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,5μm),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。

Study on determination of protamine sulfate in protamine zine insulin injection

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.