Microsampling and LC–MS/MS for Antidoping Testing of Glucocorticoids in Urine
Michele Protti, Roberto Mandrioli, Laura Mercolini
Abstract
Background. Systemic glucocorticoids are prohibited in-competition by the World Anti-Doping Agency. Here, we describe an original microsampling workflow for the quantitation of three endogenous (cortisol, corticosterone and cortisone) and three exogenous (dexamethasone, methylprednisolone and fludrocortisone) corticosteroids in 30 μl of human urine. Materials & methods. Microsampling was carried out by dried urine spot (DUS) sampling and volumetric absorptive microsampling (VAMS), followed by solvent extraction and LC–MS/MS analysis. Results & conclusion: Good linearity (r2 > 0.9989) was obtained for all analytes; extraction yields (>81%), precision (RSD < 8.6%) and matrix effect (<12%) were satisfactory. Microsample stability at room temperature was good (analyte loss <15% after 3 months). Data obtained from real urine microsample analysis were compared with those of fluid urine, providing very good agreement (r2 > 0.9991).