Simplified absolute metabolite quantification by gas chromatography–isotope dilution mass spectrometry on the basis of commercially available source material

Abstract:

In the field of metabolomics, GC–MS has rather established itself as a tool for semi-quantitative strategies like metabolic fingerprinting or metabolic profiling. Absolute quantification of intra- or extracellular metabolites is nowadays mostly accomplished by application of diverse LC–MS techniques. Only few groups have so far adopted GC–MS technology for this exceptionally challenging task. Besides numerous and deeply investigated problems related to sample generation, the pronounced matrix effects in biological samples have led to the almost mandatory application of isotope dilution mass spectrometry (IDMS) for the accurate determination of absolute metabolite concentrations. Nevertheless, access to stable isotope labeled internal standards (ILIS), which are in many cases commercially unavailable, is quite laborious and very expensive. Here we present an improved and simplified gas chromatography–isotope dilution mass spectrometry (GC–IDMS) protocol for the absolute determination of intra- and extracellular metabolite levels. Commercially available 13C-labeled algal cells were used as a convenient source for the preparation of internal standards. Advantages as well as limitations of the described method are discussed.

FAIRDOMHub ID: https://fairdomhub.org/publications/137

DOI: 10.1016/j.jchromb.2011.10.036

Projects: MOSES

Journal: Journal of Chromatography B

Citation:

Date Published: 1st Dec 2011

Authors: Oliver Vielhauer, Maksim Zakhartsev, Thomas Horn, Ralf Takors, Matthias Reuss

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Citation
Vielhauer, O., Zakhartsev, M., Horn, T., Takors, R., & Reuss, M. (2011). Simplified absolute metabolite quantification by gas chromatography–isotope dilution mass spectrometry on the basis of commercially available source material. Journal of Chromatography B, 879(32), 3859–3870. http://doi.org/10.1016/j.jchromb.2011.10.036
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Created: 1st Mar 2012 at 12:04

Last updated: 1st Mar 2012 at 12:05

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