Study On The Effective Constituents Of The Total Alkaloids From Corydalis Impatiens

Objective:We established the preparation methods of effective parts of total alkaloids in the Corydalis impatiens and separated the chemical compositions. With isolated compounds as quality control index, we determined the content of effective parts of total alkaloid compounds. And then we studied the effects of the effective parts of total alkaloid in anti-cancer experiment research. At last we improved the quality of medicinal materials standards and layed foundation for the development of effective parts of total alkaloids in the Corydalis impatiens.Methods:We got crude extracts with alkaline ethanol, then got the effective parts of total alkaloid whose content was higher than50%through purification process. Using silica gel column and recrystallization method, we separated individual compounds from the effective parts of total alkaloid, and finished the structural identification. With isolated compounds as quality control index, we established the content determination method of the effective parts of total alkaloids and finished their content determination. We studied the anticancer activity of the effective parts of total alkaloids and clarified their pharmacological activities. We established the content determination method of protopine and improved the quality of medicinal materials standard。Results:l.We established the preparation methods of total alkaloid from the Corydalis impatiens. The crude extracts was prepared through the process of alkaline ethanol extraction, enrichment and dry. The crude extract was first desolved in the1%hydrochloric acid solution, and then adjust pH to10with10%sodium hydroxide alkaline. At last we got total alkaloids whose content was higher than50%through dichloromethane extraction and silica gel column purification.2.The separation of chemical components of the total alkaloid parts was studied. Finally we got5individual compounds through silica gel column chromatography and recrystallization method. And the structures (protopine, a-bicuculline, ochotensimine, β-hydrastine,β-bicuculline) were identified according to their physical and chemical properties in the structure identification and ID-NMR, ESI-MS spectrum method. Among them, protopine, a-bicuculline,(3-hydrastine were the first time isolated from the medicinal materials.3.The content determination method of protopine, ochotensimine, a-bicuculline was studied. The methodology study suggested the method was simple and feasible, reproduced well, can be used as an effective quality control method of the total total alkaloid parts. We used this method to determine the content of the total alkaloid parts. The total amounts of protopine, ochotensimine, a-bicuculline was33%.4.The in vivo anticancer activity of total alkaloid parts was conducted. The selected tumor cells were human prostate cancer, breast cancer cells, gastric cancer cells, lung adenocarcinoma cancer, ovarian cancer cells. The results suggested that the effective parts of total alkaloids had significant inhibitory effects on breast cancer have on lung adenocarcinoma cancer cells. But the dose-effect relationship was not clear, which needed further development.5.The water, ash, insoluble ash content items were added to the medicinal materials standard. The identification item was revised. The content determination method of protopine was established, which suggested that the protopine had good linear relationship within10.45μg/ml-104.5μg/ml, R=0.9997; The average recovery was98.73%, RSD=1.06%. The method was simple and feasible, reproduced well, can be used as an effective quality control method of the medicinal materials.Conclusion:In this study, the chemical compostion of the total alkaloids preparation was clear and controllable, which also had certain anti-cancer activity. Meanwhile the quality standard of Corydalis impatiens was improved. This study was of great significance to the application of new drug development (this research had applied for national patent).