{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T08:53:22Z","timestamp":1788252802227,"version":"build-2803163510"},"reference-count":43,"publisher":"Oxford University Press (OUP)","issue":"3","license":[{"start":{"date-parts":[[2026,1,8]],"date-time":"2026-01-08T00:00:00Z","timestamp":1767830400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22306203"],"award-info":[{"award-number":["22306203"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["22276216"],"award-info":[{"award-number":["22276216"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Qing Lan Project of Jiangsu Province"},{"name":"Open Project of Key Laboratory of Forensic Science and Technology at College of Sichuan Province","award":["2023ZD002"],"award-info":[{"award-number":["2023ZD002"]}]},{"name":"Open Research Fund of State Environmental Protection Key Laboratory of Biodiversity and Biosafety"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,3,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Methyl 2-{[1-(5-fluoropentyl)-1H-indazole-3-carbonyl]amino}-3,3-dimethylbutanoate (5F-ADB), a potent synthetic cannabinoid, induces intense euphoria, hallucinations, and addiction, posing significant risks to human health. Current drug surveillance efforts lack data to identify drug abuse, and the environmental impacts of 5F-ADB entering aquatic systems via synthesis or use remain uncharacterized. To address these gaps, a multilevel assessment system (in vitro\u2013invertebrate\u2013vertebrate) was established to elucidate 5F-ADB metabolic pathways and identify robust biomarkers. Human liver microsomes (HLMs), Daphnia magna, and zebrafish were exposed to 5F-ADB, with metabolites profiled by high-performance liquid chromatography coupled with mass spectrometry. Metabolic pathways were inferred, and metabolite toxicity was evaluated. Results revealed 9, 11, and 22 metabolites in HLMs, D. magna, and zebrafish models, respectively. Dominant pathways in HLMs and zebrafish included ester hydrolysis, defluorinated hydroxylation, and combined ester hydrolysis\/defluorinated hydroxylation. Daphnia magna metabolism primarily featured defluorinated hydroxylation, depentylation, and ester hydrolysis coupled with hydroxylation. Glucuronidation metabolites were exclusive to zebrafish. Based on abundance and stability, H-M4 (ester hydrolysis), D-M1 (ester hydrolysis\/depentylation), and Z-M15 (ester hydrolysis\/condensation) were identified as key biomarkers for HLMs, D. magna, and zebrafish, respectively. Toxicity assessments indicated reduced toxicity for most metabolites versus 5F-ADB. However, H-M7, D-M7, D-M11, and Z-M15 (products of ester hydrolysis\/condensation or defluorinated hydroxylation\/oxidation) exhibited comparable toxicity to the parent compound. Critically, D-M7 (defluorinated hydroxylation\/oxidation) demonstrated heightened hydrophilicity and potentially elevated ecotoxicity in D. magna, warranting further ecological risk investigation. This study provides the first multitrophic metabolic characterization of 5F-ADB, delivering critical data for tracing illicit synthesis, monitoring drug use distribution, and evaluating environmental hazards of synthetic cannabinoids.<\/jats:p>","DOI":"10.1093\/etojnl\/vgag003","type":"journal-article","created":{"date-parts":[[2026,1,9]],"date-time":"2026-01-09T02:23:53Z","timestamp":1767925433000},"page":"719-730","source":"Crossref","is-referenced-by-count":1,"title":["Multilevel metabolic profiling of synthetic cannabinoid 5F-ADB: identifying definitive biomarkers for forensic source tracking and ecotoxicological risk assessment"],"prefix":"10.1093","volume":"45","author":[{"given":"Qinghua","family":"Liu","sequence":"first","affiliation":[{"name":"School of Engineering, China Pharmaceutical University , Nanjing,","place":["China"]},{"name":"Criminal Technology Division of the Criminal Investigation Corps, Tianjin Municipal Public Security Bureau , Tianjin,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuqing","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Engineering, China Pharmaceutical University , Nanjing,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beiya","family":"Ma","sequence":"additional","affiliation":[{"name":"School of Engineering, China Pharmaceutical University , Nanjing,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pan","family":"Yi","sequence":"additional","affiliation":[{"name":"School of Engineering, China Pharmaceutical University , Nanjing,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Runwen","family":"Shao","sequence":"additional","affiliation":[{"name":"School of Engineering, China Pharmaceutical University , Nanjing,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuhan","family":"Li","sequence":"additional","affiliation":[{"name":"School of Engineering, China Pharmaceutical University , Nanjing,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongxia","family":"Sui","sequence":"additional","affiliation":[{"name":"Donghai Street Community Health Service Center , Tianjin,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruixin","family":"Guo","sequence":"additional","affiliation":[{"name":"School of Engineering, China Pharmaceutical University , Nanjing,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianqiu","family":"Chen","sequence":"additional","affiliation":[{"name":"School of Engineering, China Pharmaceutical University , Nanjing,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanhua","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Engineering, China Pharmaceutical University , Nanjing,","place":["China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,1,8]]},"reference":[{"key":"2026032604045525800_vgag003-B1","doi-asserted-by":"publisher","first-page":"S37","DOI":"10.1002\/jcph.1871","article-title":"Natural and synthetic cannabinoids: Pharmacology, uses, adverse drug events, and drug interactions","volume":"61","author":"Brown","year":"2021","journal-title":"Journal of Clinical Pharmacology"},{"key":"2026032604045525800_vgag003-B2","doi-asserted-by":"publisher","first-page":"18445","DOI":"10.1021\/acs.analchem.4c03658","article-title":"Establishment of broad-specificity monoclonal antibody-based immunoassay for rapid detection of indole-type and indazole-type synthetic cannabinoids and metabolites","volume":"96","author":"Chen","year":"2024","journal-title":"Analytical Chemistry"},{"key":"2026032604045525800_vgag003-B3","doi-asserted-by":"publisher","first-page":"974","DOI":"10.1177\/1087057112447305","article-title":"An investigation of the bioactivation potential and metabolism profile of zebrafish versus human","volume":"17","author":"Chng","year":"2012","journal-title":"Journal of Biomolecular Screening"},{"key":"2026032604045525800_vgag003-B4","doi-asserted-by":"publisher","first-page":"52","DOI":"10.1007\/s11920-016-0694-1","article-title":"Adverse effects of synthetic cannabinoids: Management of acute toxicity and withdrawal","volume":"18","author":"Cooper","year":"2016","journal-title":"Current Psychiatry Reports"},{"key":"2026032604045525800_vgag003-B5","doi-asserted-by":"publisher","first-page":"88","DOI":"10.1016\/j.toxlet.2022.07.815","article-title":"Induction of CYP450 by illicit drugs: Studies using an in vitro 3D spheroidal model in comparison to animals","volume":"367","author":"Duan","year":"2022","journal-title":"Toxicology Letters"},{"key":"2026032604045525800_vgag003-B6","doi-asserted-by":"publisher","first-page":"152543","DOI":"10.1016\/j.scitotenv.2021.152543","article-title":"Bioconcentration of neuroactive pharmaceuticals in fish: Relation to lipophilicity, experimental design and toxicity in the aquatic environment","volume":"812","author":"Duarte","year":"2022","journal-title":"The Science of the Total Environment"},{"key":"2026032604045525800_vgag003-B7","doi-asserted-by":"publisher","first-page":"283","DOI":"10.1007\/3-540-26573-2_9","article-title":"The phylogenetic distribution and evolutionary origins of endocannabinoid signalling","author":"Elphick","year":"2005","journal-title":"Cannabinoids"},{"key":"2026032604045525800_vgag003-B8","doi-asserted-by":"publisher","first-page":"836","DOI":"10.1124\/dmd.111.043372","article-title":"Development of an in vitro system with human liver microsomes for phenotyping of CYP2C9 genetic polymorphisms with a mechanism-based inactivator","volume":"40","author":"Flora","year":"2012","journal-title":"Drug Metabolism and Disposition: The Biological Fate of Chemicals"},{"key":"2026032604045525800_vgag003-B9","doi-asserted-by":"publisher","first-page":"3561","DOI":"10.1007\/s00216-019-01837-8","article-title":"In