title: Biotransformation and metabolic profile of anemoside B4 with rat small and large intestine microflora by ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry.
abstract: Pulsatilla chinensis (Bunge) Regel is commonly used in Asia, and anemoside B4 (AB4) is its major saponin, with diverse pharmaceutical effects. Previous studies showed that intestinal flora plays an important role in the metabolism of herbs administered orally. In this study, the metabolic profile of AB4 with microflora in rat small and large intestines in vitro was investigated. Gut microflora was collected from different intestinal segments and anaerobically incubated with AB4 at 37°C for 24, 48, 72 and 96 h, respectively. A total of 10 metabolites were detected and identified by ultra- performance liquid chromatography/quadrupole time-of-flight mass spectrometry, involving the products of oxygenation and deglycosylation reactions. Gut microflora in the large intestine generated more comprehensive metabolic pathways, which appears to be attributable to the wider range of bacterial types and numbers of bacteria. Human cancer cell lines SMMC-7721, Hela and MCF-7 were treated with metabolite pools by MTT assay, together with M6 as the greatest deglycosylation product. As a result, M6 exhibited a reduction in cell viability of SMMC-7721 with an IC50value of 22.28 ± 1.26 μg/mL. The present study provided scientific evidence for AB4 metabolism in small and large intestines, which is helpful to reveal the active forms of AB4 in vivo.
authors.affiliation: College of Pharmacy, Chongqing University, Chongqing, China.|School of Pharmacy, Jiangsu University, Zhenjiang, China.
authors.forename: Jin-Yi
authors.initials: JY
authors.lastname: Wan
authors.affiliation: College of Pharmacy, Chongqing University, Chongqing, China.|Basic Medical College, Chongqing Medical University, Chongqing, China.
authors.forename: Yu-Zhen
authors.initials: YZ
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Modern Preparation of TCM, Ministry of Education, Jiangxi University of Traditional Chinese Medicine, Nanchang, China.
authors.forename: Jin-Bin
authors.initials: JB
authors.lastname: Yuan
authors.affiliation: College of Pharmacy, Chongqing University, Chongqing, China.
authors.forename: Feng-Qing
authors.initials: FQ
authors.lastname: Yang
authors.affiliation: Key Laboratory of Modern Preparation of TCM, Ministry of Education, Jiangxi University of Traditional Chinese Medicine, Nanchang, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Basic Medical College, Chongqing Medical University, Chongqing, China.
authors.forename: Lian-Di
authors.initials: LD
authors.lastname: Zhou
authors.affiliation: College of Pharmacy, Chongqing University, Chongqing, China.
authors.forename: Qi-Hui
authors.initials: QH
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001711:Biotransformation; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000069196:Gastrointestinal Microbiome; D006801:Humans; D007420:Intestine, Large / Q000187:drug effects / Q000378:metabolism / Q000382:microbiology*; D007421:Intestine, Small / Q000187:drug effects / Q000378:metabolism / Q000382:microbiology*; D008297:Male; D015394:Molecular Structure; D017207:Rats, Sprague-Dawley; D012503:Saponins / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics* / Q000494:pharmacology; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D012503:Saponins; C000620474:anemoside B4
keywords: UPLC-Q-TOF-MS; anemoside B4; gut microflora; metabolic profile; small/large intestine