目的 建立液相色谱串联质谱法测定人脑脊液中异烟肼(isoniazid, INH)、吡嗪酰胺(pyrazinamide, PZA)、乙胺丁醇(ethambutol, EMB)和利福平(rifampicin, RIF)的浓度,为指导临床用药提供帮助。方法 脑脊液样品经乙腈沉淀蛋白后进样分析,以含0.1%甲酸的水溶液-含0.1%甲酸的乙腈溶液(80∶20) 为流动相,采用ZORBAX Bonus-RP柱(2.1 mm×150 mm, 3.5 μm)分离;采用电喷雾电离源,以多反应监测(multiple reaction monitoring,MRM)方式进行正离子检测。结果 测定脑脊液中异烟肼、吡嗪酰胺、乙胺丁醇和利福平分别在20~5 000(r=0.998 6)、120~30 000(r=0.997 7)、20~5 000(r=0.999 4)、60~15 000(r=0.996 2)ng·mL-1内线性关系良好。日内和日间精密度均小于8.37%,提取回收率均处于98.84%~107.96%之间。同时该方法成功用于6例临床患者脑脊液样本分析。结论 该方法快速简便,灵敏度高,测定结果可靠,已能够服务于临床治疗药物监测。
Abstract
OBJECTIVE To develop a LC-MS method for simultaneously quantifying four anti-tuberculosis drugs [isoniazid(INH)、pyrazinamide(PZA)、 ethambutol(EMB) and rifampicin(RIF)] in human cerebrospinal fluid and to help guide the clinical medication. METHODS The drugs in cerebrospinal fluid were extracted by acetonitrile precipitation, and separated on ZORBAX Bonus-RP column(2.1 mm×150 mm,3.5 μm) with the mobile phase of aqueous solution(containing 0.1% formic acid)-acetonitrile(containing 0.1% formic acid)(80∶20) at a flow rate of 0.25 mL·min-1. Multiple reaction monitoring(MRM) mode was performed combined with electrospray ionization source operating in the positive ionization mode. RESULTS The liner calibration curves of INH、PZA、EMB and RIF were obtained in the concentration range of 20-5 000(r=0.998 6), 120-30 000(r=0.997 7), 20-5 000(r=0.999 4), 60-15 000(r=0.996 2)ng·mL-1, respectively. The intra-and inter-day precision was less than 15% and the absolute recovery was above 75%. This method was successfully applied to analyze the drugs in cerebrospinal fluid from six clinical samples. CONCLUSION The method is rapid, simple, sensitive and accurate, and proved to be suitable for clinical therapeutic drug monitoring.
关键词
异烟肼 /
利福平 /
结核 /
液相色谱串联质谱法 /
脑脊液 /
血药浓度
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Key words
isoniazid /
rifampicin /
tuberculosis /
LC-MS /
cerebrospinal fluid /
plasma concentration
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中图分类号：
R969.1
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参考文献
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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(2017ZX09009070-001-015);上海市高等级生物安全病原微生物检测专业技术服务项目资助(15DZ2290200)
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