Extraction, Separation And Purification Of Total Flavonoids From Chicory And Its Protective Effect On Acute Alcoholic Liver Injury

Cichorium intybus L.is a perennial plant of the genus chicory in compositae.It is distributed all over the world,and its main producing areas in China are Xinjiang,Inner Mongolia and other places.Chicory can not only be eaten as a vegetable,but also be used as a medicinal material,which contains polysaccharides,flavonoids,alkaloids and other biological active substances,and has good medicinal value.Flavonoids are a kind of secondary metabolites produced by plants,which have many functions such as antioxidant activities,lowering blood glucose and lipids,protecting organs,anti-tumor,inhibiting virus,anti-inflammation,improving cardiovascular and other functions.In this study,the extraction conditions of total flavonoids from chicory roots were optimized by using response surface design.The antioxidant activities of total flavonoids from roots of chicory were investigated by the effects on DPPH,ABTS and hydroxyl radical(�OH)in vitro.AB-8 macroporous resin and polyamide resin were used to isolate and purify total flavonoids from chicory roots.The separation and purification effects under different states(static and dynamic)were investigated.The flavonoids in the extract of chicory roots were identified by ultraviolet absorption spectrum and color reaction.The physiological activities and protective effects of total flavonoids from chicory roots on acute alcoholic liver injury in mice were analyzed by intragastric administration of different concentrations.1.In this study,ultrasonic technology and the extraction method of complex enzymes(cellulase:pectinase=1:1)were used to design a single factor experiment,and Box-Behnken response surface methodology was used to optimize the extraction process of total flavonoids from chicory roots.The simulated regression equation for the yield of total flavonoids was established,and the model fitted well.The optimal conditions were determined as follows:enzymatic hydrolysis time 66 min,dosage of compound enzyme 2.2%,ultrasonic power 59 W,liquid-solid ratio 37:1 m L/g,ultrasonic time 24 min.Under these conditions,the theoretical yield of total flavonoids was 5.52 mg/g.Verify the above process conditions,the yield of total flavonoids was 5.43�0.12 mg/g,which was close to the theoretical value predicted by the model.2.The antioxidant activity of total flavonoids from chicory roots in vitro was studied using vitamin C as control.Results show that when the concentration of total flavonoids of chicory roots in 0.1 mg/m L,the scavenging rates of DPPH radical,ABTS radical and�OH radical were 84.45%,98.18%and 78.61%,respectively,IC₅₀alue concentration were 0.04,0.021,0.019 mg/m L.The total flavonoids of chicory roots showed good antioxidant activity in vitro.3.AB-8 macroporous resin and polyamide resin were used to study the separation and purification efficiency of total flavonoids from chicory roots by static and dynamic adsorption and desorption tests.The dynamic adsorption capacity,adsorption rate and desorption rate of AB-8 macroporous resin were 78.53 mg/g,84.37%and 89.15%,respectively.The dynamic adsorption capacity,adsorption rate and desorption rate of polyamide resin were 70.39 mg/g,83.18%and 69.43%,respectively.The results showed that AB-8 macroporous resin had higher adsorption capacity and desorption rate than polyamide resin.In addition,both AB-8macroporous resin and polyamide resin could improve the purity of total flavonoids from chicory roots.The purity of total flavonoids purified by polyamide resin could reach 80.2%,and the purification effect was significantly better than that of AB-8macroporous resin(45.12%).The flavonoids in the extract of chicory roots were identified by UV absorption spectrum and color reaction.The results showed that there were dihydroflavonoids,flavonoids and flavonols in the total flavonoids of chicory roots after resin separation and purification.4.The animal model of acute alcoholic liver injury was established by intragastriving the total flavonoids of chicory roots(50,100,200 mg/(kg�d)in the low-dose,medium-dose and high-dose groups,respectively).The physiological activity of total flavonoids of chicory roots in vivo and the protective effect of total flavonoids on acute alcoholic liver injury in mice were analyzed by kit detection and liver tissue section analysis.The results were as follows:1)During the whole period of intragastric administration,the mice in all groups were in good growth state,and the total flavonoids of chicory roots had no significant effect on the body weight of mice,and there was no significant difference among all groups(P>0.05).2)The liver index of mice was significantly decreased by intragastric administration of high dose of total flavonoids,but there were no significant differences in spleen,thymus and kidney indexes among groups(P>0.05).3)Compared with the model group,high and medium doses of total flavonoids from chicory roots can effectively reduce the degree of damage to liver cells of mice,thereby reducing the activities of Alanine aminotransferase(ALT)and Aspartate aminotransferase(AST)in serum and the contents of Total cholesterol(TC)and Triglyceride(TG)in serum to low levels.4)Compared with the model group,the Glutathione transferase(GSH)content and the activities of Glutathione peroxidase(GSH-Px)and Superoxide dismutase(SOD)in liver of mice in each dosage group of total flavonoids from chicory roots were maintained at higher levels,the content of Malondialdehyde(MDA)was low(P<0.05or P<0.01).5)The liver tissue sections of mice showed that,compared with the model group,the liver tissue injury degree of mice in the high-dose,medium-dose and low-dose groups was reduced,especially the high-dose group had the most obvious effect.In conclusion,pre-intervention of total flavonoids from chicory roots in mice can effectively prevent and alleviate acute alcoholic liver injury in mice,enhance the antioxidant capacity of liver tissue and protect various physiological functions of liver.