It is difficult to simultaneously isolate and purify flavonoids and polyhydroxystilbenes due to their similar polarities. In this study, we isolated and purified these compounds from Trigonella foenum-graecum L. seed extracts using high-speed counter-current chromatography (HSCCC) in two steps with two different kinds of solvent systems. In the first step, n-hexane:ethyl acetate:methanol:water (1:5:1:5, v/v) was used as the two-phase solvent system, desoxy-rhaponticin and rhapontigenin were purified, and orientin, vitexin, isovitexin, and rhaponticin were eluted together. In the second step, ethyl acetate:water (1:1, v/v) was used as the two-phase solvent system, and orientin, vitexin, rhaponticin, and isovitexin were isolated and purified. Purities of the isolated compounds were determined using HPLC, while UV spectra, H- 1 -NMR, and C- 13 -NMR aided in structure identification.