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Validation of a global quantitative analysis methodology of tryptophan metabolites in mice using LC-MS

Lefèvre, Antoine, Mavel, Sylvie, Nadal-Desbarats, Lydie, Galineau, Laurent, Attucci, Sylvie, Dufour, Diane, Sokol, Harry, Emond, Patrick
Talanta 2019 v.195 pp. 593-598
blood serum, calibration, detection limit, intestines, isotope labeling, liquid chromatography, liver, mass spectrometry, metabolism, metabolites, mice, quantitative analysis, tryptophan, urine
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.