title: Targeted metabolomic approach for assessing human synthetic cannabinoid exposure and pharmacology.
abstract: Designer synthetic cannabinoids like JWH-018 and AM2201 have unique clinical toxicity. Cytochrome-P450-mediated metabolism of each leads to the generation of pharmacologically active (ω)- and (ω-1)-monohydroxyl metabolites that retain high affinity for cannabinoid type-1 receptors, exhibit Δ(9)-THC-like effects in rodents, and are conjugated with glucuronic acid prior to excretion in human urine. Previous studies have not measured the contribution of the specific (ω-1)-monohydroxyl enantiomers in human metabolism and toxicity. This study uses a chiral liquid chromatography-tandem mass spectroscopy approach (LC-MS/MS) to quantify each specific enantiomer and other nonchiral, human metabolites of JWH-018 and AM2201 in human urine. The accuracy (average % RE = 18.6) and reproducibility (average CV = 15.8%) of the method resulted in low-level quantification (average LLQ = 0.99 ng/mL) of each metabolite. Comparisons with a previously validated nonchiral method showed strong correlation between the two approaches (average r(2) = 0.89). Pilot data from human urine samples demonstrate enantiospecific excretion patterns. The (S)-isomer of the JWH-018-(ω-1)-monohydroxyl metabolite was predominantly excreted (>87%) in human urine as the glucuronic acid conjugate, whereas the relative abundance of the corresponding AM2201-(ω-1)-metabolite was low (<5%) and did not demonstrate enantiospecificity (approximate 50:50 ratio of each enantiomer). The new chiral method provides a comprehensive, targeted metabolomic approach for studying the human metabolism of JWH-018 and AM2201. Preliminary evaluations of specific enantiomeric contributions support the use of this approach in future studies designed to understand the pharmacokinetic properties of JWH-018 and/or AM2201.
authors.affiliation: Arkansas Department of Health, Public Health Laboratory , Little Rock, Arkansas 72205, United States.
authors.forename: Amy L
authors.initials: AL
authors.lastname: Patton
authors.forename: Kathryn A
authors.initials: KA
authors.lastname: Seely
authors.forename: Krishna C
authors.initials: KC
authors.lastname: Chimalakonda
authors.forename: Johnny P
authors.initials: JP
authors.lastname: Tran
authors.forename: Matthew
authors.initials: M
authors.lastname: Trass
authors.forename: Art
authors.initials: A
authors.lastname: Miranda
authors.forename: William E
authors.initials: WE
authors.lastname: Fantegrossi
authors.forename: Paul D
authors.initials: PD
authors.lastname: Kennedy
authors.forename: Paul
authors.initials: P
authors.lastname: Dobrowolski
authors.forename: Anna
authors.initials: A
authors.lastname: Radominska-Pandya
authors.forename: Keith R
authors.initials: KR
authors.lastname: McCain
authors.forename: Laura P
authors.initials: LP
authors.lastname: James
authors.forename: Gregory W
authors.initials: GW
authors.lastname: Endres
authors.forename: Jeffery H
authors.initials: JH
authors.lastname: Moran
mesh_terms: D002853:Chromatography, Liquid; D006801:Humans; D007211:Indoles / Q000378:metabolism* / Q000493:pharmacokinetics / Q000652:urine; D055432:Metabolomics; D015394:Molecular Structure; D009281:Naphthalenes / Q000378:metabolism* / Q000493:pharmacokinetics / Q000652:urine; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry; D014018:Tissue Distribution
chemical_list: C582337:1-(5-fluoropentyl)-3-(1-naphthoyl)indole; D007211:Indoles; D009281:Naphthalenes; C552597:1-pentyl-3-(1-naphthoyl)indole