LC-MS/MS法测定重症患者血浆中卡泊芬净的浓度

Determination of caspofungin in plasma of critically ill patients by HPLC-MS/MS

目的:建立LC-MS/MS法测定人血浆中卡泊芬净的浓度。方法:采用安捷伦ZORBAX Eclipse Plus C_(18)色谱柱(2.1 mm×100 mm, 3.5μm),柱温25℃,流动相为含0.1%甲酸的水和含0.1%甲酸的乙腈溶液,梯度洗脱,流速为0.3 mL·min^(-1),分析时长8 min。采用电喷雾离子化,正离子模式多反应监测扫描分析,用于定量分析的离子对分别为:m/z 547.600→137.200(卡泊芬净),m/z 749.800→591.400(内标阿奇霉素)。结果:卡泊芬净在0.06～14.27μg·mL^(-1)范围内线性关系良好,定量下限为0.06μg·mL^(-1),准确度在±15%之间,日内及日间精密度RSD均小于10%,提取回收率为96.6%～102.0%。样本在室温放置4 h、4℃放置24 h、反复冻融3次、冰冻放置5 d和处理后自动进样器(4℃)放置8 h等条件下均稳定,RSD≤7.1%。结论:本方法简便、快捷、灵敏、准确,适合临床上对卡泊芬净血药浓度的监测以及后续的药动学研究。

OBJECTIVE To establish a LC-MS/MS method for the determination of the concentration of caspofungin in human plasma.METHODS Agilent ZORBAX Eclipse Plus C18 column(2.1 mm×100 mm,3.5μm)was used with a column temperature of 25℃and a mobile phase of water containing 0.1%formic acid and acetonitrile containing 0.1%formic acid with gradient elution at a flow rate of 0.3 mL·min-1 and an analysis duration of 8 min.With electrospray ionization,positive ion mode multiple reaction monitoring scan analysis,the ion pairs used for quantitative analysis were:m/z 547.600→137.200(caspofungin),m/z 749.800→591.400(internal standard azithromycin).RESULTS The calibration curve exhibited a good linearity over the range from 0.06 to 14.27μg·mL-1,and the lower limit of quantification was 0.06μg·mL-1 for caspofungin.Accuracy was within±15%,intra-day and inter-day precision RSD were all less than 10%,and the extraction recovery was between 96.6%and 102.0%.Accuracy,precision and stability are all in accordance with the requirements of biological samples.CONCLUSION The method is simple,rapid,sensitive and accurate,and it is suitable for the clinical monitoring of caspofungin concentration in plasma and subsequent pharmacokinetic studies.