Chromatographic Separation Of Alkaloids From Coptis Chinensis And Study On The Biological Activities Of Coptis Chinensis Components

Coptis chinensis is a traditional Chinese herbal medicine.Modern pharmacological research shows that Coptis chinensis is an important source of alkaloid drugs and has a good effect in the treatment of diseases,such as diabetes and tumors.The main medicinal components of Coptis chinensis are alkaloids.Chromatographic purification of alkaloids has problems,such as tailing and low sample loading.The technical difficulties have hindered the basic research of chemical components in Coptis chinensis and the development of Coptis chinensis in the medical field.In order to solve these problems,this paper established a method for the analysis of alkaloids in Coptis chinensis based on surface electrostatic repulsion/inverse phase mixed mode chromatography with a self-developed charged C18 HCE column.The symmetrical peak shape and separation selectivity of the main alkaloids in Coptis chinensis in chromatographic analysis were realized.The main chromatographic peaks of Coptis chinensis were recognized by the established method and references,which were coptisine,epiberberine,columbamine,jatrorrhizine,Berberine and palmatine,respectively.Quantitative determination of total alkaloids was carried out on Coptis chinensis native to Hubei and Chongqing.On the basis of optimizing the High Performance Liquid Chromatography(HPLC)method for the determination of alkaloids in Coptis chinensis,preparative purification method of total alkaloids of Coptis chinensis was established with Preparative High Performance Liquid Chromatography(Prep-HPLC)and membrane separation technology.The purity of total alkaloids of Coptis chinensis could be increased from 39.62% to 88.87%,and the recovery rate of total alkaloids was 94.5%.The prepared fractions were screened for biological activity against G proteincoupled receptors including D2,Mu,M3,5-HT2 A,GPR120,FFA1.Fraction Fr.1 showed specific muscarinic acetylcholine receptor(M3)agonistic activity.This is the first reported M3 activity in Coptis chinensis extracts.Further purification of fraction Fr.1 was carried out by following M3 agonistic activity.The current research solved the difficulties of analysis,separation and purification of alkaloids from Coptis chinensis with liquid chromatography.An analytical HPLC method for the determination of alkaloids in Coptis chinensis was established.The method was able to achieve symmetrical excellent peak shape and selectivity of alkaloids in Coptis chinensis.Based on this,a Prep-HPLC method for the preparation of total alkaloids from Coptis chinensis was established.Such preparation increased the purity of total alkaloids from Coptis chinensis from 39.62% to 88.87%.The purification process met the requirements of quality conservation.A fraction exhibiting M3 agonistic activity was identified.These experimental results provide a novel separation and purification preparation method for the study of Coptis chinensis,which is of great significance for the study of the basic and pharmacological activities of Coptis chinensis.At the same time,it can provide some reference for the study of other basic compounds.