HPLC法快速测定万古霉素和去甲万古霉素的血药浓度

目的建立快速测定人血清中万古霉素和去甲万古霉素浓度的HPLC法。方法以Hypersil BDS-C₁₈（250mm×4.6mm,5μm）色谱柱为分析柱,流动相:磷酸二氢钾缓冲液（pH 3.2）-甲醇（86:14,V/V）,流速1 mL·min^-1,检测波长236nm,柱温35℃,进样量20μL,万古霉素和去甲万古霉素互为内标;血清样品经10%三氯醋酸沉淀蛋白离心后进行色谱分析。结果本法万古霉素和去甲万古霉素均在2～100 mg·L^-1间线性关系良好,两者平均回收率均为95%～105%,日内及日间RSD均<5%。结论本法操作简便、快速、准确,适用于临床血药浓度测定及人体药动学研究。

Rapid determination of the concentration of vancomycin and norvancomycin in serum by HPLC

AIM To develop a method for rapid determination of vancomycin and norvancomycin concentrations in human serum by HPLC.METHODS The Hypersil BDS C18 column（250 mm×4.6 mm,5μm）was used.The mobile phase was tribasic potassium phosphate buffer-methanol（86：14,V/V）and the flow rate was 1 mL·min^-1.The detection wavelength of UV was set at 236 nm,the column temperature was set at 35℃and the sample loading volume was of 20μL.Vancomycin and norvancomycin were used as internal standard each other.A 10%trichloroacetic acid was used for the sedimentation.Serum sample was used for chromatography after protein precipitation and centrifuging.RESULTS The peak area of vancomycin and norvancomycin against the concentrations was linear within the range of 2 - 100 mg·L^-1.The average recovery was 95%- 105%,and the RSD within-day and between-day was less than 5%.CONCLUSION This method is rapid,simple and accurate.It is usable for clinical pharmacokinetic studies and monitoring of vancomycin and norvancomycin in human serum.