Preparative Isolation and Purification of Garcinol by High-Speed Countercurrent Chromatography / 世界科学技术-中医药现代化

ABSTRACT

This study aimed at preparing the chemical reference substance of garcinol.A preparative high-speed countercurrent chromatography (HSCCC) was adopted for the isolation of garcinol from the twigs of Garcinia multifiora Champ,G.esculenta Y.H.Li and the fruits of G.xanthochymus Hook.f.ex T.Anders.The crude extracts were separated by HSCCC with two phase solvent systems composed of n-hexane-ethyl acetate-95％ ethanol-water (8∶8∶12∶4,volume ratio) using the upper stationary phase and the lower mobile phase.Under the conditions of a flow rate of 2.0 and 4.0 mL· min-1,and the apparatus rotation at 850 rpm,the column temperature at 25℃ and the detection wavelength at 254 nm,the fraction containing garcinol was purified by the Sephadex LH20 chromatography.The purity test in the extracts was determined by high-performance liquid chromatography.As a result,the contents of garcinol were 45,281 and 4080mg respectively separated from 104.765 g twigs of G.multiflora Champ,105.270 g G.esculenta Y.H.Li and 102.318 g the fruits of G.xanthochymus Hook.f.ex T.with the purity of 98.72％,98.36％ and 98.42％.Compared with the sample pretreatments and separation efficiency of the three plants,it was found that the fat-soluble extracts rapidly and efficiently extracted using n-hexane-ethyl acetate-95％ ethanol-water (5∶5∶5∶5,volume ratio) contained abundant garcinol taking fruits of G.xanthochymus Hook.f.ex T as the raw material with the combination of HSCCC and Sephadex LH-20 chromatography.In conclusion,it was indicated that the combination method is efficient with high operability and large preparation quantity.