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Free Content Elemental formula annotation of polar and lipophilic metabolites using

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Summary

The unbiased and comprehensive analysis of metabolites in any organism presents a major challenge if proper peak annotation and unambiguous assignment of the biological origin of the peaks are required. Here we provide a comprehensive multi‐isotope labelling‐based strategy using fully labelled 13C, 15N and 34S plant tissues, in combination with a fractionated metabolite extraction protocol. The extraction procedure allows for the simultaneous extraction of polar, semi‐polar and hydrophobic metabolites, as well as for the extraction of proteins and starch. After labelling and extraction, the metabolites and lipids were analysed using a high‐resolution mass spectrometer providing accurate MS and all‐ion fragmentation data, providing an unambiguous readout for every detectable isotope‐labelled peak. The isotope labelling assisted peak annotation process employed can be applied in either an automated database‐dependent or a database‐independent analysis of the plant polar metabolome and lipidome. As a proof of concept, the developed methods and technologies were applied and validated using Arabidopsis thaliana leaf and root extracts. Along with a large repository of assigned elemental compositions, which is provided, we show, using selected examples, the accuracy and reliability of the developed workflow.
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Document Type: Research Article

Affiliations: Max Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, 14476 Potsdam, Germany

Publication date: October 1, 2011

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