Extraction,Purification,Composition Analysis And Anti-hepatocellular Activity Of Scutellaria Barbata D. Don Polysaccharide

Object:To optimize the extraction process of the polysaccharide from Scutellaria barbata D.Don and to further explore the specific components of polysaccharide with anti-hepatocelluar carcinoma activity.Methods:The crud polysaccharide was extracted by hot water extraction and alcohol precipitation,and the extraction process was optimized by respose surface analysis.The crud polysaccharide were purified by ion and gel column chromatogragy after deproteinization and decolorization.The protein content,polysaccharide content,molecular weight,monosaccharid composition and infrared spectrophotometry of the purified polysaccharides were determined.Astragalus polysaccharide was used as the positive control group.The vitro inhibitory effect of polysaccharide on HepG2 cells were detected by MTT assay.Results:1.The optimal extraction conditions were as follow:solid to liquid ratio of 1:25,extraction time of 2h,temperature of 100℃.Under these conditions,the average extraction yield of polysaccharide was(3.85±0.13)%.SBP-1 and SBP-2 were obtained by ion column elution,and SBP-1A and SBP-2A were obtained by gel column elution.2.The results of chemical composition analysis showed that the polysaccharide content of SBP-1A was 93.2%,and SBP-2A was 95.5%.SBP-1A protein content was 2.87%,SBP-2A protein content was 0.87%.The monosaccharide composition of the two components were Fru,Galn,Rha,Ara,Glcn,Gal,Glc,Xyl,Man,but the molar ratio of SBP-1A was 0.6:0.3:0.6:30.6:3.3:38.4:16.1:8:1.4,SBP-2A was 0.6:0.5:0.8:36.3:4.4:42.7:9.2:3.6:0.7.Molecular weight determination of two components are presented single peak symmetry,instructions for high purified uniform polysaccharide components,molecular weight of SBP-1A was 1.15×10~5Da,and SBP-2A was 1.4×10~5Da.Infrared analysis results show that SBP-1A and SBP-2A a polysaccharide characteristic absorption peak,contain uronic acid,polysaccharide has the characteristic absorption peak of glucan,polysaccharide of sugar residue are mainly pyranose.3.In vitrol anti-tumor experiments showed that both SBP-1A and SBP-2A had inhibiting effects on HepG2 cells.Compare with the positive control group,SBP-1A high-dose group,SBP-2A medium-dose group and SBP-2A high-dose group had extremely significant differences.The cell inhibitory rate of SBP-2A was 41.94%.Conclusions:1.RSM was used to optimize the extraction conditions of polysacharides from Scutellaria barbata:solid-liquid ratio of 1:25,time of 2h,temperature of100℃,the average extraction yield was(3.85±0.13)%.2.SBP-1A and SBP-2A were isolated and purified as relatively pure homogeneous polysaccharide fractions.3.SBP-1A and SBP-2A were of higher purity,and the polysaccharide content of SBP-2A was higher.The monosaccharide composition of the two fractions was the same,but the molecular weight were different.4.Both SBP-1A and SBP-2A had inhabiting effects on HepG2 cells,and SBP-2A had more significant inhabiting effect on HepG2 cells.