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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

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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
Authors: Whiley, LW
Nye, L
Grant, I
Andreas, N
Chappell, K
Sarafian, MHS
Misra, R
Plumb, R
Lewis, M
Nicholson, J
Holmes, E
Swann, J
Wilson, I
Item Type: Journal Article
Abstract: 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.
Issue Date: 16-Apr-2019
Date of Acceptance: 8-Mar-2019
URI: http://hdl.handle.net/10044/1/69044
DOI: https://dx.doi.org/10.1021/acs.analchem.8b05884
ISSN: 0003-2700
Publisher: American Chemical Society
Start Page: 5207
End Page: 5216
Journal / Book Title: Analytical Chemistry
Volume: 91
Issue: 8
Copyright Statement: © 2019 American Chemical Society
Sponsor/Funder: UK DRI Ltd
Bill & Melinda Gates Foundation
Funder's Grant Number: N/A
PO 203378346
Keywords: 0301 Analytical Chemistry
0904 Chemical Engineering
0399 Other Chemical Sciences
Analytical Chemistry
Publication Status: Published
Embargo Date: 2020-03-08
Online Publication Date: 2019-03-08
Appears in Collections:Division of Surgery
Faculty of Medicine



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