title: Simultaneous quantitative profiling of 20 isoprostanoids from omega-3 and omega-6 polyunsaturated fatty acids by LC-MS/MS in various biological samples.
abstract: Isoprostanoids are a group of non-enzymatic oxygenated metabolites of polyunsaturated fatty acids. It belongs to oxylipins group, which are important lipid mediators in biological processes, such as tissue repair, blood clotting, blood vessel permeability, inflammation and immunity regulation. Recently, isoprostanoids from eicosapentaenoic, docosahexaenoic, adrenic and α-linolenic namely F3-isoprostanes, F4-neuroprostanes, F2-dihomo-isoprostanes and F1-phytoprostanes, respectively have attracted attention because of their putative contribution to health. Since isoprostanoids are derived from different substrate of PUFAs and can have similar or opposing biological consequences, a total isoprostanoids profile is essential to understand the overall effect in the testing model. However, the concentration of most isoprostanoids range from picogram to nanogram, therefore a sensitive method to quantify 20 isoprostanoids simultaneously was formulated and measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The lipid portion from various biological samples was extracted prior to LC-MS/MS evaluation. For all the isoprostanoids LOD and LOQ, and the method was validated on plasma samples for matrix effect, yield of extraction and reproducibility were determined. The methodology was further tested for the isoprostanoids profiles in brain and liver of LDLR(-/-) mice with and without docosahexaenoic acid (DHA) supplementation. Our analysis showed similar levels of total F2-isoprostanes and F4-neuroprostanes in the liver and brain of non-supplemented LDLR(-/-) mice. The distribution of different F2-isoprostane isomers varied between tissues but not for F4-neuroprostanes which were predominated by the 4(RS)-4-F4t-neuroprostane isomer. DHA supplementation to LDLR(-/-) mice concomitantly increased total F4-neuroprostanes levels compared to F2-isoprostanes but this effect was more pronounced in the liver than brain.
authors.affiliation: MetaToul-Lipidomic Core Facility, MetaboHUB, Inserm U1048, Toulouse, France; Institut des Maladies Métaboliques et Cardiovasculaires, Inserm U1048, Toulouse, France.
authors.forename: Aude
authors.initials: A
authors.lastname: Dupuy
authors.affiliation: MetaToul-Lipidomic Core Facility, MetaboHUB, Inserm U1048, Toulouse, France.
authors.forename: Pauline
authors.initials: P
authors.lastname: Le Faouder
authors.affiliation: Institut des Biomolécules Max Mousseron IBMM, UMR 5247 CNRS, Université de Montpellier, ENSCM, France.
authors.forename: Claire
authors.initials: C
authors.lastname: Vigor
authors.affiliation: Institut des Biomolécules Max Mousseron IBMM, UMR 5247 CNRS, Université de Montpellier, ENSCM, France.
authors.forename: Camille
authors.initials: C
authors.lastname: Oger
authors.affiliation: Institut des Biomolécules Max Mousseron IBMM, UMR 5247 CNRS, Université de Montpellier, ENSCM, France.
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Galano
authors.affiliation: Institut des Maladies Métaboliques et Cardiovasculaires, Inserm U1048, Toulouse, France.
authors.forename: Cédric
authors.initials: C
authors.lastname: Dray
authors.affiliation: School of Biological Sciences, University of Hong Kong, Hong Kong.
authors.forename: Jetty Chung-Yung
authors.initials: JC
authors.lastname: Lee
authors.affiliation: Institut des Maladies Métaboliques et Cardiovasculaires, Inserm U1048, Toulouse, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Valet
authors.affiliation: INRA, UMR1019, UNH, CRNH Auvergne, Clermont Université, Université d'Auvergne, Unité de Nutrition Humaine, BP 10448, F-63000 Clermont-Ferrand, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Gladine
authors.affiliation: Institut des Biomolécules Max Mousseron IBMM, UMR 5247 CNRS, Université de Montpellier, ENSCM, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Durand
authors.affiliation: MetaToul-Lipidomic Core Facility, MetaboHUB, Inserm U1048, Toulouse, France. Electronic address: justine.bertrand-michel@inserm.fr.
authors.forename: Justine
authors.initials: J
authors.lastname: Bertrand-Michel
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D015525:Fatty Acids, Omega-3 / Q000032:analysis* / Q000097:blood; D043371:Fatty Acids, Omega-6 / Q000032:analysis* / Q000097:blood; D006801:Humans; D028421:Isoprostanes / Q000032:analysis* / Q000097:blood; D057230:Limit of Detection; D008099:Liver / Q000737:chemistry*; D051379:Mice; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D015525:Fatty Acids, Omega-3; D043371:Fatty Acids, Omega-6; D028421:Isoprostanes
keywords: Dihomo-isoprostanes; Isoprostanes; Mass spectrometry; Neuroprostanes; Oxidative stress; Phytoprostanes; Quantification; ROS