Study On The Chemical Constituents Of Wuwei Jiangya Recipe And The Basic Research Methods Of Pharmacological Substances Based On Metabolism

The research of pharmacodynamic material basis has been the emphasis and difficulty of the study of the multicomponent traditional Chinese medicines(TCMs)all the time,is also the foundation and the only way to clarify the methanism of active action and the compatibility of formulae.Besides,the research of pharmacodynamic material of the TCMs can also provides reference for formulation optimization and making scientific technology of preparation.On the one hand,illuminating the chemical composition is the key point for the research of pharmacodynamic material of TCMs;on the other hand,the complexity of the multicomponent TCMs decides it's action mode in the body,which is different from chemical medicine with single component.Metabolism plays a important role of the existence form of the TCMs in the body.The components exposed in the body is possibly the real active components.With the development of analysis technology,the metabolism research of TCMs is springing up,and the U/HPLC-MS/MS technology has been the important tool for the analysis of TCMs metabolism.In order to find the effective method for the the research of pharmacodynamic material of the TCMs,the study analyzed the chemical components and the metabolic components,utilized the research idea of single component-single herbs-fomula at the same time.The main contents and results were described as follows:1.The study established the UHPLC-MS/MS analysis method of WUWei Jiangya Formula(WWJF)and it's herbs.At first,the concentration of sample and electrospray ionization(ESI)source ion mode of positive or negative was determined based on response;secondly,the chromatography separation method with good resolution was established.In order to fully detect the information of the chemical components of WWJF,WWJF was detect in both positive and negative ion mode.Flavonoids.diterpenoid quinines?phenolic amides had good response in positive ion mode,and phenolic acids had good response in negative ion mode.As a result,a total of 77 compounds were unambiguously identified or tentatively characterized,including 36 isoflavones,ll diterpenoid quinines,20 phenolic acids,3 phenolic amides,2 alkaloids,3 organic acids and 2 flavanols.This part developed a comprehensive understanding of the multiple chemical components in WWJF and could offer chemical basis for the further study.2.The study established the metabolism analysis model of WWJF:used in situ and in vivo model respectively to expound metabolism of WWJF.As a result,there were 25 components of the 77 identified component of WWJF in plasma samples obtained by intestinal perfusion with venous sampling model,14 components of the 77 identified component of WWJF in plasma samples obtained by intragastric administration model.The results shows that some components of WWJF are difficult to be absorbed by gut wall or absorbed by form after the metabolism of gut wall enzymes?intestinal bacteria enzymes or hepatic enzymes.The results indicates that the metabolic processes in living organisms significantly influence the composition of material basis of WWJF and the active component could be not only prototype conponents,but also metabolized components of WWJF.3.The study analyzed metabolic process and metabolites in rat of puerarin?3'-hydroxypuerarin?daidzin?daizein?genistein from Puerariae Lobatae Radix,guided by the sequential metabolism theory and the research idea of single component-single herbs-fomula,meanwhile attempted to explain the reason of effectiveness of trace amount of multiple components of TCMs from the point of view of metabolism.As a result,except for puerarin,the four single components was seriously metabolized before traveling to the systemic circulation,speculating the effective part of multicomponents of TCMs maybe the form after metabolism.In addition,daidzin?genistein could be transformed into daidzein after metabolism,3'-hydroxypuerarin could be transformed into puerarin after metabolism.There were metabolic conversion between components from the same herb,indicating the concentration of the components from TCMs can increase through metabolism,explain the mechanism of effectiveness of trace amount of multiple components of TCMs from the point of view of metabolism.