{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T13:06:56Z","timestamp":1770815216456,"version":"3.50.1"},"reference-count":36,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2019,2,5]],"date-time":"2019-02-05T00:00:00Z","timestamp":1549324800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["INST 380\/92\u20101 FUGG"],"award-info":[{"award-number":["INST 380\/92\u20101 FUGG"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000897","name":"Directorate-General for Justice","doi-asserted-by":"publisher","award":["JUST\/2013\/ISEC\/DRUGS\/AG\/6421"],"award-info":[{"award-number":["JUST\/2013\/ISEC\/DRUGS\/AG\/6421"]}],"id":[{"id":"10.13039\/501100000897","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":[[2019,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The number of newly appearing benzodiazepine derivatives on the new psychoactive substances (NPS) drug market has increased over the last couple of years totaling 23 \u2018designer benzodiazepines\u2019 monitored at the end of 2017 by the European Monitoring Centre for Drugs and Drug Addiction. In the present study, three benzodiazepines [flunitrazolam, norflurazepam, and 4\u2032\u2010chlorodiazepam (Ro5\u20134864)] offered as \u2018research chemicals' on the Internet were characterized and their main <jats:italic>in vitro<\/jats:italic> phase I metabolites tentatively identified after incubation with pooled human liver microsomes. For all compounds, the structural formula declared by the vendor was confirmed by gas chromatography\u2212mass spectrometry (GC\u2013MS), liquid chromatography\u2212tandem mass spectrometry (LC MS\/MS), liquid chromatography\u2212quadrupole time of flight\u2212mass spectrometry (LC\u2212QTOF\u2212MS) analysis and nuclear magnetic resonance (NMR) spectroscopy. The metabolic steps of flunitrazolam were monohydroxylation, dihydroxylation, and reduction of the nitro function. The detected <jats:italic>in vitro<\/jats:italic> phase I metabolites of norflurazepam were hydroxynorflurazepam and dihydroxynorflurazepam. 4\u2019\u2010Chlorodiazepam biotransformation consisted of <jats:italic>N<\/jats:italic>\u2010dealkylation and hydroxylation. It has to be noted that 4\u2032\u2010chlorodiazepam and its metabolites show almost identical LC\u2013MS\/MS fragmentation patterns to diclazepam and its metabolites (delorazepam, lormetazepam, and lorazepam), making a sufficient chromatographic separation inevitable. Sale of norflurazepam, the metabolite of the prescribed benzodiazepines flurazepam and fludiazepam, presents the risk of incorrect interpretation of analytical findings.<\/jats:p>","DOI":"10.1002\/dta.2561","type":"journal-article","created":{"date-parts":[[2018,12,22]],"date-time":"2018-12-22T10:08:01Z","timestamp":1545473281000},"page":"541-549","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Characterization and <i>in vitro<\/i> phase I microsomal metabolism of designer benzodiazepines: An update comprising flunitrazolam, norflurazepam, and 4'\u2010chlorodiazepam (Ro5\u20134864)"],"prefix":"10.1002","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1757-6359","authenticated-orcid":false,"given":"Bjoern","family":"Moosmann","sequence":"first","affiliation":[{"name":"Institute of Forensic Medicine, Forensic Toxicology Kantonsspital St. Gallen  St. Gallen Switzerland"},{"name":"Institute of Forensic Medicine, Forensic Toxicology, Medical Center \u2010 University of Freiburg, Faculty of Medicine University of Freiburg  Freiburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Philippe","family":"Bisel","sequence":"additional","affiliation":[{"name":"Institute of Pharmaceutical Sciences University of Freiburg  Freiburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Folker","family":"Westphal","sequence":"additional","affiliation":[{"name":"State Bureau of Criminal Investigation Schleswig\u2010Holstein  Kiel Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maurice","family":"Wilde","sequence":"additional","affiliation":[{"name":"Institute of Forensic Medicine, Forensic Toxicology, Medical Center \u2010 University of Freiburg, Faculty of Medicine University of Freiburg  Freiburg Germany"},{"name":"Hermann Staudinger Graduate School University of Freiburg  Freiburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J\u00fcrgen","family":"Kempf","sequence":"additional","affiliation":[{"name":"Institute of Forensic Medicine, Forensic Toxicology, Medical Center \u2010 University of Freiburg, Faculty of Medicine University of Freiburg  Freiburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3081-4609","authenticated-orcid":false,"given":"Verena","family":"Angerer","sequence":"additional","affiliation":[{"name":"Institute of Forensic Medicine, Forensic Toxicology, Medical Center \u2010 University of Freiburg, Faculty of Medicine University of Freiburg  Freiburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1883-2804","authenticated-orcid":false,"given":"Volker","family":"Auw\u00e4rter","sequence":"additional","affiliation":[{"name":"Institute of Forensic Medicine, Forensic Toxicology, Medical Center \u2010 University of Freiburg, Faculty of Medicine University of Freiburg  Freiburg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2019,2,5]]},"reference":[{"key":"e_1_2_7_2_1","unstructured":"European drug report 2018: trends and developments publications office of the European union.2018."