Study On The Activity Screening And Extraction,Isolation Of The Chemical Compounds From Radix Paeoniae Alba And Garcinia Mangastana L

Chinese natural medicines were approved over the world due to their clinical effect. However, the active ingredient in traditional Chinese medicines is not clear, the development and use of traditional Chinese medicine has been limited. Therefore, the separation, extraction and the activity evaluation of the pure compounds from Chinese medicinal herbs are necessary. The chemical compositions from the medicinal herbs are very complex, therefore, the rapid separation, extraction and structural analysis of the chemical compounds from medicinal herbs are very important. High performance liquid chromatography coupled with mass spectrometry(HPLC/MS) is a most widely used technique for the separation and identification of components from natural product. High speed counter current chromatography(HSCCC) and high performance centrifugal partition chromatography(HPCPC) are both the “liquid – liquid” partition chromatography, the superiorities of fast separation speed, large injection volume, and low sample recovery of HSCCC and HPCPC enables the fast separation and extraction of the chemical compounds from medicinal herbs. In the dissertation, the HPLC/MS, HSCCC and HPCPC were usedd for the analysis and separation methods of the chemical compounds from Paeonia lactiflora and Garcinia mangostana. The PC12 cells were used for the screening the potential active compounds for the anti-Parkinson ′s disease from P. lactiflora, and the ultrafiltration mass spectrometry was used for the screening the potential active compounds for the anti- diabetes and gout from G. mangostana.P. lactiflora is the root of the plant of Paeonia lactiflora Pall., the Latin name of Radix Paeoniae Alba. The reflux was used as the extraction method, and then the HSCCC and HPCPC were used as the separation of the chemical compounds from P. lactiflora. In the study, ethyl acetate–n-butanol–ethanol–water at a volume ratio of 1:3.5:2:4.5 was used as the two-phase solvent system for HSCCC and HPCPC, five compounds including the albiflorin, benzoylpaeoniflorin, paeoniflorin and galloylpaeoniflorin with the isolation rates of 0.86,1.82,0.85 and 1.32 mg/g were separated from P. lactiflora. The chemical structures were identified by electrospray ionization mass spectrometry(ESI/MS),(Nuclear Magnetic Resonance Spectroscopy,(NMR). High performance liquid chromatography(HPLC) was used to analysis the purities of the compounds separated. The result indicates that the purities of the fractions 2, 3 and 4 which were separated by CCC are 96.31 %, 96.46 % and 95.12 % respectively. The purities of the fractions 1, 2 and 4 which were separated by CPC are 95.84 %, 95.31 % and 97.86 % respectively. The PC 12 cells screening system were used to evaluate the protection effect of PC 12 cells induced by MPP+. The result indicates that the under the protection of the extract of P. lactiflora, PC12 cells were restore the original shape, the cell damage was weakened, and the living cells were increased. Mitochondrial succinate dehydrogenase activity were increased, the value of LDH release and Ca+ ion concentration were reduced., the results show the extract of P. lactiflora has a remarkable protective effect for PC12 cells which were induced by MPP+.G. mangostana is the peel of the plant of Garcinia mangostana L. The reflux was used as the extraction method, and then the HSCCC was used as the separation of the chemical compounds from G. mangostana. In the study, n-hexane–acetonitrile–ethyl acetate–water–methanol at a volume ratio of 5:5:2:3:1 was used as the two-phase solvent system for HSCCC, five compounds including the 3-isomangostin, 8-desoxygartanin, mangostanol, gartanin and α-mangostin with the isolation rates of 1.12,1.22,1.38,0.85 and 2.87 mg/g mg/g were separated from G. mangostana. The chemical structures were identified by electrospray ionization mass spectrometry(ESI/MS),(Nuclear Magnetic Resonance Spectroscopy, NMR). High performance liquid chromatography(HPLC) was used to analysis the purities of the compounds separated. The result indicates that the purities of the five fractions which were separated by CCC are 94.35 %, 95.45 %,94.28 %,98.68 %�'� 97.54 % respectively. The ultrafiltration mass spectrometry was employed as the active screening technique, α–glucosidase and xanthine oxidase were used the target enzyme to screening the α–glucosidase and xanthine oxidase inhibitors from G. mangostana. The result indicates that 3-isomangostin, 8-desoxygartanin, mangostanol, gartanin and α-mangostin were theα–glucosidase and xanthine oxidase inhibitors.