LC-MS/MS测定人血浆中多黏菌素B浓度及其应用

Concentration Determination of Polymyxin B in Plasma by LC-MS/MS and Its Application

关键词:  液相色谱串联质谱法  多黏菌素B  治疗药物监测

Key words:  LC-MS/MS  polymyxin B  therapeutic drug monitoring

摘要:

目的 建立测定人血浆中多黏菌素B总浓度的液相色谱串联质谱(LC-MS/MS)方法，并应用于临床多黏菌素B的治疗药物监测。方法 采用Welch Ultimate LP-C₁₈(100mm×2.1mm，5μm)色谱柱对多黏菌素B的主要成分进行色谱分离，建立时间程序为4.0min的LC-MS/MS法，以乙腈为沉淀剂进行蛋白沉淀的样品前处理，用正离子扫描以及多反应监测模式进行分析物测定。结果 多黏菌素B的总浓度线性范围为100~10000ng·mL^-1，各主要成分线性系数均>0.996，批内和批间精密度RSD均≤6%，准确度为91.88%~114.02%，测定3例使用多黏菌素B患者的体内暴露量，AUC_0-24h分别为39.73，64.15，107.75mg·L^-1·h^-1。结论 本研究建立了一种稳健的测定人血浆中多黏菌素B总浓度的LC-MS/MS方法，灵敏度和特异性好，并成功用于临床上多黏菌素B的治疗药物监测。

Abstract:

OBJECTIVE To establish a liquid chromatography tandem mass spectrometry(LC-MS/MS) method for the determination of polymyxin B concentration in human plasma, and to use it for clinical drug monitoring of polymyxin B. METHODS The Welch Ultimate LP-C₁₈(100 mm×2.1 mm, 5 μm) column was used to separate the main components of polymyxin B, and an LC-MS/MS with a time program of 4.0 min was established. Acetonitrile was used as precipitator for sample pretreatment of protein precipitation, positive ion scanning and multi-reaction detection mode were used for analyte determination. RESULTS The linear range of total polymyxin B concentration was 100-10 000 ng·mL^-1. The linear coefficients of the main components were both> 0.996. The intra-assay and inter-assay precisions were both ≤ 6%, and the accuracy was 91.88%-114.02%. The in vivo exposure of three patients using polymyxin B, AUC_{0-24 h} was 39.73, 64.15 and 107.75 mg·L^-1·h^-1, respectively. CONCLUSION This study has established a robust LC-MS/MS method for the determination of the total concentration of polymyxin B in human plasma, with good sensitivity and specificity, and has been successfully used for clinical monitoring the concentration of polymyxin B.