The Study On Pharmacodynamic Material Basis Of Shexiang Baoxin Pill

The traditional Chinese medicine (TCM) is the essence of Chinese traditional culture and world traditional medicine, which make indelible contribution to people's health in worldwide. As the complex chemical constituents in TCM and the unknown active substance in vivo, studying on the action mechanism, the process of biotransformation and metabolism will be aimless, which also has severely constrainted the modern research of TCM, and resulted in the low export rate and weak competitiveness of TCM. To change the disadvantages of TCM, the effective substances of TCM should be revealed preferentially.Serum pharmacology and serum pharmacochemistry were proposed in the 1980's, which provides a new improving method for analysis of the effective substances of TCM. Though the TCM contains complex chemical constituents, it is considered that only the compouds which can be absorbed into blood have effect on taget tissue. No matter how the constituents in TCM are directly absorbed into blood by digestive intake or by liver metabolism, which transported to taget tissue through blood circulation and show effect on target are the real effective substances. What the serum pharmacochemistry studies conformes to the theory of integrity and synergistic effect of TCM.This thesis studied the pharmacodynamic chemical basis in Shexinag Baoxin Pill (SBP) by using the serum pharmacochemistry method. The coupling technique of HPLC-DAD-ESLD, HPLC-DAD-MS/MS and GC-MS make it easier to analyze the effective substances in SBP.To provide convenience for identification of constituents in the rat plasma after oral administration of SBP, the chromatographic method should be established and the main constituents in SBP should be identified firstly. As SBP containes not only volatile constituents but also amount of nonvolatile constituents, the coupling technique such as HPLC-DAD-MS/MS and GC-MS were used to analyze the main constituents in SBP. By optimizing chromatographic conditions of HPLC-DAD-MS/MS and analyzing the obtained data, we identified 47 compounds including ginsenosides, bufadienolides, cholic acids and some other constituents in SBP. By searching the data obtained from GC-MS in NIST05 database, 49 volatile compouds were identified. By using the coupling technique HPLC-DAD-MS/MS and GC-MS, the main constituents in SBP were identified and the results lay a good foundation for further study on the effective substances in SBP.By using the established analysis method of HPLC-DAD-MS/MS and GC-MS, we optimized the administration dosage of SBP and the pretreatment conditon of plasma. Then we established the analysis method of serum pharmacochemistry for SBP. From a comprehensive analysis of the HPLC-DAD-MS/MS chromatography of SBP, controlled plasma and dosed plasma, 25 components were observed in rat's plasma. Among them, 17 of the 21 original form compounds from SBP and 3 of the metabolites were identified by comparing their retention time, MS and MS/MS spectra with the reported literature and reference standards. By using the GC-MS, 10 peaks were obseved in the dosed plasma. Through the research in NIST05 database, all of them were identified including 6 original form compounds from SBP and 4 metabolites. Most of these identified compouds were reported to have effect on cardiovascular disease, which may represent the effective substances of SBP.According to the result of serum phamacochemistry, we developed a novel and sensitive method to determine 8 main effective substances in SBP by using a HPLC/DAD/ELSD method simultaneously. The quantitative analysis was not only expected to elevate the quality control standards of SBP but also make the clinic application of SBP more effective and reliable, which also provide a new research method for quality control of TCM formlea.By using the methodology of serum pharmacochemistry, a valid method was establish for the study of the absorbed components in rat plasma after oral administration of a traditional Chinese medicine-SBP, which clarified its real effective substances. The elucidation of the effective substances of SBP provided helpful chemical information for further pharmacology and active mechanism research.