Separation of five flavonoids from tartary buckwheat (Fagopyrum tataricum (L.) Gaertn) grains via off-line two dimensional high-speed counter-current chromatography.

An off-line two dimensional (2D) high-speed counter-current chromatography (HSCCC) strategy was successfully used for preparative separation of five flavonoids from tartary buckwheat (Fagopyrum tataricum (L.) Gaertn) grains with different solvent systems for the first time in this paper. n-Hexane-ethyl acetate-methanol-water 3:5:3:5 (v/v) was selected as the first dimension solvent system to purify quercetin (4) and kaempferol (5). The second dimension solvent system, ethyl acetate-n-butanol-water 7:3:10 (v/v), was used to isolate quercetin 3-O-rutinoside-3'-O-β-glucopyranoside (1), rutin (2) and kaempferol 3-rutinoside (3). The purities of these compounds were all above 96.0% and their structures were identified through UV, MS and (1)H NMR. The results indicated that the off-line 2D HSCCC is an efficient technique to isolate flavonoids compounds from grains.