HPLC-MS/MS法同时测定重症患者血浆中多黏菌素B1和B2浓度及其临床应用

Development and validation of a liquid chromatography-tandem mass spectrometry analytical method for the therapeutic drug monitoring of polymyxin B1 and B2

目的:建立高效液相-串联质谱(HPLC-MS/MS)法快速检测血浆中多黏菌素B1(polymyxin B1,PMB1)和B2(polymyxin B2,PMB2)浓度的方法。方法:血浆样品使用含内标多黏菌素E1(polymyxin E1,PME)的甲醇-乙腈-甲酸(80∶20∶0.5,V/V)溶液进行蛋白沉淀,上清液在Phenomenex Kinetex XB-C18色谱柱(3.0 mm×5.0 mm,2.6μm)上进行梯度洗脱分离,流动相:0.1%甲酸水-乙腈溶液,流速0.3 mL·min-1,电喷雾电离源,多离子反应监测,正离子模式扫描。结果:PMB1和PMB2的多反应监测离子通道为402.1→101.1和397.5→391.5,线性范围良好(分别为0.998、0.999),定量下限分别为35 ng·mL^(-1)和15 ng·mL^(-1)。日内、日间精密度都小于15%,稳定性良好,不受基质效应影响。24例重症患者多黏菌素B用药后平均血中浓度C_(ss,avg)为(4.44±1.37)μg·mL^(-1),24 h药物浓度时间下曲线面积AUC_(0-24 h)为(87.74±27.97)μg·mL^(-1)·h。结论:该方法简单、快速、灵敏度好,可用于静脉注射多黏菌素B的重症患者的治疗药物监测。

OBJECTIVE A sensitive and rapid high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)method for simtaneous determination of polymyxin B1(PMB1)and polymyxin B2(PMB2)has been developed and validated.METHODS The plasma sample was deproteinized with methanol-acetonitrile-formic acid(80∶20∶0.5,V/V)which contained the internal standard(Polymyxin E1).Chromatographic separation was performed on a Phenomenex Kinetex XB-C18(3.0 mm×50 mm,2.6μm)column with mobile phase of acetonitrile-0.1%formic acid solution using gradient elution at a flow rate of 0.3 mL·min-1;electrospray source ion(ESI),multiple reaction monitoring(MRM)and positive ion scanning were used for the determination.RESULTS[M+3H]^(3+)was chosen as the precursor ion for both PMB1 and PMB2 and the transitions of the deprotonated molecules were 402.1→101.1 for PMB1 and 397.5→391.5 for PMB2.The lower limits of quantification for PMB1 and PMB2 were 35 ng·mL^(-1)and 15 ng·mL^(-1),respectively.The intra-day and inter-day precision of PMB1 and PMB2 were all less than 15%.The method has good stability and was not affected by matrix effect.From all 24 patients samples,the C_(ss,avg)and AUC_(0-24h)of PMB were(4.44±1.37)μg·mL^(-1),and(87.74±27.97)μg·mL^(-1)·h,respectively.CONCLUSION This method is simple,rapid and sensitive,and has been validated to simultaneous determination of plasma PMB1 and PMB2 concentrations in patients receiving intravenous polymyxin B therapy.