Study On The Chemical Components And Metabolism Of Fangji Huangqi Tang

Background: The chemical composition contained in Chinese Traditional compound medicine is the material basis for its efficacy.It is of great significance to carry out research on the chemical composition of Chinese traditional compound(CTC)and to clarify the constitution contained in CTC,to clarify the effective ingredients in CTC,and to establish the index component for evaluating the Chinese medicines.Fangji Huangqi Tang(FHT)was from Zhang Zhongjing's?Synopsis Golded Chamber?.The clinical applications of FHT in recent years have mainly focused on the diseases such as chronic heart failure,cardiogenic edema,rheumatoid arthritis,glomerulonephritis,and nephrotic syndrome.At present,the research on the efficacy of FHT is relative clearly,but there are less researches on the chemical composition and metabolic research in vivo.Therefore,we established the chemical composition spectrums of FHT in vitro and in vivo based on HPLC and UHPLC-Q-Exactive technology.It will provide the basis for studying the effective components of FHT.Objectives: To establish the chemical composition spectra of FHT in vitro and in vivo,and to make clear the chemical composition in FHT and the components that entered the body.Methods:(1)The fingerprint of FHT was established by HPLC-UV-ELSD,and the similarity evaluation software was used to calculate the similarity of the 10 batches of FHT,and the common peaks were attribution to single herb and identified.(2)Through referring the standards and related literatures,the chromatographic peaks were qualitative identified in the UHPLC-Q-Exactive chromatogram of FHT,and its chemical constituents was identified comprehensively.(3)The best concentration of the intestinal absorption of FHT was investigated through the experiment of the everted intestinal sac,and the differences of the absorbed components in different parts of the small intestine were compared.(4)The chemical components in blood,urine and feces after FHT administration were analyzed by using the established UHPLC-Q-Exactive method in vitro,and the chemical composition scombined with the metabolites entered the body were characterized.Results:(1)The HPLC-UV-ELSD fingerprint of FHT was established,and 20 common peaks were identified in the FHT HPLC-UV fingerprint.The common peaks were attribution to single herbs and identified.Among which,the common peaks were mainly from Stephania Tetrandra Radix,Astragali Radix,Glycyrrhizae Radix,and 9 components in FHT were identified by the standards.The method was convenient for the characterization of some chemical components in FHT,which provides a scientific evidence for the quality control of FHT.(2)UHPLC-Q-Exactive technology was used to establish the analytical method of chemical composition in FHT.With the help of the mass number of mass spectrometry,retention time,debris information,and combined with the standards,78 components were identified,including flavonoids,saponins,alkaloids and lactones.This study basically clarified the chemical composition of FHT,and laied a solid foundation for the study of chemical components in vivo.(3)Through the eversion gut sac experiment,the best concentration of intestinal absorption was 0.147 g/mL,and the difference of drug absorption components between different parts of the small intestine was compared.We found that the absorption components of jejunum were more compared with the other parts in the small intestine,and the absorption components were mainly alkaloids,flavonoids,saponins and lactones.(4)Through the analysis of the chemical components in the serum,urine and feces after FHT administration,the results showed that 10 prototype components,28 metabolites were identified in serum,21 prototypes were identified in urine and 12 prototype components were identified in feces.The results showed that the main chemical components in blood,urine and feces of rats after FHT administration were alkaloids,flavonoids and saponins.The main metabolic reactions were oxidation,reduction,methylation,demethylation,sulfation and glucuronization and the chemical components absorbed into serum and its metabolites may be the effective components of FHT.This study provides a basis for elucidating the effective components of FHT.Conclusion:(1)The HPLC-UV-ELSD fingerprint of FHT was established,and the main chemical components in the FHT were preliminarily speculated and identified.(2)The absorption components of the small intestine were identified,and the components in serum,urine and feces were analyzed.