HPLC法测定人血清中万古霉素和去甲万古霉素浓度

Determination of Vancomycin and Norvancomycin in human serum by HPLC

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,便于指导临床血药浓度的监测。方法色谱柱为symmetry C18(250mm×4.6mm,5μm);流动相为磷酸二氢钾缓冲液-甲醇-乙腈(85∶10∶5);流速为1.0mL.min-1;检测波长为236nm,柱温为35℃,采用乙腈-6%高氯酸=1∶6的沉淀剂处理后进样,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6.065min和4.958min;标准曲线回归方程分别为Y=28.34X+0.33,r=0.9998(n=3),Y=91.41X-0.24,r=0.9992(n=3),线性关系良好;方法回收率均>95%,绝对回收率均>95%,日内RSD均<4.0%。结论本测定方法快速、准确、简便,为万古霉素和去甲万古霉素的临床常规血药浓度监测提供依据。

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 Water 2487 column（250mm×4.6mm,5μm） with a mixture of potassium phosphate-methanol-acetonitrile（85∶10∶5） as the mobile phase.The flow rate was 1.0mL·min-1.The UV detecting wave length was 236 nm.The temperature of the column was 35℃.Injection after the treatment of precipitantacetonitrile and 6%perchloric acid（1∶6）.RESULTS The retention time of vancomycin and norvancomycin were 4.958min and 6.005min in respectively.The standard curve regression equations of vancomycin and norvancomycin were Y=28.34X＋0.33,（r=0.9998,n=3）,Y=91.41X-0.24,（r=0.9992,n=3）;The relative recovery and the objective recovery was both more than 95%,and the relative standard deviation（RSD） of intra-day is all lower than 4%.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.