Evaluation Of Germplasm Resources On Astragali Radix Based On Saccharide Mapping Of Polysaccharide And Structural Analysis Techniques

RATIONALES: Astragalus Root(RA),is one of 8 herbs from Shanxi which selected from the national authentic herbal medicine library,the amount of medicine is huge,known as "10 party eight stilbene".Currently on the market,RA can be divided into traditional RA(imitation wild cultivation)and cultured RA(nursery transplanting planted).Tu Pengfei and others make analyze of RA with different grades in accordance of the current evaluation standards.The conclusion is the quality of cultured RA was better than the traditional RA.However,the international market and some veteran TCM physicians still accept natural RA.These conflicts show that the current quality evaluation method cannot fully accurate reflect the quality of RA.Therefore,a more rational evaluation method is urgently needed for the development of genuine medicinal materials.As the main substances responsible for immunomodulatory activity,Polysaccharides was also the most abundant compounds of RA.However,due to the current phenol-sulfuric acid method to measure the total polysaccharide content of the saccharide which lacks species specificity and cannot fully reflect the characteristics of polysaccharides.In recent years,with the development of technology — saccharide mapping which is based on saccharide hydrolysis.Saccharide mapping with species specificity which can be used for qualitative and quantitative analysis of polysaccharides and also can be used for species identification and quality evaluation.OBJECTIVE: To construct the astragalus polysaccharide glycosides,to screen out differentially sugar fragments of different germplasm resources,and make the structural analysis and activity evaluation.Finally,the contentdetermination method of the peculiar active fragments and evaluation of different germplasm resources of RA.METHOD&CONTENT : Establishment the fingerprints of soluble polysaccharides and glycoconjugates based on partial acid hydrolysis and evaluation of different germplasm resources of RA:The partial acid hydrolysis of RA polysaccharides from different germplasm resources was carried out,and the hydrolyzate was analyzed by PACE and HPTLC to obtain a series of fingerprints.Through the data analysis,the differentiated sugar fragments of RA were screened out.Structure analysis of RA was determined by GC-MS,IR and NMR.And the RA peculiar fragments were obtained by comparing the active polysaccharides reported in the literature.The peculiar active fragments were obtained by vitro antioxidant capacity and immunological activity and the determination method of the content of active sugar was used to evaluate the different germplasm resources of RA.In addition,the establishment of fingerprints of soluble polysaccharides and glycoconjugates based on enzymatic and artificial gastric hydrolysis,which can used to identify different RA germplasm resources.RESULTS:The establishment of fingerprints of soluble polysaccharides and glycoconjugates based on partial acid hydrolysis and evaluation different germplasm resources of RA.Results indicated that the molecular weight of Astragalus polysaccharides(APS)range from 540 Da to 900 Da is the main segments to distinguish the growth patterns of RA and the molecular weight range from 360 Da to 540 Da is the main segments to distinguish the species of RA.Structural analysis showed that the difference was mainly composed of glucose and Mannose and xylose,and the possible structure of ASP-1 of Astragalus membranaceus was ? 2,3,6)-D-Glcp-(1 ?,D-Xylp-1 ? and? 4)-D-Manp-(1 ?.)Among them,Glc and Man constitute the main chain,the main chain configuration is ? type.Through comparison has been reported in the literature to confirm the final Astragalus difference of sugar fragment ASP-1 for Astragalus exclusive The results showed that it had a low antioxidant activity in vitro,but it could induce the inflammatory reaction and had the immunomodulatory effect.Finally,the active sugar fragment of RA was screened out by the experimental study.The results showed that DP3> 3.0 mg/mL,DP4> 3.2 mg/mL in traditional RA,DP3 <3.0 mg/mL,DP4 <3.2 mg/mL.DP3> 3.0 mg/mL,DP4> 3.2 mg/mL in traditional RA and DP4 <3.0 mg/mL,DP4 <3.2 mg/mL in cultured RA.Establishment of fingerprints of soluble polysaccharides and glycoconjugates based on enzymatic hydrolysis and identification of different germplasm resources RA: To digest the ASP from different germplasm resources with 12 kinds of enzymatic to obtain a series of PACE fingerprints,data analysis results found that each enzyme hydrolysis of ASP can produce a series of sugar fragments,which can be used to identify different RA germplasm resources.Data analysis shows that the molecular range of540-1100 Da is the main segments to distinguish the growth patterns of RA and the molecular weight range of 700-900 Da is the main segments to distinguish the species of RA.Establishment of fingerprints of soluble polysaccharides and glycoconjugates based on artificial gastric hydrolysis and identification of different germplasm RA resources: Simulated artificial gastric conditions to treat ASP obtain a series of PACE fingerprints,Data analysis shows that the molecular range of 540-1100 Da is the main segments to distinguish the growth patterns of RA and the molecular weight range of 700-900 Da is the main segments to distinguish the species of RA.CONCLUSION:In this project,we obtained the peculiar saccharide of RA by analyzing the data of the saccharide fingerprints of soluble polysaccharides and glycoconjugates based on partial acid hydrolysis.By structure analysis and pharmacological activity test of peculiar saccharide,the peculiar active saccharide of RA was screened out.The determination method of the peculiar active saccharide was established and used as an evaluation index for different RA's germplasm resources.The research results will elucidate the features structures of saccharide from different varieties of RA and improved the evaluation system of RA,which lay a foundation for the protection and development of high quality RA,as well as explore new ways for objective appraisal of plant medicinal materials' germplasm resources.