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Open AccessArticle

Metabolic Profiling of Human Plasma and Urine, Targeting Tryptophan, Tyrosine and Branched Chain Amino Acid Pathways

1
Department of Food Quality and Nutrition, Research and Innovation Centre, Fondazione Edmund Mach (FEM), Via E. Mach 1, 38010 San Michele all’ Adige, Italy
2
CIBIO, Department of Cellular, Computational and Integrative Biology, Via Sommarive 9, 38123 Povo, Italy
3
Nutritional Epidemiology, Institute of Nutrition and Food Sciences, University of Bonn, Endenicher Allee 19b, 53115 Bonn, Germany
4
Population Health Sciences, German Center for Neurodegenerative diseases (DZNE), Venusberg-Campus 1-Building 99, 53127 Bonn, Germany
5
Institute for Medical Biometry, Informatics and Epidemiology (IMBIE), Faculty of Medicine, University of Bonn, Venusberg-Campus 1-Building 11, 53127 Bonn, Germany
6
University of Trento, Department of Physics, Bioorganic Chemistry Laboratory, Via Sommarive 14, 38123 Povo, Italy
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Metabolites 2019, 9(11), 261; https://doi.org/10.3390/metabo9110261
Received: 2 October 2019 / Revised: 21 October 2019 / Accepted: 28 October 2019 / Published: 1 November 2019
(This article belongs to the Special Issue Metabolomic Profiles in Nutrition and Metabolic Health)
Tryptophan and tyrosine metabolism has a major effect on human health, and disorders have been associated with the development of several pathologies. Recently, gut microbial metabolism was found to be important for maintaining correct physiology. Here, we describe the development and validation of a UHPLC-ESI-MS/MS method for targeted quantification of 39 metabolites related to tryptophan and tyrosine metabolism, branched chain amino acids and gut-derived metabolites in human plasma and urine. Extraction from plasma was optimised using 96-well plates, shown to be effective in removing phospholipids. Urine was filtered and diluted ten-fold. Metabolites were separated with reverse phase chromatography and detected using triple quadrupole MS. Linear ranges (from ppb to ppm) and correlation coefficients (r2 > 0.990) were established for both matrices independently and the method was shown to be linear for all tested metabolites. At medium spiked concentration, recovery was over 80% in both matrices, while analytical precision was excellent (CV < 15%). Matrix effects were minimal and retention time stability was excellent. The applicability of the methods was tested on biological samples, and metabolite concentrations were found to be in agreement with available data. The method allows the analysis of up to 96 samples per day and was demonstrated to be stable for up to three weeks from acquisition. View Full-Text
Keywords: tryptophan metabolism; tyrosine metabolism; branched chain amino acids; gut microbiota metabolites; targeted metabolomics; LC-MS/MS; human plasma; urine; clinical studies tryptophan metabolism; tyrosine metabolism; branched chain amino acids; gut microbiota metabolites; targeted metabolomics; LC-MS/MS; human plasma; urine; clinical studies
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MDPI and ACS Style

Anesi, A.; Rubert, J.; Oluwagbemigun, K.; Orozco-Ruiz, X.; Nöthlings, U.; Breteler, M.M.; Mattivi, F. Metabolic Profiling of Human Plasma and Urine, Targeting Tryptophan, Tyrosine and Branched Chain Amino Acid Pathways. Metabolites 2019, 9, 261.

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