13C labeling analysis of sugars by high resolution-mass spectrometry for metabolic flux analysis

Anal Biochem. 2017 Jun 15:527:45-48. doi: 10.1016/j.ab.2017.02.005. Epub 2017 Feb 14.

Abstract

Metabolic flux analysis is particularly complex in plant cells because of highly compartmented metabolism. Analysis of free sugars is interesting because it provides data to define fluxes around hexose, pentose, and triose phosphate pools in different compartment. In this work, we present a method to analyze the isotopomer distribution of free sugars labeled with carbon 13 using a liquid chromatography-high resolution mass spectrometry, without derivatized procedure, adapted for Metabolic flux analysis. Our results showed a good sensitivity, reproducibility and better accuracy to determine isotopic enrichments of free sugars compared to our previous methods [5, 6].

Keywords: 13C-labeling; Isotopomers; LC-HRMS; Mass spectrometry; Metabolic flux analysis; Sugars.

MeSH terms

  • Carbon Isotopes
  • Chromatography, Liquid
  • Flax / chemistry
  • Flax / growth & development
  • Flax / metabolism*
  • Fructose / biosynthesis
  • Fructose / isolation & purification
  • Glucose / biosynthesis
  • Glucose / isolation & purification
  • Isotope Labeling / methods*
  • Maltose / biosynthesis
  • Maltose / isolation & purification
  • Mass Spectrometry
  • Metabolic Flux Analysis / methods*
  • Raffinose / biosynthesis
  • Raffinose / isolation & purification
  • Reproducibility of Results
  • Seeds / chemistry
  • Seeds / growth & development
  • Seeds / metabolism*
  • Sensitivity and Specificity
  • Sucrose / isolation & purification
  • Sucrose / metabolism

Substances

  • Carbon Isotopes
  • Fructose
  • Sucrose
  • Maltose
  • Glucose
  • Raffinose