论文部分内容阅读
目的:建立测定人用皮卡狂犬病疫苗和皮卡佐剂中硫酸卡那霉素含量的HPLC方法。方法:色谱柱为CAPCELL PAKC8 DD(4.6 mm×250 mm,5μm);流动相为磷酸二氢钾缓冲液(0.02 mol.L-1,pH 7.5)-甲醇-乙腈(23∶7∶40);流速为1.0mL.min-1;柱温为45℃;采用柱前衍生化法,衍生化试剂为2,4,6-三硝基苯磺酸(TNBS);检测波长为350 nm。结果:在0.01～1.0 mg.mL-1范围内,硫酸卡那霉素的浓度与峰面积呈现良好的线性关系(A=3×106C-3313.9,r=1.000,n=7);在80%,100%,120%3个浓度下的平均回收率(n=3)分别为82.5%(RSD=1.6%),90.3%(RSD=2.0%),81.6%(RSD=1.8%);方法的最低检测限为0.41μg.mL-1;最低定量限为1.39μg.mL-1;供试品溶液在8 h内稳定(RSD=1.1%)。结论:本方法准确、快速,通用性良好。
OBJECTIVE: To establish an HPLC method for the determination of kanamycin sulfate in human pickup rabies vaccines and dermatan tablets. Methods: The column was CAPCELL PAKC8 DD (4.6 mm × 250 mm, 5 μm). The mobile phase was potassium dihydrogen phosphate buffer (0.02 mol·L-1, pH 7.5) -methanol-acetonitrile The flow rate was 1.0mL.min-1. The column temperature was 45 ℃. The derivatization reagent was 2,4,6-trinitrobenzene sulfonic acid (TNBS) using the precolumn derivatization method. The detection wavelength was 350 nm. Results: In the range of 0.01 ~ 1.0 mg.mL-1, the concentration of kanamycin sulfate showed a good linear relationship with the peak area (A = 3 × 106C-3313.9, r = 1.000, n = 7) (RSD = 1.6%), 90.3% (RSD = 2.0%) and 81.6% (RSD = 1.8%) of the average recoveries (n = 3) The lowest limit of detection was 0.41μg.mL-1; the lowest limit of quantification was 1.39μg.mL-1; the test solution was stable within 8h (RSD = 1.1%). Conclusion: This method is accurate, rapid and versatile.