Isolation and Identification of Antioxidant Compounds from  Ligularia fischeri

The plant Ligularia fischeri var. spiciformis Nakai, a well-known edible medicinal herb in Korea, has been used to treat maladies such as jaundice, scarlet fever, rheumatoid arthritis, and hepatic function failure. In this research, 4 major antioxidant compounds were detected from this plant's leaves using an on-line high-performance liquid chromatography (HPLC)-ABTS screening system, which can determine the antioxidant activity based on a decrease in absorbance at 734 nm after postcolumn reaction of HPLC-separated antioxidants with the 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radicals (ABTS(*)). In order to isolate these active compounds, a preparative HPLC was applied and their chemical structures were identified as 5-O-caffeoylquinic acid (5-CQA), 3,4-di-O-caffeoylquinic acid (3,4-DCQA), 3,5-di-O-caffeoylquinic acid (3,5-DCQA), and 4,5-di-O-caffeoylquinic acid (4,5-DCQA) by ESI/MS(n) and (1)H NMR. These 4 isomers comprised over 10% of the dried leaves, with 3,5-DCQA being the most abundant compound. The radical scavenging activity of each isomer was also evaluated simultaneously through the on-line HPLC-ABTS method, which showed 94% antioxidant activity of the ethanol extract derived from caffeoylquinic acids. Among these isomers, 3,4-DCQA contained the most strong antioxidant activity while 3,5-DCQA accounted for the highest radical scavenging capacity due to having the highest content.