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

被引：60

作者：

Tam, YY ^{[1
]}

Normanly, J ^{[1
]}

机构：

[1] Univ Massachusetts, Dept Biochem & Mol Biol, Amherst, MA 01003 USA

来源：

JOURNAL OF CHROMATOGRAPHY A | 1998年 / 800卷 / 01期

关键词：

Arabidopsis thaliana; derivatization; GC; indolepyruvic acid; pentafluorophenylhydrazine; pentafluorobenzylhydroxylamine;

D O I：

10.1016/S0021-9673(97)01051-0

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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-18 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 TPA-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. (C) 1998 Elsevier Science B.V.

引用

页码：101 / 108

页数：8

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