精蛋白重组人胰岛素注射液无菌检查方法的建立

目的： 建立精蛋白重组人胰岛素注射液的无菌检查方法。方法： 参考《中华人民共和国药典》及历版美国药典精蛋白重组人胰岛素注射液无菌检查法，定性和定量比较7种试验菌在1%抗坏血酸溶液和500 IU·mL^-1肝素钠溶液中的存活状况，以肝素钠溶液为稀释剂进行精蛋白重组人胰岛素注射液无菌检查方法学适用性试验。结果： 1%抗坏血酸溶液处理20 min后，金黄色葡萄球菌、铜绿假单胞菌、枯草芽孢杆菌和生孢梭菌的回收比率分别为0.32、0.04、0和0；肝素钠溶液处理组的回收比率分别为1.08、1.01、0.88和0.85。经上述2种溶液处理后，大肠埃希菌、白色念珠菌、黑曲霉、枯草芽孢杆菌芽孢和生孢梭菌芽孢的回收比率在0.86~1.23之间。方法学适用性试验中各试验菌均生长良好。结论： 精蛋白重组人胰岛素注射液为混悬液，采用肝素钠溶液溶解结晶体，建立了合理的无菌检查方法，并对其他含结晶体胰岛素注射液产品的无菌检查具有较好的参考作用。

Objective: To establish a sterility test method for isophane protamine recombinant human insulin injection. Methods: According to sterility test methods of isophane protamine recombinant human insulin injection in the Chinese Pharmacopoeia and United States Pharmacopoeia, the living status of 7 test strains treated by 1% ascorbic acid and 500 IU·mL^-1 heparin sodium were studied in qualitative and quantitative tests. The suitability of sterility test method was taken by 500 IU·mL^-1 heparin sodium as a diluent. Results: After treatment with 1% ascorbic acid for 20 min, the recoveries of Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis and Clostridiun sporogenes were 0.32,0.04,0 and 0, respectively.The recoveries were 1.08,1.01,0.88 and 0.85, respectively while being treated by heparin sodium. The recoveries of Escherichia coli, Candida albicans,Aspergillus niger, Bacillus subtilis spores and Clostridiun sporogenes spores varied from 0.86 to 1.23 after processed by these two solutions. Six test strains required for sterility test according to Chinese Pharmacopoeia exhibited good growth. Conclution: The crystal in isophane protamine recombinant human insulin injection dissolves in Heparin sodium and then sterility test can be taken by membrane-filter procedure. This is a valuable reference for other isophane insulin suspension sterility tests.

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