In this work, flavonoid fraction from the leaves of Crataegus pinnatifida was separated into its seven main constituents using a combination of HSCCC coupled with pre-HPLC. In the first step, the total flavonoid extract was subjected to HSCCC with a two-solvent system of chloroform/methanol/water/n-butanol (4:3:2:1.5, v/v), yielding four pure compounds, namely (–)-epicatechin (1), quercetin-3-O-(2,6-di-α-l-rhamnopyranosyl)-β-d-galactopyranoside (2), 4′′-O-glucosylvitexin (3) and 2′′-O-rhamnosylvitexin (4) as well as a mixture of three further flavonoids. An extrusion mode was used to rapidly separate quercetin-3-O-(2,6-di-α-l-rhamnopyranosyl)-β-d-galactopyranoside with a big KD-value. In the second step, the mixture that resulted from HSCCC was separated by pre-HPLC, resulting in three pure compounds including: vitexin (5), hyperoside (6) and isoquercitrin (7). The purities of the isolated compounds were established to be over 98%, as determined by HPLC. The structures of these seven flavonoids were elucidated by ESI-MS and NMR spectroscopic analyses.
1 October 2019 in Journal of Pharmaceutical and Biomedical Analysis
An improved method based on HPLC-TOF/MS was developed to catalog the antioxidants in five species of Chaenomeles (Mugua). Forty-four fractions from the Mugua extracts show appreciable levels of antioxidative activity in scavenging the stable free-radical 2,2-diphenyl-1-picrylhydrazyl and the hydroxyl radicals. Twelve major antioxidant's chemical structures are identified. Antioxidant activities differ between species, but intra-species level of antioxidants, regardless of their ripeness, are similar. C. sinensis has the highest antioxidant level. A rigorous quality control procedure was implemented to ensure accuracy of antioxidant quantification. This improved procedure can be used for rapid discovery of antioxidants in other plant extracts.