HPLC-MS/MS法同时测定人血浆中美罗培南和左氧氟沙星的浓度

Determination of meropenem and levofloxacin in human plasma by HPLC-MS/MS

目的:建立同时测定人血浆中美罗培南和左氧氟沙星浓度的HPLC-MS/MS方法。方法:以环丙沙星为内标物,血浆用0.1%甲酸甲醇沉淀蛋白。色谱条件:色谱柱为Agilent HC-C_(18)柱(2)(4.6 mm×150mm,5μm),以0.1%甲酸甲醇-0.1%甲酸水溶液(50∶50)为流动相,流速0.5 mL·min^(-1),柱温30℃;质谱条件:电喷雾离子化源,正离子电离模式下多反应离子监测模式(MRM)测定,用于定量分析的离子反应分别为m/z 384.1→m/z 254.0(美罗培南)、m/z 362.0→m/z 318.0(左氧氟沙星)和m/z 3 332.0→288.0(内标,环丙沙星),扫描时间为0.1 s。结果:本试验1个分析样本的分析时间为4 min;美罗培南和左氧氟沙星的血药浓度分别在0.1～100.0和0.01～5.00μg·mL^(-1)范围内线性良好,日内精密度分别为1.8%～4.3%和2.1%～6.5%,日间精密度分别为1.3%～4.4%和2.2%～5.5%,准确度分别为98.7%～100.3%和94.8%～100.9%。美罗培南和左氧氟沙星血浆样品经处理后室温放置3 h稳定,-80℃冻融3次稳定。患者血浆样品中美罗培南浓度在0.167 8～47.678 8μg·mL^(-1)之间,左氧氟沙星浓度在0.662 0～1.575 0μg·mL^(-1)之间。结论:本法前处理简单,分析快速、灵敏、准确,适用于美罗培南和左氧氟沙星的药物浓度监测。

Objective:To establish a simple,rapid and sensitive HPLC-MS/MS quantitative detection method for the simultaneous determination of meropenem and levofloxacin in human plasma.Methods:Sample preparation was achieved by protein precipitation with 0.1% formic acid in methanol containing the ciprofloxacin as the internal standard.Chromatographic separation was achieved on a Agilent HC-C18(2)(4.6 mm×150 mm,5 μm)column with methanol and water(50 ∶50)containing 0.1% formic acid as the mobile phase at a flow rate of 0.5 mL·min^-1,and the column temperature was 30 ℃.The detection was carried out by electrospray ionization mass spectrometry in multiple reaction monitoring(MRM)mode.Meropenem and levofloxacin were detected using positive ion mode.The MRM transitions of m/z 384.1 →m/z 254.0 was for meropenem,m/z 362.0 →m/z 318.0 was for levofloxacin and m/z 332.0 →m/z 288.0 was for ciprofloxacin.Results:The overall turnaround time for the assay was 4 minutes.The linear concentration ranges of the calibration curves for meropenem and levofloxacin were 0.1-100.0μg·mL^-1 and 0.01-5.00 μg·mL^-1,respectively.Intra-assay precision for quality control samples of meropenem and levofloxacin were within the range of 1.8%-4.3% and 2.1%-6.5%,respectively.Inter-assay precision ranged within 1.3%-4.4% and 2.2-5.5%,respectively.The accuracy of meropenem was within the range of 98.7%-100.3%,levofloxacin’s were within the range of 94.8%-100.9%.The samples of meropenem and levofloxacin plasma were stable for 3 h after treated at room temperature,and the samples were also stable for frozen and thawed 3 times at -80℃.The concentration of meropenem in the patient plasma sample was between 0.167 8 and 47.678 8μg·mL^-1,and the concentration of levofloxacin was between 0.662 0 and 1.575 0 μg·mL^-1.Conclusion:This method is successfully applied to the pharmacokinetic study of meropenem and levofloxacin.