Exploration of SCFA in human multi-compartments using 3-nitrophenylhydrazine-LC-MS
Résumé
Short-chain fatty acids (SCFAs) are metabolites involved in many physiological processes. A disruption of their metabolism is suspected in diseases of the digestive system (inflammatory diseases) but also in neurodegenerative diseases such as Alzheimer's disease. Currently, one of the reference methodologies for the quantitative analysis of SCFA is based on the use of GC-MS due to its sensitivity and reproducibility. However, its technical implementation is delicate, limiting in particular the throughput of the analyzes carried out. Our objective is to propose a method for absolute quantification of SCFA that is easier to implement and for future use in a clinical environment for a large number of types of biological samples (stool, urine, serum, saliva, CSF, and dried blood spot). With this objective we favored a chemical derivatization approach in order to promote the detection and analytical separation of these metabolites. The derivatizing agent we selected, 3-nitrophenylhydrazine (3NPH), meets these criteria. In addition, the use of this molecular pattern on a complex sample will promote the detection and identification of all metabolites carrying a carboxylic acid function beyond the SCFAs targeted in this sample. After methodological validation (robustness, repeatability, matrix effect, carryover, LLOQ), its use makes it possible to propose reference values of SCFA concentrations for each type of sample. This quantitative methodology also allows longitudinal monitoring of SCFA concentrations. In particular, in saliva, our results show a circadian evolution of SCFA concentrations. Validated on several types of samples, our approach makes it possible to quantify the metabolism of SCFAs at the level of an individual and longitudinally.
Domaines
Chimie analytiqueOrigine | Fichiers produits par l'(les) auteur(s) |
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