Isolation,structure characterization and quantification of related impurities in asperosaponin VI

AIM:To study the related impurities in asperosaponin VI bulk drug and to develop a high performance liquid chromatography(HPLC) method for the determination of asperosaponin VI and its related impurities.METHODS:The related impurities were detected in asperosaponin VI bulk drug by a newly developed HPLC method,obtained by ODS column chromatography and semi-preparative HPLC methods,and the structures were elucidated by TOF-MS,IR,and NMR techniques.The HPLC method was validated according to ICH guidelines for asperosaponin VI and its related impurities.RESULTS:Seven related impurities(Imp 1-7) were isolated from asperosaponin VI bulk drug.Impurity 3 was found to be a mixture of two epimers,and was first reported in the paper.The validation results showed good sensitivity,specificity,linearity(r20.997 9),precision(RSD5.0%),accuracy(recoveries in the range of 94.61%-106.51%) and robustness.CONCLUSION:The developed HPLC method is suitable for the quality control of asperosaponin VI bulk drug.