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Rapid Profiling of Intact Glucosinolates in Arabidopsis Leaves by UHPLC‐QTOFMS Using a Charged Surface Hybrid Column
Author(s) -
Glauser Gaetan,
Schweizer Fabian,
Turlings Ted C. J.,
Reymond Philippe
Publication year - 2012
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.2350
Subject(s) - chemistry , chromatography , formic acid , mass spectrometry , arabidopsis , sample preparation , polar , elution , brassicaceae , resolution (logic) , analytical chemistry (journal) , botany , biochemistry , physics , astronomy , mutant , gene , artificial intelligence , biology , computer science
The analysis of glucosinolates (GS) is traditionally performed by reverse‐phase liquid chromatography coupled to ultraviolet detection after a time‐consuming desulphation step, which is required for increased retention. Simpler and more efficient alternative methods that can shorten both sample preparation and analysis are much needed. Objective To evaluate the feasibility of using ultrahigh‐pressure liquid chromatography coupled to quadrupole time‐of‐flight mass spectrometry (UHPLC‐QTOFMS) for the rapid profiling of intact GS. Methodology A simple and short extraction of GS from Arabidopsis thaliana leaves was developed. Four sub‐2 µm reverse‐phase columns were tested for the rapid separation of these polar compounds using formic acid as the chromatographic additive. High‐resolution QTOFMS was used to detect and identify GS. Results A novel charged surface hybrid (CSH) column was found to provide excellent retention and separation of GS within a total running time of 11 min. Twenty‐one GS could be identified based on their accurate mass as well as isotopic and fragmentation patterns. The method was applied to determine the changes in GS content that occur after herbivory in Arabidopsis . In addition, we evaluated its applicability to the profiling of other Brassicaceae species. Conclusion The method developed can profile the full range of GS, including the most polar ones, in a shorter time than previous methods, and is highly compatible with mass spectrometric detection. Copyright © 2012 John Wiley & Sons, Ltd.