Study On The Chemical Constituents Of Gandou Decoction And Metabonomics In Vivo

Objective:Gandou Decoction(GDD)is a famous formula,which has been clinically used for the treatment of HLD,initially recorded in"The diagnosis and treatment guidelines of Hepatolenticular degeneration".While the substantial foundation and pharmacological mechanism of GDD was still unclear,which hampered the systematically research and development of GDD.Our study was mainly focused on systemically analysising the chemical compounds and the majormetabolites in rats after oral administration of GDD by UPLC and UPLC-Q-Tof-MS.Our results preliminarily provided useful chemical information for further study of the pharmacodynamic material basis of GDD.Method:To establish UPLC fingerprint of GDD,which provides a new approach to assessing the quality of GDD.Furthermore,adopting UPLC-Q-TOF-MS~E technologies combined with database of UNIFI platform was established to analysis the chemical composition of GDD rapidly.In addition,we established a copper-loaded model rats by oral administration of copper.Following a three-day oral administration of GDD,we collected the plasma and liver in normal and copper-loaded rats,which were analyzed by UPLC-Q-TOF-MS~Eand processed using UNIFI and investigated the difference of components in vivo.Based on these,we established a method based on UPLC-Q-TOF-MS~Eas well as Multiple Statistical Analysis technique,which could be applied in screening and analyzing metabolites and metabolic pathway in urine by oral administration of GDD.Results:1.The UPLC fingerprinting method of different batches of liver GDD was established and 17 common peaks were selected as the common peaks,9 common Peaks were identified by standard material.The validation of methodologies such as precision,stability and repeatability were showed that the method was stable and feasible.The data were analyzed and matched with UNIFI,and then were tested by artificial check.A total of 96 compounds have been identified or tentatively characterized from the GDD,including anthraquinones and their glycosides,alkaloids,protostane triterpenoids,flavonoids,saponins,tannins,and sesquiterpenes.2.After oral administration of GDD,54 and 46 compounds were observed in the plasma of normal and copper-loaded rats,respectively.54 components were detected in the normal rats,including 36 parent components and 18 metabolites in plasma,and21 parent components and 8 metabolites in liver.46 components were detected in the copper-loaded rats,including 30 parent components and 16 metabolites in plasma,and 18 parent components and 5 metabolites in liver.3.We established a method based on UPLC-Q-TOF-MS~Eas well as Multiple Statistical Analysis technique,which could be applied in screening and identification of GDD after oral administration in urine.Based on this method,77 components of GDD were screened and identified in urine,including 45 parent components and potential metabolites and 32 potential metabolites.It was found that glucuronidation and sulfation are the major biotransformation pathways of aconitum alkaloids in vivo,and the main metabolic pathways of anthraquinones were hydroxylation,demethylation,glucuronidation and sulfation.Conclusion:In this paper,we successfully established the method to analysis the chemical composition of GDD in vitro and in vivo.We can get the target compounds and its metabolites quickly and exactly.It can provide the data support for the quality control and pharmacodynamic basis of GDD.