Study On The Metabolism Of Huangqi Jianzhong Tang In Vivo

Huangqi Jianzhong Tang(HQJZ)is derived from Synopsis of the Golden Chamber for treating cold stomach pain,and mostly used for treating gastrointestinal diseases in clinical.The effectiveness of traditional Chinese medicine is a comprehensive reflection of its effective components through certain drug delivery pathway and in vivo process,and pharmacokinetics can study the absorption,distribution,metabolism and excretion of potential effective components absorbed into the blood.The metabolism of HQJZ in the plasma of rats was characterized in the early stage of the research group.However,due to the low content of drugs and their metabolites in the plasma and the dynamic changes,some components may be lost in the collection process.The metabolites in urine,bile,feces and other biological samples are same as plasma,and are easy to be enriched.It can reverse identify the absorbed chemical components,supplement the deficiency of the analysis of plasma metabolites,and obtain complete metabolism information in vivo.Therefore,the integrated pharmacokinetics of HQJZ was studied in rats with normal and chronic atrophic gastritis.In order to understand its metabolism process in vivo more comprehensively,the metabolites in bile,urine and feces after administration were characterized and their metabolism reaction in vivo was analyzed.Combined with the previous study of plasma metabolism,the metabolism of HQJZ was characterized as a whole.Objective: To establish the quantitative analysis method of the chemical components of HQJZ,analyze the Integrated pharmacokinetics of normal and chronic atrophic gastritis rats,comprehensively characterize the metabolites in bile,urine and feces,explain the metabolism process of HQJZ,and provide scientific basis for the internal process and material basis of HQJZ.Method:(1)Rapid quantitative analysis of 18 chemical components of Huangqi Jianzhong Tang based on UHPLC-MS/MS: Based on the MRM model,a reliable LC-MS method was established for the quantitative analysis of 18 components of HQJZ.(2)Integrated pharmacokinetics of Huangqi Jianzhong Tang in normal and chronic atrophic gastritis rats: In this study,12 components and 1 metabolite of HQJZ were established by LC-MS.Then,SD rats were divided into groups and fed with high dose HQJZ.We collected the plasma samples at multiple time points and precipitated the protein with acetonitrile.After measuring the content,we used DAS 2.0 software to carry out the pharmacokinetic analysis then integrated the pharmacokinetic study on the model integration,and compared the differences between normal and CAG rats.(3)Urinary and fecal metabolism of Huangqi Jianzhong Tang in rats based on UHPLC-Q-Exactive MS: After SD rats were given high dose HQJZ,then collected the bile,urine and fecal samples.After SPE column treatment,the data were collected by UHPLC-Q-Exactive MS and imported into Compound Discover 3.0 software to obtain accurate molecular weight and speculate reaction type.Xcalibur 3.2 was extracted the characteristic fragment ions and compared with the database,analyzed the fragmentation rule,then identified the components in bile,urine and feces.Results:(1)Quantitative analysis results of 18 components of Huangqi Jianzhong Tang: The 18 components in HQJZ were analyzed,and were used to analyze the samples of Huangqi Jianzhong pill purchased in pharmacy and HQJZ prepared in the laboratory.The results showed that the method was feasible,which provided the basis for the overall quality control of HQJZ.(2)Results of integrated pharmacokinetic study on normal and chronic atrophic gastritis rats: The results showed that AUC(0-t),AUC(0-?)and Cmax of flavonoid and flavonoid glycoside decreased and T1 / 2 and MRT(0-t)increased significantly in CAG.It is speculated that HQJZ can be absorbed in pathological state reduce the body stay longer.(3)The metabolism of bile,urine and feces in Huangqi Jianzhong Tang rats were studied: The metabolism of metabolites in bile,urine and feces of rats was studied.The results showed that 94 components were identified in bile,including 6 prototypes and 88 metabolites.89 components were identified in urine,including 21 prototypes and 68 metabolites.At the same time,89 components were identified in feces,22 of which were prototypes and 67 were metabolites.Among them,the primary components are flavonoids and related glycosides,pentacyclic triterpenoid saponins and monoterpenoids.Their biotransformation methods mainly include oxidation,hydrolysis,reduction,methylation,acetylation,sulfation,glucuronide,glucoside and glycine conjugation.Conclusion: In this study,the chemical components of HQJZ in vitro were quantified based on LC-MS,and the pharmacokinetics of HQJZ in normal and model rats were compared.In addition,the metabolites in bile,urine and feces of rats after administration were characterized to obtain a comprehensive metabolic spectrum in vivo.This study provides the basis for the identification of the metabolism characteristics and the process of action in vivo of HQJZ,and also for the quality control of HQJZ and the clinical research reference for the prevention and treatment of chronic atrophic gastritis.