To study the chemical constituents of Dioscorea opposita.The chemical constituents were isolated and purified by silica gel column chromatography, and the structures were elucidated by their spectroscopic data and physico-chemical properties.Twelve compounds were obtained and identified as palmitic acid (1), beta-sitosterol (2), oleic acid (3), beta-sitosterol acetate (4), 5-(hydroxymethyl)furfural (5), nonanedioic acid (6), beta-daucosterol (7), cyclo-(Phe-Tyr) (8), cyclo-(Tyr-Tyr) (9), 6-methyl citrate (10), 1, 5-dimethyl citrate (11), trimethyl citrate (12).Compounds 3-12 were isolated from this plant for the first time and cyclodipeptides were firstly isolated from the Dioscoreae family; Compound 9 was a new natural product.