建立维生素K_1微生物限度测定方法

目的:根据计数培养基适用性检查和计数方法适用性试验结果,建立维生素K1微生物限度测定方法。方法:以聚山梨酯80作为乳化剂,使维生素K1在p H 7.0无菌氯化钠-蛋白胨缓冲液中充分乳化。取1:20供试液1 m L,按平皿法分别用营养琼脂培养基和玫瑰红钠琼脂培养基培养,以计数细菌、霉菌和酵母菌。结果:验证所用培养基的菌落平均数均大于对照培养基上的70%,且菌落形态大小一致。稀释液对照组和试验组培养基上菌落平均数的回收率均大于70%,且菌落形态大小一致。三批维生素K1及其加速和长期稳定性样品中均未见菌落。结论:所选培养基适宜大肠埃希菌、金黄色葡萄球菌和白色念珠菌生长。且含聚山梨酯80的p H 7.0无菌氯化钠-蛋白胨缓冲液和维生素K1对所含大肠埃希菌、金黄色葡萄球菌和白色念珠菌无抑菌性。

Establishment of Microbial Limit Test for Vitamin K1

Objective:To establish the microbial limit test for vitamin K1 based on the result of tests of growth promotion of the media and suitability of the counting method in the presence of product.Methods:With the aid of polysorbate 80, vitamin K1 was emulsified in pH 7. 0 sterile NaCl - peptone buffer solution. We inoculated the plates of nutrient agar and rose bengal agar with 1 mL of the test solution (1:20) and respectively counted the numbers of microorganisms.Results:For solid media, growth obtained did not differ by a factor greater than 70%fromthe calculated value for a standardized inoculum. The recovery of dilution in the control group and test group were more than 70 %. No microorganism was observed in vitamin K1 sample and its short-term and long-term test sample.Conclusion:The solid media is suitable for the growth of the microorganisms. Vitamin K1 and pH 7. 0 sterile NaCl - peptone buffer solution containing polysorbate 80 have no inhibitory role to the microorganisms.