高效液相色谱法测定人血清万古霉素浓度

Concentration Determination of Vancomycin in Human Serum by HPLC

目的建立测定人血清中万古霉素浓度的高效液相色谱法。方法色谱柱为VP-ODS C18柱(150 mm×4.6 mm,4.6μm),流动相为0.05 mol/L磷酸二氢钾缓冲溶液(p H=3.2)-甲醇(83∶17),流速为1 m L/min,紫外检测波长为205 nm,柱温为25℃,进样量为20μL,血清样品经10%三氯乙酸-乙腈(1 200μL∶300μL)蛋白沉淀剂沉淀蛋白离心后进行色谱分析。结果万古霉素的最低检测限为1.96μg/m L,万古霉素血清质量浓度在4.90～58.80μg/m L范围内与峰面积线性关系良好,线性回归方程为Y=26 385 X-19 207,r=0.999 6(n=7),日内、日间精密度均小于4%(n=5)。结论该方法提取率高、重复性好、操作简便、灵敏,适用于万古霉素的血药浓度测定。

Objective To establish an HPLC method for concentration determination of vancomycin in human serum.Methods The column was VP-ODSC18 column(150 mm×4.6 mm,4.6μm),the mobile phase was 0.05 mol/L potassium dihydrogen phosphate(pH=3.2)-methyl alcohol(87∶13),the flow rate was 1 mL/min,the UV detection wavelength was 205 nm,the column temperature was 25℃,and the sample size was 20μL.The protein precipitation of 10%trichloroacetic acid-acetonitrile(1 200μL∶20μL)was added into serum samples to precipitate protein,and the protein was analyzed by chromatography after centrifugation.Results The minimum detectable concentration of vancomycin was 1.96μg/mL,the linear range of vancomycin concentration was 4.90-58.80μg/mL,the regression equation was Y=26 385X-19 207,r=0.999 6(n=5),the RSD of intra-day and inter-day assays were less than 4%(n=5).Conclusion The method is highly efficient,reproducible,simple and sensitive,which is suitable for the concentration of vancomycin in human serum.