Ultra‐fast retroactive processing by MetAlign of liquid chromatography/high‐resolution full‐scan Orbitrap mass spectrometry data in WADA Human Urine Sample Monitoring Program

Abstract

The World Antidoping Agency (WADA) Monitoring program concentrates analytical data from the WADA Accredited Laboratories for substances which are not prohibited but whose potential misuse must be known. The WADA List of Monitoring substances is updated annually, where substances may be removed, introduced or transferred to the Prohibited List, depending on the prevalence of their use. Retroactive processing of old sample datafiles has the potential to create information for the prevalence of use of candidate substances for the Monitoring List in previous years. MetAlign is a freeware software with functionality to reduce the size of liquid chromatography (LC)/ high-resolution (HR) full-scan (FS) mass spectrometry (MS) datafiles and to perform a fast search for the presence of substances in thousands of reduced datafiles. Methods: Validation was performed to the search procedure of MetAlign applied to Anti-Doping Lab Qatar (ADLQ)-screened LC/HR-FS-MS reduced datafiles originated from antidoping samples for tramadol (TRA), ecdysterone (ECDY) and the ECDY metabolite 14-desoxyecdysterone (DESECDY) of the WADA Monitoring List. Searching parameters were related to combinations of accurate masses and retention times (RTs). Results: MetAlign search validation criteria were based on the creation of correct identifications, false positives (FPs) and false negatives (FNs). The search for TRA in 7410 ADLQ routine LC/HR-FS-MS datafiles from the years 2017 to 2020 revealed no false identification (FPs and FNs) compared with the ADLQ WADA reports. ECDY Safa Khelifi and Khadija Saad contributed equally to this work.

Olympic Anti-Doping Laboratory: The Analytical Technological Road From 2016 Rio De Janeiro to 2021 Tokyo

“…Monitoring Program is updated annually by WADA for substances to detect patterns of misuse in sport [63]. The recent application of MetAlign software for selected substances on the WADA Monitoring List, tramadol (TRA) and ecdysterone (ECDY) by using fast-retroactive processing of reduced LC HR full scan (FS) MS datafiles to reveal the prevalence of doping substances [64] has been developed.…”

“…The retroactive analysis can be greatly facilitated by the application of the MetAlign software [64,142] for fast and massive reprocessing of LC-MS/MS datafiles of samples analyzed in full-scan MS acquisition mode after electrospray ionization (ESI). The application of the MetAlign software allows the selection for reanalysis only of suspicious samples from a pool of samples under long-term storage for substances suitable to be detected by the ITP LC-MS/MS method [102,143].…”

Downregulation of CYP17A1 By 20-hydroxyecdysone: Plasma Progesterone and Its Vasodilatory Properties

“…Their most reported biological property is the ability to increase muscle mass, and therefore the potential to enhance physical performance [ 5–7 ]. This has made them attractive to elite athletes, with prevalence of their use in this cohort estimated at 0.4–5% [ 8 , 9 ]. In addition to their performance-enhancing properties, ecdysteroids have also been reported to have hormone-disruptive capability [ 10 ].…”