Greet Debruyckere

Influence of the consumption of meat contaminated with anabolic steroids on doping tests

Abstract In 1989, 3 cases out of 20 were found where gas chromatography—mass spectrometry revealed the presence of nandrolone metabolites in the urine of untreated persons. Although the origin of the nandrolone metabolites was uncertain at that time, the presumption was that such findings could be related to the consumption of meat from nandrolone-treated animals. Two years later, in a feeding experiment with volunteers, 1 out of 10 persons was positive for clostebol due to the accidental consumption of meat contaminated with clostebol acetate. In 1992, a similar random experiment was carried out ( n = 50), yielding 2 positive cases (1 nandrolone case and 1 clostebol case). The urinary concentrations found in the volunteers who had accidentally consumed contaminated meat were compared with the levels found in volunteers after a controlled intake of a minor dose of the anabolic steroid concerned. The above levels were also compared with those after intake of a therapeutic dose of the compound.

Gas chromatographic-mass spectrometric confirmation of 19-nortestosterone in the urine of untreated boars--effect of the administration of Laurabolin.

The presence of 17 beta-19-nortestosterone (nandrolone, NT, 17 beta-19-NT) and its epimer 17 alpha-19-nortestosterone (epiNT, 17 alpha-19-NT) was investigated in the urine of six untreated boars, obtained from experimental farms. The presence of 17 beta-19-nortestosterone was screened by RIA and HPTLC and confirmed by GC-MS analysis. Additionally, the two epimers (NT and epiNT) were investigated in the urine of a boar (two-year-old miniature male pig weighing 50 kg) before and after injection of 100 mg Laurabolin (nortestosterone laurate, Intervet N.V., Belgium). The isolation of the steroids was based on sample clean-up with solid phase extraction and subsequent high-performance liquid chromatography. Both gas chromatographic retention data and mass spectrometric data (selected ion monitoring and full spectrum) were used for detection and identification. The presence of 17 beta-19-nortestosterone in the urine of the boars that were not injected proves the endogenous production of the steroid. The absence of the 17 alpha-epimer in the urine of the injected boar suggests that 17 alpha-19-nortestosterone is not a major metabolite of 17 beta-19-nortestosterone.

Detection of 19-nortestosterone and its urinary metabolites in miniature pigs by gas chromatography—mass spectrometry

The metabolism of 19-nortestosterone was investigated in a miniature non-castrated male pig (boar), in a castrated pig (barrow) and in a female pig (sow). Urine samples were taken before and at regular intervals after the injection of 100 mg of Laurabolin (nortestosterone laurate). The sample clean-up consists in preliminary solid-phase extraction, followed by high-performance liquid chromatographic purification and fractionation. Finally, gas chromatography-mass spectrometry is used to identify the 19-nortestosterone metabolites.

Formation of 18-nor-13(14)-ene-17β-methyl steroids during perfluoroacylation of anabolic steroids bearing a tertiary 17-hydroxy group and compounds related to corticosteroids

The results of derivatization (for gas chromatography–mass spectrometry) of steroids bearing a tertiary 17-hydroxy group with perfluoroacyl anhydrides in acetone are presented. The reaction products are investigated by using low- and high-resolution mass spectrometry, metastable ion measurements, and proton and carbon-13 nuclear magnetic resonance spectroscopy. On the basis of the results, we conclude that all the products lose the 17-hydroxy group during derivatization and undergo a Wagner–Meerwein rearrangement to give 18-nor-13(14)-ene-17β-methyl steroids.

Gas chromatographic—mass spectrometric confirmation of a clostebol metabolite in urine

Abstract Anabolic steroids are used as doping agents in sports. Notwithstanding the total EC ban, they are also used as growth promoters in animal production. The detection of anabolic steroids in the urine of untreated persons due to the consumption of contaminated meat has been described earlier. This paper describes further confirmation of a urine sample which was clostebol positive as a result of the above described interference phenomenon. This was done by gas chromatography—mass spectrometry. Both low resolution mass spectrometry (LRMS) and high resolution mass spectrometry (HRMS) were used after gas chromatographic separation. LRMS was used to determine the relative abundances of the ions and for calculating similarity indices. With HRMS, selectivity was increased by recording exact masses instead of nominal masses.