Document Detail

Methods for the quantitation of abscisic acid and its precursors from plant tissues.
MedLine Citation:
PMID:  11161319     Owner:  NLM     Status:  MEDLINE    
Methods are given for the quantitation of the plant stress hormone, abscisic acid (ABA), and its two metabolic precursors, ketone and enolate, that are applicable to all species tested so far. The plant extract is homogenized at neutral pH, hexane-soluble neutrals are extracted and discarded, and then the free ABA and other organic acids are extracted as ion pairs. The remaining aqueous phase is acidified, allowed to stand, neutralized, and extracted to give the ABA ex ketone fraction and then the aqueous phase is treated with base and again extracted to give the ABA ex enolate fraction. Each of these three fractions, free ABA, ABA ex ketone, and ABA ex enolate, along with a deuteriated internal standard, [side-chain-(2)H(4)]ABA, is then derivatized with pentafluorobenzyl bromide and purified on an automated sample preparation system. The resulting pentafluorobenzyl abscisate samples are then quantified using electron capture negative ionization mass spectrometry with methane as the reagent gas. Using these procedures free ABA, and ABA from its precursors, can be quantified at the level of 100 fg on column. If a large volume injector is used so that the total sample is injected it should be possible to quantify ABA and its precursors in the parts per billion range on a few milligrams of plant tissue.
P H Duffield; A G Netting
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical biochemistry     Volume:  289     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  2001 Feb 
Date Detail:
Created Date:  2001-02-22     Completed Date:  2001-04-26     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  251-9     Citation Subset:  IM    
Copyright Information:
Copyright 2001 Academic Press.
School of Biochemistry and Molecular Genetics, University of New South Wales, Sydney, 2052, Australia
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MeSH Terms
Abscisic Acid / analysis*
Chemistry Techniques, Analytical / methods*
Gas Chromatography-Mass Spectrometry
Hydrogen-Ion Concentration
Lauraceae / chemistry
Lycopersicon esculentum / chemistry
Methane / chemistry
Models, Chemical
Peas / chemistry
Plants / metabolism*
Sensitivity and Specificity
Spectrometry, Mass, Electrospray Ionization / methods
Time Factors
Reg. No./Substance:
0/Ions; 21293-29-8/Abscisic Acid; 74-82-8/Methane

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