Isolation of phenolics from Rhizophora mangle by combined counter-current chromatography and gel-filtration.

Nine phenolic compounds, quercetin, epi-catechin, catechin, 4-hydroxybenzoic acid, kaempferol 3-O-β-glucopyranoside, quercetin 3-O-β-glucopyranoside, quercetin 3-O-6"-trans-coumaroyl-β-glucoside, kaempferol 3-O-β-rutinoside and quercetin 3-O-β-rutinoside, were isolated from the EtOAc leaf extract of Rhizophora mangle (Rhizophoraceae) combining counter-current chromatography (CCC) and gel-filtration. A solvent system of n-hexane-ethyl acetate-methanol-water (1.5:6:1.5:6) was employed at the preliminary stage of EtOAc extract fractionation as it was shown to contain compounds that differed highly in their hydrophobicity. The obtained fractions were further purified by either CCC or gel-filtration depending on their complexity. The isolated compounds were analyzed by NMR spectroscopy and the proposed structures were confirmed by HRES/ESI/TOF MS. Some of these compounds were isolated and/or identified for the first time in R. mangle.