An automated method for measurement of methoxetamine in human plasma by use of turbulent flow on-line extraction coupled with liquid chromatography and mass spectrometric detection

Methoxetamine is a new ketamine derivative designer drug which has recently become available via the Internet marketed as "legal ketamine". It is a new dissociative recreational drug, acting as an NMDA receptor antagonist and dopamine reuptake inhibitor. The objective of this study was to develop on-line automated sample preparation using a TurboFlow device coupled with liquid chromatography with ion-trap mass spectrometric detection for measurement of methoxetamine in human plasma. Samples (100 mu L) were vortex mixed with internal standard solution (ketamine-d4 in acetonitrile). After centrifugation, 20 mu L of the supernatant was injected on to a 50 mm x 0.5-mm C18XL Turboflow column. The retained analytes were then back-flushed on to a 50 mm x 3-mm (3 mu m) Hypersil Gold analytical column for chromatographic separation, then eluted with a formate buffer-acetonitrile gradient. Methoxetamine and the IS were ionized by electrospray in positive mode. Parent [M + H](+) ions were m/z 248.1 for methoxetamine and m/z 242.0 for the IS. The most intense product ions from methoxetamine (m/z 203.0) and the IS (m/z 224.0) were used for quantification. The assay was accurate (96.8-108.8 % range) and precise (intra and inter-day coefficients of variation < 8.8 %) over the range of 2.0 (lower limit of quantification) to 1000.0 ng mL(-1) (upper limit of quantification). No matrix effect was observed. This method has been successfully applied to determination of plasma concentrations of methoxetamine in the first French hospitalization case report after acute intoxication; the plasma concentration was 136 ng mL(-1).