Identification And Comparison Of Chemical Compounds In Commercial Tripterygium Wilfordii Genus Preparations With UHPLC-MS/MS

Objective: To identify and analyze compounds in commercial Tripterygium wilfordii genus preparations.Methods: 10 reference substances were pumped into the mass spectrometer to summarize the fragmentation behavior with different CE in positive and negative ion modes,UHPLC-QTOF-MS/MS conventional identification method and molecular networking technology were used to identify the compounds of commercial Tripterygium wilfordii genus preparations,and the metabolomics data processing technique was used to find different compounds among Tripterygium wilfordii genus preparations: the gradient elution was carried out on an Acquity UHPLC HSS T3 column with0.1% formic acid water-acetonitrile mobile phase system of high resolution mass spectrometry electrospray ion source in positive and negative ion modes,the spectrum information was manually analyzed by comparing to in-house database.Molecular networking as an assistant identification technology was built on GNPS platform and visualized by Cytoscape3.7.1.Some unknown compounds could be induced by known compounds based on the similar MS/MS spectrum.PCA,Student’s t test and OPLS-DA were used to compare the compounds among 92 Tripterygium wilfordii genus preparations in order to find common and different compounds.Results: The fragmentation rules of 10 representative compounds(4diterpenes,4 triterpenes and 2 alkaloids)of Tripterygium wilfordii genus preparations were summarized.103 compounds of Tripterygium wilfordii genus preparations were found,70 out of 103 compounds were identified,including 63 compounds identified by conventional identification method and7 compounds identified by molecular networking technology.According to PCA score plot,Tripterygium wilfordii tablets,Tripterygium hypoglaucum Hutch tablets and most of Tripterygium Glycosides tablets were gathered into one group,while the Tripterygium Glycosides tablets were grouped into three categories;compounds of three Tripterygium wilfordii genus preparations were similar,including triptolide,celsatrol,tripforidin,wilforine.While compounds of Tripterygium Glycosides tablets produced by different factories were different,including triptolide,wilforgine,wilforine,demethylzeylasteral.Conclusions: In this study,fragmentation rules of sesquiterpene alkaloids,diterpenes and triterpenes were summarized.The chemical compounds of Tripterygium wilfordii genus preparations were identified with UHPLC-QTOF-MS/MS conventional identification method and molecular networking technology.The compounds in 92 Tripterygium wilfordii genus preparations were compared,which could provide a basis for the quality control and elucidation of pharmacodynamic material of Tripterygium wilfordii genus preparations.