vitro metabolic profiling of synthetic cannabinoids by pooled human liver microsomes, cytochrome P450 isoenzymes, and Cunninghamella elegans and their detection in urine samples","volume":"411","author":"Gaunitz","year":"2019","journal-title":"Analytical and Bioanalytical Chemistry"},{"key":"2026032604045525800_vgag003-B10","doi-asserted-by":"publisher","DOI":"10.3390\/ijms23105372","article-title":"Zebrafish: A new promise to study the impact of metabolic disorders on the brain","volume":"23","author":"Ghaddar","year":"2022","journal-title":"International Journal of Molecular Sciences"},{"key":"2026032604045525800_vgag003-B11","doi-asserted-by":"publisher","first-page":"108773","DOI":"10.1016\/j.pharmthera.2024.108773","article-title":"Glucuronidation of orally administered drugs and the value of nanocarriers in strategies for its overcome","volume":"266","author":"Hervieu","year":"2025","journal-title":"Pharmacology and Therapeutics"},{"key":"2026032604045525800_vgag003-B12","doi-asserted-by":"publisher","first-page":"1237","DOI":"10.1208\/s12248-015-9788-7","article-title":"CYP3A4 mediates oxidative metabolism of the synthetic cannabinoid AKB-48","volume":"17","author":"Holm","year":"2015","journal-title":"The AAPS Journal"},{"key":"2026032604045525800_vgag003-B13","doi-asserted-by":"publisher","first-page":"62","DOI":"10.3389\/fnana.2018.00062","article-title":"Overlapping distribution of orexin and endocannabinoid receptors and their functional interaction in the brain of adult zebrafish","volume":"12","author":"Imperatore","year":"2018","journal-title":"Frontiers in Neuroanatomy"},{"key":"2026032604045525800_vgag003-B14","doi-asserted-by":"publisher","DOI":"10.3390\/ijms251910812","article-title":"Characterization and expression of the cytochrome P450 genes in Daphnia magna exposed to cerium oxide nanoparticles","volume":"25","author":"Kang","year":"2024","journal-title":"International Journal of Molecular Sciences"},{"key":"2026032604045525800_vgag003-B15","doi-asserted-by":"publisher","first-page":"3040","DOI":"10.1021\/jm5000112","article-title":"Optimization of chemical functionalities of indole-2-carboxamides to improve allosteric parameters for the cannabinoid receptor 1 (CB1)","volume":"57","author":"Khurana","year":"2014","journal-title":"Journal of Medicinal Chemistry"},{"key":"2026032604045525800_vgag003-B16","doi-asserted-by":"publisher","first-page":"100787","DOI":"10.1016\/j.cbd.2020.100787","article-title":"The genome of the marine water flea Diaphanosoma celebensis: Identification of Phase I, II, and III detoxification genes and potential applications in marine molecular ecotoxicology","volume":"37","author":"Kim","year":"2021","journal-title":"Comparative Biochemistry and Physiology Part D: Genomics and Proteomics"},{"key":"2026032604045525800_vgag003-B17","doi-asserted-by":"publisher","DOI":"10.3390\/molecules23040920","article-title":"In vitro inhibitory effects of synthetic cannabinoid EAM-2201 on cytochrome P450 and UDP-glucuronosyltransferase enzyme activities in human liver microsomes","volume":"23","author":"Kong","year":"2018","journal-title":"Molecules"},{"key":"2026032604045525800_vgag003-B18","doi-asserted-by":"publisher","first-page":"108800","DOI":"10.1016\/j.neuropharm.2021.108800","article-title":"Pharmacokinetics and pharmacodynamics of the synthetic cannabinoid, 5F-MDMB-PICA, in male rats","volume":"199","author":"Krotulski","year":"2021","journal-title":"Neuropharmacology"},{"key":"2026032604045525800_vgag003-B19","doi-asserted-by":"publisher","first-page":"145","DOI":"10.1007\/5584_2021_684","article-title":"An overview of zebrafish modeling methods in drug discovery and development","volume":"1387","author":"Larijani","year":"2022","journal-title":"Advances in Experimental Medicine and Biology"},{"key":"2026032604045525800_vgag003-B20","doi-asserted-by":"publisher","first-page":"3905","DOI":"10.1007\/s00216-022-04034-2","article-title":"Study on the metabolic process of synthetic cannabinoids 4F-MDMB-BINACA and 4F-MDMB-BICA in human liver microsome and zebrafish model via UHPLC-QE Orbitrap MS","volume":"414","author":"Li","year":"2022","journal-title":"Analytical and Bioanalytical Chemistry"},{"key":"2026032604045525800_vgag003-B21","doi-asserted-by":"publisher","DOI":"10.1002\/bmc.4113","article-title":"UPLC-HR-MS\/MS-based