Determination of indole-3-pyruvic acid levels in Arabidopsis thaliana by gas chromatography-selected ion monitoring-mass spectrometry

J Chromatogr A. 1998 Mar 20;800(1):101-8. doi: 10.1016/s0021-9673(97)01051-0.

Abstract

A rapid and simple method is described for the determination of indole-3-pyruvic acid (IPA) levels in Arabidopsis thaliana by gas chromatography-selected ion monitoring-mass spectrometry (GC-SIM-MS). The method includes derivatization of IPA with hydroxylamine in the crude extract, followed by ethyl acetate partitioning, solid-phase extraction with C18 resin, reversed-phase high-performance liquid chromatography (HPLC), and GC-SIM-MS. Three derivatizing reagents were tested; these were pentafluorobenzylhydroxylamine, pentafluorophenylhydrazine, and hydroxylamine. Hydroxylamine proved to be the most useful, as the IPA-oxime was easiest to purify from plant extracts and was the most stable. IPA was quantified in Arabidopsis seedlings ranging in age from 5 to 12 days; levels varied from 4 to 13 ng/g, peaking at 7-9 days.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Arabidopsis / chemistry*
  • Calibration
  • Free Radical Scavengers / analysis*
  • Free Radical Scavengers / chemistry
  • Gas Chromatography-Mass Spectrometry / methods*
  • Hydroxylamine / chemistry
  • Indoles / analysis*
  • Indoles / chemistry

Substances

  • Free Radical Scavengers
  • Indoles
  • Hydroxylamine
  • indol-3-yl pyruvic acid