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

• Little pharmacokinetic or pharmacodynamics data are available for AM-2201.
• We developed a method for quantifying AM-2201 and 13 metabolites in plasma.
• The most sensitive method for quantifying AM-2201 metabolites in biological samples.
• Rat plasma specimens following sc AM-2201 injection were analyzed.

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 (75 μL) consisted of an enzymatic hydrolysis and a supported liquid extraction. The method was validated with human plasma with a 0.025 or 0.050–50 μg/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 24 h, at 4 °C for 72 h, and after three freeze-thaw cycles, and for 72 h 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 ± 0.07 μg/L, respectively, 8 h after AM-2201 administration at 0.3 mg/kg (n = 5).