Simultaneous doping analysis of main urinary metabolites of anabolic steroids in horse by ion-trap gas chromatography-tandem mass spectrometry.

The research presents a new method for simultaneously testing 11 anabolic steroids, potentially applicable in doping tests for racehorses, using ion-trap gas chromatography-tandem mass spectrometry.

Objective of the Research

• The primary aim of this research was to establish a highly efficient and reliable method for simultaneous detection of 11 specific anabolic steroids that are often involved in horse doping scenarios.

Methodology

• 11 anabolic steroids including fluoxymesterone, 17alpha-methyltestosterone, mestanolone, methandienone, methandriol, oxymetholone, boldenone, furazabol, methenolone, nandrolone, and stanozolol were pinpointed for this analysis.
• 15 target substances were selected from the primary metabolites of these anabolic steroids for the doping test.
• A procedure for simultaneous analysis was developed. The urine was first hydrolyzed and then subjected to solid-phase extraction.
• The remaining extract residue was derivatized by trimethylsilylation, a process that makes the substances more volatile and hence easier to analyze by gas chromatography.
• The derivatized samples were then run through ion-trap gas chromatography-tandem mass spectrometry to obtain their mass chromatograms and product ion spectra.

Results of the Analysis

• The limit of detection for the target substances was found to be between 5-50 ng/mL.
• The average recovery, a measurement of how much of the original sample is successfully isolated for analysis, ranged from 71.3% to 104.8%.
• The coefficient of variation, which indicates the precision or reliability of the assay method, ranged from 1.1% to 9.5% showing reasonably reliable results.

Conclusion

• The proposed method achieved efficient simultaneous analysis of multiple anabolic steroids in horse urine samples. This could potentially enhance the effectiveness and comprehensiveness of doping tests in racehorses.