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  • Title: Isolation of flavanol-anthocyanin adducts by countercurrent chromatography.
    Author: Salas E, Fulcrand H, Poncet-Legrand C, Meudec E, Köhler N, Winterhalter P, Cheynier V.
    Journal: J Chromatogr Sci; 2005 Oct; 43(9):488-93. PubMed ID: 16212796.
    Abstract:
    Pigments of the flavanol-anthocyanin (F-A+) type detected earlier in wine are synthesized using a protocol adapted from the synthesis of procyanidin dimers. The F-A+ adduct thus obtained is purified by countercurrent liquid-liquid partition, currently referred to as countercurrent chromatography (CCC). The solvent system consists of tert-butyl methyl ether-n-butanol-acetonitrile-water (2:2:1:5, acidified with 0.1% trifluoroacetyl) with the light organic phase acting as a stationary phase and the aqueous phase as the mobile phase. Four fractions are recovered and analyzed by high-performance liquid chromatography coupled to a diode-array detector and electrospray ionization mass spectrometer. The multilayer CCC method allowed the separation of pigments in three different groups. The first group consists of hydrosoluble pigments present in fraction 1; the second group consists of the F-A+ adducts [catechin-malvidin 3 glucoside (Mv3glc), along with some (catechin)2-Mv3glc]; and the third group is their anthocyanin precursor, Mv3glc.
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