Journal
TALANTA
Volume 195, Issue -, Pages 593-598Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.talanta.2018.11.094
Keywords
Tryptophan pathway; Kynurenine pathway; Liver matrix; Cecal and intestinal contents; Validation methodology
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Funding
- Institut National de la Sante et de la Recherche Medicale INSERM
- University of Tours
- European Research Council (ERC) under the European Union's Horizon 2020 Research and Innovation Programme [ERC-2016-StG-71577]
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In this study, we validated a method for quantifying 20 tryptophan (Trp) catabolites by liquid chromatography coupled with high resolution mass spectrometry (LC-HRMS) in 4 different matrices (urine, serum, intestinal contents and liver). The detection limit for all metabolites ranged between 0.015 and 11.25 nmol/L and the dynamic range of the calibration curves were adjusted to allow quantification of metabolites at endogenous levels. Matrix effects were evaluated using isotope labeled internal standards. Reproducibility in the 4 matrices was characterized by CV = 6.2% with an accuracy of 6.6%. Our method has been applied to the determination and quantification of 20 metabolites concentrations in 5 different mouse compartments (plus cecal contents). Our results show that our approach allows for a global exploration of the Trp metabolism by quantifying a large number of Trp metabolites, at the individual level by multi-matrix approach.
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