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Ultrahigh-Performance Liquid Chromatography Tandem Mass Spectrometry with Electrospray Ionization Quantification of Tryptophan Metabolites and Markers of Gut Health in Serum and Plasma—Application to Clinical and Epidemiology Cohorts

[Image: see text] A targeted ultrahigh-performance liquid chromatography tandem mass spectrometry with electrospray ionization (UHPLC-ESI-MS/MS) method has been developed for the quantification of tryptophan and its downstream metabolites from the kynurenine and serotonin pathways. The assay coverag...

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Autores principales: Whiley, Luke, Nye, Leanne C., Grant, Isobelle, Andreas, Nick, Chappell, Katie E., Sarafian, Magali H., Misra, Ravi, Plumb, Robert S., Lewis, Matthew R., Nicholson, Jeremy K., Holmes, Elaine, Swann, Jonathan R., Wilson, Ian D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2019
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6503468/
https://www.ncbi.nlm.nih.gov/pubmed/30848589
http://dx.doi.org/10.1021/acs.analchem.8b05884
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author Whiley, Luke
Nye, Leanne C.
Grant, Isobelle
Andreas, Nick
Chappell, Katie E.
Sarafian, Magali H.
Misra, Ravi
Plumb, Robert S.
Lewis, Matthew R.
Nicholson, Jeremy K.
Holmes, Elaine
Swann, Jonathan R.
Wilson, Ian D.
author_facet Whiley, Luke
Nye, Leanne C.
Grant, Isobelle
Andreas, Nick
Chappell, Katie E.
Sarafian, Magali H.
Misra, Ravi
Plumb, Robert S.
Lewis, Matthew R.
Nicholson, Jeremy K.
Holmes, Elaine
Swann, Jonathan R.
Wilson, Ian D.
author_sort Whiley, Luke
collection PubMed
description [Image: see text] A targeted ultrahigh-performance liquid chromatography tandem mass spectrometry with electrospray ionization (UHPLC-ESI-MS/MS) method has been developed for the quantification of tryptophan and its downstream metabolites from the kynurenine and serotonin pathways. The assay coverage also includes markers of gut health and inflammation, including citrulline and neopterin. The method was designed in 96-well plate format for application in multiday, multiplate clinical and epidemiology population studies. A chromatographic cycle time of 7 min enables the analysis of two 96-well plates in 24 h. To protect chromatographic column lifespan, samples underwent a two-step extraction, using solvent protein precipitation followed by delipidation via solid-phase extraction (SPE). Analytical validation reported accuracy of each analyte <20% for the lowest limit of quantification and <15% for all other quality control (QC) levels. The analytical precision for each analyte was 2.1–12.9%. To test the applicability of the method to multiplate and multiday preparations, a serum pool underwent periodic repeat analysis during a run consisting of 18 plates. The % CV (coefficient of variation) values obtained for each analyte were <15%. Additional biological testing applied the assay to samples collected from healthy control participants and two groups diagnosed with inflammatory bowel disease (IBD) (one group treated with the anti-inflammatory 5-aminosalicylic acid (5-ASA) and one group untreated), with results showing significant differences in the concentrations of picolinic acid, kynurenine, and xanthurenic acid. The short analysis time and 96-well plate format of the assay makes it suitable for high-throughput targeted UHPLC-ESI-MS/MS metabolomic analysis in large-scale clinical and epidemiological population studies.
