3种咪唑类抗真菌乳膏剂微生物限度检查方法学研究  

Study on microbial limitation methodology of three imidazole antifungal creams

作　　者：李辉 杨晓莉 刘立科 马元斌 绳金房 LI Hui;YANG Xiaoli;LIU Like;MA Yuanbin;SHENG Jinfang(Shaanxi Sheng de tai lin Biological Safety Technical Testing CO.Ltd,Xi'an 710065,China;NMPA Key Laboratory for Testing Technology of Pharmaceutical Microbiology,Xi'an 710065,China;Shaanxi Institute for Food and Drug Control,Xi'an 710065,China;Shaanxi Institute of Medical Device Quality Supervision and Inspection,Xianyang 712046,China)

机构地区：[1]陕西盛德泰林生物安全技术检测有限公司,陕西西安710065 [2]国家药品监督管理局药品微生物检测技术重点实验室,陕西西安710065 [3]陕西省食品药品监督检验研究院,陕西西安710065 [4]陕西省医疗器械质量监督检验院,陕西咸阳712046

出　　处：《药物评价研究》2021年第4期721-727,共7页Drug Evaluation Research

基　　金：陕西省重点研发项目(2019SF062)。

摘　　要：目的建立硝酸咪康唑、酮康唑、曲安奈德益康唑3种咪唑类抗真菌乳膏剂的微生物限度检查方法。方法采用含5%聚山梨酯80的pH 7.0无菌氯化钠蛋白胨缓冲液制备3种乳膏剂1∶10供试液,并稀释至1∶20和1∶50,分别加入金黄色葡萄球菌、铜绿假单胞菌、枯草芽孢杆菌、白色念珠菌、黑曲霉接种无菌平皿中,按照《中国药典》2015年版四部1105考察平皿法及稀释-平皿法计数方法适用性;分别取3种乳膏剂加入无菌pH 7.0无菌氯化钠蛋白胨缓冲液和十四烷酸异丙酯两相系统中萃取,取萃取后的下层水相的上清液薄膜过滤处理后,分别加入5种试验菌,进行萃取-薄膜过滤法计数法适用性试验;分别取未经萃取-薄膜过滤处理和经过萃取-薄膜过滤的1∶10供试液,接种至不同体积的胰酪大豆胨液体培养基中,按照《中国药典》2015年版四部1106进行控制菌检查适用性试验。结果通过平皿法或稀释处理,3种乳膏剂的3种细菌类试验菌回收率即符合要求;3种乳膏剂对白色念珠菌、黑曲霉均有较强的抑菌作用,无论是平皿法还是稀释处理后的平皿法,2种试验菌回收率均远小于0.5,不符合方法适用性要求。萃取-薄膜过滤法可有效去除3种咪唑类抗真菌乳膏剂对各试验菌的抑菌作用,3种乳膏剂的试验菌回收率均在0.5~2.0,符合计数方法适用性要求。经过萃取-薄膜过滤处理的供试液控制菌检出优于未经处理的供试液。结论采用萃取-薄膜过滤法可解决样品抑菌性的去除和滤膜堵塞的问题,适用于3种咪唑类抗真菌类乳膏剂的微生物限度检查。Objective To establish the microbial limit test method for three kinds of imidazole antifungal creams,such as miconazole nitrate,ketoconazole,and triamcinolone econazole.Methods Sterile sodium chloride peptone buffer of pH 7.0 containing 5% polysorbate 80 was used to prepare three kinds of cream 1∶10 for test solution,and dilute to 1∶20 and 1∶50,respectively.Test solution was added with Staphylococcus aureus,Pseudomonas aeruginosa,Bacillus subtilis,Candida albicans,Aspergillus niger,and inoculated sterile plate.According to the 1105 of Volume four from the"Chinese Pharmacopoeia"2015 edition,the applicability of plate method and dilution plate method was investigate.Cream was added to a two-phase system of sterile pH 7.0 sterile sodium chloride peptone buffer and isopropyl myristate.Add five kinds of test bacteria and perform the counting method suitability test.The test solution of 1∶10 with and without extraction membrane filtration treatment were inoculated into different volumes of tryptic soy peptone liquid culture medium,according to 1106 of Volume four from the"Chinese Pharmacopoeia"2015 edition to carry out control bacteria test suitability test.Results Through the plate method or dilution treatment,the recovery rates of three kinds of bacteria in the three creams met the requirements;the three creams had strong antibacterial effect on Candida albicans and Aspergillus niger,and the recovery rates of the two kinds of bacteria were far less than 0.5,which did not meet the applicability requirements of the method.The recovery rate of each test bacteria for the determination of total aerobic microbial count,total combined yeasts and molds count all met the requirements of 0.5~2.0,and the detection of control bacteria in the test solution after extraction-membrane filtration treatment was better than that of the untreated Test solution.Conclusion The extraction-membrane filtration method can solve the problem of sample antibacterial removal and filter clogging.It is suitable for the microbial limit insp

关 键 词：萃取 咪唑类抗真菌药 硝酸咪康唑 酮康唑 曲安奈德益康唑 乳膏剂 微生物限度 

分 类 号：R965.3[医药卫生—药理学]