title: Validation and application of an UPLC-MS/MS method for the quantification of synthetic cannabinoids in urine samples and analysis of seized materials from the Portuguese market.
abstract: An UPLC-MS/MS method using ESI+ionization and MRM was developed and fully validated according to international guidelines for the qualitative and quantitative analysis of nine synthetic cannabinoids and/or their metabolites in urine samples (1mL). Prior to extraction the samples were subjected to an enzymatic hydrolysis using β-glucuronidase followed by a SPE procedure using Oasis(®) HLB 3cc (60mg) columns. The chromatographic separation was performed with an Acquity UPLC(®) HSS T3 (50mm×2.1mm i.d., 1.8μm) reversed-phase column using a gradient with methanol-ammonium formate 2mM (0.1% formic acid) and with a run time of 9.5min. The method was validated in terms of selectivity, capacity of identification, limits of detection (0.01-0.5ng/mL) and quantification (0.05-0.5ng/mL), recovery (58-105%), carryover, matrix effect, linearity (0.05-50ng/mL), intra-assay precision, inter-assay accuracy and precision (CV<20%). The method was applied to 80 authentic samples, five of them (6.2%) were confirmed or suspected to be positive for the metabolites JWH-018 N-hydroxypentyl and JWH-018 N-pentanoic acid of JWH-018 and for the metabolite JWH-122 N-(5-hydroxypentyl) of JWH-122, and three of them in association with THC and/or THCCOOH (substances included in the method, together with the 11-OH-THC). Additionally, 17 spice products were analyzed, for which were confirmed the presence of the following substances: AM-2201, JWH-018, JWH-022 JWH-073, JWH-122, JWH-203, JWH-210, JWH-250, HU-210 and RCS-4, according to the comparison with authentic reference material and published data. The analytical method developed allowed the analysis of synthetic cannabinoids and the notification of the first cases in Portugal.
authors.affiliation: National Institute of Legal Medicine and Forensic Sciences - South Branch, Rua Manuel Bento de Sousa n°3, 1169-201 Lisbon, Portugal. Electronic address: ssimoes@dlinml.mj.pt.
authors.forename: Susana Sadler
authors.initials: SS
authors.lastname: Simões
authors.affiliation: National Institute of Legal Medicine and Forensic Sciences - South Branch, Rua Manuel Bento de Sousa n°3, 1169-201 Lisbon, Portugal.
authors.forename: Inês
authors.initials: I
authors.lastname: Silva
authors.affiliation: National Institute of Legal Medicine and Forensic Sciences - South Branch, Rua Manuel Bento de Sousa n°3, 1169-201 Lisbon, Portugal.
authors.forename: Antonio Castañera
authors.initials: AC
authors.lastname: Ajenjo
authors.affiliation: National Institute of Legal Medicine and Forensic Sciences - South Branch, Rua Manuel Bento de Sousa n°3, 1169-201 Lisbon, Portugal.
authors.forename: Mário João
authors.initials: MJ
authors.lastname: Dias
mesh_terms: D002186:Cannabinoids / Q000652:urine*; D002853:Chromatography, Liquid / Q000379:methods*; D015198:Designer Drugs; D006801:Humans; D013287:Illicit Drugs / Q000652:urine*; D007211:Indoles / Q000652:urine; D013058:Mass Spectrometry / Q000379:methods*; D009281:Naphthalenes / Q000652:urine; D011174:Portugal; D052616:Solid Phase Extraction
chemical_list: D002186:Cannabinoids; D015198:Designer Drugs; D013287:Illicit Drugs; D007211:Indoles; D009281:Naphthalenes; C568258:JWH-073; C552597:1-pentyl-3-(1-naphthoyl)indole
keywords: Spice; Synthetic cannabinoids; UPLC–MS/MS; Urine