液相色谱串联质谱法测定人血浆中多黏菌素B及在治疗药物监测中的应用  

LC-MS/MS method for therapeutic drug monitoring of polymyxin B in human plasma

作　　者：王雨 黄晓岚 张楚晗 曲星伊 郭晨雪 陈焰 范亚新 郭蓓宁 张菁 刘笑芬 WANG Yu;HUANG Xiaolan;ZHANG Chuhan;QU Xingyi;GUO Chenxue;CHEN Yan;FAN Yaxin;GUO Beining;ZHANG Jing;LIU Xiaofen(Institute of Antibiotics,Key Laboratory of Clinical Pharmacology of Antibiotics,Ministry of Health,National Clinical Research Center for Aging and Medicine,Huashan Hospital,Fudan University,Shanghai 200040,China)

机构地区：[1]复旦大学附属华山医院抗生素研究所,国家卫生健康委员会抗生素临床药理重点实验室 [2]国家老年疾病临床医学研究中心,上海200040

出　　处：《中国感染与化疗杂志》2022年第6期694-701,共8页Chinese Journal of Infection and Chemotherapy

基　　金：上海申康医院发展中心市级医院新兴前沿技术联合攻关项目(SHDC12020106);上海市领军人才(LJ2016-01)。

摘　　要：目的建立简单快速的人血浆中多黏菌素B浓度测定的液相色谱串联质谱(LC-MS/MS)法用于治疗药物监测。方法采用AB SCIEX HPLC-API4500QTRAP进行测定,色谱柱为PhonomenexKinetex XB-C18柱(100 mm×2.1 mm,2.6μm),以0.2%甲酸-乙腈∶0.2%甲酸溶液为流动相,梯度洗脱,流速为0.4 mL/min,分析时间为3.5 min。多黏菌素B1、B2和内标多黏菌素E2的离子对分别为m/z 602.6→m/z 241.1、m/z 595.5→m/z 227.1和m/z 578.5→m/z 101.2。血样采用蛋白沉淀处理后进样。结果多黏菌素B1在0.05~10 mg/L浓度范围内线性良好(R~2>0.98),批内、批间精密度均≤14.0%,准确度在100.9%~110.8%范围内,健康者及患者的血浆、溶血、脂血及患者血清样品内标归一化基质效应因子分别为92.6%~103.0%、102.4%~112.6%、99.9%~104.7%及97.1%~106.3%,回收率为79.1%~80.6%;多黏菌素B2在0.00550~1.10 mg/L浓度范围内线性关系良好,批内、批间精密度均≤14.8%,准确度在97.6%~100.7%范围内,健康者及患者的血浆、溶血、脂血及患者血清样品内标归一化基质效应因子分别为73.6%~94.6%、85.6%~120.6%、89.7%~103.4%及82.6%~93.4%,回收率为83.2%~87.8%。结论该研究建立了一种简单、快速、灵敏度高和准确性好的LC-MS/MS法,用于人血浆中多黏菌素B中主要成分B1和B2浓度的测定,可为该药治疗药物监测提供简单、可靠的检测方法。Objective To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for therapeutic drug monitoring of polymyxin B in human plasma.Methods AB SCIEX HPLC-API4500QTRAP system was used in the method.Chromatographic separation was performed on PhonomenexKinetexXB-C18 column(100 mm×2.1 mm,2.6μm)using stepwise gradient elution of acetonitrile and water both with 0.2%formic acid.The analysis time was 3.5 min at a flow rate of 0.4 mL/min.The monitoring ion pairs of polymyxin B1,B2 and E2 were m/z 602.6→m/z 241.1,m/z 595.5→m/z 227.1 and m/z 578.5→m/z101.2,respectively.Polymyxin B was extracted from plasma by protein precipitation with acetonitrile.Results Polymyxin B1 showed a good linearity within the range of 0.05-10 mg/L(R~2>0.98).The intra-batch and inter-batch precisions were no more than 14.0%and the accuracy ranged from100.9%to 110.8%.The normalized matrix effect factor of polymyxin B1 was 92.6%-103.0%in normal and patient plasma,102.4%-112.6%in hemolytic samples,99.9%-104.7%in lipemic samples,and 97.1%-106.3%in patient serum.The recovery of polymyxin B1 was 79.1%-80.6%.Polymyxin B2 also showed a good linearity within the range of 0.00550-1.10 mg/L.The precisions of intra-and inter-batch were no more than 14.8%.The accuracy was 97.6%-100.7%.The normalized matrix effect factor of polymyxin B2 was 73.6%-94.6%in normal and patient plasma,85.6%-120.6%in hemolytic samples,89.7%-103.4%in lipemic samples,and 82.6%-93.4%in patient serum.The recovery of polymyxin B2 was 83.2%-87.8%.Conclusions A simple,rapid,sensitive and accurate LC-MS/MS method was established for determination of polymyxin B1 and B2 in human plasma,and successfully applied for therapeutic drug monitoring of polymyxin B in patients.

关 键 词：液相色谱串联质谱法 多黏菌素B 蛋白沉淀 治疗药物监测 

分 类 号：R978[医药卫生—药品]