盐酸氯胺酮注射液的质量分析与标准完善

目的 ： 考察国内盐酸氯胺酮注射液的整体质量状况，并对现行质量标准进行评价与完善。 方法：按照中国药典2010年版二部现行质量标准对样品进行检验，并针对现行标准在对特定杂质的控制、含量限度等方面存在的问题以及国内外药典标准的差异，开展多项探索性研究以完善现行标准，包括对现行标准有关物质检查项色谱方法的优化、盐酸氯胺酮原料与注射液中有关物质的研究及其主要杂质的结构确证、特定已知杂质羟亚胺与盐酸氯胺酮的体外细胞毒性和斑马鱼胚胎毒性的比较研究以及对现行标准含量测定方法的优化。 结果：优化有关物质检查色谱条件，提高了杂质的检出率；样品中检出2个未知杂质，经分析与氯胺酮的热破坏降解物有关，其中一主要杂质与USP杂质B具有相同的结构，这与羟亚胺在酸性溶液中降解相关；盐酸氯胺酮和羟亚胺对小鼠成纤维细胞L929和人神经母细胞瘤SH-SY5Y 2种试验细胞株增殖的抑制作用无显著差异，但羟亚胺对斑马鱼胚胎发育的毒性和对幼体运动系统的损伤作用均要强于盐酸氯胺酮；建议提高含量测定的限度要求，由中国药典现行版的90.0%~110.0%提高至95.0%~105.0%。 结论：盐酸氯胺酮注射液的质量基本能够符合中国药典的质量标准要求；但其质量标准尚有亟待修订和完善之处，建议在盐酸氯胺酮原料现行标准中增加对已知杂质羟亚胺的控制，在盐酸氯胺酮注射液标准中增加对降解杂质的控制，并提高盐酸氯胺酮注射液含量测定现行UV法的限度要求。

Objective: To investigate the overall quality status of domestic ketamine hydrochloride injection,and to evaluate and improve the current quality standard of ketamine hydrochloride injection. Methods: The collected samples were tested according to the current quality standard in the Chinese Pharmacopoeia 2010 edition.Several exploratory researches were carried out on the basis of the existing problems of current standard in the control of specific impurity and content limit,as well as the differences among domestic and foreign Pharmacopoeia standards.It included the optimization of the chromatographic method of related substances in the current standard,the study on related substances,structure confirmation of the main impurities for ketamine hydrochloride and its injection,and the comparative study on the in vitro cytotoxicity and the toxicity to zebrafish embryos of ketamine hydrochloride and the known impurity hydroxylamine,and the improvement of assay in the current standard. Results: The chromatographic method of related substance was optimized to improve the detection rate of impurities.Two unknown impurities detected in the samples were related with the thermal degradation products of ketamine,one of which had the same structure as the USP impurity B,and might be associated with the degradation of hydroxylamine in acidic solution.Moreover,the inhibitory effect of ketamine hydrochloride and hydroxylamine on cell proliferation of mouse fibroblast cells L929 and human neuroblastoma cells SH-SY5Y showed no significant difference,but the toxicity and damage of hydroxylamine to zebrafish embryos and larval motion systems were stronger than those of ketamine hydrochloride.And it was suggested to improve the determination limit of the assay in the current standard from 90.0%－110.0%to 95.0%－105.0%. Conclusion: The domestic ketamine hydrochloride injection can meet the requirements of the ChP standard,but the current quality standard should be revised and improved.It is suggested to control the known impurities hydroxylamine and the degradation products in the quality standard of ketamine hydrochloride and its injection,respectively,and improve the limit requirement in the current UV assay of ketamine hydrochloride injection.