Study On Extraction Of Effective Ingredients And Active Filter From Red Clover And Uncaria Tomentosa

Natural medicine are getting more and more attention in the worldwide because of the large range of activity and advantage of side effects. However, due to the complexity of natural product composition and function mechanism is not clear, to the extent that make the development of the natural medicine is restrained. It is vital that separation analysis and activity evaluation in the chemical composition of natural medicine.Based on red clover and uncaria as the research object by using high performance liquid chromatography(HPLC), electros pray mass spectrometry and high-speed countercurrent chromatography to analysis chemical components of red clover and uncaria. Ultrafiltration mass spectrometry as screening to inhibition of alpha-glycosidase and xanthine oxidase in red clover. PC12 cells as the active filter system evaluation method to analysis of chemical composition in uncaria on PC12 cell protection. The specific experiments and results are as follows:Adopting LC/ESI/MS technique to analysis identified the chemical composition of red clover ethanol extracts. Total separation identified nine chemical composition: daidzein, biochanin A, genistein, pratensein, 4’-methyoxycoumestrol, formononetin, irilone, prunetin and maackiain. N-hexane: ethyl acetate :ethanol :water : 2.5:4:4:5(v: v: v: v) as the solvent system by using high-speed countercurrent chromatography. Total separation identified five chemical composition: daidzein, prunetin, genistein, irilone and maackiain. By using high performance liquid chromatography(HPLC) to test their purity, the purity of them are : 95.65%, 97.25%, 96.35%, 97.25% and 98.65%. In addition, red clover as the research object and ultrafiltration mass spectrometry as study method to screening the inhibition of alpha-glycosidase and xanthine oxidase. It turns out that composition of red clover has strong combines ability and inhibits alpha glycosidase, such as genistein, formononetin, irilone and maackiain. Above the compositions have the potential activity of resistance to type II diabetes. Daidzein, biochanin A, genistein, irilone and maackiain combines with xanthine oxidase have strong ability, so that inhibit the xanthine oxidase activity. Above the compositions have the potential activity of gout.Adopting LC/ESI/MS technique to analysis identified the chemical composition of uncaria. Total separation identified six alkaloid compositions are ursolic acid, rhynchophylline, corynoxine B, isorhynchophylline, corynoxine and dihydrocorynatheine. Using accelerated solvent extraction and response surface analysis method to study separation and extraction method for effective components of uncaria. The best extraction condition is when ethanol concentration was 50%, extraction times 3 times, the extraction time 12 min. The extraction of rhynchophylline as the goal to extract reached to 6.121 mg/g. Chloroform-methanol-water: 5:4:6(v:v:v) as the solvent system by using high-speed countercurrent chromatography to isolated three chemical compositions, respectively, rhynchophylline,corynoxine B and corynoxine. By using the high performance liquid chromatography(HPLC) to test purity, the purity of them are 96.15%, 95.34% and 95.49%. Utilizing the PC12 cells model of MPP+ induced to evaluate the cell protection function of effective ingredients in uncaria. The results show that: the relative increment rate of three dose groups in isorhynchophylline(66.57, 98.03, 101.69%) is higher than rhynchophylline(66.85, 93.82, 100.56%), but less than the mixture of the two components(68.26,101.69, 105.05%). Respectively, corynoxine and corynoxine B of three dose groups of relative increment rate were 68.26, 84.55, 91.57% and 65.44, 92.13, 97.75%. The three dose groups of dihydrocorynatheine and cadambine of mixture fraction is relatively low, thus the high dose group have not reached 50%.