Rapid profiling of intact glucosinolates in Arabidopsis leaves by UHPLC-QTOFMS using a charged surface hybrid column.
BACKGROUND
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.


METHODS
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.


CONCLUSIONS
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.
Laboratory of Fundamental and Applied Research in Chemical Ecology, University of Neuchâtel, Rue Emile-Argand 11, 2009 Neuchâtel, Switzerland. gaetan.glauser@unine.ch
Gaetan
G
Glauser
Fabian
F
Schweizer
Ted C J
TC
Turlings
Philippe
P
Reymond
D017360:Arabidopsis / Q000737:chemistry*; D001937:Brassica / Q000737:chemistry; D005591:Chemical Fractionation / Q000295:instrumentation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D056148:Chromatography, Reverse-Phase / Q000295:instrumentation / Q000379:methods / Q000592:standards; D005947:Glucose / Q000031:analogs & derivatives / Q000737:chemistry / Q000302:isolation & purification; D005961:Glucosinolates / Q000032:analysis* / Q000737:chemistry; D007096:Imidoesters / Q000737:chemistry / Q000302:isolation & purification; D056626:Inflorescence / Q000737:chemistry; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods / Q000592:standards; D010936:Plant Extracts / Q000737:chemistry; D018515:Plant Leaves / Q000737:chemistry*; D012639:Seeds / Q000737:chemistry
D005961:Glucosinolates; D007096:Imidoesters; D010936:Plant Extracts; C119493:glucoerucin; C010330:sinigrin; D005947:Glucose