title: Identification of in vitro and in vivo metabolites of alantolactone by UPLC-TOF-MS/MS.
abstract: Alantolactone (AL), an active sesquiterpene originating from Inula helenium, is a potential anticancer and anti-inflammatory agent. However so far, studies on AL metabolism have not been reported. In the present study, we have investigated for the first time the in vivo and in vitro metabolites of AL using ultra performance liquid chromatography combined with time of flight mass spectrometry (UPLC-TOF-MS/MS). A unique on-line information-dependent acquisition (IDA) method multiple mass defect filter (MMDF) combined with dynamic background subtraction (DBS) was applied to trace all of the probable metabolites of AL. Five MMDF templates were set according to the core structure of AL and the general metabolite biotransformation patterns, and other five sulfur-containing dimer filter templates were first established on the basis of structural elucidation of AL metabolites obtained from rat intestinal bacteria biotransformation. As a result, 44 metabolites were characterized: 41 metabolites from rat urine, bile and feces after oral administration of AL, and 13 metabolites from AL biotransformation by rat intestinal bacteria. Particularly, 26 metabolites were identified as novel sulfur-containing products. The results indicated that addition of double bond at Δ((11,13)) and oxidization were the main metabolic reactions of AL. A new metabolism pathway to produce addition products of H2S to AL and further generate a series of sulfur-containing dimers of AL was revealed. This study significantly enriched our knowledge about AL metabolism, which will lead to a better understanding of the safety and efficacy of AL. At the same time, the established methodology can be widely applied for the structural determination of the metabolites of other sesquiterpene containing α-methylene-γ-lactone moiety.
authors: School of Pharmaceutical Sciences, Hebei Medical University, Shijiazhuang 050017, China; Cangzhou People⿿s Hospital, Cangzhou 061000, China.
authors: Donggui
authors: D
authors: Yao
authors: Cangzhou Medical College, Cangzhou 061001, China.
authors: Zhe
authors: Z
authors: Li
authors: School of Pharmaceutical Sciences, Hebei Medical University, Shijiazhuang 050017, China; Institute of Chinese Integrative Medicine, Hebei Medical University, Shijiazhuang 050017, China. Electronic address: rainbowhuo@126.com.
authors: Changhong
authors: C
authors: Huo
authors: School of Pharmaceutical Sciences, Hebei Medical University, Shijiazhuang 050017, China.
authors: Yufang
authors: Y
authors: Wang
authors: School of Pharmaceutical Sciences, Hebei Medical University, Shijiazhuang 050017, China.
authors: Yibing
authors: Y
authors: Wu
authors: School of Pharmaceutical Sciences, Hebei Medical University, Shijiazhuang 050017, China.
authors: Manli
authors: M
authors: Zhang
authors: School of Pharmaceutical Sciences, Hebei Medical University, Shijiazhuang 050017, China.
authors: Ligeng
authors: L
authors: Li
authors: School of Pharmaceutical Sciences, Hebei Medical University, Shijiazhuang 050017, China.
authors: Qingwen
authors: Q
authors: Shi
authors: Graduate School of Environmental & Life Sciences, Okayama University, Okayama 700-8530, Japan.
authors: Hiromasa
authors: H
authors: Kiyota
authors: School of Pharmaceutical Sciences, Hebei Medical University, Shijiazhuang 050017, China. Electronic address: shixiaowei@hebmu.edu.cn.
authors: Xiaowei
authors: X
authors: Shi
mesh_terms: D000818:Animals; D001711:Biotransformation; D066241:Carbon-13 Magnetic Resonance Spectroscopy; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007783:Lactones / Q000737:chemistry / Q000378:metabolism*; D008297:Male; D050216:Metabolic Detoxication, Phase I; D050217:Metabolic Detoxication, Phase II; D053858:Metabolic Networks and Pathways; D055442:Metabolome; D008970:Molecular Weight; D066244:Proton Magnetic Resonance Spectroscopy; D017207:Rats, Sprague-Dawley; D045787:Sesquiterpenes, Eudesmane / Q000737:chemistry / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D007783:Lactones; D045787:Sesquiterpenes, Eudesmane; C004363:alantolactone
keywords: Alantolactone; Intestinal bacteria biotransformation; Metabolites; Multiple mass defect filter; UPLC-TOF-MS/MS