Separation And Analysis Research On A Few Kinds Of Active Medicine Ingredients By Micellar Electrokinetic Capillary Chromatography

Micellar Electrokinetic Capillary Chromatography(MECC) combined capillary zone electrophoresis(CZE) with micellar solubilization possesses several advantages as rapid, high efficiency, simplicity operation and application in separation of charged particles and neutral molecules. As a result, MECC has been widely used in areas of pharmaceutical analysis, biochemistry and life science.This dissertation provided an overview on the theory of micellar electrokinetic capillary chromatography and the current studies of MECC on the analytical research of Chinese traditional medicine. Based on the overview, several MECC methods have been developed for the determination of active ingredients in Chinese traditional medicine. The results showed as followed:1. A MECC method has been developed for the separation of paciltaxel,7-Epi taxol,7-Epi-10-deacetyltaxol and cephalomannine. The separation conditions were respectively optimized by orthogonal experimental design and single factor analytical approach. The running buffer solution(pH9.36) consisted of20mmol/L sodium tetrabroate,40mmol/L SDS,40.0%(V/V) acetonitrile. Instrumental conditions are25kV separation voltage,25℃column temperature, PDA detector at211nm wavelength, injecting sample5s under0.5psi. Under the optimized conditions, These four substances were completely separated within20min. The linear range for paciltaxel is2.5-250μg/mL, and for7-Epi taxol,7-Epi-10-deacetyltaxol and cephalomannine all are0.5-250μg/mL.The determination of these four activities extracted from Taxus chinensis samples are satisfied. The relative standard deviation is less than5%and the standard recoveries is between95%and105%.2. Adopting SDS as surface active agent,a new MECC method has been developed for the separation of catechin, aesculetin, scopoletin, caffeic acid and ursolic acid after the separation conditions were respectively optimized by orthogonal experimental design and single factor analytical approach. These five substances can be completely separated within20min in22kV separation voltage, using20mmol/L sodium tetrabroate(pH9.11),20mmol/L SDS,15mmo1/L β-cyclodextrin,5.0%(V/V) acetonitrile as running buffer. The linear range for catechin, aesculetin and scopoletin all are0.5-100.0μg/mL,for caffeic acid is1.5-100.0μg/mL,and for ursolic aid is2.5-100.0μg/mL. The result of determination of real samples indicated that this new MECC method is rapid and reliable.The relative standard deviation is less than5%and the standard recoveries is between95%and105%.3. In consideration of Artemisinin is difficult to be charged as well as the light absorption capacity of Artemisinin and Dihydroartemisinin is weak, a field amplified sample accumulation MECC method with PDA detector was developed for the separation of Artemisinin and Dihydroartemisinin. Using a buffer solution(pH9.11)consisted of20mmo1/L sodium tetrabroate,20mmol/L SDS,15mmo1/L β-cyclodextrin, and20kV separation voltage,25℃column temperature, PDA detector at195nm wavelength, injecting sample5s under0.5psi after injecting high purity water10s with the same pressure to preconcentrate on line of Artemisinin and Dihydroartemisinin. Under the above conditions, Artemisinin and Dihydroartemisinin were separated with in12min, and the linear range are respectively60-300gμg/mL and65-300μg/mL.The result of determination of Artemisia apiacea from different districts are satisfied.The relative standard deviation is less than5%,and the standard recoveries is between98%and104%.4. A new MECC method has been developed for the separation of loureirin A and B at20kV separation voltage,25℃column temperature, PDA detector at211nm wavelength, injecting sample5s under0.5psi. Using a buffer solution(pH9.98)consisted of20mmol/L sodium tetrabroate,10.0%(V/V) acetonitrile,5.0%(V/V) ethyleneglycol,1.0%(V/V) butanol. The concentration and pH of run buffer, organic additive and the instrumental conditions were optimized by single factor analytical approach. After the separation condition was optimized, the separation and determination of resina draconis extracted from different districts are satisfied.The standard recoveries is between95%and105%,and the relative standard deviation is less than5%.