Untargeted liquid chromatography-mass spectrometry-based metabolomics analysis of wheat grain

Document Type

Journal Article

Publication Title

Journal of Visualized Experiments : JoVE

Publisher

NLM (Medline)

School

Centre for Integrative Metabolomics and Computational Biology / School of Science

RAS ID

32910

Comments

Abbiss, H., Gummer, J. P., Francki, M., & Trengove, R. D. (2020). Untargeted liquid chromatography-mass spectrometry-based metabolomics analysis of wheat grain. Journal of Visualized Experiments: Jove, (157), Article e60851. https://doi.org/10.3791/60851

Abstract

Understanding the interactions between genes, the environment and management in agricultural practice could allow more accurate prediction and management of product yield and quality. Metabolomics data provides a read-out of these interactions at a given moment in time and is informative of an organism's biochemical status. Further, individual metabolites or panels of metabolites can be used as precise biomarkers for yield and quality prediction and management. The plant metabolome is predicted to contain thousands of small molecules with varied physicochemical properties that provide an opportunity for a biochemical insight into physiological traits and biomarker discovery. To exploit this, a key aim for metabolomics researchers is to capture as much of the physicochemical diversity as possible within a single analysis. Here we present a liquid chromatography-mass spectrometry-based untargeted metabolomics method for the analysis of field-grown wheat grain. The method uses the liquid chromatograph quaternary solvent manager to introduce a third mobile phase and combines a traditional reversed-phase gradient with a lipid-amenable gradient. Grain preparation, metabolite extraction, instrumental analysis and data processing workflows are described in detail. Good mass accuracy and signal reproducibility were observed, and the method yielded approximately 500 biologically relevant features per ionization mode. Further, significantly different metabolite and lipid feature signals between wheat varieties were determined.

DOI

10.3791/60851

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