止痒搽剂微生物限度检查方法验证.docVIP

止痒搽剂微生物限度检查方法验证[摘要] 目的 建立止痒搽剂微生物限度检查方法。方法 用常规法、培养基稀释法、薄膜过滤法进行细菌、霉菌及控制菌计数方法的验证。结果 采用常规法后白色念珠菌及黑霉菌的试验组及稀释剂对照组的回收率均大于70% ,采用薄膜过滤法后的大肠埃希菌、金黄色葡萄球菌、枯草芽孢杆菌的试验组及稀释剂对照组的回收率也可达到70%以上;采用培养基稀释法可检出金黄色葡萄球菌,采用薄膜过滤法可检出铜绿假单胞菌。结论 经方法学验证,结果符合现行《中国药典》2005年版的要求。 [关键词] 止痒搽剂; 微生物限度; 验证 [中图分类号] R927 [文献标识码] A [文章编号] 1673-9701(2009)23-56-03 Study on the Microbial Limit Test of Zhiyangchaji QIU Kaifeng ZHU Jun The Second Affiliated Hospital of Zhongshan University,Guangzhou,510120,China [Abstract] ObjectiveTo establish a method for the determination of microbial limits in Zhiyangchaji. MethodsThe bacteria ,molds ,yeasts and controlled bacteria were counted by the common methods ,dilution method and membrane adhering method. ResultsThe recoveries of Candida albicans,Aspergillus niger in test group and diluted control group were above 70% when common method were used. The recoveries of Escherichiacoli,Staphy lococcus aureus,B acillus subtilis in test group and diluted control group were above 70% when membrane filtration method was used. Staphy lococcus aureus can be detected by dilution method and Pseudomonas aeruginosa can be detected by membrane filtration method. ConclusionThrough t he technological validation ,the result is consistent with the requirement of ChP.in force. [Key Words]Zhiyangchaji; Microbial limit; Validation 止痒搽剂为我院传统医院制剂,其主要成分为五指柑、大风艾、地肤子,此3味中药在体外有不同的抑菌作用。现根据《中国药典》2005版,对其微生物限度检查方法进行验证,以保证测定结果的可靠性。 1 材料与方法 1.1 试剂 培养基:营养琼脂培养基,玫瑰红钠琼脂培养基,胆盐乳糖培养基(BL),营养肉汤培养基,改良马丁培养基,改良马丁琼脂培养基。稀释剂:pH7.0无菌氯化钠-蛋白胨缓冲液(以上均来源于广东环凯微生物科技有限公司)。 1.2 菌种 大肠埃希菌[CMCC(B)(44102)],金黄色葡萄球菌[CMCC(B)(26003)],枯草芽孢杆菌[CMCC(B)(63501)],白色念珠菌[CMCC(F)(98001)],铜绿假单胞菌[CMCC(B)(10104)],黑曲霉菌[CMCC(F)(98003)](以上均来源于中国药品生物制品检验所)。 1.3 样品 止痒搽剂,本院自制制剂,批号分别为:0802018、0802278、0803201。 2 方法与结果 2.1 供试液制备 取样品10mL,加入pH7.0的无菌氯化钠-蛋白胨缓冲液至100mL,使成1∶10的供试液。 2.2 菌液制备[1] ①取大肠埃希菌、金黄色葡萄球菌、枯草芽孢杆菌、铜绿假单胞菌的新鲜培养物接种至营养肉汤培养基,经37℃培养18～24h,取白色念珠菌的新鲜培养物接种至改良马丁培养基,经25℃培养24～48h,上述培养物分别用0.9%无菌氯化钠溶