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

This 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 bio...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autores principales: Fernández de Luco, Julia, Prez, Gastón, Hernández Cravero, Bruno, De Mendoza, Diego, Labadie, Guillermo Roberto
Formato: article artículo acceptedVersion
Lenguaje:Inglés
Publicado: JoVE 2020
Materias:
Endocannabinoids
C. elegans
Synthesis
Deuterated analogs
2-AG
Dauer
MAGs
HPLC-MS/MS
Isotopic dilution
Quantification
Acceso en línea:http://hdl.handle.net/2133/18279
http://hdl.handle.net/2133/18279
Aporte de:
Repositorio Hipermedial de la Universidad Nacional de Rosario (UNR) de Universidad Nacional de Rosario
id I15-R121-2133-18279
record_format dspace
institution Universidad Nacional de Rosario
institution_str I-15
repository_str R-121
collection Repositorio Hipermedial de la Universidad Nacional de Rosario (UNR)
language Inglés
orig_language_str_mv eng
topic Endocannabinoids
C. elegans
Synthesis
Deuterated analogs
2-AG
Dauer
MAGs
HPLC-MS/MS
Isotopic dilution
Quantification
spellingShingle Endocannabinoids
C. elegans
Synthesis
Deuterated analogs
2-AG
Dauer
MAGs
HPLC-MS/MS
Isotopic dilution
Quantification
Fernández de Luco, Julia
Prez, Gastón
Hernández Cravero, Bruno
De Mendoza, Diego
Labadie, Guillermo Roberto
Synthesis of a deuterated standard for the quantification of 2-Arachidonoylglycerol in Caenorhabditis elegans
topic_facet Endocannabinoids
C. elegans
Synthesis
Deuterated analogs
2-AG
Dauer
MAGs
HPLC-MS/MS
Isotopic dilution
Quantification
description This 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.
format article
artículo
acceptedVersion
author Fernández de Luco, Julia
Prez, Gastón
Hernández Cravero, Bruno
De Mendoza, Diego
Labadie, Guillermo Roberto
author_facet Fernández de Luco, Julia
Prez, Gastón
Hernández Cravero, Bruno
De Mendoza, Diego
Labadie, Guillermo Roberto
author_sort Fernández de Luco, Julia
title Synthesis of a deuterated standard for the quantification of 2-Arachidonoylglycerol in Caenorhabditis elegans
title_short Synthesis of a deuterated standard for the quantification of 2-Arachidonoylglycerol in Caenorhabditis elegans
title_full Synthesis of a deuterated standard for the quantification of 2-Arachidonoylglycerol in Caenorhabditis elegans
title_fullStr Synthesis of a deuterated standard for the quantification of 2-Arachidonoylglycerol in Caenorhabditis elegans
title_full_unstemmed Synthesis of a deuterated standard for the quantification of 2-Arachidonoylglycerol in Caenorhabditis elegans
title_sort synthesis of a deuterated standard for the quantification of 2-arachidonoylglycerol in caenorhabditis elegans
publisher JoVE
publishDate 2020
url http://hdl.handle.net/2133/18279
http://hdl.handle.net/2133/18279
work_keys_str_mv AT fernandezdelucojulia synthesisofadeuteratedstandardforthequantificationof2arachidonoylglycerolincaenorhabditiselegans
AT prezgaston synthesisofadeuteratedstandardforthequantificationof2arachidonoylglycerolincaenorhabditiselegans
AT hernandezcraverobruno synthesisofadeuteratedstandardforthequantificationof2arachidonoylglycerolincaenorhabditiselegans
AT demendozadiego synthesisofadeuteratedstandardforthequantificationof2arachidonoylglycerolincaenorhabditiselegans
AT labadieguillermoroberto synthesisofadeuteratedstandardforthequantificationof2arachidonoylglycerolincaenorhabditiselegans
bdutipo_str Repositorios
_version_ 1764820410170343424

Ejemplares similares