The Study On Stability Of Tridition Chinese Medicine By Capillary Electrophoresis

Safety, effective and quality control are the basic request of pharmaceutical preparation. The stability shoud be studied for that the safety and effective are assured by it. Medicine may be influenced during the process of prepare and storage, leading to the decline of drug action and increase toxicity. The change of the property by the influence should be studied to esure the therapeutic effect and safety.Chinese Medicine is widely used in clinic, which is directed by the theory of Tridition Chinese Medicine (TCM). Nowadays, only one active component in TCM is usually used as index to exosyndrome in the study of stability of TCM, following the model of quality control of chemical drugs. However, there are multiple active components in TCM so that it's unreasonable to analyze one component to control the quality. According to TCM theory, compatibility is just the essence of TCM. The change of compatibility and effective can be reflected by the change of components content. The analysis of multi-index components not only reflects the level of the production technics of TCM and the quality of each herb, but also reflects the condtion of its compatibility. So multi-index components content and proportion should be determined in TCM to study the change of stability by the time.Under the variable mode of Capillary Electrophoresis (CE), different kinds of components in TCM could be assayed easily at the same time. In this study, CE was selected to study the method for assay of the multi-index components and the stability of TCM in order to find the change of proportion. The contents of multi-index components of some TCM were studied, such as Xiao'er Qingre Zhike oral liquid, Chailian oral liquid and Qingkailing oral liquid, etc. The change of some components content and their proportion in Shuanghuanglian oral liquid and Yinhuang oral liquid were studied by the method to find whether TCM could keep the interaction of multipe components. From the study, the conclusion can be reached that: (1) Different kinds of multipe components in TCM could be determined by the variable mode of CE technique. (2) The content of some components and their proportion in Shuanghuanglian oral liquid and Yinhuang oral liquid could be affected by high temperature. The target of the interaction of multipe components in TCM had been changed. By the theory of TCM, effective of TCM had also been changed.PART 1 The Study of the CE Method for Determination the Contents of the Multi-index Components of TCM(1) Simultaneous determination of the contents of 3 components in Xiao'er Qingre Zhike oral liquid by capillary electrophoresis Objective: A capillary electrophoresis method was established for determination the contents of ephedrine, baicalin and glycyrrhizic acid simultaneously in Xiao'er Qingre Zhike oral liquid to find whether CE method could be selected to control the quality of TCM and study the stability of TCM. Methods: An uncoated fused-silica capillary column of 50 cm×50μm (effective length: 45cm) was used. The method was developed by utilizing an injection by press (5s), a running voltage of 10kV and a 25mmol·L-1 sodium tetraborate electrolyte solution at pH 8.60. The detection wavelength was set at 192 nm.Results: The calibration curves were in good linearity in the range of 4.00×10-3~0.225mg·mL-1 for ephedrine, 2.40×10-3~1.50mg·mL-1 for baicalin and 2.10×10-2~1.33 mg·mL-1 for glycyrrhizic acid. The mean recoveries of them were 98.3%, 98.4% and 97.1% respectively.Conclusion: CE method is simple, rapid and accurate, could be used for the quality control of TCM, and could be used in the study of Stability of TCM.(2) Determination of L-ephedrine, D-pseudephedrine, rutin, and cinnamic acid in Chailian oral liquid with CZE Objective: A simple, reliable, reproducible method, based on capillary zone electrophoresis (CZE), has been established for the determination of L-ephedrine, D-pseudephedrine, rutin, and cinnamic acid in Chai-lian oral liquid to find whether CE method could be selected to control the quality of TCM and study the stability of TCM.Methods: Electrophoresis was performed in a 40cm×50μm bare fused-silica capillary column, of which 35cm was the effective length for separation under the running voltage of 10 KV at the room temperature. A 25 mmol·L-1 borax buffer and 10% absolute alcohol at pH 8.50 was applied as running buffer. The method was developed by utilizing an electrokinetic injection for 5 s and detected at 192 nm.Results: A good linearity between the peak area and the concentration was found in the range of 30.0~196μg·mL-1,51.0~328μg·mL-1,43.0×~276μg·mL-1,31.0~204μg·mL-1with all correlation coefficients more than 0.9987. With this simple and rapid method, four components in Chai-lian oral liquid were determined