紫草膏无菌检查方法的建立  

Establishment of the sterility test method for Zicao ointment

作　　者：李应超 高婷 刘佳 张小飞 周燕飞 赵茜 巩浩 LI Yingchao;GAO Ting;LIU Jia;ZHANG Xiaofei;ZHOU Yanfei;ZHAO Xi;GONG Hao(Beijing Veterinary Drug and Feed Monitoring Center,Beijing 102299,China)

机构地区：[1]北京市兽药饲料监测中心,北京102299

出　　处：《中兽医医药杂志》2023年第6期59-62,共4页Journal of Traditional Chinese Veterinary Medicine

基　　金：2022年农业行业(国家)标准项目兽药国家标准制修订计划。

摘　　要：本试验旨在建立紫草膏无菌检查法并进行验证。采用萃取-薄膜过滤相结合的方法,按照《中华人民共和国兽药典》(2020年版)附录1101要求,通过培养基灵敏度检查、方法适用性试验和无菌检查等试验,对紫草膏无菌检查方法进行了研究。将样品转移至无菌分液漏斗中,加入200 mL无菌肉豆蔻酸异丙酯,使样品充分溶解并分散摇匀,加入300 mL稀释液,充分萃取,待分层后取水层,用稀释液稀释至500 mL作为供试液,按薄膜过滤法处理,以含0.5%吐温80 NaCl-蛋白胨缓冲液(pH值7.0)为冲洗液,每张滤膜的冲洗量为500 mL,分5次进行,并在最后一次冲洗前加入小于100 cfu的菌液。结果显示,各验证菌、阳性菌对照组生长未受影响,样品组、阴性对照组均澄清,无菌落生长,说明在该检验条件下供试品的抑菌作用已被消除。紫草膏是典型的中兽药膏状制剂,具有较强的抑菌活性,本试验采用薄膜-萃取相结合的方法,建立了合理的无菌检查方法,能有效去除紫草膏中的抑菌成分,使检验结果准确、可靠,可作为该制剂的无菌检查方法。The purpose of this experiment is to create and validate a sterile test method for Zicao ointment.In accordance with the requirements of Appendix 1101 of the Veterinary Pharmacopoeia of the People’s Republic of China(2020),the sterility test method of Zicao ointment was studied by the combination of extraction and membrane filtration through the sensitivity test of culture medium,method applicability test and sterility test.Samples were placed in a sterile separatory funnel,200 mL sterile isopropyl myristate was added in order to dissolve and disperse the test article,and 300 mL diluent was added to fully extract the test product.After layering,the water phase was taken out and diluted with diluent to 500 mL as the test solution.It was treated by membrane filtration and the 0.5%polysorbate 80 sodium chloride(pH 7.0)-peptone buffer solution was used as the washing solution.The flushing volume of each filter membrane was 500 mL,divided into 5 times,and less than 100 cfu of bacterial solution was added before the last rinsing.The results showed that the growth of the validation bacteria and positive bacteria control group was not affected,while the sample group and negative control group were clear and sterile,which indicated that antibacterial activity of the substance was eliminated under the test conditions.Zicao ointment was a typical traditional Chinese veterinary medicine ointment with strong bacteriostatic activity.The reasonable sterility test method was established by the combined extraction and membrane filtration method,which could effectively remove the bacteriostatic components in Zicao ointment and make the test results more accurate and reliable.It could be used for sterility test of Zicao ointment.

关 键 词：紫草膏 薄膜过滤法 无菌检查 

分 类 号：S853.7[农业科学—临床兽医学]