title: Quantitative profiling of endocannabinoids and relatedN-acylethanolamines in human CSF using nano LC-MS/MS.
abstract: Endocannabinoids, a class of lipid messengers, have emerged as crucial regulators of synaptic communication in the CNS. Dysregulation of these compounds has been implicated in many brain disorders. Although some studies have identified and quantified a limited number of target compounds, a method that provides comprehensive quantitative information on endocannabinoids and relatedN-acylethanolamines (NAEs) in cerebrospinal fluid (CSF) is currently lacking, as measurements are challenging due to low concentrations under normal physiological conditions. Here we developed and validated a high-throughput nano LC-ESI-MS/MS platform for the simultaneous quantification of endocannabinoids (anandamide and 2-arachidonoylglycerol), ten related NAEs, and eight additional putatively annotated NAEs in human CSF. Requiring only 200 μl of CSF, our method has limits of detection from 0.28 to 61.2 pM with precisions of relative SD <15% for most compounds. We applied our method to CSF from 45 healthy humans and demonstrated potential age and gender effects on concentrations of endocannabinoids and NAEs. Notably, our results show that docosahexaenoylethanolamide concentrations increase with age in males. Our method may offer new opportunities to gain insight into regulatory functions of endocannabinoids in the context of (ab)normal brain function.
authors.affiliation: Division of Analytical Biosciences, Leiden Academic Centre for Drug Research Leiden University, Leiden, The Netherlands.
authors.forename: Vasudev
authors.identifier: 0000-0001-8541-0882
authors.initials: V
authors.lastname: Kantae
authors.affiliation: Division of Analytical Biosciences, Leiden Academic Centre for Drug Research Leiden University, Leiden, The Netherlands.
authors.forename: Shinji
authors.initials: S
authors.lastname: Ogino
authors.affiliation: Division of Analytical Biosciences, Leiden Academic Centre for Drug Research Leiden University, Leiden, The Netherlands.
authors.forename: Marek
authors.initials: M
authors.lastname: Noga
authors.affiliation: Division of Analytical Biosciences, Leiden Academic Centre for Drug Research Leiden University, Leiden, The Netherlands.
authors.forename: Amy C
authors.initials: AC
authors.lastname: Harms
authors.affiliation: Departments of Neurology Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Robin M
authors.initials: RM
authors.lastname: van Dongen
authors.affiliation: Departments of Neurology Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Gerrit L J
authors.initials: GL
authors.lastname: Onderwater
authors.affiliation: Departments of Neurology Leiden University Medical Center, Leiden, The Netherlands.|Human Genetics, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Arn M J M
authors.initials: AM
authors.lastname: van den Maagdenberg
authors.affiliation: Departments of Neurology Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Gisela M
authors.initials: GM
authors.lastname: Terwindt
authors.affiliation: Department of Bioorganic Synthesis, Leiden Institute of Chemistry, Leiden University, Leiden, The Netherlands.
authors.forename: Mario
authors.initials: M
authors.lastname: van der Stelt
authors.affiliation: Departments of Neurology Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Michel D
authors.initials: MD
authors.lastname: Ferrari
authors.affiliation: Division of Analytical Biosciences, Leiden Academic Centre for Drug Research Leiden University, Leiden, The Netherlands hankemeier@lacdr.leidenuniv.nl.
authors.forename: Thomas
authors.initials: T
authors.lastname: Hankemeier
mesh_terms: D000328:Adult; D000367:Age Factors; D001095:Arachidonic Acids / Q000134:cerebrospinal fluid*; D002853:Chromatography, Liquid / Q000379:methods; D063388:Endocannabinoids / Q000134:cerebrospinal fluid*; D004983:Ethanolamines / Q000134:cerebrospinal fluid*; D005260:Female; D005989:Glycerides / Q000134:cerebrospinal fluid*; D006801:Humans; D008297:Male; D008875:Middle Aged; D053284:Polyunsaturated Alkamides / Q000134:cerebrospinal fluid*; D012727:Sex Characteristics; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D001095:Arachidonic Acids; D063388:Endocannabinoids; D004983:Ethanolamines; D005989:Glycerides; C022203:N-acylethanolamines; D053284:Polyunsaturated Alkamides; C094503:glyceryl 2-arachidonate; C078814:anandamide
keywords: brain lipids; cerebrospinal fluid; lipidomics; liquid chromatography; miniaturization; quantitation; tandem mass spectrometry