【摘要】 目的 建立一种高效、简便的测定人血浆中多黏菌素E的液相色谱串联质谱法(LC-MS/MS)，为多黏菌素E治疗药物监测提供支持。方法 采用AB SCIEX HPLC-4500MD液质联用仪测定，色谱条件采用0.2%甲酸水和0.2%甲酸乙腈梯度洗脱，色谱柱为Phenomenex Kinetex XB-C18柱(100 mm×2.1 mm，2.6 μm)，质谱采用正离子多反应监测检出模式，待测物多黏菌素E1、E2和内标多黏菌素B1的离子对分别为m/z 390.7→101.3、m/z 386.0→101.2、m/z 402.3→101.2。血浆样本采用蛋白沉淀进行预处理。结果 多黏菌素E1、E2分别在0.031 2～6.24 mg/L、0.006 15～1.23 mg/L范围内呈良好的线性，在血浆中的批内准确度、精密度分别为89.4%～99.8%、4.9%～8.9%和91.5%～108.2%、2.8%～8.5%，批间准确度、精密度分别为91.8%～104.7%、4.1%～7.6%和95.6%～105.2%、4.2%～9.8%。在6个不同来源空白血浆中，多黏菌素E1、E2的平均内标归一化基质效应因子分别为96.9%～111.2%、106.1%～112.8%，在脂血血浆中分别为102.5%～106.8%、98.8%～105.2%，在溶血血浆中分别为107.8%～108.9%、106.9%～107.4%，精密度均小于15.0%。多黏菌素E1、E2的平均提取回收率在102.9%～107.5%范围内，内标多黏菌素B1平均回收率为107.0%，精密度<3.7%。结论 该研究成功建立一种简单、高效的测定人血浆中多黏菌素E1和E2的LC-MS/MS方法，可用于多黏菌素E治疗药物浓度监测及药代动力学研究。更多还原

【Abstract】 Objective To develop and validate an efficient and simple liquid chromatography with tandem mass spectrometry (LC-MS/MS) method for determination of polymyxin E in human plasma,and apply the established method in therapeutic drug monitoring (TDM) of polymyxin E.Methods The LC-MS/MS platform was based on AB SCIEX HPLC-4500MD system.Gradient elution was performed with 0.2% formic acid in water and 0.2% formic acid in acetonitrile.Phenomenex Kinetex XB-C18 column (100 mm × 2.1 mm,2.6 μm) were used.The analytes were detected by electrospray ionization (ESI) positive multiple reaction monitoring mode.The ion pairs for analytes (polymyxins E1,E2) and internal standard (polymyxins B1) were m/z390.7→101.3,m/z 386.0→101.2,and m/z 402.3→101.2,respectively.Plasma samples were processed with protein precipitation method.Results Polymyxin E1 and E2 showed good linearity in the range of 0.031 2-6.24 mg/L and0.006 15-1.23 mg/L,respectively.The within-run accuracy of polymyxin E1 and E2 in plasma ranged from 89.4% to 99.8% and 91.5% to 108.2%,respectively,while the between-run accuracy ranged from 91.8% to 104.7% and 95.6% to 105.2%,respectively.The within-run precision of polymyxin E1 and E2 in plasma ranged from 4.9% to 8.9% and 2.8% to 8.5%,respectively,while the between-run precision ranged from 4.1% to 7.6% and 4.2% to 9.8%,respectively.The average internal standard normalized matrix effect factors of polymyxins E1 and E2 were 96.9%-111.2% and 106.1%-112.8% in blank plasma samples from 6 different sources,102.5%-106.8% and 98.8%-105.2% in lipemic plasma,respectively,107.8%-108.9% and 106.9%-107.4% in hemolyzed plasma,respectively.The precision of matrix effects was less than 15.0%.The average recovery rate was 102.9%-107.5% for polymyxin E1 and E2,and 107.0% for internal standard polymyxin B1.The precision was less than 3.7%.Conclusions In this study,a simple and efficient LC-MS/MS method was established for determination of polymyxin E1 and E2 in human plasma,which is reliable in the therapeutic drug monitoring and pharmacokinetic study of polymyxin E.更多还原