HPLC法测定人血清中万古霉素及去甲万古霉素浓度及临床应用

目的建立同时测定血清中万古霉素及去甲万古霉素浓度的高效液相色谱法.方法：采用Diamonsil C18 色谱柱（250mm×4.6 mm,5 μm）;流动相：乙腈-磷酸二氢钾缓冲液（pH = 3.2）（7.5∶92.5）;流速：1 mL·min-1;检测波长：236 nm;柱温：30 ℃;进样量：25 μL;二极管阵列检测.内标法定量,万古霉素与去甲万古霉素互为内标.结果：去甲万古霉素低、中、高3 个浓度（3.125、25.000、50.000 μg·mL-1）的方法回收率分别为98.5%、102.2%、103.1%,万古霉素低、中、高3 个浓度（3.125、25.000、50.000 μg·mL-1）的方法回收率分别为96.2%、100.8%、103.5%,日内精密度RSD 小于3.24%,日间精密度RSD 小于6.12%.结论：本方法准确度高,灵敏度好,操作简便,符合万古霉素及去甲万古霉素血药浓度监测和药代动力学研究的要求.

Concentration determination of vancomycin and norvancomycin by HPLC as well as its clinicalapplication

Objective： To establish the HPLC method for fast, effective, simultaneous determination of vancomycinand norvancomycin in serum. Methods： The extract was separated by Diamonsil C18 column （250 mm × 4.6 mm, 5 μm）. Themobile phase consisted of acetonitrile-potassium dihydrogen phosphate buffer solution （pH = 3.2） （7.5∶92.5） with the flow rateof 1 mL·min-1. The detection wavelength was set at 236 nm. The column temperature was 30 ℃ and the sample size was 25 μL.Vancomycin and norvancomycin were used as the internal standard for each other. Results： For norvancomycin, the averagemethod recovery rates of three concentrations （3.125, 25.000, 50.000 μg·mL-1） were 98.5%, 102.2%, and 103.1% respectively. Forvancomycin, the average method recovery rates of three concentrations （3.125, 25.000, 50.000 μg·mL-1） were 96.2%, 100.8% and103.5% respectively. The within-day RSD was less than 3.24%, and between-day RSD was less than 6.12%. Conclusion： Themethod was accurate, sensitive, simple, which was suitable for the therapeutic drug monitoring （TDM） and pharmacokinetic study ofvancomycin as well as norvancomycin.