Ultra-fast retroactive processing by MetAlign of liquid chromatography/high-resolution full-scan Orbitrap mass...

Area of Science:

• Analytical Chemistry
• Mass Spectrometry
• Anti-Doping Science

Background:

• The World Antidoping Agency (WADA) Monitoring program tracks substances not currently prohibited but with potential for misuse.
• Annual updates to the WADA Monitoring List necessitate efficient methods for analyzing historical data to assess substance prevalence.
• MetAlign software offers a solution for reducing and searching large liquid chromatography/high-resolution mass spectrometry (LC/HR-FS-MS) datasets.

Purpose of the Study:

• To validate the MetAlign software's search procedure for identifying specific substances within historical antidoping sample data.
• To assess the prevalence of tramadol (TRA), ecdysterone (ECDY), and 14-desoxyecdysterone (DESECDY) in antidoping samples using MetAlign.
• To establish a validation scheme for implementing MetAlign in the WADA Monitoring program.

Main Methods:

• Validation of MetAlign's search algorithm using LC/HR-FS-MS data from the Anti-Doping Lab Qatar (ADLQ).
• Focused on identifying tramadol (TRA), ecdysterone (ECDY), and its metabolite 14-desoxyecdysterone (DESECDY).
• Search parameters combined accurate masses and retention times (RTs) for substance identification.

Main Results:

• MetAlign successfully identified TRA in 7410 ADLQ samples (2017-2020) with no false positives or negatives.
• Ecdysterone (ECDY) and its metabolite 14-desoxyecdysterone (DESECDY) were detected in approximately 5% of the analyzed samples.
• The validation confirmed MetAlign's accuracy in identifying target substances.

Conclusions:

• MetAlign is a powerful tool for rapid retrospective analysis of LC/HR-FS-MS data, enabling the determination of antidoping substance prevalence.
• The validated MetAlign search procedure can be implemented within the WADA Monitoring program to accurately identify substances.
• This approach aids in eliminating false negatives and false positives, enhancing the reliability of the WADA Monitoring program.