Study On Cefathiamidine Impurity

Objective: Analyzing the impurities produced by the destruction of cefathiamidine,study the preparation,separation and purification methods of the impurities of the cefathiamidine,analyze and confirm the structure of impurities,the cytotoxicity of the impurities were detected.It provides a reference for the determination of the type and content of impurities in cefathiamidine and provides method and data support for the quality control and quality standards of the drug.Methods:(1)Destruction of cefathiamidine raw materials by acid,alkali,light,heat and oxidation;detect and analyze the impurities by LC-MS;(2)The corresponding pure impurities were obtained after isolation and purification.The precise structure of the impurities were characterized by LC-MS/MS and NMR methods.(3)The cytotox--icity on the three cell lines of HUVEC,Ha Cat and Vero were detected by MTT method.Results: Through the destruction of cefathiamidine and detection by LC-MS,it was found that the raw material produce many impurity fragments under alkaline conditions,3 more obvious impurities were detected,6 impurities can be detected by treating cefathiamidine with acid,9 impurities were detected by heat treated cefathiamidine aqueous solution,2 impurities were detected after irradiation with254 nm UV light,5 kinds of impurities were detected after treatment with H2O2,6impurities were isolated and purified.The structures of the six impurities were determined by LC-MS and NMR methods.LM-A was desacetyl cefathiamidine,LM-D was cefathiamidine lactone,and LM-O was the sulfur atom on the cefathiamidine six-membered ring is oxygenated,and the LM-?3 is the product of the transfer of the double bond on the six-membered ring of cefathiamidine.The LM-7S is the configuration change of the cefathione C-7 and the LM-AO is sulfur atoms on the LM-A six-membered ring were oxygenated;cytotoxicity experiments showed that six impurities concentrations in the 1.56 ?g/m L-100 ?g/m L range had minimal effect on cell viability,and it can be speculated that the six impurities are non-cytotoxic on HUVEC,Ha Cat and Vero cells in this concentration range.Conclusion: The separation and purification method established in this project can effectively purify the target impurities,and can produce impurity products with higher purity more quickly and efficiently than the traditional separation method.Six chemical structures of cefathiamidine impurities were identified and the six impurities were found to have no apparent toxicity to HUVEC,Ha Cat and Vero cells lines.Cephathiamidine are easy to produce impurities under the conditions of acid,alkali,and high temperature.Therefore attention should be paid to in the production and storage process in order to reduce the generation of impurities.The results of this study can provide valuable reference for the optimization of the production process and impurity control of cefathiamidine drugs.