| Eucommia ulmoides is mainly produced in Hunan,Guangxi,Guizhou,Sichuan,Shanxi,Shaanxi and other places.The bark has a history of more than 2,000 years.In recent years,it was found that Eucommia ulmoides leaf has the pharmacological effects of lowering blood pressure,blood lipid,anti-inflammatory and antibacterial.Ducommia leaves had similar chemical composition and pharmacological effects,and total flavonoids content is higher in eucommia leaves.Eucommia ulmoides leaf was included in the 2005 edition of the Pharmacopoeia,and the 2020 edition of the Pharmacopoeia leaf only included the contents of chlorogenic acid.Therefore,this study aims to fully develop and utilize eucommia leaf resources to provide reference for its food and drug development.The main work completed is as follows:Firstly,by the method of HPLC,the method of the total flavonoids,and the method of mobile phase of methanol:0.4%water phosphoric acid（50:50）;flow rate:1 m L·min-1;wavelength:360 nm;column temperature:50℃.Secondly,using eucommia leaves as raw material,by using a one-factor binding orthogonal assay,the best extraction process parameters of total flavonoids were determined:the extraction solvent was 50%ethanol;the extraction temperature was70℃;the extraction time was 2.5 h,the extraction number is 3 times;material-liquid ratio is 1 g:10 m L;static adsorption experiment were used to screen polyamide resin with D101,HPD-826,HPD-417,LK-17,BS-75,and ADS-17 large well resin,it was found that HPD-826 and polyamide resin had a better adsorption effect;HPD-826large hole resin and polyamide resin for dynamic column chromatography,compare the parameters,the purification of euchum was determined by double column chromatography,that is,HPD-826 large-hole resin was used for primary column chromatography,the loading flow rate is 1.5 BV·h-1,The eluent was 60%ethanol,The eluate dosage was 3 BV,a flow rate of 2 BV·h-1,the optimal loading quantity is 22.14mg·g-1;select the polyamide resin for the secondary column chromatography,the loading flow rate is 1.5 BV·h-1,The eluent was 60%ethanol,Elution dosage was 3times the column volume,a flow rate of 2 BV·h-1,The optimal loading quantity is15.81 mg·g-1;The purity of total flavonoids was 65.19%.Thirdly,using the solid-liquid ratio was 1:10;the number was 1;the extraction solvent was 95%ethanol for solid-liquid extraction of the column chromatography extract,Obtained 78.88%total flavonoid extract,recovery rate was 80.84%;3 batches of pilot batches of extraction and purification,The average yield of the pilot product was 1.06%,the mean recovery of total flavonoids was 81.71%,the average content of total flavonoids in the pilot product was 78.78%;Detection of the process waste liquid was found to contain a large amount of chlorogenic acid,and purified by concentration and phase extraction,the chlorogenic acid extract of 83.62%,Total amount of chlorogenic acid reached 65.17 g,the recovery rate was 74.17%.Quercetin-3-O-biercetin,isocercetin,97%.The test of the influencing factors of the extract was investigated,and it have suggested that the packaging of flavonoids was dried and protected from light.Fourthly,the anti-inflammatory activity was evaluated by measuring the inhibition effect of NO generation.The results showed that 18.14%crude extract,78.78%,rutin,isquercetin-3-O-sambu disglycoside,and NO production of 78.78%,while monomer compound was>milquercetin-3-O-Sambu disglycoside>rutin>isoquercetin.To sum up,this paper determined the content of flavonoids,and studied the pilot process,the process is stable and feasible,finally to the flavonoids extract and monomer flavonoids anti-inflammatory activity test,found that the flavonoids extract has a good anti-inflammatory,will provide data support. |
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