液相色谱串联质谱法测定人血浆中替考拉宁的血药浓度

目的:建立测定人血浆中替考拉宁浓度的液相色谱串联质谱(LC-MS/MS)测定方法。方法:选用Welch Ultimate XB-C₁₈(2.1 mm×50 mm,5.0 μm)色谱柱,以达托霉素为内标,以含0.1%甲酸的水溶液和乙腈为流动相进行梯度洗脱,柱温为40 ℃。利用乙腈进行沉淀蛋白样品前处理,采用电喷雾离子化多反应监测正离子模式,测定替考拉宁A_2-1、A_2-2、A_2-3、A_2-4和A_2-5 5种主要成分的总浓度。结果:替考拉宁的总浓度线性范围为1~60 μg·mL^-1,定量下限为1 μg·mL^-1,日内和日间精密度RSD均小于15%,提取回收率80.04%~96.02%,内标归一化基质效应为90.12%~104.04%。研究测定了16例给予替考拉宁后患者的血药谷浓度,浓度范围从定量下限至31.2 μg·mL^-1,达标率56.25%,患者个体差异大且初始达标率低。结论:该研究建立了一种快速、简便、稳定的LC-MS/MS测定人血浆中替考拉宁血药浓度的方法,具有较高准确度、精密度和回收率,为临床给药方案的优化奠定基础。

Quantification of teicoplanin in human plasma by liquid chromatography tandem-mass spectrometry

OBJECTIVE To establish a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the determination of teicoplanin in human plasma.METHODS A Welch Ultimate XB-C₁₈ (2.1 mm×50 mm, 5.0 μm) column was used with gradient elution of acetonitrile-water (containing 0.1% formic acid). The internal standard was daptomycin and the column temperature was 40 ℃. Acetonitrile was used to remove protein for sampling pretreatment. The five components of teicoplanin, A_2-1, A_2-2, A_2-3, A_2-4 and A_2-5, were determined by multiple reaction monitoring in an ESI ion source and positive ion mode.RESULTS The linear range of total teicoplanin was 1-60 μg·mL^-1 and the lower limit of quantification was 1 μg·mL^-1. The intra-day and inter-day precision measurements showed the RSDs were lower than 15%. The recoveries were around 80.04%-96.02%, and the normalized matrix effect was 90.12%-104.04%. The trough concentration of 16 patients after teicoplanin administration was measured. The concentration range was from the lower limit of quantitation to 31.2 μg·mL^-1, and the compliance rate was 56.25%.CONCLUSION A rapid, simple and stable LC-MS/MS method was established for the determination of teicoplanin concentration in human plasma, which has high accuracy, precision and recovery, and lays foundation for the optimization of clinical dosing regimen.