Article(id=1291707299458998502, tenantId=1146029695717560320, journalId=1246415772164075586, issueId=1291707045514858818, articleNumber=null, orderNo=null, doi=10.13699/j.cnki.1001-6821.2025.23.010, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1751472000000, receivedDateStr=2025-07-03, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1785896618952, onlineDateStr=2026-08-05, pubDate=1765900800000, pubDateStr=2025-12-17, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1785896618952, onlineIssueDateStr=2026-08-05, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1785896618952, creator=13701087609, updateTime=1785896618952, updator=13701087609, issue=Issue{id=1291707045514858818, tenantId=1146029695717560320, journalId=1246415772164075586, year='2025', volume='41', issue='23', pageStart='3301', pageEnd='3450', issueExtLink='null', onlineDate='null', pubDate='1765900800000', pubDateStr='2025-12-17', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1785896558407, creator='13701087609', updateTime=1785896663780, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1291707487678390577, tenantId=1146029695717560320, journalId=1246415772164075586, issueId=1291707045514858818, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1291707487682584882, tenantId=1146029695717560320, journalId=1246415772164075586, issueId=1291707045514858818, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=3366, endPage=3373, ext={EN=ArticleExt(id=1291707300193001703, articleId=1291707299458998502, tenantId=1146029695717560320, journalId=1246415772164075586, language=EN, title=Uncertainty evaluation for the determination of 10,11-dihydro-10-hydroxycarbazepine in human plasma by UHPLC-MS/MS, columnId=1246531413395263850, journalTitle=Chinese Journal of Clinical Pharmacology, columnName=Research Method, runingTitle=null, highlight=null, articleAbstract=
Objective To evaluate the uncertainty for the determination of 10,11-dihydro-10-hydroxycarbazepine (MHD) in human plasma by ultra performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS).
Methods An isotope-labeled internal standard was employed. Chromatographic separation was achieved within 3 min on a BEH C18 column (2.1 mm×50.0 mm, 1.7 μm) using a gradient elution at a flow rate of 0.4 mL·min-1, with a mobile phase consisting of methanol containing 0.1% formic acid and water containing 5% methanol plus 0.1% formic acid. Plasma sample concentrations were determined by UHPLC-MS/MS with electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode. The method was validated for selectivity, calibration curve, lower limit of quantification, precision, recovery, matrix effect, and stability. The influencing factors in the quantification of MHD were analyzed and evaluated to calculate the individual uncertainties, combined uncertainty, and finally the expanded uncertainty.
Results MHD demonstrated good linearity within the range of 0.03 to 6.00 mg·L-1. The accuracy (bias from the theoretical concentration) for within-run and between-run analyses was within ±15%, and the precision (coefficient of variation) was within 13.32%. Plasma samples showed good stability under the following conditions: 9 hours at 25 ℃, 5 hours at 4 ℃, three freeze-thaw cycles, and storage at -80 ℃ for 18 days. Based on the calculated uncertainty values for each contributing factor, repeatability, matrix effect, and recovery represented the major sources of uncertainty for the low-concentration quality control (QC) sample, with values of 1.96×10-2, 5.49×10-2, and 7.25×10-2, respectively. For the high-concentration QC sample, matrix effect and recovery contributed substantially to the overall uncertainty, with values of 4.92×10-2 and 2.01×10-2, respectively. The expanded uncertainties (P=95%, k=2) for MHD at low (0.09 mg·L-1) and high (4.50 mg·L-1) concentrations in human plasma were 1.62×102 mg·L-1 and 0.48 mg·L-1, respectively.
Conclusion The measurement uncertainty of MHD concentration in human plasma determined by UHPLC-MS/MS is primarily contributed by repeatability, matrix effect and recovery at low concentration levels, while dominated by matrix effect and recovery at high concentrations
, authors=Wei LI
1, Xue-qiang PAN
1, Zhi-gang ZHAO
2, Sheng-hui MEI
2, authorsList=Wei LI, Xue-qiang PAN, Zhi-gang ZHAO, Sheng-hui MEI, authorCompany=null, correspAuthors=Sheng-hui MEI, 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=1291707300507574504, articleId=1291707299458998502, tenantId=1146029695717560320, journalId=1246415772164075586, language=CN, title=UHPLC-MS/MS法测定人血浆中10,11-二氢-10-羟基卡马西平浓度的不确定度评定, columnId=1246531413600784769, journalTitle=中国临床药理学杂志, columnName=研究方法, runingTitle=null, highlight=null, articleAbstract=
目的 评定超高效液相色谱-串联质谱法（UHPLC-MS/MS）测定人血浆中10,11-二氢-10-羟基卡马西平（MHD）浓度的不确定度。
方法 采用同位素内标，流动相为含0.1%甲酸的甲醇和含5%甲醇与0.1%甲酸的水溶液，BEH C18色谱柱（2.1 mm×50.0 mm，1.7 μm），在0.4 mL·min-1流速下进行梯度洗脱，3分钟内实现分离。利用电喷雾电离（ESI），在多反应监测（MRM）模式下，使用UHPLC-MS/MS分析测定血浆样品浓度。考察该方法的专属性、标准曲线和定量下限、精密度、回收率、基质效应、稳定性。对MHD浓度测定过程中的各影响因素进行分析评定，计算各因素的不确定度和合成不确定度，最终计算扩展不确定度。
结果 MHD在0.03至6.00 mg·L-1范围内具有良好线性，批内、批间样品检测的准确度与理论浓度的偏差在±15％以内，精密度变异系数均在13.32%以内。血浆样品25 ℃下放置9 h，4 ℃下放置5 h，3次冻融循环，-80 ℃下储存18 d等条件下均表现出良好的稳定性。根据各因素的不确定度计算结果，低浓度质控的重复性、基质效应和回收率的不确定度较大，分别为1.96×10-2，5.49×10-2和7.25×10-2；高浓度质控的基质效应和回收率不确定度较大，分别为4.92×10-2和2.01×10-2。人血浆中MHD低浓度（0.09 mg·L-1）和高浓度（4.50 mg·L-1）的扩展不确定度分别为1.62 ×10-2 mg·L-1和0.48 mg·L-1（P = 95%，k = 2）。
结论 UHPLC-MS/MS法测定人血浆中MHD浓度的不确定度在低浓度时主要由重复性、基质效应和回收率引入，高浓度时主要由基质效应与回收率引入。
, authors=李伟
1, 潘学强
1, 赵志刚
2, 梅升辉
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李伟（1986-）女，副教授，主要从事药物分析的相关研究
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