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

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-C₁₈柱（2）（4.6 mm×150 mm，5 μm），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0.01~5.00 μg·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 8 μg·mL^-1之间，左氧氟沙星浓度在0.662 0~1.575 0 μg·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 0.1-100.0 μg·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 8 μg·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.

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