摘要
目的:研究注射用替加环素质量标准中无菌检查项的现存问题并对检验方法进行优化。方法:分析7家企业质量标准中无菌检查项的内容,并对样品溶液pH范围、不同缓冲液对样品中微生物的影响、检查用样品溶液的稀释浓度、缓冲液中Mg^2+浓度对替加环素及杂质抑菌性的消除、冲洗液用量等问题展开试验。结果:不同配制方法所得注射用替加环素溶液pH范围在4.70~6.93,符合大多数微生物生长范围;不同缓冲液对注射用替加环素溶液的无菌检查无明显影响,考虑到pH 7.0氯化钠-蛋白胨溶液更适宜偏酸或偏碱的样品、0.9%氯化钠溶液会在后续试验中增加离子干扰,故0.1%蛋白胨水溶液为最适宜的基础性缓冲液;7家企业中仅有2家企业在无菌检查中使用Mg^2+作为中和剂,但所用Mg^2+浓度均不能最大限度地消除注射用替加环素的抑菌作用,经研究发现,0.10 mol·L^-1为缓冲液中最佳Mg^2+浓度,与替加环素和杂质(替加环素差向异构体、9-氨基米诺环素、9-硝基米诺环素)的金属络合反应可瞬间发生并完成,可消除部分抑菌性;1.25 mg·mL^-1注射用替加环素浓度为稀释数值的拐点,此浓度下污染微生物可在较低稀释级生长;300 mL的0.1%蛋白胨水溶液为适宜的冲洗液用量,此用量下所有人工污染的样品均可24 h检出阳性。结论:新缓冲液体系减少了缓冲液和冲洗液的整体用量,特别是冲洗液的用量,同时缓冲液中Mg^2+浓度适宜,适合本次国抽的注射用替加环素无菌检查,可为该品种质量标准中无菌检查项的修订和统一提供一定的参考。
Objective:To study the existing problems of sterility test item in quality standards of tigecycline for injection and to optimize the testing methods. Method:The sterility test in quality standards of 7 companies was analyzed with the experiments on deficiencies such as the pH range of the sample solution,the influence of different buffers on the microorganisms in the sample,and diluting concentration of the sample solution for examination,the concentration of Mg^2+ in the buffer to eliminate antimicrobial of tigecycline and impurities,buffer time and antimicrobial elimination effect and the amount of rinsing liquid. Results:The pH of tegacycline solution for injection obtained by different preparation methods ranged from pH 4.70 to 6.93 with the best condition of growth range of most microorganisms;Different buffers had no significant effect on the sterility test of tetracycline solution for injection. The pH 7.0 sodium-peptone solution was more suitable for acid or alkali samples while 0.9% sodiumchloride solution increased ion interference in subsequent tests. 0.1% peptone aqueous solution was the most suitable basic buffer;Only two of the seven enterprises used Mg^2+ as the neutralizer in the sterility test,but the concentration of Mg^2+ could not eliminate the antimicrobial effect of the injection tigecycline completely. According to the study,the optimal 0.10 mol·L^-1 of Mg^2+ concentration in the buffer react instantaneously with tigecycline and impurities(tetracycline differential isomer,9-aminocycline,9-nitrominocycline),which can eliminate part of the antimicrobial;the concentration of 1.25 mg·mL^-1 tigecycline is the inflection point of the dilution value,the contaminated microorganism could grow at a lower dilution level at this concentration;300 mL of 0.1% peptone aqueous solution was a suitable volume of rinsing liquid,all artificially contaminated samples under this dosage could be tested positive at 24 h. Conclusion:The new buffer system reduces the overall amount of buffer and flushing amount,especially the amount of rinsing liquid,and makes the Mg^2+ concentration suitable in buffer and easy to operate. It is suitable for the sterility test of tigecycline for injection in this national evaluative sampling inspection,provides the reference of revision and harmonization of sterility test in quality standards.
作者
刘洪祥
刘海玲
解慧
曹晓云
白海娇
LIU Hong-xiang;LIU Hai-ling;XIE Hui;CAO Xiao-yun;BAI Hai-jiao(Tianjin Institute for Drug Control,Tianjin 300070,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2020年第8期1443-1450,共8页
Chinese Journal of Pharmaceutical Analysis
关键词
注射用替加环素
无菌检查
抑菌性
Mg2+浓度
缓冲液体系
方法优化
tigecycline for injection
sterility test
antimicrobial
Mg2+concentration
buffer system
method optimization
