Document Detail

Rapid extraction of abscisic acid and its metabolites for liquid chromatography-tandem mass spectrometry.
MedLine Citation:
PMID:  14503817     Owner:  NLM     Status:  MEDLINE    
We have described a simple, reliable and rapid method of extracting and partially purifying the phytohormone (+)-abscisic acid and its catabolites for liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS-MS) analysis. Lyophilized tissue samples were powdered by high-speed agitation with ceramic beads for 5 s. Metabolites were extracted from the tissue powder using acetone-water-acetic acid (80:19:1, v/v) with the addition of deuterated internal standards for quantification. Essentially all endogenous hormones were recovered by three successive tissue extractions. However we demonstrated that, with the use of internal standards, one extraction with vigorous vortexing was sufficient to obtain accurate results (recovery 65-90%). Solvents were optimized for partial purification of abscisic acid and related compounds by solid-phase extraction using Oasis HLB cartridges. The eluted metabolites were then analyzed by LC-MS-MS. To illustrate the applicability of these techniques, we analyzed the levels of abscisic acid and metabolites in seeds and valves of Brassica napus siliques at two stages of development. We detected abscisic acid, phaseic acid, 7'-hydroxyabscisic acid, dihydrophaseic acid and abscisic acid glucose ester. In both tissues, dihydrophaseic acid was the major accumulating product, reaching 97300 pmol/g dry mass in valves at 24 days after anthesis. The amount of abscisic acid in seeds was high at 24 days after anthesis (23300 pmol/g dry mass), but low in the other samples (292-447 pmol/g dry mass).
Rong Zhou; Timothy M Squires; Stephen J Ambrose; Suzanne R Abrams; Andrew R S Ross; Adrian J Cutler
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of chromatography. A     Volume:  1010     ISSN:  0021-9673     ISO Abbreviation:  J Chromatogr A     Publication Date:  2003 Aug 
Date Detail:
Created Date:  2003-09-23     Completed Date:  2004-04-22     Revised Date:  2009-01-15    
Medline Journal Info:
Nlm Unique ID:  9318488     Medline TA:  J Chromatogr A     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  75-85     Citation Subset:  IM    
Plant Biotechnology Institute, 110 Gymnasium Place, National Research Council of Canada, Saskatoon S7N 0W9, Canada.
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MeSH Terms
Abscisic Acid / isolation & purification*,  metabolism
Brassica / metabolism*
Chromatography, Liquid / methods*
Spectrometry, Mass, Electrospray Ionization / methods*
Reg. No./Substance:
0/Solutions; 21293-29-8/Abscisic Acid

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