HPLC法测患者血清万古霉素和去甲万古霉素浓度

Determination of Vancomycin and Norvancomycin in human serum by HPLC

目的建立高效液相色谱法同时检测人血清中万古霉素和去甲万古霉素浓度。方法色谱柱为Hypersil ODS2（250mm×4.6 mm,5μm）,流动相为甲醇-乙腈-磷酸二氢钾缓冲液（9.5∶2.5∶88）;流速为1.0 m L/min;检测波长为280 nm,柱温为35℃,万古霉素和去甲万古霉素互为内标物质。结果万古霉素和去甲万古霉素的血药浓度在1.25~100μg/m L内线性关系良好,R2=0.9996;万古霉素低、中、高浓度的平均方法回收率分别为96.88%、99.50%、105.01%;去甲万古霉素低、中、高浓度的平均方法回收率分别为94.40%、103.90%、103.81%;日内、日间精密度RSD均〈5%。结论该方法灵敏度高、简单、采血量少且成本低,适于万古霉素及去甲万古霉素血药浓度监测和临床药代动力学研究。

Objective To establish HPLC method for fast,effective,simultaneous determination of Vancomycin and Norvancomycin in serum.Methods The separation was achieved on the Hypersil ODS2（ 250 mm × 4. 6 mm,5 μm） with methanol-acetonitrile-a mixture of potassium phosphate（ 9. 5∶2. 5∶88） as the mobile phase. The flow rate was 1. 0 m L / min. The UV detecting wavelength was 280 nm. The temperature of the column was 35 ℃.Vancomycin and Norvancomycin were used as the internal standard for each other. Results Vancomycin and Norvancomycin blood concentration were in well linearity in the range of 1. 25-100 μg / m L,R2= 0. 9996. The average method recovery rate of three concentrations were 96. 88%,99. 50%,105. 01%respectively,the average method recovery rate of three concentrations were 94. 40%,103. 90,103. 81% respectively,the intra-day and inter-day RSD were less than 5%. Conclusion The HPLC method is proved to be simple,sensitive,accurate and precise in which little blood sample is taken,thus it is suitable for the therapeutic drug monitoring（ TDM） and pharmacokinetic study of Vancomycin and Norvancomycin.