},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11419-013-0187-4"},{"key":"e_1_2_7_4_1","unstructured":"European drug report 2016: trends and developments publications office of the European union.2016."},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/dta.1628"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1002\/jms.3840"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1002\/wps.20236"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.3109\/15563650.2015.1112907"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1515\/cclm-2016-0831"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/dta.2003"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jchromb.2016.08.047"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/jms.3279"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-016-9439-6"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1002\/dta.2146"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1208\/s12248-016-0040-x"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.forsciint.2016.10.021"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.1002\/dta.2099"},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11419-015-0277-6"},{"key":"e_1_2_7_19_1","volume-title":"Disposition of toxic drugs and chemicals in man","author":"Baselt RC","year":"2011"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/dta.1484"},{"key":"e_1_2_7_21_1","volume-title":"Drugs affecting the central nervous system","author":"Sternbach LH","year":"1968"},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.scijus.2017.12.004"},{"key":"e_1_2_7_23_1","doi-asserted-by":"publisher","DOI":"10.1097\/00004714-198304000-00036"},{"issue":"1","key":"e_1_2_7_24_1","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/S0026-895X(25)15026-8","article-title":"Characterization of peripheral\u2010type benzodiazepine binding sites in brain using [3H] Ro 5\u20104864","volume":"22","author":"Marangos PJ","year":"1982","journal-title":"Mol Pharmacol"},{"key":"e_1_2_7_25_1","first-page":"139","article-title":"Pharmacological, electrophysiological, and neurochemical actions of the convulsant benzodiazepine Ro 5\u20104864 (4'\u2010chlordiazepam)","volume":"38","author":"Weissman BA","year":"1983","journal-title":"Adv Biochem Psychopharmacol"},{"issue":"4","key":"e_1_2_7_26_1","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1016\/S0031-6997(24)01424-8","article-title":"Enigma of the peripheral benzodiazepine receptor","volume":"51","author":"Gavish M","year":"1999","journal-title":"Pharmacol Rev"},{"key":"e_1_2_7_27_1","doi-asserted-by":"publisher","DOI":"10.1021\/jm00293a015"},{"key":"e_1_2_7_28_1","doi-asserted-by":"publisher","DOI":"10.1093\/jat\/bky012"},{"key":"e_1_2_7_29_1","doi-asserted-by":"publisher","DOI":"10.1002\/dta.2480"},{"key":"e_1_2_7_30_1","unstructured":"Cayman spectral library.https:\/\/www.caymanchem.com\/app\/template\/SpectralLibrary. Accessed April 2018."},{"key":"e_1_2_7_31_1","volume-title":"Mass Spectral and GC Data of Drugs, Poisons, Pesticides, Pollutants and Their Metabolites","author":"Maurer HH","year":"2011"},{"key":"e_1_2_7_32_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.forsciint.2014.04.035"},{"key":"e_1_2_7_33_1","doi-asserted-by":"publisher","DOI":"10.1002\/dta.2201"},{"key":"e_1_2_7_34_1","doi-asserted-by":"publisher","DOI":"10.1016\/0731-7085(92)80019-J"},{"key":"e_1_2_7_35_1","unstructured":"Recommended methods for the detection and assay of barbiturates and benzodiazepines in biological specimens. Manual for use by National laboratories United Nations Office on Drugs and Crime.1997."},{"key":"e_1_2_7_36_1","doi-asserted-by":"publisher","DOI":"10.1002\/cpt1976194443"},{"key":"e_1_2_7_37_1","doi-asserted-by":"publisher","DOI":"10.1520\/JFS16081J"}],"container-title":["Drug Testing and Analysis"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fdta.2561","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/dta.2561","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/dta.2561","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/dta.2561","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T20:19:41Z","timestamp":1760386781000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/dta.2561"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,2,5]]},"references-count":36,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2019,3]]}},"alternative-id":["10.1002\/dta.2561"],"URL":"https:\/\/doi.org\/10.1002\/dta.2561","archive":["Portico"],"relation":{},"ISSN":["1942-7603","1942-7611"],"issn-type":[{"value":"1942-7603","type":"print"},{"value":"1942-7611","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,2,5]]},"assertion":[{"value":"2018-11-12","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2018-12-17","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2019-02-05","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}