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Metabolomics

Title:		Volatile apocarotenoid discovery and quantification in Arabidopsis thaliana: optimized sensitive analysis via HS-SPME-GC/MS
Author(s):		Rivers JY; Truong TT; Pogson BJ; McQuinn RP;
Address:		"Australian Research Council Centre of Excellence in Plant Energy Biology, Research School of Biology, The Australian National University, Canberra, Australia. Joint Mass Spectrometry Facility, Research School of Chemistry, The Australian National University, Canberra, Australia. Australian Research Council Centre of Excellence in Plant Energy Biology, Research School of Biology, The Australian National University, Canberra, Australia. ryan.mcquinn@anu.edu.au"
Journal Title:		Metabolomics
Year:		2019
Volume:		20190513
Issue:		5
Page Number:		79 -
DOI:		10.1007/s11306-019-1529-y
ISSN/ISBN:		1573-3890 (Electronic) 1573-3882 (Linking)
Abstract:		"INTRODUCTION: In the field of carotenoid metabolism researchers' focus has been directed recently toward the discovery and quantification of carotenoid cleavage products (i.e. apocarotenoids, excluding the well-studied carotenoid-derived hormones abscisic acid and strigolactones), due to their emerging roles as putative signaling molecules. Gas chromatography mass spectrometry (GC/MS) and sample preparation via headspace solid phase micro-extraction (HS-SPME) are widely used analytical techniques for broad untargeted metabolomics studies and until now, no optimized quantitative targeted HS-SPME-GC/MS method has been developed specifically for volatile apocarotenoids (VAs) in planta. OBJECTIVES: Optimization and subsequent validation of the HS-SPME technique for extracting and quantifying volatile apocarotenoids in planta. METHODS: Factors considered during method optimization were HS-SPME parameters; vial storage conditions; different adsorbent SPME fibre coating chemistries; plant tissue matrix effects; and fresh tissues to be analyzed. RESULTS: Mean linear regression in planta calibration correlation coefficients (R(2)) for VAs was 0.974. The resultant method mean limits of detection (LOD) and lower limits of quantification (LLOQ) for VAs using in planta standard additions were 0.384 +/- 0.139 and 0.640 +/- 0.231 microg/L, respectively. VAs remained stable at elevated SPME incubation temperatures, with no observable effects of thermal and photo-stereoisomerisation and oxidation. The bipolar 50/30 microm divinylbenzene/carboxen on polydimethylsiloxane (PDMS/DVB/CAR) was identified as the optimal fibre for broad molecular weight range VA analysis. CONCLUSIONS: An optimized HS-SPME-GC/MS method for VA detection and quantification was validated in vitro and in planta: based on biological replicates and stringent QA/QC approaches, thereby providing robust detection and quantification of VAs across a broad range of Arabidopsis tissues, fifteen of which were identified for the first time in Arabidopsis"
Keywords:		Arabidopsis/*chemistry Carotenoids/*analysis/metabolism *Drug Discovery Gas Chromatography-Mass Spectrometry Solid Phase Microextraction Volatile Organic Compounds/*analysis/metabolism Apocarotenoid Arabidopsis thaliana Gc/ms Spme Volatiles;
Notes:		"MedlineRivers, John Y Truong, Thy T Pogson, Barry J McQuinn, Ryan P eng Research Support, Non-U.S. Gov't 2019/05/16 Metabolomics. 2019 May 13; 15(5):79. doi: 10.1007/s11306-019-1529-y"