Studies On Separation And Purification Of Chemical Constituents From Traditional Chinese Medicine Evodia Rutaecarpa (Juss.) Benth, Andrographis Paniculata (Burm.f.) Nees And So On

There are various types of traditional Chinese medicine in China.The chemical compositions of traditional Chinese medicine are very complicated,and the content of chemical compositions is less.Therefore,separation and purification of effective components from traditional Chinese medicine has a far-reaching influence,such as the research of pharmacology and the determination of content.In my master’s thesis,separation and purification of chemical constituents from Evodia rutaecarpa（Juss.）Benth,Andrographis paniculata（Burm.f.）Nees,Oroxylum indicum,and Rhizoma anemarrhenal by supercritical fluid chromatography（SFC）and preparative high performance liquid chromatography（PHPLC）.The purity of chemical constituents was determined by high performance liquid chromatography（HPLC）.Their chemical structures were identified by mass spectrometry（MS）,ultraviolet spectrum（UV）and nuclear magnetic resonance（NMR）analysis.1.Preparative isolation and purification of two kinds of alkaloids from Chinese medicinal herb Evodia rutaecarpa（Juss.）Benth by SFCIn my master’s thesis,a SFC method for isolating and purifing of evodiamine and rutaecarpine from Chinese traditional medicine Evodia rutaecarpa（Juss.）Benth was established.SFC always regards supercritical carbon dioxide（SC-CO₂）as mobile phase,and adds some organic solvents such as methanol,ethanol,isopropanol and acetonitrile to SC-CO₂ to get better separation effect.On an analytical scale,the experiment parameters were optimized including the type of the stationary phase,the type and concentration of the modifier,the flow rate of the mobile phase,the column temperature and the column pressure.Finally,the suitable conditions were obtained through a series of experiment parameters.The purity of two compounds was determined as 99.5% and 99.8% by HPLC.And their chemical structures were identified as evodiamine and rutaecarpine by UV and NMR analysis.The thermodynamics of chromatographic separation process was also studied,and it was illustrated that the chromatographic separation process was controlled by enthalpy.2.Isolation and purification of diterpene lactones from Andrographis paniculata（Burm.f.）Nees by SFCIn my master’s thesis,a SFC method for isolating and purifing of diterpene lactones from Andrographis paniculata（Burm.f.）Nees was established.SFC always regards SC-CO₂ as mobile phase,and adds some organic solvents such as methanol,ethanol,isopropanol and acetonitrile to SC-CO₂ to get better separation effect.The experiment factors such as the type of the stationary phase,the type of the modifier,the percentage of the modifier in the mobile phase,the flow rate of mobile phase,the back pressure and the column temperature were first investigated on analytical SFC.The suitable conditions were obtained through a series of experiment parameters.Then the chemical structures of the two diterpene lactones were identified as andrographolide and dehydroandrographolide by UV and NMR analysis,and their purity was measured as 99.1% and 98.5% by HPLC.Lastly,the thermodynamics of SFC separation process was studied,and the result indicated that the SFC separation process was controlled by enthalpy.3.Separation and purification of two kinds of flavonoids from Oroxylum indicum by PHPLCIn my master’s thesis,a PHPLC method for separation and purification of two kinds of flavonoids from Oroxylum indicum was established.A C18 column（250 mm×25.4 mm I.D.,10 μm）was used as separation column and methanol-water（45:55,V/V）was used as the mobile phase with the flow rate of 30 m L/min,the sampling amount was 20 mg,and the detection wavelength was set at 280 nm.Under the above conditions,two major peaks were obtained through one-step separation,and the purity was determined as 98.1% and 98.9% by HPLC.They were identified as baicalein-7-O-diglucoside and baicalein-7-O-glucoside by UV,MS and NMR analysis.4.Isolation and purification of two kinds of flavonoids from Rhizoma Anemarrhenal by PHPLCIn my master’s thesis,a PHPLC method was established for separation and purification of the chemical constituents from Rhizoma Anemarrhenal.The composition and flow rate of the mobile phase and the sample amount were optimized and the suitable conditions were listed as follows: a C18 column（250 mm×25.4 mm I.D.,10 μm）was used using methanol-water（35:65,V/V）as the mobile phase with the flow rate of 35 m L/min,the sampling amount was 27.6 mg,and the detection wavelength was 254 nm.Under the above conditions,two kinds of chemical constituents were obtained from Rhizoma Anemarrhenal,and the purity was determined as 98.4% and 99.1% by HPLC.The chemical structures were identified as neomangiferin and mangiferin by UV,MS and NMR analysis.