/epitestosterone (T/E) ratio was elevated only in some of the subjects. The rapid increases in the concentrations of An and Etio, as well as in T/E ratio for some subjects could provide indicators for initiating IRMS analysis only for a short time period, 2-22 h post-administration. However, IRMS could provide positive determinations for up to 55 h post-administration. This study demonstrated that, 5β-diol concentration or Etio/An ratio could be utilized as useful indicators for initiating IRMS analysis during 2-36 h post-administration. Lastly, Etio, with slower clearance, could be more effectively used than An for the confirmation of androstenedione doping using IRMS.
Keywords:5α-diol  5α-androstane-3α  17β-diol  5β-diol  5β-androstane-3α  17β-diol  An  androsterone  DHEA  dehydroepiandrosterone  ERC  endogenous reference compound  ET  epitestosterone  Etio  etiocholanolone  GC/C/IRMS  gas chromatography/combustion/isotope ratio mass spectrometry  GC-MSD  gas chromatography-mass spectrometry detection  HPLC  high-performance liquid chromatography  IRMS  isotope ratio mass spectrometry  LSD  Least-Significant Difference  PDB  Pee Dee Belemnite  PD  5β-pregnane-3α  20α-diol  RL  reference limit  T  testosterone  T/E  testosterone/epitestosterone  WADA  World Anti-Doping Agency
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