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Screening for Wound‐induced Oxylipins in Arabidopsis thaliana by Differential HPLC‐APCI/MS Profiling of Crude Leaf Extracts and Subsequent Characterisation by Capillary‐scale NMR
Author(s) -
Thiocone Aly,
Farmer Edward E.,
Wolfender JeanLuc
Publication year - 2008
Publication title -
phytochemical analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 72
eISSN - 1099-1565
pISSN - 0958-0344
DOI - 10.1002/pca.1051
Subject(s) - chemistry , chromatography , high performance liquid chromatography , metabolome , oxylipin , repeatability , atmospheric pressure chemical ionization , metabolomics , mass spectrometry , sample preparation , biochemistry , enzyme , organic chemistry , chemical ionization , ion , ionization
A simple non‐targeted differential HPLC‐APCI/MS approach has been developed in order to survey metabolome modifications that occur in the leaves of Arabidopsis thaliana following wound‐induced stress. The wound‐induced accumulation of metabolites, particularly oxylipins, was evaluated by HPLC‐MS analysis of crude leaf extracts. A generic, rapid and reproducible pressure liquid extraction procedure was developed for the analysis of restricted leaf samples without the need for specific sample preparation. The presence of various oxylipins was determined by head‐to‐head comparison of the HPLC‐MS data, filtered with a component detection algorithm, and automatically compared with the aid of software searching for small differences in similar HPLC‐MS profiles. Repeatability was verified in several specimens belonging to different series. Wound‐inducible jasmonates were efficiently highlighted by this non‐targeted approach without the need for complex sample preparation as is the case for the ‘oxylipin signature’ procedure based on GC‐MS. Furthermore this HPLC‐MS screening technique allowed the isolation of induced compounds for further characterisation by capillary‐scale NMR (CapNMR TM ) after HPLC scale‐up. In this paper, the screening method is described and applied to illustrate its potential for monitoring polar and non‐polar stress‐induced constituents as well as its use in combination with CapNMR for the structural assignment of wound‐induced compounds of interest. Copyright © 2008 John Wiley & Sons, Ltd.