Metabolite identification of triptolide by data-dependent accurate mass spectrometric analysis in combination with online hydrogen/deuterium exchange and multiple data-mining techniques
Article first published online: 26 SEP 2011
Copyright © 2011 John Wiley & Sons, Ltd.
Rapid Communications in Mass Spectrometry
Volume 25, Issue 20, pages 3167–3177, 30 October 2011
How to Cite
Du, F., Liu, T., Liu, T., Wang, Y., Wan, Y. and Xing, J. (2011), Metabolite identification of triptolide by data-dependent accurate mass spectrometric analysis in combination with online hydrogen/deuterium exchange and multiple data-mining techniques. Rapid Commun. Mass Spectrom., 25: 3167–3177. doi: 10.1002/rcm.5211
- Issue published online: 13 SEP 2011
- Article first published online: 26 SEP 2011
- Manuscript Accepted: 4 AUG 2011
- Manuscript Revised: 29 JUL 2011
- Manuscript Received: 3 JUL 2011
- National Natural Science Foundation of China. Grant Numbers: 30901829, 31071146
- Shandong Natural Science Foundation of China. Grant Number: BS2009SW013
- Excellent Young Teachers Program of Southeast University. Grant Number: 3231001201
Triptolide (TP), the primary active component of the herbal medicine Tripterygium wilfordii Hook F, has shown promising antileukemic and anti-inflammatory activity. The pharmacokinetic profile of TP indicates an extensive metabolic elimination in vivo; however, its metabolic data is rarely available partly because of the difficulty in identifying it due to the absence of appropriate ultraviolet chromophores in the structure and the presence of endogenous interferences in biological samples. In the present study, the biotransformation of TP was investigated by improved data-dependent accurate mass spectrometric analysis, using an LTQ/Orbitrap hybrid mass spectrometer in conjunction with the online hydrogen (H)/deuterium (D) exchange technique for rapid structural characterization. Accurate full-scan MS and MS/MS data were processed with multiple post-acquisition data-mining techniques, which were complementary and effective in detecting both common and uncommon metabolites from biological matrices. As a result, 38 phase I, 9 phase II and 8 N-acetylcysteine (NAC) metabolites of TP were found in rat urine. Accurate MS/MS data were used to support assignments of metabolite structures, and online H/D exchange experiments provided additional evidence for exchangeable hydrogen atoms in the structure. The results showed the main phase I metabolic pathways of TP are hydroxylation, hydrolysis and desaturation, and the resulting metabolites subsequently undergo phase II processes. The presence of NAC conjugates indicated the capability of TP to form reactive intermediate species. This study also demonstrated the effectiveness of LC/HR-MSn in combination with multiple post-acquisition data-mining methods and the online H/D exchange technique for the rapid identification of drug metabolites. Copyright © 2011 John Wiley & Sons, Ltd.