复方氟康唑乳膏微生物限度检查方法的建立

Validation of Microbial Limit Test Method for Compound Fluconazole Cream

为了建立复方氟康唑乳膏的微生物限度检查方法,试验采用2020年版《中国兽药典(一部)》附录1105、1106和1107。以5种菌为试验菌,选择平皿法和薄膜过滤法进行微生物计数法方法适用性检查。以金黄色葡萄球菌、铜绿假单胞菌为试验菌,采用培养基稀释法和薄膜膜过滤法进行控制菌方法适用性检查。结果表明:缓冲液中加入1%聚山梨酯80,并预热至40℃,制备1∶10供试品溶液,可使供试品均匀分散。微生物计数法:常规平皿法不能消除其抑菌活性,方法不成立,进一步采用薄膜过滤法,总冲洗量为800 mL,进行微生物计数时,5种菌的菌液回收比值均在0.5~2之间,方法符合要求。控制菌检查:常规培养基稀释法,培养基量增大至800 mL,药物加菌组没有检出对应的试验菌,进一步使用薄膜过滤法,总冲洗量为500 mL,供试液加菌组和菌液对照组均检出对应的试验菌。说明复方氟康唑乳膏具有较强的抑菌活性,进行微生物计数和控制菌检查时,供试液需采用薄膜过滤法消除其抑菌活性,按照2020年版《中国兽药典》规定,成功建立了该产品的微生物检查方法。

In order to establish the microbial limit checking method of compound fluconazole cream,the test was conducted using the 2020 edition of Chinese Veterinary Pharmacopoeia(Ⅰ),Appendix 1105,1106 and 1107.Five kinds of bacteria were selected as test bacteria,and the plate method and membrane filtration method were selected to check the suitability of microbial enumeration method.Staphylococcus aureus and Pseudomonas aeruginosa were used as test bacteria,and the suitability of control bacteria method was examined by medium dilution method and membrane filtration method.The results showed that a 1∶10 solution of the test material was prepared by adding 1%polysorbate 80 to the buffer and preheating to 40℃,which resulted in a homogeneous dispersion of the test material.Microbial counting method:conventional flat dish method could not eliminate its bacteriostatic activity,the method was not valid,further membrane filtration method was used,the total rinsing volume was 800 mL,when microbial counting was carried out,the bacterial recovery ratio of 5 bacteria were between 0.5 and 2,the method meets the requirements.Control bacteria test:conventional medium dilution method,increase the volume of medium to 800 mL,no corresponding test bacteria were detected in the drug bacteria adding group,membrane filtration method was further used,the total washing volume was 500 mL,corresponding test bacteria were detected in the test solution bacteria adding group and bacteria solution control group.It indicates that compound fluconazole cream has strong bacteriostatic activity,and for microbial counting and control bacteria examination,the test solution needs to be eliminated by membrane filtration to eliminate its bacteriostatic activity.In accordance with the 2020 edition of the Chinese Veterinary Pharmacopoeia,the microbial examination method of this product was successfully established.