In order to study the differences between Viticis Negundo Fructus and Viticis Fructus,the characteristic chromatogram and multi-index content determination method was established.Based on the UPLC characteristic chromatogram of Viticis Negundo Fructus, nine chemical components were identified by high resolution mass spectrometry(HRMS) then determined,which were protocatechuic acid A,protocatechualdehyde,4-hydroxybenzoic acid,vanillic acid,isoorientin,orientin,agnuside,vitex and isovitex.The determination of 15 batches of Viticis Negundo Fructus and 12 batches of Viticis Fructus showed that there were differences among these nine components.The means of protocatechuic acid,protocatechualdehyde,4-hydroxybenzoic acid,vanillic acid and agnuside for Viticis Negundo Fructus were higher than Viticis Fructus that isoorientin and orientin were lower,while vitexin and isovitexin were not detected in Viticis Fructus.The UPLC characteristic chromatogram combined with multi-index content determination was applied to compare the differernces between Viticis Negundo Fructus and Viticis Fructus,which provide a reference for their quality control and clinical medication.