液-质联用法测定人体血浆中万古霉素的浓度

Determination of vancomycin in human plasma by LC-MS/MS

目的:运用液-质联用技术(LC-MS/MS),建立测定人体血浆中万古霉素浓度的方法,监测万古霉素血药浓度。方法:以替洛福韦为内标,乙腈沉淀蛋白后,经LC-MS/MS方法分析。采用色谱柱为安捷伦Poroshell 120 EC-C18柱(4.6 mm×100mm,2.7μm),流动相为混合有机相(乙腈∶甲醇-2∶1)-0.1%甲酸水溶液(10∶90);流速0.4 mL·min-1;采用质谱电喷雾电离源(ESI),正离子模式,多重反应监测方式(MRM)检测;待测物质万古霉素用于定量的母子离子对:m/z 725.6→m/z 144.2,内标替洛福韦用于定量的母子离子对:m/z288.0→m/z176.2。结果:万古霉素在0.2~100μg·mL^(-1)范围内线性关系良好,最低定量限为0.2μg·mL^(-1);日内RSD均在6%以下,日间RSD均在12%以下。提取回收率为58.01%~66.58%。结论:该分析方法快速、准确、灵敏度高、专属性强、重复性好,可用于医疗机构检测人体血浆中万古霉素的药物浓度。

OBJECTIVE To develop an LC-MS/MS method for determination of vancomycin concentration in plasma.METHODS Plasma samples were precipitated by acetonitrile.Tenofovirdisoproxil was used as internal standard.Agilent Poroshell 120 EC-C18column（4.6 mm×100 mm,2.7μm）was adopted,mixed organic phase（acetonitrile∶methanol-2∶1）-0.1%formic acid water solution（10∶90）was mobile phase,at a flow rate of 0.4 mL·min^-1.Analytes were determined by using positive ion electrospray ionization（ESI）in multiple reaction monitoring（MRM）mode.Ion pair for quantitative analysis was m/z725.6→m/z 144.2 for vancomycin and m/z 288.0→m/z 176.2 for tenofovirdisoproxil.RESULTS Vancomycin showed good linearity in range of 0.2–100μg·mL^-1,with a lower limit of quantitation at 0.2μg·mL^-1,intra-day relative standard deviation lower than 6%,inter-day relative standard deviation lower than 12%,and average recovery in range of 58.01%-66.58%.CONCLUSION The method is rapid,sensitive,accurate,specific and reliable,and suitable for determination of vancomycin in human plasma.