摘要
目的:建立人血清中万古霉素和去甲万古霉素浓度的HPLC测定方法。方法:色谱柱为Hypersil BDS C_(18)(250 mm×4.6 mm,5μm)柱;流动相为磷酸二氢钾缓冲液-甲醇(85:15),流速1 ml·min^(-1);检测波长236 nm;以甲硝唑为内标,采用高氯酸直接沉淀蛋白后进样。结果:万古霉素、去甲万古霉素血清浓度与峰面积线性关系良好,万古霉素回归方程为Y=0.0106+ 0.0202X,r=0.999 7;去甲万古霉素回归方程为Y=-0.0164+0.0286X,r=0.999 9;线性范围2.0—100.0 mg·L^(-1)。日内RSD小于7.2%,日间RSD小于6.9%;万古霉素高、中、低浓度的平均方法回收率分别为97.73%,97.58%,103.91%,平均提取回收率分别为86.80%,85.50%,87.63%;去甲万古霉素高、中、低浓度的平均方法回收率分别为100.23%,92.63%,97.57%,平均提取回收率分别为90.84%,87.82%,89.68%。结论:方法操作简便快速,回收率稳定,适于临床血药浓度测定及相关科研的应用。
Objective： To develop a method for the determination of vancomycin and norvancomycin concentration in human serum by HPLC. Method： The Hypersil BDS C18 column （250 mm × 4.6 mm, 5 μm）was used. The mobile phase was tfibasie potassium phosphate buffer-methanol （85：15 ） and flow rate was 1 ml ·min^-1. The detecting wavelength of UV was at 236 nm. Arilin was used as the internal standard. The sample was deposited by perchloric acid and then injectied. Result： Standard curve was liner in the range of 2.0 - 100.0 mg· L^- 1. The linear equation of vancomycin was Y = 0. 0106 ＋ 0. 0202X, r = 0. 999 7 ; the linear equation of norvancomy- cin was Y = - 0. 0164 ＋ 0. 0286X,r = 0. 999 9. The within-day RSD less than 7.2% , and between-day RSD less than 6.9%. As for vancomycin, the average method recovery rate of three concentration were 97.73% ,97.58% and 103.91% respectively, the average extraction recovery rate of three concentration were 86.80% ,85.50% and 87.63% respectively. As for norvancomycin, the average method recovery rate of three concentration were 100.23% ,92.63% and 97.57% respectively, the average extraction recovery rate of three concentration were 90.84% ,87.82% and 89.68% respectively. Conclusion： The operation was simple. The method had stable rate of extraction recovery. It is applicable convention determination of vancomycin and norvancomycin concentration in human serum in clinical practice and other fields.
出处
《中国药师》
CAS
2008年第3期278-280,共3页
China Pharmacist
