Carotenoid Isotopolog Profiling in 13C-Labeled Leaf Extracts by LC-MS and LC-FTICR-MS

Methods Mol Biol. 2020:2083:263-277. doi: 10.1007/978-1-4939-9952-1_20.

Abstract

Mass spectrometry (MS)-based metabolite analysis combined with stable isotope labeling offers a powerful tool to study dynamic regulation of metabolic pathways and metabolite fluxes in biological systems. Here we describe a method to analyze the composition of carotenoid isotopologs in 13C-labeled leaf extracts by using liquid chromatography (LC)-MS and LC-Fourier transform ion cyclotron resonance (FTICR)-MS. High mass resolution of the latter enables unambiguous assignment of observed mass to a unique chemical formula. Based on peak intensity the relative abundance and the degree of 13C labeling are calculated for individual carotenoid isotopologs.

Keywords: 13C labeling; Carotenoid metabolism; Degree of labeling; FTICR-MS; Isotopolog profiling; LC-MS.

MeSH terms

  • Carbon Isotopes*
  • Carotenoids / analysis
  • Carotenoids / chemistry*
  • Chromatography, Liquid*
  • Isotope Labeling*
  • Pigments, Biological / analysis
  • Pigments, Biological / chemistry
  • Pigments, Biological / isolation & purification
  • Plant Extracts / analysis
  • Plant Extracts / chemistry*
  • Plant Extracts / isolation & purification
  • Plant Leaves / chemistry*
  • Tandem Mass Spectrometry*

Substances

  • Carbon Isotopes
  • Pigments, Biological
  • Plant Extracts
  • Carotenoids