The Study Of In Vivo And In Vitro Metabolism Of Four Flavonoids Based On LC-MS Technology

Farrerol,acacetin,biochanin A and oroxin B are flavonoid monomer compounds,all of which exhite wide biological activities,causing people's attention.Drugs went througt a series of complex biological transformations after entering into the human body by oral,and generated ineffective,biologically active or toxic compounds.Rat plasma,bile,urine and feces were used as biological samples for in vivo metabolism.Rat liver microsomes and intestinal flora were used as in vitro metabolic models.In order to ensure the safe use of the above four flavonoids,it was necessary to comprehensively and systematically study the metabolism in vitro and in vivo.UHPLC-Q-TOF-MS/MS has high accuracy,high resolution and high sensitivity,and is widely used for qualitative research,such as drug metabolism research.At precent,the data on farrerol,acacetin,biochanin A and oroxin B metabolism is unavailable.Hence,the purpose of this study was to systematically study the in vitro and in vivo metabolism of farrerol,acacetin,biochanin A and oroxin B using liquid chromatography-mass spectrometry(LC-MS),which could guide clinical medicinal use and provide a basis for new drug development.Objective: To study the metabolism of farrerol,acacetin,biochanin A and oroxin B in rat plasma,bile,urine and feces,and the metabolism in rat liver microsomes and intestinal flora based on UHPLC-Q-TOF-MS/MS technology.Methods: After oral administration,the rat blood,bile,urine and feces were collected.The incubation models of liver microsomes and intestinal flora were established,and the mixures after incubation were collected.Then,the collected samples were analyzed.Through UHPLC-Q-TOF-MS/MS,online full scan data was acquired relying on MMDF and DBS settings in IDA pattern.XIC,MDF,PIF and NLF technologies in Metabolite Pilot software were combined for data post-processing;metabolites are identified based on their accurate masses,the cleavage pathway of parent drugs,and related biotransformation information.In addition,metabolites were further determined by comparion with standards or reference to related literatures.Results: In this study,the metabolites of farrerol,acacetin,biochanin A,oroxin B were identified in vivo and in vitro by UHPLC-Q-TOF-MS/MS.In the study of farrerol metabolism,42 metabolites were identified in rats and 15 metabolites were identified in liver microsomes.In the study of acacetin metabolism,31 metabolites were identified in rats and 25 metabolites in liver microsomes.Among them,4 metabolites were identified in comparison with the reference stanndards.In the study of biochanin A metabolism,43 metabolites in rats,22 metabolites in liver microsomes and 18 metabolites in intestinal flora were identified,and 5 metabolites were determined by comparison the elution times and MS data of standards.In the study of oroxin B metabolism,30 metabolites in rats,8 metabolites in liver microsomes and 18 metabolites in intestinal flora were identified,and 9 metabolites were characterized by comparison with standards.In addition,the metabolic pathways of farrerol,acacetin,biochanin A and oroxin B in vitro and in vivo were summarized.Conclusions: In this study,the in vivo and in vitro metabolism of farrerol,acacetin,biochanin A and oroxin B were studied by UHPLC-Q-TOF-MS/MS technology,which enriched their corresponding metabolic transformation information bases and provided the basis for clinical pharmacological research.