In this study, a preparative countercurrent chromatography (CCC) method for isolation and purification of the bioactive component rutin directly from the ethanol extract of Boenninghausenia sessilicarpa was successfully established by using n-butanol-ethyl acetate-water as the two-phase solvent system. The upper phase of n-butanol-ethyl acetate-water (4:1:5, v/v) was used as the stationary phase of CCC. Under the optimum conditions, 112mg of rutin at 98.6% purity was obtained from 2.0g of the crude extract in a single CCC separation. The peak fraction of CCC was identified by negative ESI, 1H NMR, and 13C NMR.