determination study on the metabolism of four synthetic cannabinoids, ADB-FUBICA, AB-FUBICA, AB-BICA and ADB-BICA, by human liver microsomes","volume":"32","author":"Li","year":"2018","journal-title":"Biomedical Chromatography: BMC"},{"key":"2026032604045525800_vgag003-B22","doi-asserted-by":"publisher","first-page":"459","DOI":"10.12116\/j.issn.1004-5619.2021.310602","article-title":"Research progress on metabolite identification of synthetic cannabinoid new psychoactive substances","volume":"37","author":"Li","year":"2021","journal-title":"Fa yi Xue za Zhi"},{"key":"2026032604045525800_vgag003-B23","doi-asserted-by":"publisher","first-page":"2298","DOI":"10.1093\/etojnl\/vgaf135","article-title":"Aquatic toxicity of synthetic cannabinoid: Multisystem impacts of 5F-ADB on neurodevelopment in Daphnia magna","volume":"44","author":"Liu","year":"2025","journal-title":"Environmental Toxicology and Chemistry"},{"key":"2026032604045525800_vgag003-B24","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1515\/dmdi.1982.4.2-3.123","article-title":"Species differences in cytochromes P450 and their corresponding messenger RNA\u2019s","volume":"4","author":"Makowski","year":"1982","journal-title":"Quarterly Reviews on Drug Metabolism and Drug Interactions"},{"key":"2026032604045525800_vgag003-B25","doi-asserted-by":"publisher","first-page":"2603","DOI":"10.2174\/1381612827666211115170521","article-title":"Fourth generation of synthetic cannabinoid receptor agonists: A review on the latest insights","volume":"28","author":"Malaca","year":"2022","journal-title":"Current Pharmaceutical Design"},{"key":"2026032604045525800_vgag003-B26","doi-asserted-by":"publisher","first-page":"555","DOI":"10.1007\/s00438-007-0207-3","article-title":"A shifted repertoire of endocannabinoid genes in the zebrafish (Danio rerio)","volume":"277","author":"McPartland","year":"2007","journal-title":"Molecular Genetics and Genomics: MGG"},{"key":"2026032604045525800_vgag003-B27","doi-asserted-by":"publisher","first-page":"907","DOI":"10.1111\/j.1440-1681.1997.tb02718.x","article-title":"Structure and function of uridine diphosphate glucuronosyltransferases","volume":"24","author":"Meech","year":"1997","journal-title":"Clinical and Experimental Pharmacology and Physiology"},{"key":"2026032604045525800_vgag003-B28","doi-asserted-by":"publisher","first-page":"261","DOI":"10.1007\/978-1-0716-3625-1_13","article-title":"Daphnia magna as a model organism to predict the teratogenic effect of different compounds","volume":"2753","author":"Mishra","year":"2024","journal-title":"Methods in Molecular Biology"},{"key":"2026032604045525800_vgag003-B29","doi-asserted-by":"publisher","first-page":"594","DOI":"10.1002\/dta.3195","article-title":"Metabolism of third generation synthetic cannabinoids using zebrafish larvae","volume":"14","author":"Morales-No\u00e9","year":"2022","journal-title":"Drug Testing and Analysis"},{"key":"2026032604045525800_vgag003-B30","doi-asserted-by":"publisher","first-page":"195","DOI":"10.1002\/dta.2697","article-title":"Simultaneous analysis of 29 synthetic cannabinoids and metabolites, amphetamines, and cannabinoids in human whole blood by liquid chromatography-tandem mass spectrometry\u2014A New Zealand perspective of use in 2018","volume":"12","author":"Ong","year":"2020","journal-title":"Drug Testing and Analysis"},{"key":"2026032604045525800_vgag003-B31","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1016\/j.toxlet.2019.01.010","article-title":"Tools for studying the metabolism of new psychoactive substances for toxicological screening purposes\u2014A comparative study using pooled human liver S9, HepaRG cells, and zebrafish larvae","volume":"305","author":"Richter","year":"2019","journal-title":"Toxicology Letters"},{"key":"2026032604045525800_vgag003-B32","doi-asserted-by":"publisher","first-page":"1467","DOI":"10.1007\/s00414-021-02561-8","article-title":"Involuntary 5F-ADB-related intoxication following e-cigarette use","volume":"135","author":"Salle","year":"2021","journal-title":"International Journal of Legal Medicine"},{"key":"2026032604045525800_vgag003-B33","doi-asserted-by":"publisher","first-page":"1495","DOI":"10.1002\/etc.4436","article-title":"When fluorescence is not a