HPLC法测定患者血浆中达托霉素浓度

Determination of the concentration of daptomycin in human plasma by high performance liquid chromatography

目的:建立HPLC法测定人血浆中达托霉素浓度,并检测临床使用达托霉素患者的血药浓度。方法:以卡马西平为内标,乙腈沉淀蛋白法处理样品。色谱柱为Diamonsil C_(18)柱(150 mm×4.6 mm,5μm);流动相为乙腈-磷酸二氢铵溶液(38∶62,v/v,pH=3.5);流速1.0 mL·min^(-1);柱温30℃;检测波长214 nm;进样量10μL。结果:人血浆中达托霉素浓度在2~80μg·mL^(-1)内线性关系良好(r=0.9998),定量下限为1μg·mL^(-1)。低、中、高浓度达托霉素的提取回收率均>90%,方法回收率>80%,日内、日间精密度RSD均<10%。结论:本研究建立的方法具有操作简单、专属性强、重现性好等优点,适用于临床患者达托霉素血药浓度的检测。

Objective:To develop a method for determining the concentration of daptomycin in human plasma by high performance liquid chromatography(HPLC)in order to detect the plasma concentration of daptomycin in patients with clinical use.Methods:Carbamazepine was selected as the internal standard and the plasma samples were precipitated by acetonitrile.Chromatographic column Diamonsil C_(18)(150 mm×4.6 mm,5μm)was used.The mobile phase was acetonitrile-ammonium dihydrogen phosphate(38∶62,v/v,pH=3.5).The flow rate was 1.0 mL·min^(-1).The column temperature was 30℃.The detection wavelength was 214 nm and the injection volume was 10μL.Results:The linear range of daptomycin in human plasma was 2-80μg·mL^(-1)(r=0.9998),and the lower limit of quantitation was 1μg·mL^(-1).The extraction recovery rates of low,medium and high concentration of daptomycin were more than 90%,and the method recovery rates were more than 80%.The intra-day and interday relative standard deviations were both below 10%.Conclusion:The method was simple,specific and reproducible,which was suitable for therapeutic drug monitoring of daptomycin in clinic.