Rapid Analysis Of Migrating Components And Metabolites Of Siwei Tumuxiang San In The Blood Based On HPLC-Q-Exactive-MS/MS

Objective: 1)Using high performance liquid chromatography with quadrupoleelectrostatic field orbitrap high resolution mass spectrometry(HPLC-Q-ExactiveMS/MS)to establish a precise,efficient and rapid methoud for the analysis and identification of complex components of Siwei Tumuxiang San.2)HPLC-Q-ExactiveMS/MS technique was used to analyze the blood migration components of Mongolian medicine compound preparation Siwei Tumuxiang San,and to find out the index components of its true blood.3)HPLC-Q-Exactive-MS/MS high-resolution mass spectrometry was used to analyze the metabolites of the single-flavored Sophora flavescens.Ait in the blood,urine and bile of rats,which laid a material foundation for its metabolic process.This paper provides an experimental basis for elucidating the material basis and pharmacokinetics of Mongolian medicine Siwei Tumuxiang San.Methods: 1)The experiment used Agilent Zorbax SB-Aq(4.6 × 150 mm,5 ?m)column with 5 mmol ammonium acetate / 0.1% formic acid aqueous solution(A)-acetonitrile(B)as mobile phase and using thermoelectric spray(HESI)mass spectrometry ion Source.Data were collected in positive and negative ion mode,and the molecular formula and structural formula of the compound were determined according to the exact mass and Chemspider database.By comparing the retention time,the mass spectrum and the secondary fragment ion reported in the literature,combined with the fragment pathways of compounds.The compounds in Siwei Tumuxiang San were accurately identified.2)HPLC-Q-Exactive-MS/MS technique was used to analyze the serum of rats after oral administration ofSiwei Tumuxiang San,by comparing the blank serum and the administered serum in positive and negative ion mode,and combining the phases I and II.Phase metabolites,as well as the Compound Discover software for structural identification of metabolites.3)Mixing serum,urine and bile biological samples at different time points by oral administration of oral Sophora flavescens in rats,using HPLC-Q-Exactive-MS/MS technique for blank serum,urine and bile.Analysis of blank serum,blank urine,blank bile,administration serum,administration of urine and administration of bile by HPLCQ-Exactive-MS/MS technique.Then combined with a variety of adducts,the metabolic rules of the metabolites of the oral extract of Sophora flavescens Ait in rats in different organs were identified.Results: 1)For the first time,a total of 110 compounds were identified from the Siwei Tumuxiang San by HPLC-Q-Exactive-MS/MS technology,including 3 sesquiterpene compounds from Inula helenium L,16 alkaloid compunds and 38 isoprene flavonoid compounds from Sophora flavescens Ait,12 triterpenoid saponin compounds,1 catechin compounds and 4 flavonoid glycoside compounds from Rubus sachalinensis Leveille.The fragment pathways of the main types of compounds were summarized.Among them,31 compounds of mass spectrometry information were reported for the first time.This study summarizes the cleavage pattern of each class of compounds.2)A total of 63 transition components,including 43 absorption prototype components and 20 metabolites,were identified from rat serum by oral administration of four flavors of soil fragrance,and 23 compounds were the first to report mass spectrometry information for this study.3)HPLC-Q-Exactive-MS/MS technology and Discover Compound data analysis software were used to analyze and identify the metabolites of oral Sophora flavescens Ait.in serum,urine and bile,and a total of 171 compounds were identified.Including 89 prototype products and 82 metabolites,including 46 matrine alkaloids and 125 isoprene flavonoids,69 compounds for the first time reported mass spectrometry information in rats.Conclusion: 1)The method can systematically,accurately and quickly identify the complex chemical constituents of Siwei Tumuxiang San,providing an experimental basis for clarifying the in vivo analysis of the compound and establishing quality standards.2)Using liquid chromatography-mass spectrometry(HPLC-Q-ExactiveMS/MS)technology to establish a method for the in vivo analysis of Siwei Tumuxiang San,and laid the foundation for the search for index components and dynamic metabolic processes in vivo.3)HPLC-Q-Exactive-MS/MS was used to determine the metabolites in serum,urine and bile after oral administration of singleflavored medicinal herbs in rats,which laid a foundation for the clarification of metabolic processes in vivo.