{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T13:01:04Z","timestamp":1770814864987,"version":"3.50.1"},"reference-count":31,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2017,2,10]],"date-time":"2017-02-10T00:00:00Z","timestamp":1486684800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Drug Testing and Analysis"],"published-print":{"date-parts":[[2017,8]]},"abstract":"<jats:p>Flubromazolam is a triazole benzodiazepine with high potency and long\u2010lasting central nervous system depressant effects; however, limited data about its pharmacokinetics are available. Here, we report <jats:italic>in vitro<\/jats:italic> studies of the human flubromazolam metabolism analyzed by liquid chromatography high\u2010resolution mass spectrometry (LC\u2010HRMS). <jats:italic>In vitro<\/jats:italic> investigations were carried out in pooled human liver microsomes (pHLM) and recombinant cytochrome P450 (CYP)\u2010enzymes. To confirm those metabolites detected <jats:italic>in vitro<\/jats:italic>, authentic samples obtained from two forensic cases were also analyzed by LC\u2010HRMS. Additionally, determination of the unbound fraction of flubromazolam in pHLM and in plasma was performed by equilibrium dialysis with subsequent prediction of its hepatic clearance (<jats:italic>CL<jats:sub>H<\/jats:sub><\/jats:italic>) using well\u2010stirred and parallel\u2010tube models. Additional findings obtained by routine screening methods of these forensic cases are also reported. Studies using incubations with nicotinamide adenine dinucleotide phosphate\u2010fortified pHLM with or without uridine 5'\u2010diphosphoglucuronic acid and incubations with CYP\u2010enzymes identified the main metabolic pathway of flubromazolam as hydroxylation on the \u03b1\u2010 and\/or 4\u2010position mediated by CYP3A4 and CYP3A5, with subsequent glucuronidation of the hydroxylated metabolites as well as of the parent drug. Further, \u03b1\u2010hydroxy\u2010flubromazolam and its corresponding glucuronide were detected <jats:italic>in vivo<\/jats:italic> together with the <jats:italic>N<\/jats:italic>\u2010glucuronide of flubromazolam. The predicted <jats:italic>CL<\/jats:italic><jats:sub><jats:italic>H<\/jats:italic><\/jats:sub> of flubromazolam using the well\u2010stirred and parallel\u2010tube models were 0.42 and 0.43\u2009mL\/min\/kg, respectively. Based on the data presented here, flubromazolam is primarily metabolized by CYP3A4\/5 with a high protein\u2010binding and a predicted low clearance. Analysis of authentic samples suggested that analytical targets for flubromazolam should be the compound itself and \u03b1\u2010hydroxy\u2010flubromazolam. Copyright \u00a9 2016 John Wiley &amp; Sons, Ltd.<\/jats:p>","DOI":"10.1002\/dta.2146","type":"journal-article","created":{"date-parts":[[2016,12,9]],"date-time":"2016-12-09T08:39:15Z","timestamp":1481272755000},"page":"1182-1191","source":"Crossref","is-referenced-by-count":36,"title":["<i>In vitro<\/i> studies on flubromazolam metabolism and detection of its metabolites in authentic forensic samples"],"prefix":"10.1002","volume":"9","author":[{"given":"Carolina","family":"Noble","sequence":"first","affiliation":[{"name":"Section of Forensic Chemistry, Department of Forensic Medicine, Faculty of Health and Medical Sciences University of Copenhagen  Frederik V's Vej 11, 3 DK\u20102100 Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marie","family":"Mardal","sequence":"additional","affiliation":[{"name":"Section of Forensic Chemistry, Department of Forensic Medicine, Faculty of Health and Medical Sciences University of Copenhagen  Frederik V's Vej 11, 3 DK\u20102100 Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Niels","family":"Bjerre Holm","sequence":"additional","affiliation":[{"name":"Section of Forensic Chemistry, Department of Forensic Medicine, Faculty of Health and Medical Sciences University of Copenhagen  Frederik V's Vej 11, 3 DK\u20102100 Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sys","family":"Stybe Johansen","sequence":"additional","affiliation":[{"name":"Section of Forensic Chemistry, Department of Forensic Medicine, Faculty of Health and Medical Sciences University of Copenhagen  Frederik V's Vej 11, 3 DK\u20102100 Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kristian","family":"Linnet","sequence":"additional","affiliation":[{"name":"Section of Forensic Chemistry, Department of Forensic Medicine, Faculty of Health and Medical Sciences University of Copenhagen  Frederik V's Vej 11, 3 DK\u20102100 Denmark"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2017,2,10]]},"reference":[{"key":"e_1_2_5_2_1","unstructured":"Emcdda. 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