title: Analysis of Omega-3 Fatty Acid Derived N-Acylethanolamines in Biological Matrices.
abstract: The adequate quantification of endocannabinoids can be complex due to their low endogenous levels and structural diversity. Therefore, advanced analytical approaches, such as LC-MS, are used to measure endocannabinoids in plasma, tissues, and other matrices. Recent work has shown that endocannabinoids that are synthesized from n-3 fatty acids, such as docosahexaenoylethanolamide (DHEA) and eicosapentaenoylethanolamide (EPEA), have anti-inflammatory and anti-tumorigenic properties and stimulate synapse formation in neurites. Here, an LC-MS based method for the quantification of n-3 endocannabinoids DHEA and EPEA which is also suited to measure a wider spectrum of endocannabinoids is described. The chapter contains a step-by-step protocol for the analysis of n-3 endocannabinoids in plasma, including sample collection and solid phase extraction, LC-MS analysis, and data processing. Modifications to the protocol that allow quantifying n-3 endocannabinoids in tissues and cell culture media will also be discussed. Finally, conditions that alter endocannabinoid concentrations are briefly discussed.
authors.affiliation: Division of Human Nutrition, Wageningen University, 8129, 6700 EV, Wageningen, The Netherlands.
authors.forename: Renger F
authors.initials: RF
authors.lastname: Witkamp
authors.affiliation: Division of Human Nutrition, Wageningen University, 8129, 6700 EV, Wageningen, The Netherlands. michiel.balvers@wur.nl.|Clinical Chemistry and Haematology Laboratory, Gelderse Vallei Hospital, 9025, 6710 HN, Ede, The Netherlands. michiel.balvers@wur.nl.
authors.forename: Michiel G J
authors.initials: MG
authors.lastname: Balvers
mesh_terms: D002853:Chromatography, Liquid; D063388:Endocannabinoids / Q000737:chemistry; D004983:Ethanolamines / Q000097:blood / Q000737:chemistry* / Q000378:metabolism; D015525:Fatty Acids, Omega-3 / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry
chemical_list: D063388:Endocannabinoids; D004983:Ethanolamines; D015525:Fatty Acids, Omega-3; C022203:N-acylethanolamines; C561061:N-docosahexaenoylethanolamide
keywords: Docosahexaenoylethanolamide; Eicosapentaenoylethanolamide; Endocannabinoids; LC-MS; Solid phase extraction; n-3 Fatty acids