目的 建立UPLC-MS/MS定量分析人脑脊液中的依非韦伦(efavirenz,EFV)和杜鲁特韦(dolutegravir, DTG)浓度的方法,为指导临床用药提供帮助。方法 脑脊液样品经纯乙腈蛋白质沉淀后,以乙腈/0.1%甲酸水溶液为流动相,Venusil ASB C18(2.1 mm×50 mm,5 μm)柱分离;采用电喷雾电离的方法,以多反应监测(MRM)进行正离子检测,用于定量分析的离子对为316.1→244.0(EFV),420.3→277.1(DTG)。结果 测定脑脊液中EFV和DTG的线性范围分别为2~200和1~100 ng·mL-1。日内、日间精密度都低于15%,准确度87%~115%。提取回收率、基质效应和稳定性85%~115%。该方法成功用于34例获得性免疫缺陷综合征患者脑脊液分析,10例患者脑脊液中检测到了EFV,2.30~150.0 ng·mL-1,所有的受试者均未检测到DTG。结论 本实验建立了人脑脊液中EFV和DTG的检测方法,该方法快速简便,灵敏度高,重现性好,能够同时准确测定人脑脊液中EFV、DTG的浓度,并且节约检测时间和样本用量,可以应用于临床EFV和DTG药物浓度的监测。
Abstract
OBJECTIVE To establish an UPLC-MS/MS method for simultaneously quantifying efavirenz and dolutegravir in human cerebrospinal fluid (CSF), so as to provide help for guiding clinical medication. METHODS CSF samples were treated with acetonitrile for protein precipitated. Compounds were separated by Venusil ASB C18 (2.1 mm×50 mm, 5 μm) column with acetonitrile/0.1% formic acid aqueous solution as mobile phase. The positive ions were detected by multi-reaction monitoring (MRM) with electrospray ionization. The ion pairs used for quantitative analysis were 316.1→244.0 for efavirenz (EFV) and 420.3→277.1 for dolutegravir (DTG). RESULTS The linear ranges of EFV and DTG were 2-200 and 1-100 ng·mL-1 in cerebrospinal fluid. The inter-or intra-day precision RSD was less than 15%, and the accuracy was between 87% and 115%. The extraction recovery, matrix effect and stability data were all from 85% to 115%. The method was successfully performed to detect the concentrations of 34 clinical samples. EFV was detected in cerebrospinal fluid of 10 patients with concentration of 2.30 to 150.0 ng·mL-1,while DTG was not detected in all of the subjects. CONCLUSION In this study, a method for the detection of EFV and DTG in human cerebrospinal fluid was established, and the method is rapid, simple, sensitive and accurate, and can be used to determine the concentrations of EFV and DTG in human CSF simultaneously, which can save testing time and sample size. The method can be used to monitor the concentrations of EFV and DTG in clinic.
关键词
依非韦伦 /
杜鲁特韦 /
高效液相色谱-串联质谱法 /
获得性免疫缺陷综合征 /
脑脊液
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Key words
efavirenz /
dolutegravir /
UPLC-MS/MS /
acquired immunodeficiency syndrome /
cerebrospinal fluid
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中图分类号：
R969
R917
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脚注
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基金
上海市卫生和计划生育委员会项目资助(201640005);十三五重大新药创制—抗艾滋病病毒新药临床评价技术平台建设项目资助(2017ZX09304027);国家自然科学基金项目资助(81271834)
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