Screening Analysis of Highly Polar Doping Agents in Urine using 2DLC-MS/MS

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The use of performance-enhancing drugs known as “doping” has been recognized for decades, and since 1999 the World Anti-Doping Agency (WADA) has been harmonizing the worldwide sports drug testing efforts. The continuing discovery of new doping strategies means that a range of testing methods are required to cover the target compounds comprehensively. For instance, emerging doping strategies include androgenic steroids, pro-hormones, peptide hormones and designer drugs.

In this application note, we report the simultaneous analysis of highly polar doping agents including meldonium, synephrine, norfenefrine, etilefrine, oxilofrine and octopamine by two-dimensional LC-MS/MS system (Nexera X2 coupled to LCMS-8060) using HILIC mode separation. This system could successfully quantify all doping agents at low ng/mL in urine. Moreover, fast polarity-switching MRM analysis enabled simultaneous detection of sulphated metabolites of each compound. Furthermore, norfenefrine and octopamine, which are closely resembling isomers, were baseline-resolved by monitoring their sulphated metabolites.

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