Chemical Constituents And Bioactivities From Pholidota Cantonensis Of Fujian

Objective:(1)Separates and purificates the extracts from Pholidota cantonensis dissolvede by petroleum ether and EtoAc in the way of column chromatography and HSCCC.Then indenty chemical and physical componention and structure;(2)Preparation of refined crud polycaccharide and characterization;To assess the biological activity of refined polycaccharide;To elucidate new pharmacological action and new medicinal value and furnish the theoretical basis for developing and exploiting new drug from Pholidota cantonensisMethods:(1)Silica gel column chromatography was developed for the preparative separation on petroleum ether extract.High-speed countercurrent chromatography(HSCCC)combined with silica gel column chromatography was developed for the preparative separation on a combination of ethyl acetate extracts.(2)The materials after separation and purification are identified by HPLC,are determinated melts point and are analysised structure by Infrared Chromatography. Mass Spectrometry determinate its molecular weight.After that,investigated the structure of compounds;(3)After the crude polycaccharide was extracted with hot water and precipitated with ethanol,protein and pigment were also removed.DEAE-cellulose chromatography was used to separate crude polycaccharide,further purification was carried on by SephadexG-200 chromatography.Finally,it's homogeneity was proved by HPLC.(4)The chemical structure of polycaccharide was analyzed by using GC,IR spectrum.The monosaccharide and linkage of polycaccharide were through chemical analyzation.(5)Through in vitro antioxidant and anti-lipid peroxidation experimental liver, the immune function in vitro experiments,inhibition experiments and experimental mice analgesic,anti-oxidation of the polysaccharide,enhance immunity,such as pain and inhibit bacteria biological activity.Results:(1)After purified by silica gel column chromatography,extract from petroleum ether has accept two compounds.The solvent system was ether:ethyl acetate=20:1.After purified by silica gel column chromatography and HSCCC, extract from ethyl acetate has accept-ted two compounds.(2)Melting point detector,IR spectrum,MS spectrum will be carried out.The four compounds are identified,They are Eicosane,Abundance,4,4'- dimethoxy-2,2'-dimethoxy(biphenyl) and 1,3-diamine 5,6-dihydro-7-fennel.They are gotted from Pholidota cantonensis first time.(3)The content of total sugar is 14.01%.The crude polysaccharide was fractionated by column chromatography DEAE-52,Sephadex G-200,and the refined polysaccharide was obtained.It was proved to be homo- geneous polysaccharide on molecular weight.(4)IR spectrum analysis showed that it has characteristic absorption peaks of polysaccharide,the GC analysis of its component monosaccharides for xylose, mannose,glucose,galactose,the molar ratio are 13.33:24.13:5.59:3.10.Periodate oxidation and Simth degradation showed that the polysaccharide must has 1→6 connect, 1→2 and 1→3 connections.and the highest ratio is 1→2connection.(5)The study of antioxidant activity of polysaccharides in vitro showed effect, when polysaccharide concentration is 3.0 mg/ml,the hydroxyl radical scavenging rate is 33.7%which showed a dose-dependent manner;when the polysaccharide concentration is 2.0 mg/ml,the superoxide anion clearance rate is the highest value of 70.7%.When polysaccharide concentration is 1.5 mg/ml of lipid peroxidation inhibition rate is 97.06%.Cell culture in vitro showed that the polysaccharide could promote transformation of spleen lymphocyte.Polysaccharide possesses the analgesic effect.Conclusion:HSCCC and column chromatography are used to separate and purify extracts from Pholidota cantonensis dissolved by petroleum ether and ethyl acetate. Four compounds have been got.After purification,homogeneous heterosaccharide was obtained.It has good anti-oxidation action and hepatoprotective.activities and can also enhance the immunity activities.