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LC–MS analysis of low molecular weight organic acids derived from root exudation

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Abstract

A sensitive method for quantification of citric, fumaric, malic, malonic, oxalic, trans aconitic, and succinic acid in soil- and root-related samples is presented. The method is based on a novel, fast, and simple esterification procedure and subsequent analysis via liquid chromatography–mass spectrometry. Derivatization comprises in situ generation of HCl, which catalyzes the Fischer esterification with benzyl alcohol. As a key advance, the esterification with the aromate allows reversed-phase separation and improves electrospray ionization efficiency. The method provided procedural detection limits of 1 nM for citric, 47 nM for fumaric, 10 nM for malic, 10 nM for malonic, 16 nM for oxalic, 15 nM for succinic, and 2 nM for aconitic acid utilizing 500 μL of liquid sample. The working range was 3 nM to 10 μM for citric acid, 158 nM to 10 μM for fumaric acid, 34 nM to 10 μM for malic acid, 33 nM to 10 μM for malonic acid, 53 nM to 10 μM for oxalic acid, 48 nM to 10 μM for succinic acid, and 6 nM to 10 μM for aconitic acid. Quantification of the analytes in soil-related samples was performed via external calibration of the entire procedure utilizing 13C-labeled oxalic and citric acid as internal standards. The robustness of the method was tested with soil extracts and samples from hydroponic experiments. The latter concerned the regulation of phosphorus solubilization via plant root exudation of citric, malic, and oxalic acid.

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Acknowledgement

We are grateful for financial support from the Austrian Science Fund (Project P20069-B16).

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Authors and Affiliations

  1. Department of Chemistry, University of Natural Resources and Applied Life Sciences – BOKU Vienna, Muthgasse 18, 1190, Vienna, Austria

    Leonhard Jaitz, Bernhard Mueller, Gunda Koellensperger, Daniela Huber & Stephan Hann

  2. Department of Forest and Soil Science, University of Natural Resources and Applied Life Sciences—BOKU Vienna, Peter-Jordanstraße 82, 1190, Vienna, Austria

    Eva Oburger & Markus Puschenreiter

Authors
  1. Leonhard Jaitz
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  2. Bernhard Mueller
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  3. Gunda Koellensperger
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  4. Daniela Huber
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  5. Eva Oburger
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  6. Markus Puschenreiter
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  7. Stephan Hann
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Correspondence to Stephan Hann.

Additional information

Published in the special issue Analytical Sciences in Austria with guest editors G. Allmaier, W. Buchberger, and K. Francesconi.

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Jaitz, L., Mueller, B., Koellensperger, G. et al. LC–MS analysis of low molecular weight organic acids derived from root exudation. Anal Bioanal Chem 400, 2587–2596 (2011). https://doi.org/10.1007/s00216-010-4090-0

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  • DOI: https://doi.org/10.1007/s00216-010-4090-0

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