摘要
目的 优化硫酸新霉素含量管碟法测定方法,增大新霉素抑菌圈直径大小,提高检测结果的准确性.方法 按照《中国兽药典》2020版一部附录1200抗生素微生物(检定法)的实验基本要求,设置菌液麦氏浊度(0.5、1、2、3和＞4 mcf)、培养基pH(7.6、7.8、8.0、8.2和8.4)、磷酸盐缓冲液中NaCl添加比例(0、3％、4％、5％、7％和9％)等变量进行含量测定,比较抑菌圈直径大小和可信限率等指标.结果 在添加0.5％的菌液麦氏浊度为3 mcf的金黄色葡萄球菌液、培养基pH为8.0、缓冲液添加5％的NaCl条件下,高剂量抑菌圈直径扩大至20 mm左右,抑菌圈边缘清晰,且符合《中国兽药典》可信限率的要求(不大于5％).通过多产品多厂家的实样验证此条件稳定可靠.结论 优化后的方法可用于硫酸新霉素常规效价含量的测定.
Abstract
Objective To optimize the determination method of neomycin content,increase the diameter of the antibacterial ring in the tube disc method,and improve detection efficiency.Method According to the experimental requirements of Appendix 1200 Antibiotic Microorganisms(Assay Method)in the 2020 edition of the Chinese Veterinary Pharmacopoeia,different bacterial concentrations(0.5,1,2,3,and＞4 mcf),medium pH(7.8,8.0,8.2,and 8.4),and NaCl addition ratios(0,3％,4％,5％,7％,and 9％)in phosphate buffer were selected based on the indicators,such as diameter of the antibacterial zone and confidence limits.Results The results showed that under the conditions of adding 0.5％of the bacterial solution with a concentration of 3 mcf of Staphylococcus aureus,a culture medium pH of 8.0,and adding 5％NaCl to the buffer,the diameter of the antibacterial zone expanded to about 20 mm,and the edge of the antibacterial zone was clear.The confidence limit rate met the requirements of the Veterinary Pharmacopeia of the People's Republic of China(not more than 5％).This condition was stable and reliable,and it had been verified through actual samples from multiple products and manufacturers.Conclusion The optimized method can be used for the determination of conventional potency content of neomycin.
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