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‘Wrong‐way‐round ionization’ and screening for doping substances in human urine by high‐performance liquid chromatography/orbitrap mass spectrometry
Author(s) -
Virus E. D.,
Sobolevsky T. G.,
Rodchenkov G. M.
Publication year - 2012
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.2055
Subject(s) - chemistry , orbitrap , chromatography , mass spectrometry , direct electron ionization liquid chromatography–mass spectrometry interface , urine , high performance liquid chromatography , ionization , chemical ionization , ion , organic chemistry , biochemistry
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. Copyright © 2012 John Wiley & Sons, Ltd.