Metabolic Profiling of Human Plasma and Urine, Targeting Tryptophan, Tyrosine and Branched Chain Amino Acid Pathways
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...
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doaj-9fcc07aa27c047cbb7af64f5aee6fd222020-11-24T21:50:05ZengMDPI AGMetabolites2218-19892019-11-0191126110.3390/metabo9110261metabo9110261Metabolic Profiling of Human Plasma and Urine, Targeting Tryptophan, Tyrosine and Branched Chain Amino Acid PathwaysAndrea Anesi0Josep Rubert1Kolade Oluwagbemigun2Ximena Orozco-Ruiz3Ute Nöthlings4Monique M.B. Breteler5Fulvio Mattivi6Department of Food Quality and Nutrition, Research and Innovation Centre, Fondazione Edmund Mach (FEM), Via E. Mach 1, 38010 San Michele all’ Adige, ItalyCIBIO, Department of Cellular, Computational and Integrative Biology, Via Sommarive 9, 38123 Povo, ItalyNutritional Epidemiology, Institute of Nutrition and Food Sciences, University of Bonn, Endenicher Allee 19b, 53115 Bonn, GermanyPopulation Health Sciences, German Center for Neurodegenerative diseases (DZNE), Venusberg-Campus 1-Building 99, 53127 Bonn, GermanyNutritional Epidemiology, Institute of Nutrition and Food Sciences, University of Bonn, Endenicher Allee 19b, 53115 Bonn, GermanyPopulation Health Sciences, German Center for Neurodegenerative diseases (DZNE), Venusberg-Campus 1-Building 99, 53127 Bonn, GermanyCIBIO, Department of Cellular, Computational and Integrative Biology, Via Sommarive 9, 38123 Povo, ItalyTryptophan 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 (<i>r</i><sup>2</sup> > 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.https://www.mdpi.com/2218-1989/9/11/261tryptophan metabolismtyrosine metabolismbranched chain amino acidsgut microbiota metabolitestargeted metabolomicslc-ms/mshuman plasmaurineclinical studies |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Andrea Anesi Josep Rubert Kolade Oluwagbemigun Ximena Orozco-Ruiz Ute Nöthlings Monique M.B. Breteler Fulvio Mattivi |
spellingShingle |
Andrea Anesi Josep Rubert Kolade Oluwagbemigun Ximena Orozco-Ruiz Ute Nöthlings Monique M.B. Breteler Fulvio Mattivi Metabolic Profiling of Human Plasma and Urine, Targeting Tryptophan, Tyrosine and Branched Chain Amino Acid Pathways Metabolites tryptophan metabolism tyrosine metabolism branched chain amino acids gut microbiota metabolites targeted metabolomics lc-ms/ms human plasma urine clinical studies |
author_facet |
Andrea Anesi Josep Rubert Kolade Oluwagbemigun Ximena Orozco-Ruiz Ute Nöthlings Monique M.B. Breteler Fulvio Mattivi |
author_sort |
Andrea Anesi |
title |
Metabolic Profiling of Human Plasma and Urine, Targeting Tryptophan, Tyrosine and Branched Chain Amino Acid Pathways |
title_short |
Metabolic Profiling of Human Plasma and Urine, Targeting Tryptophan, Tyrosine and Branched Chain Amino Acid Pathways |
title_full |
Metabolic Profiling of Human Plasma and Urine, Targeting Tryptophan, Tyrosine and Branched Chain Amino Acid Pathways |
title_fullStr |
Metabolic Profiling of Human Plasma and Urine, Targeting Tryptophan, Tyrosine and Branched Chain Amino Acid Pathways |
title_full_unstemmed |
Metabolic Profiling of Human Plasma and Urine, Targeting Tryptophan, Tyrosine and Branched Chain Amino Acid Pathways |
title_sort |
metabolic profiling of human plasma and urine, targeting tryptophan, tyrosine and branched chain amino acid pathways |
publisher |
MDPI AG |
series |
Metabolites |
issn |
2218-1989 |
publishDate |
2019-11-01 |
description |
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 (<i>r</i><sup>2</sup> > 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. |
topic |
tryptophan metabolism tyrosine metabolism branched chain amino acids gut microbiota metabolites targeted metabolomics lc-ms/ms human plasma urine clinical studies |
url |
https://www.mdpi.com/2218-1989/9/11/261 |
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