A screening method for the simultaneous detection of glucocorticoids,diuretics, stimulants,anti-oestrogens,beta-adrenergic drugs and anabolic steroids in human urine by LC-ESI-MS/MS |
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Authors: | Monica Mazzarino Xavier de la Torre Francesco Botrè |
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Institution: | (1) Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti, 1, 00197 Rome, Italy;(2) Dipartimento per le Tecnologie, le Risorse e lo Sviluppo, Università “La Sapienza”, Via del Castro Laurenziano 9, 00161 Rome, Italy |
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Abstract: | This paper presents a general screening method, based on liquid chromatography/mass spectrometry (LC/MS), for the simultaneous
detection in human urine of 72 xenobiotics (21 diuretics, 16 synthetic glucocorticoids, 17 beta-adrenergic drugs, 10 stimulants,
5 anti-oestrogens and 3 anabolic steroids), excreted free or as glucuro-conjugates in urine. Although the method has been
specifically designed and evaluated in view of its potential application to anti-doping analyses, it can also be effective
in other areas of analytical toxicology. Sample preparation was based on two liquid/liquid separation steps (performed at
alkaline and at acid pH, respectively) of hydrolyzed human urine, and then an assay by LC/MS-MS in positive and negative ionization
mode using an electrospray ionization source (ESI) and multiple reaction monitoring (MRM) as the acquisition mode. The overall
time needed for an LC run was less than 15 minutes. All compounds showed good reproducibility in terms of both the retention
times (CV%<1) and the relative abundances of the diagnostic transitions (CV%<10). The limits of detection (LOD) were in the
range of 1–50 ng/mL for glucocorticoids, anti-oestrogens and steroids, and 50–500 ng/mL for diuretics, beta-adrenergic drugs
and stimulants, thus satisfying the minimum required performance limits (MRPL) set by the World Anti-Doping Agency (WADA)
for the accredited anti-doping laboratories. |
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Keywords: | Glucocorticoids Anti-oestrogens Designer steroids Stimulants Diuretics Beta-adrenergic drugs LC-ESI-MS/MS Anti-doping analysis |
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