Jean-Marc Routaboul, Christian Dubos, Gilles Beck, Catherine Marquis, Przemyslaw Bidzinski,
Olivier Loudet and Loıc Lepiniec. INRA, UMR1318, Institut Jean-Pierre Bourgin, RD10, F-78026 Versailles, France.
AgroParisTech, Institut Jean-Pierre Bourgin, RD10, F-78026 Versailles, France.

Overview

  • Key words: Arabidopsis, flavonoids, metabolite profiling, natural variation, quantitative trait loci
  • Aim of the study: Identification of genes controlling flavonoid metabolism
  • Application: Flavonoid extraction and analysis by LC-MS
  • Sample name: Arabidopsis
  • Sample type: Seeds
  • Material: FastPrep-24™ instrument
  • Buffers: Acetonitrile/Water (3/1; v/v) or Methanol/Acetone/water/Trifluoroacetic acid (30/42/28/0.05; v/v/v/v)

Protocols & Parameters

  1. Three representative seed aliquots from the three biological repeats were pooled before flavonoid extraction.
  2. All seed samples were ground for 90 s at maximum speed with a FastPrep-24™ instrument in 1 ml of solvent mixe.
  3. A 4 μg aliquot of apigenin was added as an internal standard.
  4. Following centrifugation, the pellet was extracted further with 1 ml of the same solvent mixes overnight at 4°C.

Conclusion

  • The use of FastPrep-24™ instrument succeeded in full homogenization of seeds, allowing flavonoid extraction, quantification and complete analysis of their metabolism by LC-MS method.