盐酸头孢噻呋乳房注入剂无菌检查方法的建立

Establishment of the Sterility Test Method for Ceftiofur Hydrochloride Intramammary Infusion

为建立盐酸头孢噻呋乳房注入剂的无菌检查方法,本试验按照《中华人民共和国兽药典》2015年版一部附录1101无菌检查法中方法适用性试验的有关要求,对取样量、稀释液种类、冲洗方式、加酶量等进行了考察。取供试品置于无菌分液漏斗中,加300 mL无菌十四烷酸异丙酯,摇匀,再加200 mL含1%聚山梨酯80的0.1%无菌蛋白胨溶液,充分振摇,静置,取水层,按照薄膜过滤法处理,用pH 7.0无菌氯化钠-蛋白胨缓冲液为冲洗液,每张滤膜每次冲洗量为100 mL,冲洗4次,每管培养基中加入600万单位的青霉素酶溶液。结果显示,方法适用性试验中,供试品6种阳性菌试验组与阳性菌对照组相比均生长良好,供试品组、阴性对照组均无菌生长,说明供试品的检验量在该检验条件下已消除其抑菌作用。盐酸头孢噻呋具有较强的抑菌活性,本试验采用薄膜过滤法,建立了合理的无菌检查方法,能有效去除盐酸头孢噻呋乳房注入剂中的抑菌成分,使检验结果更准确、可靠,可作为该制剂的常规无菌检查方法。

In order to establish an sterility test method for ceftiofur hydrochloride intramammary infusion,the sampling amount,diluent type,washing method and enzyme dosage were investigated in accordance with the relevant requirements of the method suitability test in appendix 1101 of Chinese Veterinary Pharmacopoeia 2015 edition.Samples were placed in a sterile funnel,isopropyl tetradecanoate was added to the funnel,the 0.1%sterile peptone solution containing 1%polysorbide 80 was added to the funnel,the water layer was shaken and placed quietly,the water was taken out and treated by membrane filtration and the pH 7.0 sterile sodium chloride-peptone buffer solution was used as the washing solution.Each filter membrane was washed four times with a washing capacity of 100 mL.Penicillinase solution of not less than 6 million units was added to each tube of culture medium.The results showed that the six positive bacteria test groups grew as well as the positive bacteria control group,the test group and the negative control group grew aseptically.These indicated that the test quantity of the test samples had eliminated its bacteriostasis under the test conditions.Ceftiofur hydrochloride had strong bacteriostasis activity.In this experiment,a reasonable method of sterility test was established by membrane filtration method,which could effectively remove the bacteriostasis ingredients in ceftiofur hydrochloride intramammary infusion and make the test results more accurate and reliable.It could be used as a routine aseptic test method for the preparation.