Bioactivity And Separation Of Polyphenols Of Kudingcha Made From Ilex Kudingcha C.J.Tseng

Kudingcha is a kind of tea drank by people from north and east part of China Ilex kudingcha C.J. Tseng is a one of the main breed varieties of kudingcha and its active ingredients as well as health protection functions have been more and more concerned in recent years. The source of Ilex kudingcha C.J. Tseng is considerable in China, but can hardly been popularize as daily beverage because of its bitter taste. If we can make further processing of the plants, extract the bioactive compounds, increase the add value, there will be important economic value and practical significance.The bioactivity of kudingcha in antioxidant (in vitro and in vivo), anti-obesity and anticancer were studied. Macroporous resins were employed to purify polyphenolic compounds, also the constituents were charicteristiced by HPLC and LC-MS. The results were as follows:In vitro antioxidant activities of Ilex kudingcha C.J. Tseng were determined by the DPPH (1,1-diphenyl-2-picrylhydrazyl free radical), ABTS (2,2’-azino-bis(3-ethylben zothiazolin-6-sulfonate) diammonium salt) and reducing activity. The results showed that polyphenols from Ilex kudingcha C.J. Tseng is a potential antioxidant. Antioxidant activities in vivo of kudingcha polyphenols were determined by using the animal model experiments. Results indicted that administration of low dose polyphenols could overcome oxidant injury induced by D-Gal, and treatment of high dose could inhibit significantly the formation of MDA, enhance significantly the activities of antioxidant enzymes (SOD, CAT and GSH-Px) and increase significantly the value of TAOC in livers and serums in a dose-dependent manner.The anti-obesity activity of polyphenols extracted from Ilex kudingcha CJ. Tseng was evaluated through its inhibitory ability towards α-amylase, pancreatic lipase and a-glucosidase. The results showed that kudingcha polyphenols can strongly inhibit activity of α-amylase (IC50=18μg/mL), followed by lipase. The inhibitory ability towards a-glucosidase was relatively lower.Proliferation inhibition of human liver cancer Hep-G2and gastric cancer BGC-823cells (tumor inhibition rate,%) were studied in vitro by MTT assay. Different concentration of Ilex kudingcha C.J. Tseng was employed to test the effects of polyphenols towards cancer cells. The results showed that the tumor inhibition rate depended on the dose of polyphenols provided, and under the same condition, with the increasing of the concentration, the inhibition rate increased.750μg/mL kudingcha polyphenols could inhibit80%Hep-G2cells and lmg/mL could inhibit88.94%BGC-823cells.Macroporous resins were employed to purify polyphenolic compounds from Ilex kudingcha C.J. Tseng. HP20has been chosen from7different resins. According to Langmuir and Freundlich absorption curves, conclusion could be observed that HP20serves as suitable resin for polyphenolic compounds purification. The optimized pH values for sample and slovet were4and10. The best volume for elution solvent is6BV. The polyphenol of Ilex kudingcha C.J. Tseng was analyzed by HPLC and LC-MS. The results indicate that the main phenolic substances in kudingcha are caffeyol qunic acid (CQA) derivatives, including3-CQA,3-CQA,5-CQA (CHA),3,4-diCQA,3,5-diCQA and4,5-diCQA. And after macroporous purify,3-CQA,3-CQA,5-CQA (CHA) were obtained eluting by water, while3,4-diCQA,3,5-diCQA and4,5-diCQA were gained eluting by ethanol.