Qualitative Analysis Of Triterpenoid Saponins From Achyranthes Bidentata And Preliminary Study On Metabolism In Vitro And In Vivo

Objective:Rapid qualitative analysis of Achyranthes bidentata triterpenoid saponins was carried out by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS)combined with UNIFI data processing platform.The metabolites of Achyranthes bidentata triterpenoid saponins in intestinal microflora,liver microsomes and plasma were determined,and their metabolic characteristics were discussed.It may provide experimental basis for the study of the pharmacodynamic basis of Achyranthes bidentata triterpenoid.Method:The UNIFI database of Achyranthes bidentata triterpenoid saponins was established;Firstly,the characteristic ions of four triterpenoid saponins reference standards were investigated and the mass spectrometry fragmentation behaviors of triterpenoid saponins was summarized;then the samples were analyzed by UPLC-Q-TOF/MS~Eunder the negative ion mode,and the collected MS~Eraw data were imported into the UNIFI data processing platform;Finally,the ion peaks were identified by preset parameters and automatically matched with the established target database;establishment of intestinal microflora incubation system,by measuring the metabolites of three representative compounds in intestinal microflora,to explore the metabolites and metabolic characteristics of Achyranthes bidentata triterpenoid saponins in intestinal microflora;establishment of in vitro liver microsome incubation system,to explore the transformation of Achyranthes bidentata triterpenoid saponins in liver microsome;the metabolic regularity of Achyranthes bidentata triterpenoid saponins in plasma was investigated by analyzing the plasma of the rats treated with Achyranthes bidentata.Results:1.A total of 40 triterpenoid saponins were isolated and identified.Among them,there are 13 compounds with one substituent in C3 or C28 on the mothernucleus of oleanolic acid,25 compounds with substituents at C3 and C28,and 2 compounds from other mother nuclei.2.The intestinal flora incubation system was constructed.The intestinal floraincubation solution of Achyranthes bidentata triterpenoid saponins wasdetermined and analyzed by UPLC-Q-TOF/MS~Ecombined with UNIFI dataprocessing platform.39 metabolites were identified.The main metabolicreactions were deglycosylation,dehydrogenation,oxidation andglycosylation.3.The rat liver microsome incubation system was constructed,and 26metabolites of Achyranthes bidentata triterpenoid saponins in livermicrosome incubation solution were detected by UPLC-Q-TOF/MS~Ecombined with UNIFI data processing platform.Metabolites mainly undergooxidation,dehydrogenation and other metabolism reaction.4.Collecting the plasma of rats with intragastric administration,21 metabolitesin rat plasma were detected.Metabolites mainly undergo metabolic reactionssuch as double oxidation and dehydrogenation.Conclusion:In this study,UPLC-Q-TOF/MS combined with UNIFI data processing platform can quickly identify the components of Achyranthes bidentata triterpenoid saponins and their metabolites in intestinal microflora,liver microsomes and plasma.The chemical composition and metabolism of Achyranthes bidentata triterpenoid saponins are preliminarily clarified,which may provide experimental basis for its subsequent basic material research.