Application of Solid-Phase Extraction for the Quantification of Urinary AICAR by Ultra-High Performance Liquid Chromatography-Tandem Mass-Spectrometry

A procedure for the determination of AICAR (5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside, Acadesine), an AMP-activated protein kinase agonist prohibited by the World Anti-Doping Agency since 2009, in urine has been proposed including sample clean-up from matrix components by means of solid-phase extraction and detection of the analyte by ultra-high performance liquid chromatography-tandem mass-spectrometry with heated electrospray ionization ion source. Due to the endogenous nature of AICAR, standard additions approach was applied to quantify AICAR and evaluate matrix effects. The limit of detection was 5 ng/mL, calibration curves were linear in the concentration range of 50-5000 ng/mL. The proposed procedure was applied to the analysis of real samples.