国产注射用硫酸核糖霉素质量评价

目的：采用法定检验与探索性研究相结合的方法对国产注射用核糖霉素质量现状及存在问题进行评价。方法：采用现行标准对抽验样品进行法定检验；建立并完善用于探索性研究的多个方法：（1）建立ELSD、CAD和PAD法测定有关物质及含量；采用在线膜抑制脱盐技术辅助LC-Orbitrap HRMS研究本品杂质谱；采用豚鼠模型和斑马鱼胚胎模型，以及纸片扩散和常量肉汤稀释法分别测定主药和相关物质的毒性以及体外抗菌活性；（2）制备精制品，采用质量平衡法测得其纯度，管碟法测得其效价，建立量效转换桥接关系；（3）建立HPLC-ELSD法及HPIC-CD法测定硫酸核糖霉素中硫酸盐含量；（4）综合应用多种现代分析技术优化干燥失重（或水分）测定条件；等等。结果：法定检验显示，本次抽验样品所有批次均符合标准规定；新建立的3种有关物质和含量测定方法，在灵敏度、专属性、重复性及检验经济学等方面均优于法定方法；杂质谱研究结果显示，注射用硫酸核糖霉素有个10潜在杂质，结合毒理实验与抗菌活性显示，有必要在标准中对新霉胺单独控制；测定得到量效转换系数为991 u·mg^-1，验证结果显示，该系数可实现量效方法转移的要求；HPLC-ELSD法测定硫酸盐结果被HPIC-CD法验证，酸碱度与硫酸盐含量呈负相关，分子式中硫酸个数为约为1.5；干燥失重研究显示，60℃减压干燥6 h较科学。结论：目前国产注射用硫酸核糖霉素制剂工艺较为稳定，总体质量较好；现行注射用硫酸核糖霉素质量标准存在缺陷，有待完善。

Objective: To evaluate the current status and existing problems of domestic ribostamycin sulfate for injection using a combination of statutory testing methods and exploratory research methods. Methods: The current statutory testing methods were used to test samples. Several methods for exploratory research were established and improved,including:(1) ELSD,CAD and PAD methods were established to determine the related substances and content. On-line membrane suppression desalination technology was used to assist LC-Orbitrap HRMS for illustration of impurities profile of this product. The guinea pig model and the zebrafish embryo model,as well as disk diffusion method and broth dilution method were established respectively to determine the toxicity and in vitro antibacterial activity of the main drug and related substances. (2) The refined product was prepared,the purity was measured by the mass balance method,the potency was measured by the tube dish method,and the dose-effect conversion bridging relationship was established. (3) HPLC-ELSD method and HPIC-CD method were established for the determination of sulfate content in ribostamycin sulfate. (4) A variety of modern analytical techniques were introduced to optimize loss on drying (or water) measurement conditions,and so on. Results: According to statutory tests,all batches of samples met the standard requirements.The three newly established methods for related substances and content determination were superior to the statutory methods in terms of sensitivity,specificity,repeatability and test economics. Results of impurities profile study showed that there were 10 potential impurities in ribostamycin sulfate for injection. According to toxicological experiments and antibacterial activity,neomycin was necessary to be controlled separately in the standard. The measured dose-effective conversion coefficient of 991 u·mg^-1 had been verified to be the transfer of the dose-effect method. The results of HPLC-ELSD determination of sulfate were verified by HPIC-CD method. The pH was negatively correlated with the sulfate content,and the number of sulfuric acid in the molecular formula was about 1.5. The results of loss on drying showed that decompression drying at 60℃ for 6 hours was more scientific. Conclusion: The domestic ribostamycin sulfate for injection has a relatively stable process,and the overall quality is relatively good. The current quality standard of ribostamycin sulfate for injection shall be improved.

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