Staying ahead of regulations for the analysis of steroids in urine with GC-MS/MS

Significance of the Topic

The reliable detection of performance enhancing steroids in athletes urine is essential to uphold fair play and comply with evolving World Anti-Doping Agency regulations. Advanced analytical methods enable laboratories to meet stricter detection limits and ensure accurate, reproducible results under routine and competition conditions.

Study Objectives and Overview

This technical evaluation demonstrates how a TRACE 1610 gas chromatograph coupled with a TSQ 9610 triple quadrupole mass spectrometer can achieve required sensitivity and robustness for steroid screening and confirmation at recently lowered MRPLs and MRLs defined by WADA. Key performance aspects including detection limits, quantitation accuracy, carry-over, and long-term stability were assessed.

Applied Methodology and Instrumentation

The method uses a TRACE 1610 GC with an iConnect split/splitless injector fitted with LinerGOLD precision liners, a TraceGOLD TG-5SilMS column and helium carrier gas managed via HeSaver-H2Safer technology. Samples are introduced by an AI/AS 1610 autosampler and ionized in an Advance Electron Ionization source on the TSQ 9610. A two-point calibration and Chromeleon CDS automate data acquisition, processing and reporting while ensuring full traceability.

Main Results and Discussion

Sensitivity testing at or below new MRPLs showed signal-to-noise ratios above 3 for all target steroids, often greatly exceeding that threshold. Quantitation based on two calibration points yielded deviations within ±30 percent for most analytes; minor matrix effects impacted testosterone and etiocholanolone (5B-diol) at the lowest QC level. Carry-over evaluation confirmed no residual peaks in blank injections following concentrated standards. Robustness trials involving over 120 urine injections demonstrated stable response with no significant loss in sensitivity. Implementation of the HeSaver-H2Safer module reduced helium usage by 4.5× without method modification.

Benefits and Practical Applications

• Meets or exceeds current WADA MRPL and MRL requirements for key anabolic steroids
• High throughput capability and long-term stability support routine anti-doping workflows
• Automated data handling enhances efficiency, sample integrity and audit readiness
• Significant cost savings through carrier gas conservation

Future Trends and Potential Applications

Next-generation approaches may integrate high-resolution MS for non-targeted screening of emerging substances, supercritical fluid chromatography to reduce solvent use and AI-driven data interpretation for rapid flagging of atypical profiles. Continued optimization of carrier gas strategies and further miniaturization of instrumentation could expand routine applications to biological passport monitoring and trace-level detection of novel doping agents.

Conclusion

The combined TRACE 1610 GC and TSQ 9610 GC-MS/MS platform provides robust, sensitive and accurate analysis of urinary steroids in alignment with updated anti-doping requirements. Its automation, long-term stability and gas savings deliver reliable performance for both single-sample screening and longitudinal testing paradigms.

References

• World Anti-Doping Agency List of Prohibited Substances and Methods 2023
• WADA Technical Document TD2023IDCR Identification Criteria for GC-MS and LC-MS Methods
• WADA International Standard for Laboratories 2021
• Thermo Fisher Scientific Technical Note on HeSaver-H2Safer Carrier Gas Technology