Chemical constituents from Curcuma xanthorrhiza Roxb. (Zingiberaceae) rhizome were isolated and purified by column chromatography (silica gel, RPsilica gel MPLC, HPLC) and their structures were elucidated on the basis of spectral analyses. Six compounds were identified by MS and NMR spectroscopy as zederone(Ⅰ), βsitosterol(Ⅱ), Guai-1(10),3,5,7(11),8-pentaen-2-on-11,8-olide(Ⅲ), Curcumadione(Ⅳ), Curcumenone (Ⅴ), Cuparenol (Ⅵ). Compound Ⅲ and Ⅵ were isolated for the first time from the plants of Zingiberaceae, and compound Ⅳand Ⅴ were isolated from Curcuma xanthorrhiza Roxb. for the first time. Meanwhile, the acetylcholinesterase inhibitory activity of the isolated compounds was evaluated. Compounds Ⅳ and Ⅵ showed certain inhibitory activity, the minimum inhibitory quantity of Ⅳ on TLC bioautography test was 95μg.