Simultaneous quantification of the concentration and carbon isotopologue distribution of polar metabolites in a single analysis by gas chromatography and mass spectrometry
Evers B, Gerding A, Boer T, Heiner-Fokkema MR, Jalving M, Reijngoud DJ, Bakker BM, Wahl SA (2021)
Publication Type: Journal article
Publication year: 2021
Journal
Book Volume: 93
Pages Range: 8248-8256
Journal Issue: 23
DOI: 10.1021/acs.analchem.1c01040
Abstract
13C-isotope tracing is a frequently employed approach to study metabolic pathway activity. When combined with the subsequent quantification of absolute metabolite concentrations, this enables detailed characterization of the metabolome in biological specimens and facilitates computational time-resolved flux quantification. Classically, a 13C-isotopically labeled sample is required to quantify 13C-isotope enrichments and a second unlabeled sample for the quantification of metabolite concentrations. The rationale for a second unlabeled sample is that the current methods for metabolite quantification rely mostly on isotope dilution mass spectrometry (IDMS) and thus isotopically labeled internal standards are added to the unlabeled sample. This excludes the absolute quantification of metabolite concentrations in 13C-isotopically labeled samples. To address this issue, we have developed and validated a new strategy using an unlabeled internal standard to simultaneously quantify metabolite concentrations and 13C-isotope enrichments in a single 13C-labeled sample based on gas chromatography−mass spectrometry (GC/MS). The method was optimized for amino acids and citric acid cycle intermediates and was shown to have high analytical precision and accuracy. Metabolite concentrations could be quantified in small tissue samples (≥20 mg). Also, we applied the method on 13C-isotopically labeled mammalian cells treated with and without a metabolic inhibitor. We proved that we can quantify absolute metabolite concentrations and 13C-isotope enrichments in a single 13C-isotopically labeled sample.
Authors with CRIS profile
Involved external institutions
Delft University of Technology (TU Delft)
Netherlands (NL)
University of Groningen / Rijksuniversiteit Groningen
Netherlands (NL)
Netherlands (NL)
University of Groningen / Rijksuniversiteit Groningen
Netherlands (NL)
How to cite
APA:
Evers, B., Gerding, A., Boer, T., Heiner-Fokkema, M.R., Jalving, M., Reijngoud, D.-J.,... Wahl, S.A. (2021). Simultaneous quantification of the concentration and carbon isotopologue distribution of polar metabolites in a single analysis by gas chromatography and mass spectrometry. Analytical Chemistry, 93(23), 8248-8256. https://dx.doi.org/10.1021/acs.analchem.1c01040
MLA:
Evers, Bernard, et al. "Simultaneous quantification of the concentration and carbon isotopologue distribution of polar metabolites in a single analysis by gas chromatography and mass spectrometry." Analytical Chemistry 93.23 (2021): 8248-8256.
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