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

Tipo de material: TextoTextoSeries ; Journal of Chromatography A, 800(1), p.101-108, 1998Trabajos contenidos:
  • Tam, Y.Y
  • Normanly, J
Tema(s): Recursos en línea: Resumen: 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 C resin, reversed-phase high-performance liquid chromatography (HPLC), and GC-SIM-MS. Three derivatizing 18 reagents were tested; these were entafluorobenzylhydroxylamine, 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.
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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 C resin, reversed-phase high-performance liquid chromatography (HPLC), and GC-SIM-MS. Three derivatizing 18 reagents were tested; these were entafluorobenzylhydroxylamine, 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.

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