Ultra‐fast retroactive processing of liquid chromatography high‐resolution full‐scan Orbitrap mass spectrometry data in anti‐doping screening of human urine

Abstract

Rationale Retroactive analysis of previously tested urine samples has become an important sports anti‐doping tool. Retroactive reprocessing of old data files acquired from a generic screening procedure can reveal detection of initially unknown substances, like illegal drugs and newly identified metabolites. Methods To be able to efficiently search through hundreds to thousands of liquid chromatography high‐resolution full‐scan Orbitrap mass spectrometry data files of anti‐doping samples, a combination of MetAlign and HR_MS_Search software has been developed. MetAlign reduced the data size ca 100‐fold making possible local storage of a massive volume of data. Results The newly developed HR_MS_Search module can search through the reduced data files for new compounds (mass or isotope pattern) defined by mass windows and retention time windows. A search for 33 analytes in 940 reduced data files lasted 10 s. The output of the automatic search was compared to the standard manual routine evaluation. The results of searching were evaluated in terms of false negatives and false positives. The newly banned b2‐agonist higenamine and its metabolite coclaurine were successfully searched in reduced data files originating from a testing period for which these substances were not banned, as an example of retroactive analysis. Conclusions The freeware MetAlign software and its automatic searching module HR_MS_Search facilitated the retroactive reprocessing of reduced full‐scan high‐resolution liquid chromatography/mass spectrometry screening data files and created a new tool in anti‐doping laboratories' network.

Annual banned‐substance review – Analytical approaches in human sports drug testing

“…Overall, the GC‐Q/Orbitrap data were described as superior but both systems were found fit‐for‐purpose in anti‐doping analysis, and the employed full scan HRMS measurements also allowed for data reprocessing where desirable. Proof‐of‐concept concerning the retroactive analysis of datasets (and corresponding long‐term stored urine samples) was provided by Lommen et al, who demonstrated the practicality of data reduction and subsequent automated evaluation . Employed in the context of screening ITP data, false‐negative results were virtually excluded and algorithm settings tailored to individual analytes allowed for reducing the frequency of false warnings to an acceptable minimum.…”

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

“…[7][8][9][10] MetAlign has been studied in Anti-Doping Lab Qatar (ADLQ) for Orbitrap LC/HR-FS-MS data processing. 11 MetAlign performs datafile digital information reduction and subsequently searches for substances using the HR_MS_search module. MetAlign has the potential to be used as an antidoping tool for preventive and fast retroactive reprocessing of old datafiles for the discovery of designer drugs or new long-term metabolites for prohibited substances without reporting limits, like anabolic androgenic steroids (AAS), as defined in the WADA Technical Document for the Minimum Required Performance Limits (MRPL).…”

Discovery of chemical markers for improving the quality and safety control of Sinomenium acutum stem by the simultaneous determination of multiple alkaloids using UHPLC-QQQ-MS/MS

“…Cheng, and Asarum heterotropoides F. Schmidt 9 , as well as in S. acutum stem. Coclaurine could also be a potential stimulant because it is a metabolite of higenamine with a similar chemical structure 10 – 12 . Coclaurine has also been identified in rat urine by gas chromatography/mass spectrometry after higenamine administration 13 .…”