Isolation and Purification of Six Bioactive Compounds from the Seeds of Trigonella foenum-graecum L. using High-Speed Counter-Current Chromatography

Abstract

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, ¹ H-NMR, and ¹³ C-NMR aided in structure identification.

Keywords:

• flavonoids
• high-speed counter-current chromatography
• polyhydroxystilbenes
• preparative isolation
• Trigonella foenum-graecum L