Simultaneous analysis of 210 prohibited substances in human urine by ultrafast liquid chromatography/tandem mass spectrometry in doping control

RATIONALE Doping analysis is a two‐step process consisting of a screening step for prohibited substances and a confirmation step to verify the presence of specific substances found during the screening. The entire process must be performed within a limited time period, but traditional screening procedures commonly employ separate analytical methods for each class of prohibited substances being screened and thus require a great deal of human resources and instrumentation. A single simple and rapid multiresidue analytical method that could accommodate multiple classes of prohibited substances would be extraordinarily useful in doping analyses. METHODS Urine samples were extracted via two consecutive liquid–liquid extractions at different pH values following enzymatic hydrolysis. Analyses were performed by ultrafast liquid chromatography/triple‐quadrupole mass spectrometry with polarity switching and time‐dependent selected reaction monitoring. RESULTS We developed a rapid multiresidue screening and confirmation method for efficient high‐throughput doping analyses. The present method was validated with regard to the limits of detection (0.01–100.0 ng/mL for screening analyses and 0.2–500.0 ng/mL for confirmation assays), matrix effects (48.9–118.9%), recovery (20.6–119.7%) and intra‐ (0.6–17.6%) and inter‐day (4.0–20.0%) precision. CONCLUSIONS A multiresidue analytical method was developed and validated for screening and confirming the presence of performance‐enhancing drugs. A total of 210 substances from diverse classes of prohibited substances were successfully identified with an analytical run time of 10 min. Copyright © 2015 John Wiley & Sons, Ltd.