LONG-TERM DETECTION AND IDENTIFICATION OF METANDIENONE AND STANOZOLOLABUSE IN ATHLETES BY GAS-CHROMATOGRAPHY HIGH-RESOLUTION MASS-SPECTROMETRY

The misuse of anabolic androgenic steroids (AAS) in human sports is co ntrolled by gas chromatography-mass spectrometric analysis of urine sp ecimens obtained from athletes. The analysis is improved with modern h igh-resolution mass spectrometry (HRMS). The detection and identificat ion of metabolites of stanozolol (I) [3'-hydroxystanozolol (II) and 4 beta-hydroxystanozolol (III)] and metandienone (IV) [17 beta-methyl-5 beta-androst-1-ene-3 alpha, 17 alpha-diol (V) and 18-nor-17,17-dimethy l-5 beta-androsta-1,13-dien-3 alpha-ol (VI)] with GC-HRMS at 3000 reso lution yielded a large increase in the number of positive specimens. A total of 116 anabolic steroid positives were found in this laboratory in 1995 via GC-MS and GC-HRMS screening of 6700 human urine specimens collected at national and international sporting events and at out-of -competition testing. Of the 116 positive cases, 41 were detected usin g conventional (quadrupole) GC-MS screening. The ether 75 positives we re identified via GC-HRMS screening. To confirm the HRMS screening res ult, the urine sample was reanalyzed using a specific sample workup pr ocedure to selectively isolate the metabolites of the identified subst ance. II and III were selectively isolated via immunoaffinity chromato graphy (IAC) using an antibody which was prepared for methyltestostero ne and shows high cross reactivity to II and III. V and VI were isolat ed using high-performance liquid chromatography (HPLC) fractionation.