优化UPLC-MS/MS法测定人血浆中万古霉素的浓度

Optimization of determination of vancomycin in human plasma by UPLC-MS/MS

目的建立超高效液相色谱-质谱联用法测定人血浆中万古霉素浓度,用于临床万古霉素血药浓度监测。方法以对乙酰氨基酚为内标,血浆样品经甲醇沉淀后,经UPLC-MS/MS分离-分析。采用Agilent Eclipse XDB-C18(2.1×150mm,5μm),以甲醇(含0.1%甲酸)∶水(含0.1%甲酸)=50∶50为流动相;流速:0.2m L·min-1,采用电喷雾离子源(ESI),以多离子反应监测方式(MRM)进行正离子监测,万古霉素和内标对乙酰氨基酚的定量分析离子对分别为m/z 725.0→144.0和m/z 151.8→109.8。结果万古霉素的线性范围为0.2~100.0mg·L-1,定量下限为0.2mg·L-1。批内和批间精密度均小于12%。结论该方法操作简便、灵敏、准确,适用于万古霉素的治疗药物监测。

OBJECTIVE To establish an UPLC-MS/MS method to determine vancomycin in human plasma,and apply to the clinical vancomycin monitoring. METHODS Plasma samples were precipitated with acetonitrile and determined by UPLC-MSMS using acetaminophen as the internal standard. The separation was carried out on a Agilent Eclipse XDB-C18（2. 1 mm × 150 mm,5μm） with a mobile phase of methanol（including 0. 1% formic acid）-water（ including 0. 1% formic acid）（ 50∶ 50） and the flow rate was 0. 2 m L·min^-1. ESI was performed in the MRM positive mode with using target ions at m/z725. 0→144. 0（ vancomycin） and m/z 151. 8→109. 8（ acetaminophen）.RESULTS The calibration curve of vancomycin was obtained in the range of 0. 2 ~ 100. 0 mg·L^-1 with good linearity（ r〉 0. 99）. The LLOQ of vancomycin was 0. 2μg · L^-1. The within and between run RSDs were less than12%. CONCLUSION This method is simple,of good sensitivity and precision,which is suitable for therapeutic drug monitoring of vancomycin.