simultaneously with recoveries of 98.3%~99.4%.Conclusion: A capillary electrophoresis method was developed for the simultaneous determination the contents of L-ephedrine, D-pseudephedrine, rutin, and cinnamic acid in Chailian oral liquid. Under the optimize conditions, the four analytes were completely separated within 12 minutes. CE method is simple, rapid and accurate, could be used for the quality control of TCM, and could be used in the study of Stability of TCM. (3) Simultaneous determination of Baicalin and Chlorogenic acid in Qingkailing oral liquid by CEObjective: A capillary electrophoresis method was established for determination the contents of ephedrine, baicalin and glycyrrhizic acid simultaneously in Qingkailing oral liquid to find whether CE method could be selected to control the quality of TCM and study the stability of TCM.Methods: An uncoated fused-silica capillary column of 40 cm×50μm (effective length: 35cm) was used. The method was developed by utilizing an injection by press (5s) with a running voltage of 15kV and a 25mmol·L-1 sodium tetraborate electrolyte solution at pH 8.50. The detection wavelength was set at 254nm.Results: The calibration curves were in good linearity in the range of 0.119~7.62mg·mL-1 and 3.40×10-2~2.18 mg·mL-1 with all correlation coefficients more than 0.9992. The mean recoveries of them were 100.2% and 97.1% respectively.Conclusion: A capillary electrophoresis method was developed for the simultaneous determination Baicalin and Chlorogenic acid in Qingkailing oral liquid. CE method is simple, rapid and accurate, could be used for the quality control of TCM, and could be used in the study of Stability of TCM. PART 2 The Study of the CE Method for the stability of TCM(1) The Study of the CE Method for the stability of Shuanghuanglian oral liquidObjective: A capillary electrophoresis method was established for determination Baicalin, Chlorogenic acid and forsythin in Shuanghuanglian oral liquid. The method was used to inspect the effect of high temperature on the content of 3 components in Shuanghuanglian oral liquid.Methods: 3(1) item in the stability principle of TCM was followed: Three batch samples were put in an airtight clean container and the temperature was kept at 60 centigrade for 10 days. The samples were taken out at zeroth, fifth and tenth day. Baicalin, Chlorogenic acid and forsythin were determined to inspect the change of the 3 componets content and their proportion.Electrophoresis condition An uncoated fused-silica capillary column of 40cm×50μm (effective length: 35cm) was used. The method was developed by utilizing an injection by press (6s) with a running voltage of 10kV and a 40mmol·L-1 sodium tetraborate-40mmol·L-1 SDS electrolyte solution at pH 8.30. The detection wavelength was set at 192nm.Results: The content of Baicalin, Chlorogenic acid and forsythin in 3 batch samples were all decreased. The proportionality of 3 componets was also changed.Conclusion: TCM is used in clinic by the interaction of multipe components by the theory of TCM. After put in 60℃for 10 days, The content of Baicalin, Chlorogenic acid and forsythin in Shuanghuanglian oral liquid were all decreased. The proportion of 3 componets was also changed. The TCM application was not consistent with the theory of TCM and could not insure the safety and effective.(2) The Study of the CE Method for the stability of Yinhuang oral liquidObjective: A capillary electrophoresis method was established for determination Baicalin and Chlorogenic acid in Yinhuang oral liquid. The method was used to inspect the effect of high temperature on the content of 2 components in Yinhuang oral liquid.Methods: 3(1) item in the stability principle of TCM was followed: Three batch samples were put in an airtight clean container and the temperature was kept at 60 centigrade for 10 days. The samples were taken out at zeroth, fifth and tenth day. Chlorogenic acid and Baicalin were determined to inspect the change of the 2 componets content and their proportion.Electrophoresis condition An uncoated fused-silica capillary column of 40cm×50μm (effective length: 35cm) was used. The method was developed by utilizing an injection by press (6s) with a running voltage of 20kV and a 40mmol·L-1 sodium tetraborate electrolyte solution at pH 8.30. The detection wavelength was set at 310nm.Results: The content of Baicalin and Chlorogenic acid in 3 batch samples were all decreased. The proportionality of 2 componets was also changed.Conclusion: TCM is used in clinic by the interaction of multipe components by the theory of TCM. After put in 60℃for 10 days, The content of Chlorogenic acid and Baicalin in Yinhuang oral liquid were all decreased. The proportion of 2 componets was also changed. The TCM application was not consistent with the theory of TCM and could not insure the safety and effective.