title: Quantification of [1-(5-fluoropentyl)-1H-indol-3-yl](naphthalene-1-yl)methanone (AM-2201) and 13 metabolites in human and rat plasma by liquid chromatography-tandem mass spectrometry.
abstract: AM-2201 is a popular synthetic cannabinoid first synthesized in 2000. AM-2201 pharmacokinetic and pharmacodynamic data are scarce, requiring further investigation. We developed a sensitive method for quantifying AM-2201 and 13 metabolites in plasma to provide a tool to further metabolic, pharmacokinetic and pharmacodynamic studies. Analysis was performed by liquid chromatography-tandem mass spectrometry. Chromatographic separation was performed by gradient elution on a biphenyl column with 0.1% formic acid in water/0.1% formic acid in acetonitrile:methanol 50:50 (v/v) mobile phase. Sample preparation (75μL) consisted of an enzymatic hydrolysis and a supported liquid extraction. The method was validated with human plasma with a 0.025 or 0.050-50μg/L working range, and cross-validated for rat plasma. Analytical recovery was 88.8-110.1% of target concentration, and intra- (n=30) and inter-day (n=30) imprecision<11.9% coefficient of variation. Method recoveries and matrix effects ranged from 58.4-84.4% and -62.1 to -15.6%, respectively. AM-2201 and metabolites were stable (±20%) at room temperature for 24h, at 4°C for 72h, and after three freeze-thaw cycles, and for 72h in the autosampler after extraction. The method was developed for pharmacodynamic and pharmacokinetic studies with controlled administration in rats but is applicable for pre-clinical and clinical research and forensic investigations. Rat plasma specimen analysis following subcutaneous AM-2201 administration demonstrated the suitability of the method. AM-2201, JWH-018 N-(5-hydroxypentyl), and JWH-018 N-pentanoic acid concentrations were 4.8±1.0, 0.15±0.03, and 0.34±0.07μg/L, respectively, 8h after AM-2201 administration at 0.3mg/kg (n=5).
authors.affiliation: Clinical Pharmacology & Therapeutics Research Branch, Chemistry and Drug Metabolism Section, Intramural Research Program, National Institute on Drug Abuse (NIDA), National Institutes of Health (NIH), 251 Bayview Boulevard Suite 200, Baltimore, MD 21224, USA.
authors.forename: Jeremy
authors.initials: J
authors.lastname: Carlier
authors.affiliation: Clinical Pharmacology & Therapeutics Research Branch, Chemistry and Drug Metabolism Section, Intramural Research Program, National Institute on Drug Abuse (NIDA), National Institutes of Health (NIH), 251 Bayview Boulevard Suite 200, Baltimore, MD 21224, USA. Electronic address: KScheidweiler@intra.nida.nih.gov.
authors.forename: Karl B
authors.initials: KB
authors.lastname: Scheidweiler
authors.affiliation: Clinical Pharmacology & Therapeutics Research Branch, Chemistry and Drug Metabolism Section, Intramural Research Program, National Institute on Drug Abuse (NIDA), National Institutes of Health (NIH), 251 Bayview Boulevard Suite 200, Baltimore, MD 21224, USA.
authors.forename: Ariane
authors.initials: A
authors.lastname: Wohlfarth
authors.affiliation: Designer Drug Research Unit, Intramural Research Program, National Institute on Drug Abuse (NIDA), National Institutes of Health (NIH), 333 Cassell Drive Suite 4400, Baltimore, MD 21224, USA.
authors.forename: Bonita D
authors.initials: BD
authors.lastname: Salmeron
authors.affiliation: Designer Drug Research Unit, Intramural Research Program, National Institute on Drug Abuse (NIDA), National Institutes of Health (NIH), 333 Cassell Drive Suite 4400, Baltimore, MD 21224, USA.
authors.forename: Michael H
authors.initials: MH
authors.lastname: Baumann
authors.affiliation: Clinical Pharmacology & Therapeutics Research Branch, Chemistry and Drug Metabolism Section, Intramural Research Program, National Institute on Drug Abuse (NIDA), National Institutes of Health (NIH), 251 Bayview Boulevard Suite 200, Baltimore, MD 21224, USA.
authors.forename: Marilyn A
authors.initials: MA
authors.lastname: Huestis
mesh_terms: D000818:Animals; D002186:Cannabinoids / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D007211:Indoles / Q000008:administration & dosage / Q000097:blood* / Q000378:metabolism* / Q000493:pharmacokinetics; D009281:Naphthalenes / Q000008:administration & dosage / Q000097:blood* / Q000378:metabolism* / Q000493:pharmacokinetics; D010421:Pentanoic Acids / Q000097:blood; D051381:Rats; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: C582337:1-(5-fluoropentyl)-3-(1-naphthoyl)indole; D002186:Cannabinoids; D007211:Indoles; D009281:Naphthalenes; D010421:Pentanoic Acids; C552597:1-pentyl-3-(1-naphthoyl)indole; C038780:n-pentanoic acid
keywords: AM-2201; Liquid chromatography-tandem mass spectrometry; Metabolites; Plasma; Synthetic cannabinoid