Study on Ecological Technology of Efficient Utilization of Chinese Medicine Resources Eucommia Ulmoides

Author:Cui Guo Qiang

Supervisor:yang lei


Degree Year:2019





Eucommia ulmoides is a endemic plant of China and is widely distributed throughout China.However,currently the E.ulmoides resources have not been fully developed,resulting in environmental pollution and waste of resources.In this paper,the barks of E.ulmoides as a renewable resource was used as experimental material to innovate the separation process of target components,and established a process route for comprehensive utilization of resources to achieve high efficiency,environmental protection and multi-level utilization of resources.The four active compounds in E.ulmoides barks were extracted by tea saponin synergistic cycle ultrasonic extraction method.The extracted active compounds were enriched and purified by high-speed countercurrent chromatography.The gutta percha was extracted from residues of E.ulmoides barks by using limonene as extraction solvent.DEAE-cellulose ion exchange column chromatography and glucose gel column chromatography were used to classify and purify polysaccharides of E.ulmoides barks and glycan structure analysis was carried out on polysaccharide of E.ulmoides barks components.Silica gel immobilized acidic ionic liquid catalysts catalyzes the hemicellulose of E.ulmoides barks to synthesize furfural,and the physicochemical properties and catalytic activity of the catalyst are characterized.The sulfonated carbon is prepared by the secondary residue of E.ulmoides barks as raw material to catalyze the conversion of oleuropein to hydroxytyrosol.Physical and chemical properties and their catalytic activities were characterized.This study realized the efficient use of E.ulmoides resources,and provided theoretical and data support for the ecological technology research of E.ulmoides.The main contents are as follows:The four active compounds(gentipinic acid,geniposide,genipin and pinorsinol diglucoside)in E.ulmoides barks were extracted by tea saponin synergistic cycle ultrasonic method.Tea saponin,as a natural nonionic surfactant,is non-toxic,degradable and increasing the solubility of target compounds.Cycle ultrasonic method increases the fluidity of the extraction solvent and promotes rupture of cell wall.Applying single factor experiments and Box-behnken design optimization to get the optimal process conditions:0.3%tea saponin concentration,49℃extraction temperature,10 mL/g liquid to solid ratio,490 W ultrasonic power,21 min ultrasonic time,1000 r/min stirring speed.Under the optimal conditions,the total yield of the target compounds in E.ulmoides barks was 6.62 ± 3.15 mg/g,which was significantly improved compared with the Soxhlet extraction method.This method has no similar reports by searching domestic and foreign literature.The macroporous resin and high speed countercurrent chromatography were used to prepare the four target compounds extract from E.ulmoides barks.From the data of adsorption amount and desorption rate,HPD-417 was judged to be the optimized macroporous adsorption resin,and the dynamic adsorption and elution conditions of the HPD-417 are optimized:2 BV/h loading flow rate,13 BV loading amount,40%ethanol volume fraction and 3 BV/h elution flow rate.The solvent system of high-speed countercurrent chromatography was screened.The final solvent system was:two-phase solvent system ethyl acetate-n-butanol-water(0.7:1.1:2,v/v)and(1:4:5,v/v)were used to prepare four target compounds.At the same time,the preparation conditions were also optimized.The optimized results were a mobile phase flow rate of 1.5 mL/min,a coil speed of 1000 rpm/min,and a system temperature of 35℃.The purity of the four compounds was finally obtained as geniposide(94.53%±1.05%),rosinol diglucoside(91.24%±1.23%),and geniposide.(92.14%±2.15%)and genipin(91.46%±3.24%)and the four target compounds were identified by HPLC-MS and 1H-NMR.The waste liquid produced by the preparation is ecologically treated,and the solvent is recovered by distillation to achieve the purpose of recycling.The gutta-percha was extracted from the extraction residue using limonene as the extraction solvent.Applying single factor experiments and Box-behnken design optimization to get the optimal process conditions:liquid to solid ratio of 25 mL/g,extraction temperature of 83℃,heating time of 1 h and soaking time of 4 h.The actual yield of gutta percha was 80.46±2.55 mg/g.The physicochemical properties of gutta percha extracted from petroleum ether and limonene were characterized,including GPC,FTIR,1H-NMR,TG,DSC analysis,and the kinetic curve was established.The gutta percha was extracted under the same optimized conditions,indicating that the extraction of limonene reached equilibrium.The point consumes much less time than petroleum ether extraction,and the shorter extraction time will inevitably consume lower energy and reduce energy waste.Therefore,limonene is superior to petroleum ether as solvent for extracting gutta percha.The recovered limonene was subjected to a reusable experiment.During the 6 times recycling,the average yield of gutta percha was 80±4 mg/g,which proved that the recovered limonene had good stability and can be applied to extract gutta percha repeatedly.After consulting domestic and foreign literature,there is no similar report.The deproteinization result of polysaccharides of Eucommia ulmoides barks indicated that papain combined with sevage reagent extraction performed better.DEAE-cellulose ion exchange chromatography column and sephadex gel chromatography column were applied to classify and purify the polysaccharides of Eucommia ulmoides barks.Three component polysaccharides were Obtained.High performance gel permeation chromatography was applied to determine the purity and molecular weight of polysaccharides(1,2 and 3)of Eucommia ulmoides barks indicating that the three polysaccharide components had high purity.After hydrolysis and pre-column derivatization,the glycosyl composition of high purity polysaccharide components was analyzed by HPLC,similar glycosyl composition but different contents were found.Finally,structural analysis of the three polysaccharide components was conducted by UV,FTIR and 1H-NMR.After consulting domestic and foreign literatures,there is no report to the glycosyl composition of polysaccharides of Eucommia ulmoides barks.This research lays a certain foundation for the purification,advanced structure analysis and activity study of polysaccharides of E.ulmoides in the future.Furfural was synthesized by preparing silica gel immobilized acidic ionic liquid catalyst,and the synthesized catalyst was analyzed by FTIR,TG,SEM,EDX energy spectrum.Glycosyl structure analysis of hemicellulose of Eucommia ulmoides barks was carried out by acid hydrolysis and pre-column derivatization.Ultrasonic microwave synergistic reaction extractor was used as catalyst to catalyze the preparation of furfural in Eucommia ulmoides hemicellulose.The optimal conditions for the preparation of furfural were microwave irradiation power of 500 W,microwave irradiation time of 40 min,reaction temperature of 140℃,ultrasonic irradiation power of 50 W and catalyst amount of 400 mg.Under the optimized conditions,the yield of furfural was 581.94±28.32 mg/g.When catalyst was repeatedly recovered 6 times the yield of furfural is 87.09%±3.9%of the first time.Therefore,the recovered catalyst has good stability and can be reused to prepare furfural.Secondary extraction residues of E.ulmoides barks were applied to preprare sulfonated carbon and was analyzed by FTIR,TG,SEM and EDX energy spectrum.Microwave synergistic extractor as reaction apparatus for conversion of oleuropein to hydroxytyrosol.The optimal reaction conditions for the conversion of oleuropein to hydroxytyrosol were optimized and as followed:microwave irradiation power of 500 W,microwave irradiation time of 30 min and sulfonated carbon addition of 9%.When catalyst was repeatedly recovered 6 times the yield of furfural is 89.37%±3.66%of the first time.It is indicated that the recovered sulfonated carbon has good stability and can be reused.