Rapid and Reliable Quantification of Glycerol-3-phosphate Using Gas Chromatography-coupled Mass Spectrometry.
Arabidopsis thaliana
GC-MS
Glycerol-3-phosphate
N-Methyl-N-(trimethylsilyl)trifluoroacetamide
Plant defense
Journal
Bio-protocol
ISSN: 2331-8325
Titre abrégé: Bio Protoc
Pays: United States
ID NLM: 101635102
Informations de publication
Date de publication:
05 Apr 2023
05 Apr 2023
Historique:
received:
19
05
2022
revised:
17
08
2022
accepted:
22
02
2023
medline:
15
4
2023
entrez:
14
4
2023
pubmed:
15
4
2023
Statut:
epublish
Résumé
Glycerol-3-phosphate (G3P) is a conserved precursor of glycerolipids that also plays an important role in plant defense. Its levels and/or metabolism are also associated with many human disorders including insulin resistance, diabetes, obesity, and cancer, among others. In plants, G3P accumulates upon pathogen infection and is a critical component of systemic acquired resistance, which confers broad spectrum disease resistance against secondary infections. G3P also plays an important role in root-shoot-root signaling in soybean that regulates incompatible interactions with nitrogen-fixing bacteria. Thus, accurate quantification of G3P is key to drawing a valid conclusion regarding its role in diverse processes ranging from lipid biosynthesis to defense. G3P quantification is further compounded by its rapid degradation in extracts prepared at room temperature. Here, we describe a simplified procedure for accurate quantitative analysis of G3P from plant tissues. G3P was extracted along with the internal standard ribitol, derivatized with N-Methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA) and analyzed by gas chromatography-coupled mass spectrometry using selective ion mode. This procedure is simple, economical, and efficient, and does not involve isotopic internal standards or multiple-step derivatizations.
Identifiants
pubmed: 37056243
doi: 10.21769/BioProtoc.4645
pii: e4645
pmc: PMC10086549
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e4645Informations de copyright
Copyright © 2023 The Authors.
Déclaration de conflit d'intérêts
Competing interestsThe authors declare no conflict of interest.
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