2020-05-282020-05-282019-09-211940-087Xhttp://hdl.handle.net/2133/18279This 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.application/pdfengembargoedAccessEndocannabinoidsC. elegansSynthesisDeuterated analogs2-AGDauerMAGsHPLC-MS/MSIsotopic dilutionQuantificationarticleFernández de Luco, JuliaPrez, Gastón M.Hernández Cravero, BrunoDe Mendoza, DiegoLabadie, Guillermo RobertoThe Journal of Visualized Experiments “JoVE”Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y FarmacéuticasAtribución-NoComercial-SinDerivadas 3.0 No portada (CC BY-NC-ND 3.0)