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Clandestine laboratories constantly produce new synthetic cannabinoids to circumvent legislative efforts,complicating toxicological analysis. No extensive synthetic cannabinoid quantitative urinary methodsare reported in the literature. We developed and validated a liquid chromatography–tandem mass spec-trometric (LC–MS/MS) method for simultaneously quantifying JWH-018, JWH-019, JWH-073, JWH-081,JWH-122, JWH 200, JWH-210, JWH-250, JWH-398, RCS-4, AM-2201, MAM-2201, UR-144, CP 47,497-C7,CP 47,497-C8 and their metabolites, and JWH-203, AM-694, RCS-8, XLR-11 and HU-210 parent com-pounds in urine. Non-chromatographically resolved alkyl hydroxy metabolite isomers were consideredsemi-quantitative. Glucuronidase hydrolyzed urine was extracted with 1 ml Biotage SLE+ columns.Specimens were reconstituted in 150 L mobile phase consisting of 50% A (0.01% formic acid in water)and 50% B (0.01% formic acid in 50:50 methanol:acetonitrile). 4 and 25 L injections were performed toacquire data in positive and negative ionization modes, respectively. The LC–MS/MS instrument consistedof a Shimadzu UFLCxr system and an ABSciex 5500 Qtrap massspectrometer with an electrospray source.Gradient chromatographic separation was achieved utilizing a Restek Ultra Biphenyl column with a0.5 ml/min flow rate and an overall run time of 19.5 and 11.4 min for positive and negative mode methods,respectively. Quantification was by multiple reaction monitoring with CP 47,497 compounds and HU-210ionized via negative polarity; all other analytes were acquired in positive mode. Lower and upper limitsof linearity were 0.1–1.0 and 50–100 g/l (r2> 0.994).
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