Comprehensive multiresidue analysis of 220 prohibited substances in doping control analyses using ultra-fast liquid chromatography/tandem mass spectrometry with polarity switching and EZ-method

Abstract

It is of great importance to screen a wide range of drug in a limited time and to confirm adverse finding in doping control laboratories. In this study, a comprehensive multiresidue method based on ultra-fast liquid chromatography (UFLC) coupled to triple-quadrupole mass spectrometry has been developed and validated for the simultaneous screening and generic confirmation of 220 prohibited substances in human urine. The present method was used for the simultaneous determination of eight different classes of prohibited substances; stimulants (75 drugs), narcotics (16 drugs), beta-2-agonists (7 drugs), hormone antagonists and modulators (14 drugs), beta-blockers (21 drugs), diuretics and other masking agents (26 drugs), glucocorticosteroids (24 drugs), and anabolic steroids (37 drugs). The samples were extracted by liquid-liquid extraction after enzymatic hydrolysis. The supernatant was taken and evaporated to dryness under nitrogen gas. The dried residue was dissolved in 100 μL of reconstitution solvent and analyzed by UFLC-MS/MS system with the EZ-method including a retention time-dependent SRM mode and fast polarity switching. Total run time was 10 min and the good resolution of isomers was achieved on Kinetex C18 (100 × 2.1 mm, 2.6 μm) column using gradient elution at 500 μl/min. The developed method was successfully applied to screening and confirmation of real urine samples.