Analysis Of The Impurities In Cephalosporin By Two Dimensional HPLC-QTOF-MS

Investigation indicates the allergy caused by ceftriaxone is the top among cephalosporin drugs,so we choose ceftriaxone for our study.Liquid chromatography coupled with mass spectrometry was applied to explore related substance and the mechanism of impurities,thus providing reference for new drugs development.To ensure safe and effective medication is the ultimate aim of our study.The main research contents are as follows:(1)A new related substance method was developed by liquid chromatography coupled with mass spectrometry.Eleven impurities,including the high-molecular weight impurities,were found in the degradation solutions.By MS and MS2,the structures and generation pathways of these impurities were figured out.(2)The resolution of each peaks was greater than 1.0,the LOD was 0.02 μg·ml-1,the average recovery was 99.4% with RSD of 1.2%.Methodology validation of the new method showed high sensitivity,good accuracy,reproducibility,and met the detection requirement.Through comparison of the optimized method and existing methods,the HPLC and BP method for related substances results were basically consistent,and higher than that of Ch.P method.Thus the HPLC method performed the same analysis effect and better than Ch.P method for related substance.(3)By cascading the gel chromatography,developed on Xtimate SEC-120 column,with HPLC method,a system of two dimensional chromatography was established.After the analysis of all the polymer peaks in gel chromatography,it was found that the polymer peaks were mixtures.According to MS and MS2 spectra,combined with the established impurity profile,we identified the polymer impurity peak position and find the polymer pointer which was relates to the polymer level.(4)Amoxicillin polymer was quantified by gel exclusion chromatography method in Ch.P.Experiment indicated that the excipients effected the result of amoxicillin granules.So related substance method was used for the qualitative and quantitative.Without changing the related substances in the Pharmacopoeia,two dimensional chromatography was applied to identify the dipolymer in the amoxicillin systemsuitability and amoxicillin capsule.