Extraction And Purification Process,Activity Determination And Composition Analysis Of Polyphenols And Flavonoids From Cup Plant

Stems、leaves and flowers of flowering cup plant(Silphium perfolialum L.)are characterized by rich flavonoids and polyphenols.As a widely available alternative bioenergy plant,it was still used as a forage.The purification process of polyphenols and flavonoids was rarely studied.Therefore,it was of great significance for the development of clinical therapeutics in the future to extract polyphenols and flavonoids from the purified strychnos and to identify their chemical constituents.Using response surface method the best extraction technology conditions were determined to investigate cup plant extracts in vitro antioxidant properties.The extracted polyphenols and flavonoids were purified by resins,and the main purified components were analyzed by ultra-high performance liquid phase.(1)The optimal extraction conditions of polyphenols were liquid-to-solid ratio of29∶1(mL/g),extraction time of 94 min,extraction temperature at 39 ℃ for 60% with alcohol concentration.The mean value of polyphenols was 24.60 mg/g.The optimal extraction conditions of flavonoids were liquid-to-solid ratio of 31 ∶1(mL/g),extraction time of 130 min,extraction temperature at 38 ℃ for 42% with alcohol concentration.The mean value of flavonoids was 38.81mg/g.(2)Antioxidant activity of polyphenols and flavonoids in cup plant test showed that extracts of the group were better able to eliminate DPPH free radicals and ABTS free radicals.Therefore the cup plant had strong radical scavenging capacity and a certain ferric reducing power.(3)Polyphenols test results indicated that the best macroporous resin was AB-8,adsorption time was 4h;adsorption capacity was 2.72 mg/g.The upper sample concentration was 3mg/mL,the pH was 4,the volume of the upper column was 100 mL,the sample velocity was 2BV/h,the eluent was 60% ethanol concentration solution,the elution rate was 2BV/h,and the volume of washout was 200 mL.We measured under this condition the concentration of the polyphenols from cup plant was increased from 9.5% to 22.4% after purification.(4)Flavonoids test results showed that he best macroporous resin was D101,sorption time was 2h;adsorption capacity was 13.12 mg/g.The upper sample concentration was 3 mg/mL,the pH was 4,the volume of the upper column was 180 mL,the sample velocity was 2BV/h,the eluent was 80% ethanol concentration solution,the elution rate was 2BV/h,and the volume of washout volume was 200 mL.We measured under this condition the concentration of the flavonoids from cup plantwas increased from 34.1% to 45% after purification.(5)The combination of polyphenols and flavonoids lyophilized powder from cup plant by UPLC-Triple-TOF/MS was determined to contain 10 components and the content proportion:Ⅰ neochlorogenic acids(2.66%);Ⅱ chlorogenic acid(10.45%);Ⅲimplicit chlorogenic acid(3.7%);Ⅳ 3-coumaroyl acid(1.38%);Ⅴ isochlorogenic acid B(0.91%);Ⅵ isochlorogenic acid C(2.27%);Ⅶ kaempferol3-O-apiofuranosyl-rhamnopyranosyl-galactopyranoside(1.96%);Ⅷ kaempferol3-O-rhamnopyranosyl(1-2)-galactopyranoside(3.32%);Ⅸ kaempferol 3-Orhamnopyranosyl(1-6)-galactopyranoside(3.09%);Ⅹ astragalin(1.91%)。...