Isolation and Preparation of Teasaponin Monomers from Tea Seed Cake by High-Speed Counter-current Chromatography

Objective:To develop a rapid and efficient method for preparing teasaponin monomers from tea seed cake using preparative high-speed counter-current chromatography (HSCCC). Methods:A crude extract rich in teasaponin monomers was obtained from tea seed cake by microwave-assisted extraction,and preliminarily purified by D-101 macroporous resin chromatography,followed by further HSCCC purification/fractionation with a binary phase system made up of ethyl acetate,n-butanol and 3% acetate (1:4:4,V/V) at a flow rate of 1.5 mL/min with rotation at 800 r/min under the conditions of 267 nm detection wavelength and 100 mg sample load. The final purification product was analyzed by HPLC. Results:Two teasaponin monomers with 99.1% and 94.5% purities were obtained after the HSCCC separation,and their dry weights were 11 mg and 15 mg,respectively. Conclusion:The proposed HSCCC method has the characteristics of simplicity,rapidity and high product purity,thereby providing a novel strategy for the separation of teasaponin monomers.