Detection of non-steroidal anti-inflammatory drugs in equine plasma and urine by gas chromatography-mass spectrometry

Abstract

A gas chromatographic-mass spectrometric (GC-MS) procedure for the detection of seventeen non-steroidal anti-inflammatory drugs (NSAIDs) in equine plasma and urine samples is described. The extraction of the compounds from the biological matrix was performed at acidic pH (2–3) with diethyl ether. Ethereal extracts were washed with a saturated solution of sodium hydrogencarbonate (urine) or treated with a solid mixture of sodium carbonate and sodium hydrogencarbonate (plasma). The ethereal extracts were dried and derivatized by incubation at 60°C with methyl iodide in acetone in the presence of solid potassium carbonate. Mono- or bismethyl derivatives of the NSAIDs were obtained. After derivatization kinetic studies, 90 min was the incubation time finally chosen for screening purposes for adequate methylation of all the compounds under study. For individual confirmation analyses, shorter incubation times can be used. The chromatographic analysis of the derivatives was accomplished by GC-MS with a run time of 13 min. In general, extraction recoveries ranged from 23.3 to 100% in plasma and from 37.5 to 83.8% in urine samples. Detection limits from less than 5 to 25 ng/ml were obtained for both plasma and urine samples using selected-ion monitoring. The procedure was applied to the screening and confirmation of NSAIDs in routine doping control of equine samples.