title: Profiling a gut microbiota-generated catechin metabolite's fate in human blood cells using a metabolomic approach.
abstract: The microbial catechin metabolite δ-(3,4-dihydroxy-phenyl)-γ-valerolactone (M1) has been found in human plasma samples after intake of maritime pine bark extract (Pycnogenol). M1 has been previously shown to accumulate in endothelial and blood cells in vitro after facilitated uptake and to exhibit anti-inflammatory activity. The purpose of the present research approach was to systematically and comprehensively analyze the metabolism of M1 in human blood cells in vitro and in vivo. A metabolomic approach that had been successfully applied for drug metabolite profiling was chosen to detect 19 metabolite peaks of M1 which were subsequently further analyzed and validated. The metabolites were categorized into three levels of identification according to the Metabolomics Standards Initiative with six compounds each confirmed at levels 1 and 2 and seven putative metabolites at level 3. The predominant metabolites were glutathione conjugates which were rapidly formed and revealed prolonged presence within the cells. Although a formation of an intracellular conjugate of M1 and glutathione (M1-GSH) was already known two GSH conjugate isomers, M1-S-GSH and M1-N-GSH were observed in the current study. Additionally detected organosulfur metabolites were conjugates with oxidized glutathione and cysteine. Other biotransformation products constituted the open-chained ester form of M1 and a methylated M1. Six of the metabolites determined in in vitro assays were also detected in blood cells in vivo after ingestion of the pine bark extract by two volunteers. The present study provides the first evidence that multiple and structurally heterogeneous polyphenol metabolites can be generated in human blood cells. The bioactivity of the M1 metabolites and their contribution to the previously determined anti-inflammatory effects of M1 now need to be elucidated.
authors.affiliation: Universität Würzburg, Institut für Pharmazie und Lebensmittelchemie, Würzburg, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Mülek
authors.affiliation: Universität Würzburg, Julius-von-Sachs-Institut für Biowissenschaften, Würzburg, Germany.
authors.forename: Agnes
authors.initials: A
authors.lastname: Fekete
authors.affiliation: Universität Würzburg, Institut für Pharmazie und Lebensmittelchemie, Würzburg, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Wiest
authors.affiliation: Universität Würzburg, Institut für Pharmazie und Lebensmittelchemie, Würzburg, Germany.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Holzgrabe
authors.affiliation: Universität Würzburg, Julius-von-Sachs-Institut für Biowissenschaften, Würzburg, Germany.
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Mueller
authors.affiliation: Universität Würzburg, Institut für Pharmazie und Lebensmittelchemie, Würzburg, Germany. Electronic address: hogger@pharmazie.uni-wuerzburg.de.
authors.forename: Petra
authors.initials: P
authors.lastname: Högger
mesh_terms: D001711:Biotransformation; D001773:Blood Cells / Q000378:metabolism*; D002392:Catechin / Q000737:chemistry*; D005419:Flavonoids / Q000737:chemistry*; D000069196:Gastrointestinal Microbiome; D005978:Glutathione / Q000737:chemistry; D006801:Humans; D007783:Lactones / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D055432:Metabolomics / Q000379:methods*; D028223:Pinus; D024301:Plant Bark / Q000378:metabolism; D010936:Plant Extracts / Q000737:chemistry; D059808:Polyphenols / Q000737:chemistry; D012016:Reference Values; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D005419:Flavonoids; D007783:Lactones; D010936:Plant Extracts; D059808:Polyphenols; C024070:pycnogenols; D002392:Catechin; D005978:Glutathione; C037556:gamma-valerolactone
keywords: LC–MS/MS; Metabolism; NMR; Pine bark extract; Polyphenols