Separation and purification of coumarins from Heracleum millefolium by high-speed counter-current chromatography / 中草药

ABSTRACT

Objective: To separate and purify coumarins from Heracleum millefolium by high-speed counter-current chromatography (HSCCC). Methods: HPLC was used to analyze and optimize the solvent system and hexane-ethyl acetate-water-methanol (1.52.521.5) was chosen as the two-phase solvent system of HSCCC, in which the upper phase was used as the stationary phase, while the lower phase was used as the mobile phase with flow rate of 3 mL/min, the revolution speed was set at 850 r/min, and detected at 323 nm. Three compounds were obtained as crystals by vacuum concentration at 50℃, and determined through HPLC, then their structures were identified by IR, ESI-MS, 1H-NMR, and 13C-NMR. Results: Columbianetin acetate (6.3 mg), osthole (10.6 mg), and columbianadin (6.4 mg) were obtained from crude sample (300 mg) extracts and their purity were above 96.0%. Conclusion: HSCCC is a powerful technique for the rapid separation and purification of coumarins from crude extract of H. millefolium.