The measurement and mass spectral identification of indole-3-pyruvate from tomato shoots.
Endogenous indole-3-pyruvate has been identified by full-scan combined gas chromatography-mass spectrometry and measured using gas chromatography with an electron capture detector. High specific-activity [5-3H]indole-3-pyruvate was synthesized from [5-3H]tryptophan and used as an internal standard. In order to allow purification of the labile indole-3-pyruvate it was stabilised by the formation in the crude extract of its pentafluorobenzyl oxime derivative. This derivative also allowed sensitive detection and measurement of indole-3-pyruvate in the picogram range using a gas chromatograph with an electron capture detector. Endogenous levels were found to be between 8-10 ng/g f.wt. of tomato shoots which is comparable to that of the indole-3-acetic acid pool size, 11 ng/g f.wt., in this tissue.
Cooney
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TP
Department of Biochemistry, Auckland University, New Zealand.
Nonhebel
H M
HM
D002851:Chromatography, High Pressure Liquid; D008401:Gas Chromatography-Mass Spectrometry; D006898:Hydroxylamines; D007202:Indicators and Reagents; D007211:Indoles / Q000032:analysis*; D013058:Mass Spectrometry; D015394:Molecular Structure; D010944:Plants / Q000032:analysis*; D005466:Fluorocarbons
D006898:Hydroxylamines; D007202:Indicators and Reagents; D007211:Indoles; C008122:indol-3-yl pyruvic acid; C010277:Florox Reagent; D005466:Fluorocarbons