国内外精蛋白重组人胰岛素注射液的有关物质比较

Comparison of related substances of protamine recombinant human insulin injection at home and abroad

目的精蛋白重组人胰岛素适用于2型糖尿病患者的治疗,现行标准中仅对本品有关物质A_(21)脱氨人胰岛素进行控制,未进行B_(3)和B_(3)iso脱氨人胰岛素限度的控制。采用两种方法对精蛋白重组人胰岛素注射液中的有关物质进行测定并比较国内外共17批样品的质量。方法采用高效液相色谱法梯度洗脱,色谱柱:Waters Xbridge C_(18)4.6 mm×250 mm(5μm),检测波长:214 nm,柱温:40℃,流速:1.0 ml/min,进样体积:20μl;《中国药典》2020年版方法:流动相A为0.2 mol/L硫酸钠溶液(pH值2.3)-乙腈(82∶18),流动相B为乙腈-水(50∶50);某企业自拟方法:流动相A为0.2 mol/L硫酸钠溶液(pH值3.6)-乙腈(90∶10),流动相B为乙腈-水(50∶50)。结果国内外样品的A_(21)脱氨人胰岛素含量差异不大、国内一家企业B_(3)和B_(3)iso脱氨人胰岛素含量低于其他企业,国内另一家企业有关物质总和高于其他企业;两种方法检出A_(21)脱氨人胰岛素的含量差异无统计学意义(P>0.05),企业自拟方法能检测出更多的有关物质。结论两种方法均快速、简便,可用于比较国内外样品中的有关物质,同时为企业在制定杂质控制限度时提供依据。

Objective Protamine recombinant human insulin injection is suitable for the treatment of type 2 diabetes patients,and the current standard only controls the content of A_(21) desamido human insulin which is related to the product,but does not control the content of B_(3) and B_(3)iso desamido human insulin.To determine the related substances in 17 batches of protamine recombinant human insulin injection by two methods,and compare the quality at home and abroad.Methods An HPLC method was used with a Waters Xbridge C_(18) column 250 mm×4.6 mm(5μm).The detection wavelength was 214 nm,the column temperature was 30℃,the flow rate was 1.0 ml/min and the injection volume was 20μl.The mobile phase A of Chinese Pharmacopoeia 2020 edition was 0.2 mol/L sodium sulfate solution(pH 2.3)-acetonitrile(82∶18),mobile phase B was acetonitrile-water(50∶50)with gradient elution.The mobile phase A of enterprise self-designed method was 0.2 mol/L sodium sulfate solution(pH 3.6)-acetonitrile(90∶10),mobile phase B was acetonitrile-water(50:50)with gradient elution.Results There was no significant difference in the content of A_(21) desamido human insulin at home and abroad,the content of B_(3) and B_(3)iso desamido human insulin in one domestic enterprise was lower than that of other enterprises,and the total amount of related substances in another domestic enterprise was higher than that of other enterprises.There was no significant difference in the content of A_(21) desamido human insulin detected by the two methods(P>0.05),and more related substances can be detected by the enterprise self-designed method.Conclusion Both methods are fast and simple,and can be used to compare the related substances in domestic and foreign samples,and provide a basis for enterprises to establish impurity control limits.