High-speed counter-current chromatography (HSCCC) was successfully used for the isolation and purification of 1,2,3,4,6-penta-O-galloyl-beta-D-glucose from the ethyl acetate extract of the leaves of Acer truncatum Bunge using a two-phase system composed of n-hexane-ethyl acetate-methanol-water at a volume ratio of (0.25:5:1:5, v/v/v/v) for the first time. Each injection of 80 mg crude extract yielded 7.25 mg of pure 1,2,3,4,6-penta-O-galloyl-beta-D-glucose. High-performance liquid chromatography (HPLC) analyses of the CCC fraction revealed that the purity of 1,2,3,4,6-penta-O-galloyl-beta-D- glucose was over 95%.