Rapid and Reliable Quantification of Glycerol-3-phosphate Using Gas Chromatography–coupled Mass Spectrometry

Huazhen Liu, Keshun Yu, Pradeep Kachroo

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Article numbere4645
JournalBio-protocol
Volume13
Issue number7
DOIs
StatePublished - Apr 5 2023

Bibliographical note

Publisher Copyright:
Copyright: © 2023 The Authors; exclusive licensee Bio-protocol LLC.

Keywords

  • Arabidopsis thaliana
  • GC-MS
  • Glycerol-3-phosphate
  • N-Methyl-N-(trimethylsilyl)trifluoroacetamide
  • Plant defense

ASJC Scopus subject areas

  • General Neuroscience
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology
  • Plant Science

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