Tripterygium wilfordii Hook F (TwHF) has a long history of clinical use for treating various immune and inflammatory diseases including rheumatoid arthritis, systemic lupus erythematous, nephritis and skin diseases in China. Meanwhile, it is well known that this herb shows certain toxic effects. Terpenoids such as triptolide and tripterine were found to be the main bioactive constituents responsible for these pharmacological effects. However, the mechanism of their pharmacological and toxic effects is not very clear due to the lack of corresponding pure compounds. Separation and detection of all these compounds is a key task to further elucidate all the effective/toxic components and control the quality of TwHF products because these terpenoids are numerous but quite low content and weak UV absorption. Detection of these components may suffer serious interference from other components with high content and strong UV absorption, especially from sesquiterpene alkaloids, another kind of constituents of TwHF. Though various analytical methods, including TLC, UV, GC, CE, HPLC and HPLC-MS have been developed for the determination of triptolide, tripterine or wilforlide A in extracts, there still remains the problem to specific simultaneous analysis of terpenoids in TwHF extracts.In this study, a rapid, specific and selective method for the simultaneous analysis and identification of terpenoids in extracts of TwHF was developed. By optimizing the solid-phase extraction (SPE) and UHPLC procedure, the alcoholic extracts of TwHF were separated well on an Agilent Poroshell 120 SB-C18 column (150 � 3.0 mm, 2.7 �m) eluted with acetonitrile-water gradient program after they were extracted with ethyl acetate and then enriched and cleaned up on a C18 SPE cartridge. Dozens of terpenoids in TwHF extracts were identified by determining accuracy molecular weight, matching the empirical molecular formula with those of the reported compounds and/or their retention time with those of reference compounds, and further elucidated by targeted MS/MS. Our results indicate that SPE is a useful pretreatment technique and UHPLC-QTOF-MS is a rapid and effective tool for specific simultaneous separation and identification of terpenoids at trace level. In addition, it lays a good foundation for further pharmacological, toxic and quality control studies.

Identification of terpenoids in extracts of Tripterygium wilfordii Hook F by SPE-UHPLC-QTOF-MS

248th National Meeting (2014)

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