Simultaneous Quantification of Kynurenine-related Metabolites in Biological Matrices Using UHPLC–MS/MS

A new study published in the Journal of Chromatography B presents how ultrahigh-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) can be used to quantify kynurenine-related metabolites in diverse biological matrices (1). Researchers from Technische Universität Dresden explored the kynurenine pathway in an effort to learn more about its relevance in various diseases, including neurodegenerative/neuropsychiatric disorders, diabetes mellitus type 2, and cancer. Through using UHPLC–MS/MS, the researchers discovered various advantages to employing this approach, including its versatility, interday precision (<14.8%), mean accuracy, and a linear detection range spanning more than three orders of magnitude (1).

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The kynurenine pathway, responsible for the metabolism of tryptophan, an essential amino acid, and the neurotransmitter serotonin, has long been a focus of scientific investigation due to its implications in health and disease. Dysregulations in this pathway have been linked to a range of medical conditions. Consequently, accurate measurements of kynurenine-related metabolites have the potential to shed light on disease pathogenesis.

To achieve this, the researchers developed a simple and sensitive UHPLC–MS/MS method capable of quantifying up to ten kynurenine-related metabolites in diverse biological matrices. Notably, this method allowed for the simultaneous quantification of these metabolites without the need for derivatization.

With a remarkably short run time of 6.5 min, the researchers successfully separated kynurenine-related metabolites, even those with similar structures, such as nicotinic acid and picolinic acid (1). The method's versatility was further established through its application in quantifying native metabolite concentrations in murine tissues and cellular systems. It allowed for the monitoring of pathway shifts in response to treatment with the tryptophan-2,3-dioxygenase-inhibitor 680C91, opening new avenues for research into the kynurenine pathway's role in disease pathogenesis.

Moreover, the UHPLC–MS/MS method was found to be suitable for integration into multi-omics approaches, streamlining metabolite extraction from a single sample for comprehensive analyses. This capability enhances its utility in exploring the broader landscape of biological processes (1).

The potential applications of this novel method are vast. It offers researchers a valuable tool for investigating the kynurenine pathway's involvement in various diseases, providing a deeper understanding of the mechanisms at play. Furthermore, its compatibility with a wide range of physiological matrices ensures its adaptability to different experimental settings.

These findings have opened the door to more in-depth research into the kynurenine pathway and its implications for health and disease. To summarize, the research team was able to demonstrate LC–MS/MS analysis of up to kynurenine-related metabolites––a method that could be integrated into multi-assay analyses using one sample extract (1).

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Reference

(1) Frobel, D.; Stanke, D.; Langner, M.; et al. Liquid chromatography-tandem mass spectrometry based simultaneous quantification of tryptophan, serotonin and kynurenine pathway metabolites in tissues and cell culture systems. J. Chromatogr. B. 2023, 1229, 123870. DOI: j.jchromb.2023.123870