Extraction, Purification And Antioxidation Of Flavonoids From Artemisia Argyi

Artemisia argyi is a kind of Composite family plant widely spread in China, and is a kind of herbal medicine which combines food nutritive value and medical value together. Artemisia argyi contains many kinds of chemical composition with physiological activation, the flavonoids among them usually has very strong antioxidant ability. The flavonoids which has a highly hydroxyl structure. It can act as an antioxidant, removes free radicals, and reduces risk of chronic diseases such as cardiovascular disease and cancer.(1) My studies firstly focused on measuring the main nutrition contents in the Artemisia argyi Leaves. The study results illustrated that a certain type of Artemisia argyi, which grows in Wuhan of Hubei Province in our country has a high rate of flavonoids. Therefore this type of Artemisia argyi is chosen as my researching material.(2) The extraction technics of flavonoids of Artemisia argyi (FA) was studied . The extraction conditions were optimized by orthorhombic analysis. The optimal conditions of extraction were as follows: the extraction time 149min, the concentration of ethanol 56%, the temperature 72℃, the rate of solution/material rate20:l, the extraction yield was more than 91.2%.AB-8 type macroporous resin is the best for separating and purificating Artemisia argyi flavonoids; the optimun conditions is: the loaded amount of Artemisia argyi extract is 16 mL , wash with water (pH 5.0), the concentration of desorption agent is 50 % (pH 5.0) ethanol. The AB-8 type resin can be used for 3 times repeatedly. The content of flavonoids in the 50% ethanol eluant is 69.37% after purification with AB-8 type resin.The purity of Ethyl Acetate extraction of FA was 78.28% ; the yields of procyanidins was 2.64% . The purity of sephadex extraction of FA was 99.16%;the yields of procyanidins was 1.89% .(3) The purity of sephadex extraction of FA were further studied by such means as UV and IR and HPLC and HPLC-ESI/MS. Study results indicate that apigenin, quercetin and chrysoeriol are the main contents in the flavonoids of Artemisia argyi(4) The antioxidant effects, abilities of scavenging and antibacteria activity of different extracts in vitro and invivo were studied.①The antioxidant study results exposes that sephadex extraction of FA is very strong antioxidant, The study showed that FA2 and FAS could effectively scavenge superoxide anion, hydroxyl radical, hydrogen peroxide and inhibit DNA oxidative damage. Compared with Flavonoids of Artemisia argyi and Vc,we could see that they had significant antioxidant activity higher than V_C. The results showed that the scavenging rate on superoxide anion of FA2 and FAS was 1.5 times and 2.7 times respectively stronger than that of Vc; The scavenging rate on hydroxyl radical of FAS was 1.3 times respectively stronger than that of Vc; The scavenging rate on hydrogen peroxide of FA2 and FAS was 1.8 times and 4.5 times respectively stronger than that of Vc; The inhibiting rate on DNA oxidative damage of FA2 and FAS was 1.3 times and 1.6 times respectively stronger than that of Vc.②The content of MDA inliver mitochondria and microsome was measured by TBA assay; hemolysis of RBC and the swelling of mitochondria were detected by spectrophotometric methods. Results: FA (0.5-10.0mg/mL) evidently reduced the level of MDA in mice liver and mitochondria, and inhibit the erythrocatalysis and the swelling extent of mitochondria of liver. It suggests that FA maybe used as an antioxidant and might effectively protect mice cell, cell from the injury of oxidation.③The experiment of mice proved that the flavonoids can enhance the activity of superoxide dismutase (SOD) in red blood cell and reduce the MDA level in liver homogenate and had a good anti-oxidation function.