Study And Application Of Capillary Electrophoresis In Tibetan Medicines Active Constituents

Particular tableland plants in Qinghai and Tibetan altiplano are substantial foundation of Tibetan medicines. Strong radiation and long sunlight of Tibetan medicines plants leads to doughty photosynthesis, high accumulation of the effective components and strong bioactivities. At present, it exist obvious confine in analysis of quality and distinguishing of Tibetan medicines, which can't assure their qualities and exploit new Tibetan medicines. Capillary electrophoresis (CE) is a new analysis technology. It provides advantages in terms of higher efficiency, faster separation time, less injected sample volume and more practicable operating modes. But CE has seldom been applied to the analysis of real Tibetan medicines, because the detected sensitivity of CE is lower than HPLC and the content of active constituents in Tibetan medicines is very small. So, Correlative study is developed in theory and methods about application of capillary electrophoresis in active components of Tibetan medicines.In the second part of this dissertation, the advantages of CE were made use of to separate and determinate active components in Gentianopsis paludosa (Ma). On the basis of the previous literatures, in this dissertation, some original studies are carried out as followings:1. A novel micellar electrokinetic chromatography(MEKC) method has been developed for the simultaneous separation of 1,7-dihydroxy-3,8-dimethoxyxanthone and 1,8-dihydroxy-3,7-dimethoxyxanthone, and the proposed method has been successfully applied to determine the two xanthones in Gentianopsis paludosa (Ma).2. A novel nonaqueous capillary electrophoresis method has been applied to separate and determinate the content of ursolic acid and oleanolic acid in Gentianopsis paludosa (Ma)This dissertation consists of three parts and four chapters.Part I Review of the development of capillary electrophoresis and Tibetan medicine and application of CE in xanthone and terpenoids compounds.Chapter 1: The principle and characteristics of CE were simply described. Study actuality of Tibetan medicines and application of CE in xanthone and terpenoids compounds were reviewed and discussed in detail.Part II Study and application of CE in Tibetan medicine Gentianopsis paludosa (Ma).Chapter 2: A novel micellar electrokinetic chromatography(MEKC) method was developed for the simultaneous separation and determination of 1,7-dihydroxy-3,8-dimethoxyxanthone and 1,8-dihydroxy-3,7-dimethoxyxanthone in flower and stem of Tibetan medicine Gentianopsis paludosa (Ma). The optimum conditions were: 20 mM borate - 20 mM SDS at pH 9.80, 260 nm detected wavelength and 17.5 kV applied voltage. The proposed method was not only applied for analying the two xanthone of Tibetan medicine Gentianopsis paludosa, but also an efficient technology for the preliminary study on them in other Gentianopsis paludosa plants.Chapter3: A novel nonaqueous capillary electrophoresis method was developed for the separation and determination of ursolic acid and oleanolic acid in Tibetan medicine Gentianopsis paludosa (Ma). The optimum conditions were: 25 mM Tris -10% acetonitrile (v/v) in methanol - 1.5% acetic acid (v/v) at 214 nm UV detected wavelength and 25 kV applied voltage. It was found that flowers are the most effective parts of Gentianopsis paludosa in extracting the effective compounds.Part III Study and application of CE in Yinhuang preparations.Chapter 4: A novel, simple, sensitive, fast free solution capillary electrophoresis (FSCE) method was developed for analysis of baicalin and chlorogenic acid in Shuanghuang lian oral fluid, Yinhuang granule and Yinhuang tablet. The optimun conditions were composed of n_borate: n_NaH2PO4=1:1 (pH 8.0) at 334 nm UV detected wavelength and 15 kV applied voltage. In optimum conditions, baicalin and chlorogenic acid in three Yinhuang preparations were separated and identified within 4.3 minutes.