用HPLC-MS/MS法同时测定人血浆中左氧氟沙星和头孢他啶的浓度

Simultaneous determination of Levofloxacin and ceftazidime in human plasma by HPLC-MS/MS

目的建立超高液相色谱串联质谱同时测定人血浆中左氧氟沙星和头孢他啶浓度的方法。方法用甲醇沉淀蛋白后,以环丙沙星为内标,色谱柱:Agilent Zorbax Eclipse Plus C18(2.1 mm×100.0 mm,1.8μm),流动相∶水(含0.1%甲酸)-乙腈(含0.1%甲酸)=90∶10,等度洗脱,流速:0.2 mL·min^(-1),柱温:40℃,进样量:1μL。电喷雾离子源,多离子反应监测,正离子扫描模式进行检测。考察该方法的专属性、标准曲线和定量下限、精密度与回收率、基质效应及稳定性。结果血浆样品中左氧氟沙星在0.05~25.72μg·mL^(-1)线性关系良好,标准曲线为y=4.53x+0.02(r=0.9996),定量下限为0.05μg·mL^(-1);头孢他啶在0.21~107.25μg·mL^(-1)线性关系良好,标准曲线为y=0.40x+0.02(r=0.9997),定量下限为0.21μg·mL^(-1);2种药物的平均提取回收率均在88.66%~108.21%,日内和日间的精密度相对标准偏差均小于10%,基质均不影响测定,样品的稳定性均良好。结论本方法简便快速、准确度高、灵敏度好,适用于人血浆中左氧氟沙星和头孢他啶浓度的同时测定。

Objective To establish a high performance liquid chromatography tandem mass spectrometry(HPLC-MS/MS)method for the determination of levofloxacin and ceftazidime in human plasma.Methods Plasma were processed with methanol to precipitate protein and the chromatographic separation1 was performed on a Agilent ZorbaxEclipse Plus C18(2.1 mm×100.0 mm,1.8μm)with ciprofloxacin as internal standard.Mobile phase was water(containing 0.1%formic acid)-acetonitrile(containing 0.1%formic acid)90:10 by isocratic elution at a flow rate of 0.2 mL·min^(-1).C.olumn temperature was 40℃.Injection volume was 1μL.MS detection was carried out using an electrospray ionization source operated in multiple reaction monitoring(MRM)mode.The specificity,standard curve,lower limit of quantification,precision,recovery,stability and matrix effect of the method were investigated.Results The calibration curves of levofloxacin and ceftazidime were y=4.53x+0.02(r=0.9996)and y=0.40x+0.02(r=0.9997),with the linear ranges of 0.05-25.72μg·mL^(-1)and 0.21-107.25μg·mL^(-1),and the lower limits of quantification were 0.05μg·mL^(-1)and 0.21μg·mL^(-1),respectively.The average extraction recoveries of the two drugs were 88.66%-108.21%,and the standard deviations of intra-day and inter-day precisions were less than 10%.The experiment has good stability and is not affected by matrix effect.Conclusion This method is accurate,rapid and sensitive,suitable for the determination of levofloxacin and ceftazidime in human plasma.