{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,18]],"date-time":"2026-01-18T16:28:52Z","timestamp":1768753732056,"version":"3.49.0"},"reference-count":28,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2024,12,12]],"date-time":"2024-12-12T00:00:00Z","timestamp":1733961600000},"content-version":"vor","delay-in-days":11,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100006261","name":"Taif University","doi-asserted-by":"publisher","award":["TU\u2010DSPP\u20102024\u2010175"],"award-info":[{"award-number":["TU\u2010DSPP\u20102024\u2010175"]}],"id":[{"id":"10.13039\/501100006261","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Luminescence"],"published-print":{"date-parts":[[2024,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Clonazepam, a high\u2010potency benzodiazepine widely prescribed for seizure and panic disorders, carries a risk of abuse and dependency. This study developed a sensitive and selective spectrofluorimetric method for determining 7\u2010aminoclonazepam, the major metabolite of clonazepam, in human urine. A 2<jats:sup>6\u20102<\/jats:sup> factorial design was employed to screen the optimal conditions for derivatization with NBD\u2010Cl as the fluorescent label, considering factors such as pH, reagent volumes, temperature, and reaction time. A significant model was attained (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.0001) revealing alkaline pH (9), elevated temperature (80\u00b0C), and high reagent concentrations as crucial for maximizing fluorescence intensity. The method demonstrated excellent linearity from 10 to 500\u2009ng\/mL (<jats:italic>R<\/jats:italic><jats:sup>2<\/jats:sup>\u2009=\u20090.9997), with limits of detection and quantitation of 3.3 and 10\u2009ng\/mL, respectively. Intra\u2010 and inter\u2010day precision (% RSD) were less than 4%, and recoveries ranged from 97.59% to 106.12%. The method also showed no significant interference from endogenous compounds, pharmaceutical excipients, or the parent drug. Applicability of the method was validated in human subjects receiving clonazepam therapy; 7\u2010aminoclonazepam was first detected after 12\u2009h, peaked at 24\u2009h (54.61\u2009\u00b1\u20099.870\u2009ng\/mL), and remained detectable up to 72\u2009h post\u2010dose, offering a simple, cost\u2010effective spectrofluorimetric method for monitoring clonazepam metabolism in clinical and forensic settings.<\/jats:p>","DOI":"10.1002\/bio.70049","type":"journal-article","created":{"date-parts":[[2024,12,12]],"date-time":"2024-12-12T20:06:13Z","timestamp":1734033973000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Spectrofluorimetric Determination of 7\u2010Aminoclonazepam, a Major Clonazepam Metabolite, in Human Urine: A Factorial Design of Experiment Approach to Optimize Method Development"],"prefix":"10.1002","volume":"39","author":[{"given":"Maram\u00a0H.","family":"Abduljabbar","sequence":"first","affiliation":[{"name":"Department of Pharmacology and Toxicology, College of Pharmacy Taif University  Taif Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yusuf\u00a0S.","family":"Althobaiti","sequence":"additional","affiliation":[{"name":"Department of Pharmacology and Toxicology, College of Pharmacy Taif University  Taif Saudi Arabia"},{"name":"Addiction and Neuroscience Research Unit, Health Science Campus Taif University  Taif Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Reem\u00a0M.","family":"Alnemari","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutics and Industrial Pharmacy, College of Pharmacy Taif University  Taif Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Majed\u00a0A.","family":"Algarni","sequence":"additional","affiliation":[{"name":"Department of Clinical Pharmacy, College of Pharmacy Taif University  Taif Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ahmed\u00a0K.","family":"Bamaga","sequence":"additional","affiliation":[{"name":"Pediatric Neurology Unit, Department of Pediatrics, Faculty of Medicine King Abdulaziz University  Jeddah Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Adnan","family":"Alharbi","sequence":"additional","affiliation":[{"name":"Pharmaceutical Practices Department, College of Pharmacy Umm Al\u2010Qura University  Makkah Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1559-8079","authenticated-orcid":false,"given":"Ahmed","family":"Serag","sequence":"additional","affiliation":[{"name":"Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy Al\u2010Azhar University  Cairo Egypt"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Atiah\u00a0H.","family":"Almalki","sequence":"additional","affiliation":[{"name":"Addiction and Neuroscience Research Unit, Health Science Campus Taif University  Taif Saudi Arabia"},{"name":"Department of Pharmaceutical Chemistry, College of Pharmacy Taif University  Taif Saudi Arabia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,12,12]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1016\/S0031-6997(25)00468-5","article-title":"Benzodiazepines: Use, Abuse, and Consequences","volume":"44","author":"Woods J. 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