Article(id=1292126458256847580, tenantId=1146029695717560320, journalId=1246415772164075586, issueId=1292126409196064779, articleNumber=null, orderNo=null, doi=10.13699/j.cnki.1001-6821.2026.02.012, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1764000000000, receivedDateStr=2025-11-25, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1785996554196, onlineDateStr=2026-08-06, pubDate=1769529600000, pubDateStr=2026-01-28, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1785996554196, onlineIssueDateStr=2026-08-06, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1785996554196, creator=13701087609, updateTime=1785996554196, updator=13701087609, issue=Issue{id=1292126409196064779, tenantId=1146029695717560320, journalId=1246415772164075586, year='2026', volume='42', issue='2', pageStart='151', pageEnd='300', issueExtLink='null', onlineDate='null', pubDate='1769529600000', pubDateStr='2026-01-28', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1785996542500, creator='13701087609', updateTime=1786014671444, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1292202447569707674, tenantId=1146029695717560320, journalId=1246415772164075586, issueId=1292126409196064779, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1292202447569707675, tenantId=1146029695717560320, journalId=1246415772164075586, issueId=1292126409196064779, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=224, endPage=230, ext={EN=ArticleExt(id=1292126458596586205, articleId=1292126458256847580, tenantId=1146029695717560320, journalId=1246415772164075586, language=EN, title=Application of LC-MS/MS method based on NBD-Cl derivatization reagent for determining blood concentrations of amino/mercaptan drugs, columnId=1246531413395263850, journalTitle=Chinese Journal of Clinical Pharmacology, columnName=Research Method, runingTitle=null, highlight=null, articleAbstract=
Objective This study used 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) as a derivatization reagent to establish an LC-MS/MS analytical method for the quantitative detection of a class of amine drugs, mesalazine(5-ASA) and two thiol drugs, methimazole (MMI) and captopril (CAP).
Methods Plasma samples were processed using derivatization and protein precipitation methods. Optimal derivatization conditions (derivatization reagent concentration, reaction temperature, and derivatization time) were investigated for different drugs. The analytical columns used were Thermo AccucoreTM C18 (2.4 μm, 30 mm×4.6 mm), Agilent Poroshell 120 EC-C18 (4 μm, 50 mm×4.6 mm) as analytical columns, with 0.1% formic acid-acetonitrile, 0.1% formic acid-methanol, and 5 mM ammonium acetate-methanol: acetonitrile (V∶V=50∶50) as the gradient mobile phase, with flow rates of 0.5 mL·min-1 and 0.6 mL·min-1, respectively, using an ESI source for positive/negative ion detection in multiple reaction monitoring (MRM) mode.
Results 5-ASA, MMI, and CAP all undergo nucleophilic substitution reactions with NBD-Cl, reacting rapidly and completely under suitable derivatization conditions. The established MMI analytical method has a linear range of 1.00-500 ng·mL-1, with good linearity (r=0.998 9). The linear ranges of the CAP and 5-ASA analytical methods are both 2.00-1 000 ng·mL-1, with good linear relationships (CAP: r=0.999 3, 5-ASA: r=0.999 6). The intra- and inter-batch precision (CV) and accuracy (RE) were both less than 15%, making the methods suitable for the analysis of actual samples of the corresponding drug formulations and successfully applied to the pharmacokinetic studies of MMI.
Conclusion The established NBD-Cl derivatization LC-MS/MS analytical methods for the three drugs exhibit advantages such as high sensitivity, strong specificity, and low background noise, and successfully address the issues of low mass spectrometry response, poor spectral retention, and the tendency of CAP to oxidize. The methods require minimal plasma sample volume and have short analysis times, providing a feasible solution for the analysis of biological samples of the three drugs. They also demonstrate the applicability of NBD-Cl derivatization technology in the analysis of amine- and thiol-containing drugs.
, authors=Jing SHI
1, Ji-Fen GUO
2, Ji-Jun TANG
2, Yan-Bin HE
1, authorsList=Jing SHI, Ji-Fen GUO, Ji-Jun TANG, Yan-Bin HE, authorCompany=null, correspAuthors=Yan-Bin HE, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1292126460408525534, articleId=1292126458256847580, tenantId=1146029695717560320, journalId=1246415772164075586, language=CN, title=基于NBD-Cl衍生化试剂的LC-MS/MS法在测定胺基/巯基类药物血药浓度中的应用, columnId=1246531413600784769, journalTitle=中国临床药理学杂志, columnName=研究方法, runingTitle=null, highlight=null, articleAbstract=
目的 本研究以4-氯-7-硝基苯并-2-氧杂-1，3-二唑（NBD-Cl）为衍生化试剂，建立LC-MS/MS分析方法，用于定量检测胺基类药物美沙拉秦（mesalazine，5-ASA）以及巯基类药物甲巯咪唑（methimazole，MMI）与卡托普利（captopril，CAP）。
方法 血浆样品采用衍生化法和蛋白沉淀法。对不同药物进行衍生化条件考察（衍生化试剂浓度、反应温度、衍生化时间），以Thermo AccucoreTM C18（2.4 μm，30 mm×4.6 mm）、Agient Poroshell 120 EC-C18（4 μm，50 mm×4.6 mm）为分析柱，0.1%甲酸水-乙腈、0.1%甲酸水-甲醇和5 mM乙酸铵-甲醇∶乙腈（V∶V=50∶50）为梯度流动相，流速分别为0.5 mL·min-1和0.6 mL·min-1，采用ESI源在多反应监测方式（MRM）下进行正/负离子检测。
结果 5-ASA、MMI和CAP均与NBD-Cl发生亲核取代反应，在适宜衍生化条件下反应迅速且彻底。所建立的MMI分析方法线性范围为1.00~500 ng·mL-1，线性关系良好（r=0.998 9）。CAP和5-ASA分析方法线性范围均为2.00~1 000 ng·mL-1，且线性关系良好（CAP：r=0.999 3，5-ASA：r=0.999 6），批内、批间精密度（CV）和准确度（RE）均<15%，能够满足相关药物剂型的实际样品检测。
结论 所建立的3种药物的NBD-Cl衍生化LC-MS/MS分析方法具有高灵敏度、专属强、背景噪音小等优势，并成功解决了MMI和5-ASA质谱响应低、色谱保留差以及CAP易被氧化等方面的缺陷，所需血浆样品量少，分析时间短，为3种药物的生物样品分析提供了可行的方案，同时也表明NBD-Cl衍生化技术在胺基类和巯基类药物分析中具有一定的适用性。
, authors=石景
1, 郭继芬
2, 唐吉军
2, 贺艳斌
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石景（1999-），男，硕士研究生，主要从事药物分析研究
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