| This dissertation was divided into four chapters.In Chapter 1,Literature review.This part summarized the latest research progress of modern research and resource utilization of Abelmoschus manihot.Based on the content of polysaccharides in the plant,the study mostly stopped at the crude polysaccharide stage.Therefore,the progress of separation and purification of plant polysaccharides and the main biological activity were summarized.In order to improve the use of polysaccharides resources of A.manihot.In addition,Based on the analysis of the A.manihot of the main chemical composition,distribution of resources,utilization status and so on,it can improve the overall utilization rate of A manihot,and provide reference and guidance for the utilization of waste during the production and processing.In Chapter 2,Analysis and utilization value discussion of the multiple chemical composition in different tissues of A.manihot.To analyze the resourceful chemical composition in different tissues(root,stem,leaf and flower)and the dreg of flower of A manihot and evaluate their utilizing value.The flavonoids,soluble polysaccharides,cellulose,nucleosides and amino acids in the different tissues of A.manihot were determined by HPLC,UV-Vis spectrophotpmetry,Weende and UPLC-TQ/MS.The flowers are rich in the resourceful chemical compositions of flavonoids which mainly consist of hyperoside,isoquercitrin,cotton-8-O-glucuronide,myricetin,quercetin-3'-O-glucoside,rutin and quercetin.The total content of these flavonoids is 25.450 mg/g in the flowers,while they are trace in the other tissues.Different tissues of A.manihot are rich in soluble polysaccharides and celluloses,and the stems have the highest content(10.86%)of soluble polysaccharides,while the roots have the highest content(29.88%)of cellulose.Total of 21 amino acids and 9 nucleosides were detected in the plant,and the flowers have the highest content of amino acids(4.737 mg/g),while the leaves have the highest content of nucleosides(1.474 mg/g).A.manihot is rich in the resourceful chemical compositions,and its constituents and contents are various in different tissues of this plant.There is also a small amount of ingredient in the dreg of flower,which may be related to insufficient extraction The results provided a scientific basis for the utilization and industrial development of A.manihot plants.In Chapter 3,Studies on the chemical constituents of the seeds of A.manihot.The chemical constituents the seeds of A.manihot(L.)Medic.were determined by GC-MS,UV-Vis spectrophotpmetry,Weende,BCA and UPLC-TQ/MS.The seed of A.manihot is rich in fatty acids,soluble polysaccharides,celluloses,soluble protein,amino acids and nucleosides,The total content of the fatty acids is 10.12%,and the unsaturated fatty acids accounted for 78.01%-79.40%of all these fatty acids.The contents of soluble polysaccharides,celluloses and soluble protein are 6.53%?6.68%,12.77%?14.26%,and 10.36%?14.51%,respectively.Total of 19 amino acids and 7 nucleosides were detected in the seed,and the content of amino acids is 10.08%?10.15%,in which essential amino acids are accounted for 3 8.42%?39.40%.The content of nucleosides in the seed was determined as 3.01?3.11 mg/g.The results provided a theoretical basis for the resourceful utilization of the seed of A.manihot.In Chapter 4,Study on the resource recycling of A.manihot.Based on the study of the previous chapter,found that the existence of the use of the value of the resource components in the different tissues and organs of A.manihot.In particular,the content of polysaccharides in the stems and leaves of A.manihot is higher.And the market also has A.manihot gun(a part of polysaccharide)for sale,used as food additives.However,most of the studies on polysaccharides remain in the crude polysaccharide stage.In addition,the residue of A.manihot were mostly abandoned in the actual production,resulting in a great waste,in order to make better use of polysaccharide and residue resources in the A.manihot,do the following research:(1)Isolation and purification of stem and leaf polysaccharide from A.manihot and its structural characterization.Isolation and purification of polysaccharides from stem and leaf ofAmanihot.Study on the bestextraction process of polysaccharide from stem and leaf of A manihot by orthogonal experiment.In the single factor test,it was found that the extraction temperature,extraction time and ratio of material to liquid had the greatest effect on polysaccharide extraction rate.Therefore,the best extraction process of polysaccharide was obtained by three factors and three levels table.,the time of extraction was 3 h,the ratio of material to liquid was 1:20,the temperature of extraction was 100?,twice extraction,and verified in the follow-up experiment.On the basis of the best extraction process,the polysaccharides were extracted and purified by macroporous resin,on-line membrane interception system and DEAE-52 anion exchange resin.And finally get a homogeneous molecular weight,high purification of polysaccharide of A.manihot,named HSK-JT,the average molecular weight,weight average molecular weight and distribution coefficient were calculated by GPC molecular weight distribution software with HPGPC.The weight average molecular weight is 13821 Da and the distribution coefficient is 1.9654.The separation and purification of the whole process are based on the in vitro immunological activity,and select the best activity of the polysaccharide segment for the next step separation.(2)Evaluation of the immune activity of polysaccharides from stem and leaf of A.manihot.The immunological activity of polysaccharide was described by two methods,including in vitro and in vivo,on the basis of KSK-JT,and the introduction of crude polysaccharide to compare.Based on the immunoprecipitation of mouse splenic lymphocytes,the purified polysaccharide can promote the proliferation of mouse spleen cells in vitro and show a certain two-way dose relationship.The experimental results show that it can show some immune-enhancing activity in vitro.In addition,purified polysaccharides can also promote the production of NO with mouse monocyte macrophages RAW264.7,and with the increase in concentration,the production of NO also increased,its promotion and concentration was positively correlated,indicating that purified polysaccharides can be directly applied to RAW264.7 cells to activate macrophages,improve the phagocytosis,and promote theproduction of NO.It is suggested that the purified polysaccharide of A.manihot has immunomodulatory activity and it is a good immunomodulator.In the intervention and treatment of mouse immunodeficiency model,the mice were immunized by cyclophosphamide and then treated with three doses of crude polysaccharide and purified polysaccharide,and the positive drug was used as contrast,and the immune organ index,peripheral blood leukocyte project,serum cytokines as a reference index.In vivo experiments show that cyclophosphamide can cause mouse immunodeficiency model,and purified polysaccharides and crude polysaccharides have immune promoting effect,but there are differences.At the three dose levels,the effect of purified polysaccharides on immunodeficient mice was superior to that of crude polysaccharides,which may be related to the improvement of purity.Which for the development of immune-enhanced drugs or health food to provide a certain basis,but to be further studied.(3)Evaluation of the resourceful material of the residues of A.manihot and study on its bio-carbonization.Based on the correlation analysis of various chemical constituents in residues of A.manihot.The results showed that the residues of A.manihot contains a certain amount of cellulose,total in 13.40%.Combined with the existing bio-carbon platform in the laboratory,we found that residues of A.manihot has the potential to produce bio-carbon.So it is prepared into bio-carbon,and the relevant physical indicators were measured.In order to make better use and provide data support of the residues of A.manihot and make it into bio-carbon. |
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