HPLC法测定万古霉素和去甲万古霉素血清药物浓度

目的建立人血清中万古霉素和去甲万古霉素的HPLC测定方法 ,用于万古霉素和去甲万古霉素血药浓度临床监测。方法色谱柱为Shim-pack（150×4.6 mm,5μm）,流动相为甲醇∶磷酸二氢钾缓冲液（pH=3.2）=19∶81,检测波长为236 nm,流速1 ml/min,柱温为35℃,万古霉素和去甲万古霉素互为内标。结果万古霉素和去甲万古霉素的保留时间分别为4.835 min和4.020 min;标准曲线回归方程分别为Y=32.16624X-2.117683,r=0.9997（n=3）,Y=51.53402X-2.958621,r=0.9992（n=3）;线性范围均为2.5~100μg/ml;方法回收率〉95%,绝对回收率〉80%,日内和日间RSD〈6.2%。结论本测定方法快速、准确、简便,适用于万古霉素和去甲万古霉素的临床常规血药浓度的监测。

Determination of Vancomycin and Norvancomycin in Human Serum by HPLC

Objective To establish a method for the clinical determination of vancomycin and norvancomycin in human serum by HPLC.Methods The separation was achieved on the Shim-pack column 150×4.6 mm,5 μm.The mobile phase consisted of methanol-tribasic potassium phosphate buffer（pH=3.2）=19∶81,the detection wavelength was 236 nm,the flow rate was lmL/min,and column temperature was 35℃.Vancomycin and norvancomycin were mutually used as the internal standard.Results The retention time of vancomycin and norvancomycin were 4.835 min and 4.020 min respectively.The standard curve regression equations of Vancomycin and norvancomycin were Y=32.16624X-2.117683 r=0.9997（n=3）,Y=51.53402X-2.958621,r=0.9992（n=3） respectively.The calibration curves of vancomycin and norvancomycin were linear over the range of 2.5100 μg/ml.The relative recovery was more than 95%,the objective recovery was more than 80%,and the relative standard deviation（RSD） of intra-day and inter-day were all less than 6.2%.Conclusion Thanks to its rapidity,accuracy and simplicity,it is suitable to apply this method to measure the concentration of vancomycin and norvancomycin in human serum