This project belongs to a new method and new technology for traditional Chinese medicine research. It is an applied basic research in the field of traditional Chinese medicine pharmaceutical technology and in vivo process research. After oral administration of traditional Chinese medicine decoction, in addition to the original version of the drug, which is absorbed by the body, some of the active ingredients need to be converted into active compounds under the action of intestinal flora before being absorbed, and further undergo Phase I and Phase II metabolism in the liver, thereby exerting pharmacological effects or producing certain toxic and side effects. These metabolic processes in vivo are affected by individual differences in metabolic enzymes and transporters in the body under different pathological and physiological states. In the past ten years, our research group has focused on systematic research on the metabolic transformation laws of active ingredients of traditional Chinese medicine in vivo, and used in vitro biological enzymatic transformation technology to simulate the metabolic activation mechanism of drugs in the intestine of the body, explore the targeted preparation process of active ingredients of traditional Chinese medicine, and screen out A batch of candidate compounds with stronger pharmacological activity will lay the foundation for the research of new traditional Chinese medicine drugs. The following series of achievements have been achieved: (1) A research method for the intestinal transformation and metabolism process of active ingredients in traditional Chinese medicine has been established. Using UHPLC-Q/TOF technology combined with pharmacokinetic research methods, the in vivo metabolic process of active ingredients in more than 10 kinds of traditional Chinese medicines such as gardenia, gentian, and anemarrhenae were studied, and the intestinal enzymatic activation and in vivo metabolism laws of various traditional Chinese medicine glycoside components such as gentiopicroside, icariin and anemarrhena saponin were initially clarified. This technology was applied to the study of the in vivo metabolic activation mechanism of iridoid glycosides in traditional Chinese medicine for clearing heat and promoting bile. Comprehensive use of analytical chemistry and computational chemistry techniques, it was proved for the first time that iridoid glycosides were deglycosylated in the intestine. After, they need to undergo secondary metabolism or rearrangement to produce a pharmacodynamic structure, laying a theoretical foundation for the discovery of the pharmacodynamic structure of active ingredients of traditional Chinese medicine. (2) A method for determining the content of phase II metabolites in active ingredients of traditional Chinese medicine was established. A method for the determination of the content of garniposide, icariin, and phase II metabolites of the active ingredient in compound Yinchenhao Decoction was established by using biological enzymatic hydrolysis combined with HPLC-MS/MS detection technology, and the active ingredient phase II metabolites was completed for the first time. The in vivo pharmacokinetic research has solved the technical problems in this field and provided new technologies and methods for related research by colleagues at home and abroad. (3) Based on the laws of metabolic transformation and in vitro biological enzymatic hydrolysis technology, a targeted preparation process for the active ingredients of traditional Chinese medicine was established. Simulating the metabolic and transformation process of glycoside components of traditional Chinese medicine in vivo, in vitro biological enzymatic hydrolysis and bi-phase immobilized enzyme technology have been used to realize the directional preparation of its secondary glycosides (Anemarrhena saponin A III, Baohuoside I) and aglycones (Anemarrhena sapogenin, Icariaglycone) and other active compounds, which has a high degree of orientation and specificity compared with conventional acid-base hydrolysis processes. At present, the preparation process and activity screening research of the above-mentioned traditional Chinese medicine extracts and monomer compounds have been completed, meeting the pilot production requirements, and a series of candidates with high active pharmacological effects such as liver protection and gallbladder, antifungal, and antitumor have been screened out. drug. (4) To study the correlation between the metabolic process of active ingredients of traditional Chinese medicine and bilirubin metabolic enzymes and transporters, and clarify the cholagenic mechanism of traditional Chinese medicine compound Yinchenhao Tang. Based on the metabolic laws in vivo, combined with techniques such as metabolomics and molecular information, we studied the phase II metabolic process and endogenous substances of the active ingredients of traditional Chinese medicine.(Bilirubin metabolic enzymes and transporters, etc.) proves for the first time that the mechanism of Yinchenhao Tang promoting bilirubin metabolism is related to up-regulating the levels of liver metabolic enzymes Ugt1a1 and transporters Mrp2, Oct1, Ntcp and other proteins. Its main component, garniposide and its phase II metabolites, may be the pharmacological basis for its cholagogic effect, clarifying its cholagic effect from the perspective of drug metabolism. The research results have been used in the research of new traditional Chinese medicine drugs. Based on the study of effective substances in traditional Chinese medicine compounds in vivo, the preparation process of the compound has been optimized and the quality standards of traditional Chinese medicine have been improved.