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An ultra fast and sensitive detection of 165 drugs of abuse in human urine using polarity switching ultra performance liquid chromatography tandemmass spectrometry

Sachin Dubey, ShobhaAhi,Alka Beotra, TejinderKaur, Shila Jain

The screening of wide variety of prohibited substances in a time bound manner by adhering to latest World Anti-Doping Agency (WADA) guidelines is a challenging task for doping control laboratories. The revised criterion of detection limits (WADATD2013MRPL) has further required the doping laboratories to reviewtheir testing procedures. The presentwork was aimed at developing a fast, sensitive and robust analytical method based on solid phase cleanup (SPE) and ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) to achieve the required detection levels. The method development involved optimization of deconjugation of phase II metabolites, SPE usingmixed-mode ion cartridges for extraction of analytes of wider chemistries; and fast polarity switching UPLC-MS/MS detection. The developed method was validated to detect approximately 165 compounds and/or metabolites prohibited by WADA. The eight minutes runtime allowed testing of approximately 180 samples in 24 hours at the limit of detection (LOD) of 50%belowrequired detection levels.