液质联用法同时测定人血浆中5种抗菌药物及其在重症患者中的应用

Simultaneous determination of five antimicrobial agents in human plasma by LC-MS/MS and its clinical application in critically ill patients

目的:建立并验证一种人血浆中5种常见抗菌药物的检测方法,并将其用于重症患者治疗药物监测。方法:血浆样品经有机溶剂沉淀蛋白后,采用液相色谱串联质谱联用仪测定。色谱柱为Kinetex五氟苯基柱,以含0.1%甲酸水溶液-0.1%甲酸甲醇溶液为流动相梯度洗脱,流速为0.5 m L·min^(-1),柱温为40℃,分析时间为3.5 min,电喷雾离子源,正/负离子多反应监测模式扫描。结果:伏立康唑、利奈唑胺、美罗培南、头孢哌酮、舒巴坦分别在0.2~40μg·m L^(-1)(r=0.9974)、0.5~100μg·m L^(-1)(r=0.9994)、0.5~100μg·m L^(-1)(r=0.9994)、1.5~300μg·m L^(-1)(r=0.9936)、1.5~300(r=0.9979)μg·m L^(-1)内线性关系良好;定量下限的准确度97.44%~100.72%、精密度≤9.35%;批内和批间精密度≤13.88%;方法回收率91.35%~106.08%;基质效应95.48%~107.42%;稀释效应相对偏差<15.00%;在多种储存条件下,处理前后的样品稳定性良好。统计了苏北人民医院2022年11月至2023年3月共138例血药浓度检测结果,伏立康唑、利奈唑胺、美罗培南、头孢哌酮、舒巴坦达标率分别为53.84%、27.27%、59.10%、20.00%、25.00%。纳入了20例重症患者基本信息及血药浓度,发现在相同给药方案下,个体间差异较大。结论:该法操作简便,准确度、灵敏度、重复性好,可同时检测重症患者5种抗菌药物的血药浓度。经临床实践证明,该法可为重症患者个体化用药提供重要依据。

OBJECTIVE To establish and validate a method of simultaneously detecting 5 common antibiotics in human plasma and implement therapeutic drug monitoring in 138 patients in intensive care unit(ICU).METHODS The plasma samples were precipitated with organic solvent and then examined by liquid chromatography-tandem mass spectrometry.Flow rate was 0.5 mL·min^(-1),column temperature 40℃and analysis time 3.5 min.Electrospray ion source was scanned in positive/negative ion multiple reaction monitoring mode.RESULTS The linear relationship of voriconazole,linezolid,meropenem,cefo⁃perazone and sulbactam were decent at 0.2—40μg·mL^(-1)(r=0.9974),0.5—100μg·mL^(-1)(r=0.9994),0.5—100μg·mL^(-1)(r=0.9994),1.5—300μg·mL^(-1)(r=0.9936)and 1.5—300(r=0.9979)μg·mL^(-1).The accuracy of lower quantification limit was 97.44%—100.72%with a precision of≤9.35%and a within/between-run precision≤13.88%.The recovery rates were 91.35%—106.08%and matrix effects 95.48%—107.42%.The relative deviation of dilution effect was<15.00%.The stabil⁃ity of samples before and after treatment was decent under a variety of storage conditions.From November 2022 to March 2023,as for blood drug concentration test,the compliance rates of voriconazole,linezolid,meropenem,cefoperazone and sulbactam were 53.84%,27.27%,59.10%,20.00%and 25.00%.Basic profiles and blood drug concentrations of 20 critically ill patients were recorded.Under the same dosing regimen,great differences existed between individuals.CONCLUSION This simple and fast method offers decent profiles of accuracy,sensitivity and repeatability.It may be used for detecting the blood levels of 5 kinds of antibiotics in critically ill patients.It provides rationales for individualized dosing for patients with severe infection.