Separation of Regaloside A£¬Acetylregaloside C and Regaloside B in Bulbs of Lilium Species by High Speed Counter-current Chromatography and Research on Their Biological Activities

KeyWord:separation  Lilium  phenolic glycerol glycosides  high speed counter-current chromatography(HSCCC)  activities

Abstract:

      A method of high speed counter-current chromatography was developed for the separation and preparation of three active compounds£¬phenolic glycerol glycosides from the bulbs of Lilium species used as Chinese tradition medicine.The ethylacetate-n-butanol-water(0.5% acetic acid£¬3¡Ã1.5¡Ã5£¬by volume) was used as a two-phase solvent system£¬in which the upper phase was stationary phase and the lower phase was mobile phase for the isolation of the compounds from the extract of Lilium brownii F.E.Brown var.viridulum Baker by high speed counter-current chromatography(HSCCC).Additionally£¬the antioxidant activities on free radical scavenging and lipid peroxidation£¬and the effects on ¦Á-amylase activity were evaluated.Results showed that 4.2 mg of compound 1(96.2%£¬purity)£¬2.3 mg of compound 2(95.1%) and 5.8 mg of compound 3(98.8%) were obtained from the 250 mg extract by one-step using high-performance liquid chromatography(HPLC).The chemical structures of the compound 1-3 were identified as regaloside A£¬acetylregaloside C and regaloside B£¬respectively by mass spectrometry(MS) and nuclear magnetic resonance imaging(NMR).Biological activity test showed that acetylregaloside C exhibited a strong capacity scavenging DPPH and inhibiting lipid peroxidation£¬while regaloside A and regaloside B had weak capacity scavenging DPPH and definite inhibitory effects on lipid peroxidation.The antioxidant effects of the compounds were as follows:acetylregaloside C>regaloside A¡Öregaloside B.The promoting effects of the compounds on the activity of ¦Á-amylase were different in different concentrations£¬but no significant difference existed among the compounds