particle: The tissue translocation of microplastics in Daphnia magna seems an artifact","volume":"38","author":"Sch\u00fcr","year":"2019","journal-title":"Environmental Toxicology and Chemistry"},{"key":"2026032604045525800_vgag003-B34","doi-asserted-by":"publisher","first-page":"323","DOI":"10.1007\/164_2019_298","volume-title":"Substance use disorders: From etiology to treatment","author":"Schurman","year":"2020"},{"key":"2026032604045525800_vgag003-B35","doi-asserted-by":"publisher","first-page":"143038","DOI":"10.1016\/j.scitotenv.2020.143038","article-title":"Daphnia magna model in the toxicity assessment of pharmaceuticals: A review","volume":"763","author":"Tkaczyk","year":"2021","journal-title":"The Science of the Total Environment"},{"key":"2026032604045525800_vgag003-B36","doi-asserted-by":"publisher","first-page":"412","DOI":"10.1089\/zeb.2020.1930","article-title":"Zebrafish as an emerging model system in the global south: Two decades of research in Brazil","volume":"17","author":"Trigueiro","year":"2020","journal-title":"Zebrafish"},{"key":"2026032604045525800_vgag003-B37","doi-asserted-by":"publisher","first-page":"127","DOI":"10.1002\/dta.2688","article-title":"5F-MDMB-PICA metabolite identification and cannabinoid receptor activity","volume":"12","author":"Truver","year":"2020","journal-title":"Drug Testing and Analysis"},{"key":"2026032604045525800_vgag003-B38","doi-asserted-by":"publisher","first-page":"247","DOI":"10.1016\/j.taap.2004.10.019","article-title":"Constitutive and xenobiotics-induced expression of a novel CYP3A gene from zebrafish larva","volume":"205","author":"Tseng","year":"2005","journal-title":"Toxicology and Applied Pharmacology"},{"key":"2026032604045525800_vgag003-B39","author":"U.S. Environmental Protection Agency","year":"2024"},{"key":"2026032604045525800_vgag003-B40","doi-asserted-by":"publisher","first-page":"278","DOI":"10.1007\/s11419-022-00612-2","article-title":"Comparison between human liver microsomes and the fungus Cunninghamella elegans for biotransformation of the synthetic cannabinoid JWH-424 having a bromo-naphthyl moiety analysed by high-resolution mass spectrometry","volume":"40","author":"Watanabe","year":"2022","journal-title":"Forensic Toxicology"},{"key":"2026032604045525800_vgag003-B41","doi-asserted-by":"publisher","first-page":"109866","DOI":"10.1016\/j.forsciint.2019.06.024","article-title":"Detection and quantification of 5F-ADB and its methyl ester hydrolysis metabolite in fatal intoxication cases by liquid chromatography-high resolution mass spectrometry","volume":"302","author":"Yeter","year":"2019","journal-title":"Forensic Science International"},{"key":"2026032604045525800_vgag003-B42","doi-asserted-by":"publisher","first-page":"516","DOI":"10.1089\/can.2020.0161","article-title":"Synthetic cannabinoid-related deaths in England, 2012\u20132019","volume":"7","author":"Yoganathan","year":"2022","journal-title":"Cannabis and Cannabinoid Research"},{"key":"2026032604045525800_vgag003-B43","doi-asserted-by":"publisher","first-page":"1223","DOI":"10.1002\/dta.3019","article-title":"Metabolism of 4F-MDMB-BICA in zebrafish by liquid chromatography\u2013high resolution mass spectrometry","volume":"13","author":"Yue","year":"2021","journal-title":"Drug Testing and Analysis"}],"container-title":["Environmental Toxicology and Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/etc\/advance-article-pdf\/doi\/10.1093\/etojnl\/vgag003\/66313429\/vgag003.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/etc\/article-pdf\/45\/3\/719\/66313429\/vgag003.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/etc\/article-pdf\/45\/3\/719\/66313429\/vgag003.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T08:05:03Z","timestamp":1774512303000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/etc\/article\/45\/3\/719\/8417629"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,8]]},"references-count":43,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,1,8]]},"published-print":{"date-parts":[[2026,3,1]]}},"URL":"https:\/\/doi.org\/10.1093\/etojnl\/vgag003","relation":{},"ISSN":["0730-7268","1552-8618"],"issn-type":[{"value":"0730-7268","type":"print"},{"value":"1552-8618","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2026,3]]},"published":{"date-parts":[[2026,1,8]]}}}