改进培养基制备的平皿法应用于芬太尼透皮贴剂微生物限度检查

Application of the plate-count methods of improved medium preparation on fentanyl transdermal patches microbial limit examination

目的建立一种芬太尼透皮贴剂的微生物限度检查新方法。方法按照《中国药典》2015年版要求,分别取2种规格相当于供试品100 cm^2芬太尼透皮贴剂,加入100 mL供试液中制成1∶10供试液(按10 cm^2计)。分别加入不大于100 cfu的铜绿假单胞菌、金黄色葡萄球菌、枯草芽孢杆菌、白色念珠菌和黑曲霉试验菌,取供试液10mL倾注直径150 mm平皿,按培养基处方量的1.25倍制备胰酪大豆胨琼脂培养基和沙氏葡萄糖琼脂培养基,倾注平皿进行计数法的适用性试验,另取供试液10 mL加入100 mL胰酪胨大豆液体培养基中进行金黄色葡萄球菌、铜绿假单胞菌检查方法适用性试验。结果需氧菌总数、霉菌和酵母菌总数计数方法适用性试验中各试验菌回收率均不小于0.70,控制菌方法适用性试验均检出金黄色葡萄球菌和铜绿假单胞菌,符合《中国药典》要求。结论所建立方法准确、简单、科学、经济,适用于芬太尼透皮贴剂的微生物限度检查。

Objective To establish a kind of new method for fentanyl transdermal patches microbial limit examination.Methods According to Chinese Pharmacopoeia 2015 edition,2 kinds of test samples were taken equivalently 100 cm^2 fentanyl transdermal patches separately,added to 100 mL test solution,and a 1 ∶ 10 dilution was prepared（ calculated according to 10 cm^2）.To add no more than 100 cfu Pseudomonas aeruginosa,Staphylococcus aureus,Bacillus subtilis,Candida albicans and Aspergillus niger test bacterium separately,took 10 mL test liquid to pour into diameter of 150 mm plates. According to 1.25 times culture prescription,soybean-casein digest agar and sabouraud dextrose agar culture medium were prepared and used for count method applicability test.Another 10 mL test liquid was took to 100 mL pancreatic cheese peptone soybean liquid medium for the operational test of Staphylococcus aureus,Pseudomonas aeruginosa.Results The recoveries in the operational test of total aerobic microbial count,total combined yeasts and molds count counting method were less than 0.70,Staphylococcus aureus and Pseudomonas aeruginosa were detected in control bacteria examination,and all met the requirements of Chinese Pharmacopoeia.Conclusion The established method was accurate,simple,scientific,economic,applicable to the microbial limit examination of fentanyl transdermal patches.