title: 'Wrong-way-round ionization' and screening for doping substances in human urine by high-performance liquid chromatography/orbitrap mass spectrometry.
abstract: To free analytical resources for new classes of doping substances, such as banned proteins, maximization of the number of compounds that can be determined with high sensitivity in a single run is highly urgent. This study demonstrates an application of 'wrong-way-round ionization' for the simultaneous detection of multiple classes of doping substances without the need to switch the polarity. A screening method for the detection of 137 compounds from various classes of prohibited substances (stimulants, diuretics, β(2)-agonists, β-blockers, antiestrogens, glucocorticosteroids and anabolic agents) has been developed. The method involves an enzymatic hydrolysis, liquid-liquid extraction and detection by liquid chromatography/orbitrap mass spectrometry with wrong-way-round ionization. Up to 64% of compounds had a 10-fold lower limit of detection (LOD) than the minimum required performance limit. To compare the efficiency of conventional ionization relative to wrong-way-round ionization of doping substances in + ESI, a fortified blank urine sample at the minimum required performance limit was analyzed using two ESI approaches. All compounds were detected with markedly better S/N in a high-pH mobile phase, with the exception of acetazolamide (minimal change in S/N, < 20%).The method was validated by spiking 10 different blank urine samples at five different concentrations. Validation parameters included the LOD, selectivity, ion suppression, extraction recovery and repeatability.
authors.affiliation: Moscow Antidoping Center, 105005, Moscow, Elizavetynsky10. virus@dopingcontrol.ru
authors.forename: E D
authors.initials: ED
authors.lastname: Virus
authors.forename: T G
authors.initials: TG
authors.lastname: Sobolevsky
authors.forename: G M
authors.initials: GM
authors.lastname: Rodchenkov
mesh_terms: D000319:Adrenergic beta-Antagonists / Q000652:urine; D045930:Anabolic Agents / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004232:Diuretics / Q000652:urine; D004300:Doping in Sports; D005938:Glucocorticoids / Q000652:urine; D006801:Humans; D057230:Limit of Detection; D059625:Liquid-Liquid Extraction; D004364:Pharmaceutical Preparations / Q000737:chemistry / Q000652:urine*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000319:Adrenergic beta-Antagonists; D045930:Anabolic Agents; D004232:Diuretics; D005938:Glucocorticoids; D004364:Pharmaceutical Preparations