二维高效液相色谱法检测达托霉素血药浓度的可行性考察

A feasibility study to evaluate the use of 2D-HPLC for measuring the blood concentration of daptomycin

目的:建立二维高效液相色谱结合中间柱拦截技术测定达托霉素血药浓度的方法。方法:样品通过一维色谱柱Aston SRC(4.6 mm×50 mm,5μm)初步分离后,被中间柱ASTON-PLI(20 mm×4.6 mm,3μm)截取到二维色谱柱Aston SCB(4.6 mm×250 mm,5μm)进行二次分离;一维流动相为LC1移动相,流速:0.6 mL·min^(-1)。二维流动相为LC2移动相,流速:1.2 mL·min^(-1),检测波长260 nm,温度40°C,进样量200μL。结果:达托霉素在1.00~40.06μg·mL^(-1)范围内线性关系良好,r=0.9999,最低检测限为0.03μg·mL^(-1),绝对回收率为99.2%~101.9%,日间、日内RSD均小于5.94%。结论:该方法新颖,自动化程度高,样本检测简单快速、准确稳定,适用于临床达托霉素血药浓度的测定。

OBJECTIVE To establish method for the determination of the plasma concentration of daptomycin by using twodi⁃mensional high-performance liquid chromatography coupled with intermediate column interception.METHODS The analytes were preliminarily separated in the first-dimensional chromatographic column Aston SRC(4.6 mm×50 mm,5μm),intercepted by the intermediate column Aston PLI(20 mm×4.6 mm,3μm),and transferred to the second dimensional chromatographic col⁃umn Aston SCB(4.6 mm×150 mm,5μm)for further separation.The first-dimensional mobile phase was LC1 mobile phase at a flow rate of 0.6 mL·min^(-1).The second-dimensionmobile phase was LC2 mobile phase at a flow rate of 1.2 mL·min^(-1).The plasma samples were detected at a wavelength of 260 nm and a temperature of 40℃,with an injection volume of 200μL.RESULTS The linearity of daptomycin was good in the range of 1.00~40.06μg·mL^(-1)(r=0.9999).The lowest detection limit was 0.03μg·mL^(-1),the absolute recovery was 99.2%~101.9%,and the intra-day and inter-day RSD were less than 5.94%.CONCLUSION This novel method is automaticand simple,with high efficiency as well as accurate and stable perfor⁃mance,which is suitable for determining the clinical plasma concentration of daptomycin.