Xie Weiwei, Jin Yiran, Hou Ludan, Ma Yinghua, Xu Huijun, Zhang Kerong, Zhang Lantong, Du Yingfeng
Department of Pharmaceutical Analysis, School of Pharmacy, Hebei Medical University, Shijiazhuang, Hebei, 050017, PR China.
Department of Pharmaceutical Analysis, School of Pharmacy, Hebei Medical University, Shijiazhuang, Hebei, 050017, PR China; The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, 050000, PR China.
J Pharm Biomed Anal. 2017 Oct 25;145:865-878. doi: 10.1016/j.jpba.2017.08.003. Epub 2017 Aug 11.
Ponicidin is an active natural ent-kaurane diterpenoid ingredient originating from many Isondon herbs and is expected to become a new anticancer agent. In this study, a practical strategy was developed for the identification of ponicidin metabolites in vivo and in vitro utilizing ultra-high-performance liquid chromatography coupled with hybrid triple quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS). The analytical strategy was as follows: potential ponicidin metabolites were detected by a novel on-line data acquisition approach, i.e., sequential window acquisition of all theoretical fragment-ion spectra (SWATH™). Compared to the traditional information-dependent acquisition (IDA) method, SWATH™ significantly improved the hit rate of low-level or trace metabolites because it could obtain all MS/MS spectra. Moreover, many data post-processing methods were used to deduce the metabolites structures. As a result, a total of 20 metabolites were characterized in vivo and in vitro. The results showed that ponicidin could undergo general metabolic reactions, such as oxidation, reduction, hydrolysis, methylation and glucuronidation. Furthermore, there was an obvious difference in the ponicidin metabolites among four species in vitro. This is the first time that the SWATH™ data acquisition mode has been used to characterize ponicidin metabolites in trace amounts or in a biological matrix. These results not only provided a better understanding of the safety and efficacy of ponicidin but also showed a valuable methodology for the identification of other ent-kaurane diterpenoid metabolites.
冬凌草甲素是一种活性天然对映-贝壳杉烷二萜类成分，源自多种香茶菜属植物，有望成为一种新型抗癌药物。在本研究中，开发了一种实用策略，利用超高效液相色谱与混合三重四极杆飞行时间质谱联用技术（UHPLC-Q-TOF-MS）鉴定冬凌草甲素的体内外代谢产物。分析策略如下：通过一种新型在线数据采集方法，即所有理论碎片离子谱的顺序窗口采集（SWATH™）来检测潜在的冬凌草甲素代谢产物。与传统的信息依赖采集（IDA）方法相比，SWATH™显著提高了低水平或痕量代谢产物的命中率，因为它可以获取所有的二级质谱图。此外，还使用了许多数据后处理方法来推导代谢产物的结构。结果，在体内和体外共鉴定出20种代谢产物。结果表明冬凌草甲素可发生普遍的代谢反应，如氧化、还原、水解、甲基化和葡萄糖醛酸化。此外，四种体外物种的冬凌草甲素代谢产物存在明显差异。这是首次使用SWATH™数据采集模式来鉴定痕量或生物基质中的冬凌草甲素代谢产物。这些结果不仅有助于更好地理解冬凌草甲素的安全性和有效性，还展示了一种鉴定其他对映-贝壳杉烷二萜类代谢产物的有价值方法。
