LC-MS/MS法同时测定人血浆中亚胺培南和西司他丁浓度  

Simultaneous determination of imipenem and cilastatin in human plasma by LC-MS/MS

作　　者：袁智龙 张文静 刘梦丹 王卓 马涛 YUAN Zhi-long;ZHANG Wen-jing;LIU Meng-dan;WANG Zhuo;MA Tao(School of Pharmacy,Bengbu Medical University,Bengbu 233000,China;Department of Pharmacy,The First Affiliated Hospital of Naval Medical University,Shanghai 200433,China;College of life sciences and Biopharmaceuticals,Shenyang Pharmaceutical University,Shenyang 110000,China)

机构地区：[1]蚌埠医科大学药学院,蚌埠233000 [2]海军军医大学第一附属医院药剂科,上海200433 [3]沈阳药科大学生命科学与生物制药学院,沈阳110000

出　　处：《药物分析杂志》2025年第2期228-236,共9页Chinese Journal of Pharmaceutical Analysis

基　　金：海军军医大学第一附属医院“十四五”学科固海计划项目(GH145-38)。

摘　　要：目的:建立LC-MS/MS法测定人血浆中亚胺培南和西司他丁的浓度,用于临床治疗药物浓度监测,并考察样品预处理时添加稳定剂对质谱信号强度的影响。方法:样品经蛋白沉淀后,采用Agilent TC-C_(18)(2)(150 mm×4.6mm,5μm)色谱柱,以0.15%甲酸水-甲醇为流动相,进行梯度洗脱。质谱采用电喷雾离子源(ESI),正离子模式下多反应监测(MRM)扫描,亚胺培南、西司他丁和内标(美罗培南)的定量离子对分别为m/z300.1→141.9、m/z359.7→97.0和m/z 384.1→141.1。将含有和不含稳定剂3-(N-吗啉基)丙磺酸(MOPS)的样品分别进行预处理后,连续进样分析,比较质谱信号强度的变化。结果:亚胺培南和西司他丁质量浓度均在0.1~100.0μg·mL^(-1)范围内线性关系较好(r≥0.99),其日内和日间准确度均在95.3%~108.5%,精密度(RSD)均<9.3%,提取回收率在77.4%~84.3%,基质效应在97.1%~111.2%。血浆样品中亚胺培南在室温下放置3 h、4℃下放置6 h和-80℃下放置12 d内均稳定,在-20℃下放置12 d降解显著。西司他丁在多种条件下均稳定。方法在柱温±5℃、流速±0.1 mL·min^(-1),水相中甲酸浓度土0.025%、离子源温度土50℃的条件改变范围内具有良好的耐用性。添加稳定剂的样品在连续进样后质谱信号强度出现明显降低,而不添加稳定剂方法对质谱信号强度无明显影响。结论:本方法操作简单,结果准确,可用于亚胺培南和西司他丁的临床治疗药物监测。MOPS等非挥发性盐类稳定剂会降低质谱灵敏度,不添加这类稳定剂更适合LC-MS/MS法长期使用。Objective:To establish an LC-MS/MS method for determining the concentrations of imipenem and cilastatin in human plasma,for monitoring clinical therapeutic drug concentrations,and to investigate the effects of adding stabilizers during the sample pretreatment on mass spectrometry signal intensity.Methods:After protein precipitation,the sample was subjected to gradient elution using an Agilent TC-C_(18)(2)(150 mm×4.6 mm,5μm)column with a mobile phase system of 0.15%formic acid in water and methanol.The electrospray ionization(ESI)mass spectrometer was operated in positive ion mode using multiple reaction monitoring(MRM):m/z 300.1→141.9(imipenem),m/z 359.7→97.0(cilastatin)and m/z 384.1→141.1(meropenem,internal standard).The samples containing and without 3-(N-morpholino)propane sulfonic acid(MOPS)as stabilizers were pretreated and continuously analyzed to compare the changes in mass spectrometry signal intensity.Results:Both imipenem and cilastatin showed good linearities in the concentration ranges of 0.1-100.0μg mL^(-1)(r>0.99),The intra-day and inter-day accuracy ranges from 95.3%to 108.5%,the precision(RSDs)were less than 9.3%,the extraction recovery rate ranges from 77.4%to 84.3%,and the matrix effect ranges from 97.1%to 111.2%.Imipenem in plasma samples was stable at room temperature for 3 h,at 4℃for 6 h,and at-80℃for 12 d,while it was significantly degraded at-20℃for 12 d.Cilastatin was stable under a variety of conditions.The method was robust to changing conditions of column temperature±5℃,flow rate±0.1 mL·min^(-1),formic acid concentration in the aqueous phase±0.025%,and ion source temperature±50℃.The samples containing stabilizers exhibited significant ion inhibition on mass spectrometry after continuous injection,while samples without stabilizers had no significant effect on the signal intensity of mass spectrometry.Conclusion:The method is simple and accurate and can be used for clinical drug monitoring of imipenem and cilastatin.Nonvolatile salt stabilizers such as MOPS can red

关 键 词：亚胺培南 西司他丁 液相色谱串联质谱 治疗药物监测 检测灵敏度 

分 类 号：R917[医药卫生—药物分析学]