Synthesis of a deuterated standard for the quantification of 2-Arachidonoylglycerol in Caenorhabditis elegans

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dc.citation.titleJournal of Visualized Experimentses
dc.citation.volume151es
dc.creatorFernández de Luco, Julia
dc.creatorPrez, Gastón M.
dc.creatorHernández Cravero, Bruno
dc.creatorDe Mendoza, Diego
dc.creatorLabadie, Guillermo Roberto
dc.date.accessioned2020-05-28T15:28:36Z
dc.date.available2020-05-28T15:28:36Z
dc.date.issued2019-09-21
dc.descriptionThis work presents a method to prepare an analytical standard to analyze 2-arachidonoyl glycerol (2-AG) qualitatively and quantitatively by liquid chromatography-electrospray Ionization-tandem mass spectrometry (LC-ESI-MS/MS). Endocannabinoids are conserved lipid mediators that regulate multiple biological processes in a variety of organisms. In C. elegans, 2-AG has been found to possess different roles, including modulation of dauer formation and cholesterol metabolism. This report describes a method to overcome the difficulties associated with the costs and stability of deuterated standards required for 2-AG quantification. The procedure for the synthesis of the standard is simple and can be performed in any laboratory, without the need for organic synthesis expertise or special equipment. In addition, a modification of Folch's method to extract the deuterated standard from C. elegans culture is described. Finally, a quantitative and analytic method to detect 2-AG using the stable isotopically labeled analog 1-AG-d5 is described, which provides reliable results in a fast-chromatographic run. The procedure is useful for studying the multiple roles of 2-AG in C. elegans while also being applicable to other studies of metabolites in different organisms.es
dc.description.filFil: Fernández de Luco, Julia. Universidad Nacional del Rosario. Facultad de Ciencias Bioquímicas y Farmaceúticas. Instituto de Química Rosario (IQUIR-CONICET); Argentina.es
dc.description.filFil: Prez, Gastón M. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario. Laboratorio de Fisiología Microbiana (IBR-CONICET); Argentina.es
dc.description.filFil: Hernández Cravero, Bruno. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario. Laboratorio de Fisiología Microbiana (IBR-CONICET); Argentina.es
dc.description.filFil: De Mendoza, Diego. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario. Laboratorio de Fisiología Microbiana (IBR-CONICET); Argentina.es
dc.description.filFil: Labadie, Guillermo Roberto. Universidad Nacional del Rosario. Facultad de Ciencias Bioquímicas y Farmaceúticas. Instituto de Química Rosario (IQUIR-CONICET); Argentina.es
dc.description.sponsorshipAgencia Nacional de Promoción Científica y Tecnológica (ANPCyT, Argentina): PICT 2014-3693es
dc.formatapplication/pdf
dc.format.extent
dc.identifier.issn1940-087Xes
dc.identifier.urihttp://hdl.handle.net/2133/18279
dc.language.isoenges
dc.publisherJoVEes
dc.relation.publisherversionhttps://dx.doi.org/10.3791/59882es
dc.relation.publisherversionhttps://www.jove.com/video/59882/synthesis-deuterated-standard-for-quantification-2es
dc.rightsembargoedAccesses
dc.rights.holderFernández de Luco, Juliaes
dc.rights.holderPrez, Gastón M.es
dc.rights.holderHernández Cravero, Brunoes
dc.rights.holderDe Mendoza, Diegoes
dc.rights.holderLabadie, Guillermo Robertoes
dc.rights.holderThe Journal of Visualized Experiments “JoVE”es
dc.rights.holderUniversidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticases
dc.rights.textAtribución-NoComercial-SinDerivadas 3.0 No portada (CC BY-NC-ND 3.0)es
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/3.0/deed.es*
dc.subjectEndocannabinoidses
dc.subjectC. eleganses
dc.subjectSynthesises
dc.subjectDeuterated analogses
dc.subject2-AGes
dc.subjectDaueres
dc.subjectMAGses
dc.subjectHPLC-MS/MSen
dc.subjectIsotopic dilutionen
dc.subjectQuantificationen
dc.titleSynthesis of a deuterated standard for the quantification of 2-Arachidonoylglycerol in Caenorhabditis eleganses
dc.typearticle
dc.typeartículo
dc.typeacceptedVersion
dc.type.collectionarticulo
dc.type.versionacceptedVersiones

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