超高效液相色谱法测定替考拉宁血药浓度

Determination of teicoplanin in human plasma by UPLC

目的:建立超高效液相色谱测定人血浆中替考拉宁的方法,为临床监测替考拉宁血药浓度调整给药方案提供方法学依据。方法:色谱柱为Zorbax SB-C18柱(2.1 mm×100 mm,1.8μm),流动相为乙腈-0.1%三氟乙酸水溶液(24.2:75.8,v/v),流速0.4 m L·min-1,柱温30℃,检测波长220 nm。取患者稳态谷浓度血样,经乙腈沉淀蛋白,三氯甲烷萃取,Agilent1290超高效液相色谱仪进行测定。结果:替考拉宁五个成分10 min内全部出峰,保留时间分别为:3.2 min、4.4 min、5.2 min、8.9 min、9.9 min,峰形对称与血浆中其它组分分离良好,线性范围2.0~80.0μg·m L-1(r=0.999 8,n=7),平均提取回收率88.53%,平均方法回收率97.3%,日内和日间精密度的相对标准偏差(RSD)均小于6%。测定14例应用替考拉宁的患者,其中4例低于有效血药浓度范围。结论:该检测方法简单、快速,具有较高的灵敏性和准确性,可满足临床替考拉宁血药浓度的监测和药动学研究的要求。同时,该法有效避免了内标法合并用药时内标干扰及内标不稳定,磷酸盐作流动相对色谱柱损害大、清洗耗时等问题。

Objective:To establish a method for the determination of teicoplanin in human plasma by ultra performance liquid chromatography,in order to provide a methodological basis for clinical monitoring of teicoplanin plasma concentration for adjusting the dosage regimen.Methods:The column was a Zorbax SB-C18 column(2.1 mm×100 mm,1.8μm)with a mobile phase of acetonitrile-0.1%trifluoroacetic acid(24.2:75.8,v/v)at a flow rate of 0.4 mL·min-1,the column temperature was 30℃,and detection wavelength was 220 nm.Blood samples of steady state trough concentration were taken from patients using teicoplanin.The plasma was precipitated with acetonitrile,extracted with chloroform,and analyzed with an Agilent 1290 ultra performance liquid chromatography.Results:The five components of teicoplanin can all be detected within 10 minutes,with retention times of 3.2 min,4.4 min,5.2 min,8.9 min,and 9.9 min,respectively.The peaks were symmetrical and well separated from other components in the plasma.The calibration curve was linear through the range of 2.0– 80.0μg·mL-1(r=0.999 8,n=7).The average extraction recovery rate and average method recovery rate were 88.53%and 97.3%,respectively.The relative standard deviations(RSDs)of intraand inter-day precision were all less than 6%.We measured 14 patients who used teicoplanin,while 4 of them were lower than the effective range of plasma concentration.Conclusion:Because of its high sensitivity and accuracy,our UPLC method can meet the requirements of clinical monitoring and pharmacokinetic study of teicoplanin.At the same time,the method is easy and fast,which effectively avoids interference and instability of internal standard,and the damage of column and time-consuming cleaning when using phosphate buffer as the mobile phase.