精蛋白锌胰岛素注射液中硫酸鱼精蛋白含量测定的方法研究

目的: 建立高效液相色谱(HPLC)法测定精蛋白锌胰岛素注射液中硫酸鱼精蛋白的含量。 方法: 采用Grace Vydac C₁₈色谱柱(250 mm×4.6 mm,5 μm),以A:乙腈-0.2 mol·L^-1硫酸钠溶液(4∶96);B:乙腈-水(50∶50)为流动相进行梯度洗脱(0~8 min时B0%→70%,8~15 min时B70%,15~16 min时B70%→0%,16~30 min时B0%);流速1.0 mL·min^-1;柱温40 ℃;检测波长:214 nm。 结果: 硫酸鱼精蛋白线性范围为0.05~2.5 mg·mL^-1(r=0.9994),方法平均回收率(n=9)为99.7%。 结论: 本方法准确、简便、快速,重复性好,可用于精蛋白锌胰岛素注射液中硫酸鱼精蛋白的含量测定。

Objective: To approach an assay method of determination for protamine sulfate in protamine zine insulin injection by HPLC method. Methods: The assay was performed on Grace Vydac C₁₈ column(250 mm×4.6 mm,5 μm)with a gradient program (0-8 min B 0%→70%,8-15 min B 70%,15-16 min B 70%→0%,16-30 min B 0%)of A:acetonitrile-0.2 mol·L^-1sodium sulfate(4∶96) and B:acetonitrile-water(50∶50)as the mobile phase;The column temperature was set up at 40 ℃ and the flow rate was 1.0 mL·min^-1.The absorbance was monitored at 214 nm. Results: The line ranges of protamine sulfate was 0.05-2.5 mg·mL^-1(r=0.9994).The average recoveries (n=9) of protamine sulfate was 99.7%. Conclusions: The method is proved to be rapid,accurate and convenient,and provided a reliable way for protamine sulfate the quality of.protamine zine insulin injection.

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