, 15:79 | Cite as

Volatile apocarotenoid discovery and quantification in Arabidopsis thaliana: optimized sensitive analysis via HS-SPME-GC/MS

  • John Y. Rivers
  • Thy T. Truong
  • Barry J. Pogson
  • Ryan P. McQuinnEmail author
Original Article
Part of the following topical collections:
  1. Plant metabolomics and lipidomics



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.


Optimization and subsequent validation of the HS-SPME technique for extracting and quantifying volatile apocarotenoids in planta.


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.


Mean linear regression in planta calibration correlation coefficients (R2) 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 µg/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 µm divinylbenzene/carboxen on polydimethylsiloxane (PDMS/DVB/CAR) was identified as the optimal fibre for broad molecular weight range VA analysis.


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.


Arabidopsis thaliana Apocarotenoid SPME Volatiles GC/MS 



John Y. Rivers acknowledges the support of the Grains Research and Development Corporation (GRDC) Grains Industry Research Scholarship (GRS10687) and the Australian Government Research Training Program. John Y. Rivers, Ryan P. McQuinn and Barry J. Pogson acknowledge the support of the Australian Research Council Centre of Excellence in Plant Energy Biology (CE140100008).

Supplementary material

11306_2019_1529_MOESM1_ESM.xlsx (121 kb)
Electronic supplementary material 1 (XLSX 121 kb) (1.2 mb)
Electronic supplementary material 2 (AI 1221 kb)


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Australian Research Council Centre of Excellence in Plant Energy Biology, Research School of BiologyThe Australian National UniversityCanberraAustralia
  2. 2.Joint Mass Spectrometry Facility, Research School of ChemistryThe Australian National UniversityCanberraAustralia

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