Study On The Extraction, Separation Technology And Bioactivity Of Effective Components From Eucalyptus Grandis X E.Urophylla

In this thesis, eucalyptus oil, saponin and tannin from Eucalyptus grandis x E.urophylla were extracted by water. Analysis of the composition of the extraction, and their activities were studied. The main research contents and results as follows:(1) The eucalyptus oil of leaves from Eucalyptus grandis x E.urophylla was obtained through steam distillation, and identified by GC-MS method. The results showed that oil extraction of Eucalyptus grandis x E.urophylla was0.39%(mass fraction),56compounds of eucalyptus oil were identified. The major constituents in oil were1,8-cineole36.36%, followed by a-pinene27.04%, a-terpineyl acetate9.65%, a-terpineol5.07%, limonene2.21%, camphene2.14%,2-the camphene alcohol2.04%, perillyl alcohol1.09%, fenchol1.00%.(2) The extraction and separation process of saponin were studied. Based on the results of single factor experiment, response surface method was used to optimize the extraction process of saponin. The optimum extraction conditions were obtained as follows:extraction temperature61.8℃, time5.02h and ratio of liquid to solid as20.09:1, the yield of saponin was5.49%. (3) The crude extract saponins were purified by n-butanol alcohol separation, macroporous resin adsorption and the macroporous resin adsorption combined with acetone precipitation method. The results showed that three kinds of separation method obtained the purity of20.11%,31.64%and50.36%, respectively.(4) The antioxidant activity of the saponin from the residue of Eucalyptus grandis x E.urophylla were first studied. The results demonstrated that the maximum scavenging rates of crude saponins for1,1-Diphenyl-2-picrylhydrazyl radical, hydrogen peroxide and superoxide anion radical were83.02%,80.10%and16.62%, respectively. In the same systems, the maximum scavenging rates of the purified saponins of the macroporous resin separation were84.20%,55.02%and13.43%, and the maximum scavenging rates of the purified saponins of the macroporous resin adsorption combined with acetone precipitation were85.62%,48.21%and10.01%, respectively.(5) The extraction and separation process of tannin were studied. Based on the results of single factor experiment, response surface method was used to optimize the extraction process of tannin. The optimum extraction conditions were obtained as follows:extraction temperature52.46℃, ultrasonic power279.14w and extraction time41.94min, the yield of tannin was4.12%. Tannin in crude extract was4.01%and increased to53.12%when was chromatograhied by macroporous resin. (6) The antimicrobial activities of tannin on multiple common bacteria were studied by using filter paper method. Tannins had strong inhibition on the7kinds of bacteria activities but no distinct inhibition on Excherichia coli. Minimun antimicrobial concentration of the purified tannins on Staphyloccocus aureus、Yeast and Shigella was1.0mg·mL-1, and the antimicrobial activities against Live Bacillus Cereus Preparation and Salmonella were stronger than their effects against positive control.(7) The maximum scavenging rates of crude tannins and purified tannins for H2O2were80.00%,89.12%when mass concentration was0.50mg·mL-1, and their IC50were0.11mg·mL-1and0.12mg·mL-1, respectively. The maximum scavenging rates of Vc, crude tannins and purified tannins for DPPH·were98.43%,88.97%and92.21%. When mass concentration was4.00mg·mL-1, the highest scavenging rate of crude tannins and purified tannins for·OH were87.52%and94.32%.