高效液相色谱法测定人血清中万古霉素和去甲万古霉素浓度

目的建立人血清中万古霉素和去甲万古霉素浓度的高效液相色谱法（HPLC）测定方法。方法色谱柱为Hypersil BDS C18（250mm×4.6mm,5μm）柱;流动相为磷酸二氢钾缓冲液-甲醇（87∶13）,流速1ml/min;紫外检测波长236nm;以甲硝唑为内标,采用乙腈和硫酸锌沉淀蛋白后直接进样。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=0.0873X-0.0061,r=0.9999;去甲万古霉素回归方程为Y=0.1000X-0.0004,r=0.9997;线性范围1.0～50.0mg/L。日内相对标准编差（RSD）小于4.8%,日间RSD小于6.6%;万古霉素低、中、高浓度的平均方法回收率分别为102.41%,100.44%,104.69%,平均提取回收率分别为87.54%,89.86%,88.13%;去甲万古霉素低、中、高浓度的平均方法回收率分别为99.46%,98.88%,99.47%,平均提取回收率分别为78.13%,77.41%,76.45%。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。

Concentration Determination of Vancomycin and Norvancomycin in Human Serum by HPLC

Objective To develop a method tor the determination of vancomycin and norvancomycm concentration in human serum by high-performance liquid chromatography （HPLC）. Methods The chromatographic bar was Hypersil BDS C/8 （4. 6 minx 250 mm, 5 μm）. The mobile phase was tribasie potassium phosphate buffer-methanol （87：13） and the flow rate was 1 ml.min^-1. The detecting wavelength of ultraviolet was 236 nm. Arilin was used as the internal standard. The sample was deposited by acetonitrile and zinc sulfate then was injected. Results Standard curve was liner in the range of 1.0-50. 0 mg/L. The linear equation of vancomyein was Y= 0. 0873320. 0061, r= 0. 9999; the linear equation of norvaneomycin was Y = 0. 1000X 0. 0004, r = 0. 9997. The within-day relative standard deviation （RSD） was less than 4. 8%, and between-day RSD less than 6.6%. As for vaneomycin, the average method recovery rate of three concentrations were 102. 41%, 100. 44% and 104.69% respectively, and the average extraction recovery rate were 87. 54%, 89. 86% and 88.13% respectively. As for norvancomyein, the average method recovery rate of three concentrations were 99.46%, 98.88% and 99. 47% respectively, and the average extraction recovery rate were 78. 13%, 77. 41% and 76. 45% respectively. Conclusion The operation is simple. The method has stable rate of extraction recovery. It is applicable in concentration determination of vancomycin and norvancomycin in human serum in clinic and other fields.