管碟法测定硫酸新霉素效价的方法优化  

Optimization of the tube-plate method for determining the potency of neomycin sulphate

作　　者：吴雨珊 孙冰清 张妤 姜芹 顾欣[1] 张文刚[1] 曹莹[1] Wu Yushan;Sun Bingqing;Zhang Yu;Jiang Qin;Gu Xin;Zhang Wengang;Cao Ying(Shanghai Animal Disease Control Center,Shanghai 201103)

机构地区：[1]上海市动物疫病预防控制中心,上海201103

出　　处：《中国抗生素杂志》2024年第6期667-672,共6页Chinese Journal of Antibiotics

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p H为8.0、缓冲液添加5%的Na Cl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。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.

关 键 词：新霉素 抗生素微生物检定法 效价测定 金黄色葡萄球菌 

分 类 号：R978.1[医药卫生—药品]