蔗糖铁注射液过滤除菌前配制液的微生物限度检查方法研究

Study on the Method of Microbial Limit Test for Preparation Liquid of Iron Sucrose Injection before Filtration and Sterilization

目的:建立蔗糖铁注射液过滤除菌前配制液的微生物限度检查方法。方法:按照2015版《中国药典》(四部)"通则1005"和"通则1006"微生物限度检查方法,分别采用平皿法和薄膜过滤法检查需氧菌(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)数、霉菌及酵母菌(白色念珠菌、黑曲霉)数,通过比较试验菌回收率来确定适宜的方法,并进行方法验证。结果:以平皿法进行试验,铜绿假单胞菌和枯草芽孢杆菌的回收率分别为2%、5%,用p H 7.0氯化钠-蛋白胨缓冲液稀释样品原液10倍后,试验菌回收率范围为88%～96%,但培养基颜色较深。以薄膜过滤法不加冲洗液进行试验,试验菌回收率范围为88%～95%;加入冲洗液后,试验菌回收率范围为91%～103%。经验证,加入冲洗液的薄膜过滤法回收率为50%～200%,符合规定。结论:本试验建立的薄膜过滤法较平皿法试验菌回收率高且加入冲洗液后,试验菌回收率更高,滤膜表面均无颜色较深的物质残留影响计数,适用于蔗糖铁注射液过滤除菌前配制液的微生物限度检查。

OBJECTIVE： To establish a method of microbial limit test for liquid preparation of Iron sucrose injection before fil- tration and sterilization, METHODS： According to the microbial limit test in the ＂1005＂ and ＂general rules 1006＂ of 2015 edition of Chinese Pharmacopoeia （vol. V ）, plate method and membrane filtration method were used to measure total number of aerobic bacteria （Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Candida albicans, Aspergillus niger） and total num- ber of molds and yeasts （C. albicans, A. niger）. The optimal test method was obtained by comparing the bacterial recoveries. RE- SULTS： By plate method, the recoveries of P. aeruginosa and B. subtilis were 2% and 5%. The test sample was diluted 10 times with pH 7.0 sodium chloride-peptone buffer solution, and the bacterial recoveries were in the range of 88% to 96% ; but he medi- um was dark in color. By membrane filtration method, without rinse solution, the bacterial recoveries in the range of 88% to 95%. Add rinse solution, the bacterial recoveries were in the range of 91% to 103 %. After validated, the recoveries of menbrane filtra- tion method with tlushing tluid ranged 50%-200%, which was in line with the requnements. CONCLUSIONS： The membrane filtra- tion method established in this experiment has higher bacterial recovery rate than the plate method. The bacterial recoveries rate were higher after adding rinse solution, and no dark substance in the surface of filter membrane affect the account. It can be used as the microbial limit test method for preparation liquid of Iron sucrose iniection before filtration and sterilization.