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spelling pubmed-65034682019-05-08 Ultrahigh-Performance Liquid Chromatography Tandem Mass Spectrometry with Electrospray Ionization Quantification of Tryptophan Metabolites and Markers of Gut Health in Serum and Plasma—Application to Clinical and Epidemiology Cohorts Whiley, Luke Nye, Leanne C. Grant, Isobelle Andreas, Nick Chappell, Katie E. Sarafian, Magali H. Misra, Ravi Plumb, Robert S. Lewis, Matthew R. Nicholson, Jeremy K. Holmes, Elaine Swann, Jonathan R. Wilson, Ian D. Anal Chem [Image: see text] A targeted ultrahigh-performance liquid chromatography tandem mass spectrometry with electrospray ionization (UHPLC-ESI-MS/MS) method has been developed for the quantification of tryptophan and its downstream metabolites from the kynurenine and serotonin pathways. The assay coverage also includes markers of gut health and inflammation, including citrulline and neopterin. The method was designed in 96-well plate format for application in multiday, multiplate clinical and epidemiology population studies. A chromatographic cycle time of 7 min enables the analysis of two 96-well plates in 24 h. To protect chromatographic column lifespan, samples underwent a two-step extraction, using solvent protein precipitation followed by delipidation via solid-phase extraction (SPE). Analytical validation reported accuracy of each analyte <20% for the lowest limit of quantification and <15% for all other quality control (QC) levels. The analytical precision for each analyte was 2.1–12.9%. To test the applicability of the method to multiplate and multiday preparations, a serum pool underwent periodic repeat analysis during a run consisting of 18 plates. The % CV (coefficient of variation) values obtained for each analyte were <15%. Additional biological testing applied the assay to samples collected from healthy control participants and two groups diagnosed with inflammatory bowel disease (IBD) (one group treated with the anti-inflammatory 5-aminosalicylic acid (5-ASA) and one group untreated), with results showing significant differences in the concentrations of picolinic acid, kynurenine, and xanthurenic acid. The short analysis time and 96-well plate format of the assay makes it suitable for high-throughput targeted UHPLC-ESI-MS/MS metabolomic analysis in large-scale clinical and epidemiological population studies. American Chemical Society 2019-03-08 2019-04-16 /pmc/articles/PMC6503468/ /pubmed/30848589 http://dx.doi.org/10.1021/acs.analchem.8b05884 Text en Copyright © 2019 American Chemical Society This is an open access article published under a Creative Commons Attribution (CC-BY) License (http://pubs.acs.org/page/policy/authorchoice_ccby_termsofuse.html) , which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.
spellingShingle Whiley, Luke
Nye, Leanne C.
Grant, Isobelle
Andreas, Nick
Chappell, Katie E.
Sarafian, Magali H.
Misra, Ravi
Plumb, Robert S.
Lewis, Matthew R.
Nicholson, Jeremy K.
Holmes, Elaine
Swann, Jonathan R.
Wilson, Ian D.
Ultrahigh-Performance Liquid Chromatography Tandem Mass Spectrometry with Electrospray Ionization Quantification of Tryptophan Metabolites and Markers of Gut Health in Serum and Plasma—Application to Clinical and Epidemiology Cohorts
title Ultrahigh-Performance Liquid Chromatography Tandem Mass Spectrometry with Electrospray Ionization Quantification of Tryptophan Metabolites and Markers of Gut Health in Serum and Plasma—Application to Clinical and Epidemiology Cohorts
title_full Ultrahigh-Performance Liquid Chromatography Tandem Mass Spectrometry with Electrospray Ionization Quantification of Tryptophan Metabolites and Markers of Gut Health in Serum and Plasma—Application to Clinical and Epidemiology Cohorts
title_fullStr Ultrahigh-Performance Liquid Chromatography Tandem Mass Spectrometry with Electrospray Ionization Quantification of Tryptophan Metabolites and Markers of Gut Health in Serum and Plasma—Application to Clinical and Epidemiology Cohorts
title_full_unstemmed Ultrahigh-Performance Liquid Chromatography Tandem Mass Spectrometry with Electrospray Ionization Quantification of Tryptophan Metabolites and Markers of Gut Health in Serum and Plasma—Application to Clinical and Epidemiology Cohorts
title_short Ultrahigh-Performance Liquid Chromatography Tandem Mass Spectrometry with Electrospray Ionization Quantification of Tryptophan Metabolites and Markers of Gut Health in Serum and Plasma—Application to Clinical and Epidemiology Cohorts
title_sort ultrahigh-performance liquid chromatography tandem mass spectrometry with electrospray ionization quantification of tryptophan metabolites and markers of gut health in serum and plasma—application to clinical and epidemiology cohorts
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6503468/
https://www.ncbi.nlm.nih.gov/pubmed/30848589
http://dx.doi.org/10.1021/acs.analchem.8b05884
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