Studies On The Application Of Capillary Electrophoresis With Electrochemical Detection In The Traditional Chinese Medicine

Capillary electrophoresis(CE) is a new-style separation technique which was established and quickly developed in the early 1980s. Nowdays,CE has been widely used in the fields of chemistry,life science,pharmaceutics,envioronment analysis and so on, due to the advantages of higher separation power,shorter analysis time,less sample and reagent consumption and simplicity. Capillary electrophoresis-electrochemical detection(CE-ED) is an important complement technique of CE and expanded its application to many fields. Many medicaments and its effective components are electrochemical active,therefore,CE-ED method can be used to separation and analysis of medicaments and its effective components. In the dissertation,the methods to determine the components of pharmaceuticals by capillary electrophoresis with high frequency conductivity were studied. The contents of this dissertation include three parts.In the first part,the development,principles,detection techniques and applications of CE to the analysis of pharmaceuticals were review. In this part,we also highlights the different analysis methods of oleanolic acid and ursolic acid in the traditional chinese medicine. The research strategy and direction were made out based on the review of the development in this field.In the second part,the determination of oleanolic acid and ursolic acid in different chinese herbs by capillary electrophoresis with high frequency conductivity detection was studied. The chinese herbs include prunella vulgaris,Ligustrum lucidum Ait,cornus officinalis sieb. et zucc,Eriobotrya japonica Oldenlandia diffusa(Willd.)Roxb.,Hedyotis corymbosa ( L.) Lam. , Clematis chinensis Osbeck. The effect of buffer , buffer concentration,pH andβ-CD concentration,separation voltage and length of capillary were investigated. In research about prunella vulgaris,Good sepration was achieved using a 1.2mmol/L Triethylamine(TEA) buffer containing 0.08mmol/Lβ-cyclodextrin and pH adjusted to10.60 with Hydrochloric acid. Operating voltage was 12.5kV and temperature was maintained at 25℃. The oleanolic acid and the ursolic acid have a good linearity relationship(r1=0.9995,r2=0.9996,n=5). Average recovery of oleanolic acid was 96.7% with RSD of 2.48%;ursolic acid was 98.2% with RSD of 3.10%. It was proved that the method was reliable,simple,rapid and accurate.In the third part , the determination of ursolic acid and hesperidin in jian-wei-xiao-shi-pian by capillary electrophoresis with high frequency conductivity detection was studied. The effect of buffer concentration,β-CD concentration and length of capillary were investigated. Good sepration was achieved using a 1.2 mmol·L^-1 Triethylamine(TEA) buffer containing 0.24mmol·L^-1β-cyclodextrin and pH adjusted to10.60 with Hydrochloric acid. Operating voltage was 12.5kV. The ursolic acid and hesperidin have a good linearity relationship(r1=0.9992,r2=0.9993,n=5). Average recovery of ursolic acid was 97.7 with RSD of 2.08%;hesperidin was 97.6 with RSD of 3.21%. The method has been successfully applied to analysis of real sample,with satisfactory results.