{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T14:40:25Z","timestamp":1786977625172,"version":"build-2736575974"},"reference-count":23,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2023,2,19]],"date-time":"2023-02-19T00:00:00Z","timestamp":1676764800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003945","name":"Link\u00f6pings Universitet","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003945","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Drug Testing and Analysis"],"published-print":{"date-parts":[[2023,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Acetylbenzylfentanyl, benzoylbenzylfentanyl, 3\u2010fluoro\u2010methoxyacetylfentanyl, and 3\u2010phenylpropanoylfentanyl are fentanyl analogs that have been reported to the European Monitoring Centre for Drugs and Drug Addiction in recent years. The aim of this study was to identify metabolic pathways and potential biomarker metabolites of these fentanyl analogs. The compounds were incubated (5\u00a0\u03bcM) with cryopreserved hepatocytes for up to 5\u00a0h in vitro. Metabolites were analyzed with liquid chromatography\u2013quadrupole time of flight\u2013high\u2010resolution mass spectrometry (LC\u2010QTOF\u2010HRMS). The experiments showed that acetylbenzylfentanyl, benzoylbenzylfentanyl, and 3\u2010phenylpropanoylfentanyl were mainly metabolized through\n                    <jats:italic>N<\/jats:italic>\n                    \u2010dealkylation (forming nor\u2010metabolites) and 3\u2010fluoro\u2010methoxyacetylfentanyl mainly through demethylation. Other observed metabolites were formed by mono\u2010\/dihydroxylation, dihydrodiol formation, demethylation, dehydrogenation, amide hydrolysis, and\/or glucuronidation. The experiments showed that a large number of metabolites of 3\u2010phenylpropanoylfentanyl were formed. The exact position of hydroxy groups in formed monohydroxy metabolites could not be established solely based upon recorded MSMS spectra of hepatocyte samples. Therefore, potential monohydroxy metabolites of 3\u2010phenylpropanoylfentanyl, with the hydroxy group in different positions, were synthesized and analyzed together with the hepatocyte samples. This approach could reveal that the \u03b2 position of the phenylpropanoyl moiety was highly favored; \u03b2\u2010OH\u2010phenylpropanoylfentanyl was the most abundant metabolite after the nor\u2010metabolite. Both metabolites have the potential to serve as biomarkers for 3\u2010phenylpropanoylfentanyl. The nor\u2010metabolites of acetylbenzylfentanyl, benzoylbenzylfentanyl, and 3\u2010fluoro\u2010methoxyacetylfentanyl do also seem to be suitable biomarker metabolites, as do the demethylated metabolite of 3\u2010fluoro\u2010methoxyacetylfentanyl. Identified metabolic pathways and formed metabolites were in agreement with findings in previous studies of similar fentanyl analogs.\n                  <\/jats:p>","DOI":"10.1002\/dta.3454","type":"journal-article","created":{"date-parts":[[2023,2,9]],"date-time":"2023-02-09T03:17:55Z","timestamp":1675912675000},"page":"711-729","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["In vitro metabolite identification of acetylbenzylfentanyl, benzoylbenzylfentanyl, 3\u2010fluoro\u2010methoxyacetylfentanyl, and 3\u2010phenylpropanoylfentanyl using LC\u2010QTOF\u2010HRMS together with synthesized references"],"prefix":"10.1002","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0857-6261","authenticated-orcid":false,"given":"Tobias","family":"Rautio","sequence":"first","affiliation":[{"name":"Department of Physics, Chemistry and Biology Link\u00f6ping University  Link\u00f6ping Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daan","family":"Vangerven","sequence":"additional","affiliation":[{"name":"Department of Forensic Genetics and Forensic Toxicology National Board of Forensic Medicine  Link\u00f6ping Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johan","family":"Dahl\u00e9n","sequence":"additional","affiliation":[{"name":"Department of Physics, Chemistry and Biology Link\u00f6ping University  Link\u00f6ping Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5211-3283","authenticated-orcid":false,"given":"Shimpei","family":"Watanabe","sequence":"additional","affiliation":[{"name":"Department of Forensic Genetics and Forensic Toxicology National Board of Forensic Medicine  Link\u00f6ping Sweden"},{"name":"Division of Drug Research, Department of Biomedical and Clinical Sciences Link\u00f6ping University  Link\u00f6ping Sweden"},{"name":"Forensic Science Group, Photon Science Research Division RIKEN SPring\u20108 Center  Sayo\u2010gun Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert","family":"Kronstrand","sequence":"additional","affiliation":[{"name":"Department of Forensic Genetics and Forensic Toxicology National Board of Forensic Medicine  Link\u00f6ping Sweden"},{"name":"Division of Drug Research, Department of Biomedical and Clinical Sciences Link\u00f6ping University  Link\u00f6ping Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Svante","family":"Vikingsson","sequence":"additional","affiliation":[{"name":"Department of Forensic Genetics and Forensic Toxicology National Board of Forensic Medicine  Link\u00f6ping Sweden"},{"name":"Division of Drug Research, Department of Biomedical and Clinical Sciences Link\u00f6ping University  Link\u00f6ping Sweden"},{"name":"Center for Forensic Science RTI International  Research Triangle Park North Carolina USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Konradsson","sequence":"additional","affiliation":[{"name":"Department of Physics, Chemistry and Biology Link\u00f6ping University  Link\u00f6ping Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiongyu","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Physics, Chemistry and Biology Link\u00f6ping University  Link\u00f6ping Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8015-5728","authenticated-orcid":false,"given":"Henrik","family":"Gr\u00e9en","sequence":"additional","affiliation":[{"name":"Department of Forensic Genetics and Forensic Toxicology National Board of Forensic Medicine  Link\u00f6ping Sweden"},{"name":"Division of Drug Research, Department of Biomedical and Clinical Sciences Link\u00f6ping University  Link\u00f6ping Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2023,2,19]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.drugalcdep.2016.11.033"},{"key":"e_1_2_7_3_1","unstructured":"SpencerMR WarnerM BastianBA TrinidadJP HedegaardH.Drug overdose deaths involving fentanyl 2011\u20102016 National Vital Statistic Report201968:3.3."},{"key":"e_1_2_7_4_1","unstructured":"United Nations Office on Drugs and Crime (UNODC).The growing complexity of the opioid crisis. Global SMART update;2020.Global_SMART\u20102020\u2010Vol_24_web.pdf(unodc.org) Accessed January 9 2023."},{"key":"e_1_2_7_5_1","unstructured":"European Monitoring Centre for Drugs and Drug Addiction (EMCDDA).EMCDDA\u2010europol 2017 annual report on the implementation of council decision 2005\/387\/JHA. 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