High-speed counter-current chromatography (HSCCC) was used for the separation of anthocyanins on a preparative scale from bilberry fruit crude extract ( Vaccinium myrtillus, Ericaceae) with a biphasic solvent system composed of methyl tert-butyl ether– n-butanol–acetonitrile–water–trifluoroacetic acid (1:4:1:5:0.01, v/v). Each injection of 500 mg crude extract yielded 130 mg of pure delphinidin-3-O-sambubioside, and 77 mg of pure cyanidin-3-O-sambubioside. The two anthocyanin disaccharides with a glucose (2 → 1) xylose unit are novel compounds for bilberry fruit, and were elucidated by means of electrospray ionization MS–MS, 1 H , 13 C , distortion less enhancement by polarization transfer NMR, and two-dimensional HC-correlation experiments (heteronuclear multiple bond correlation and heteronuclear multiple quantum correlation).