Multiplexed analysis of amino acids in mice brain microdialysis samples using isobaric labeling and liquid chromatography-high resolution tandem mass spectrometry

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Heininen , J , Julku , U , Myöhänen , T , Kotiaho , T & Kostiainen , R 2021 , ' Multiplexed analysis of amino acids in mice brain microdialysis samples using isobaric labeling and liquid chromatography-high resolution tandem mass spectrometry ' , Journal of Chromatography. A , vol. 1656 , 462537 . https://doi.org/10.1016/j.chroma.2021.462537

Title: Multiplexed analysis of amino acids in mice brain microdialysis samples using isobaric labeling and liquid chromatography-high resolution tandem mass spectrometry
Author: Heininen, Juho; Julku, Ulrika; Myöhänen, Timo; Kotiaho, Tapio; Kostiainen, Risto
Other contributor: University of Helsinki, Division of Pharmaceutical Chemistry and Technology
University of Helsinki, Division of Pharmacology and Pharmacotherapy
University of Helsinki, Faculty of Pharmacy
University of Helsinki, Department of Chemistry
University of Helsinki, University Management










Date: 2021-10-25
Language: eng
Number of pages: 10
Belongs to series: Journal of Chromatography. A
ISSN: 0021-9673
DOI: https://doi.org/10.1016/j.chroma.2021.462537
URI: http://hdl.handle.net/10138/335776
Abstract: We developed a new multiplexed reversed phase liquid chromatography-high resolution tandem mass spectrometric (LC-MS/MS) method. The method is based on isobaric labeling with a tandem mass tag (TMT10-plex) and stable isotope-labeled internal standards, and was used to analyze amino acids in mouse brain microdialysis samples. The TMT10-plex labeling of amino acids allowed analysis of ten samples in one LC-MS/MS run, significantly increasing the sample throughput. The method provides good chromatographic performance (peak half-width between 0.04-0.12 min), allowing separation of all TMTlabeled amino acids with acceptable resolution and high sensitivity (limits of detection typically around 10 nM). The use of stable isotope-labeled internal standards, together with TMT10-plex labeling, ensured good repeatability (relative standard deviation 0.994), indicating good quantitative performance of the multiplexed method. The method was applied to study the effect of d-amphetamine microdialysis perfusion on amino acid concentrations in the mouse brain. All amino acids were reliably detected and quantified, indicating that the method is sensitive enough to detect low concentrations of amino acids in brain microdialysis samples. (c) 2021 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY license ( http://creativecommons.org/licenses/by/4.0/ )
Subject: Multiplexing
Isobaric labeling
Isotope dilution
Metabolites
Amino acids
High resolution tandem mass spectrometry
D-AMPHETAMINE
QUANTITATIVE PROTEOMICS
EXTRACELLULAR LEVELS
LC-MS
RAT
GLUTAMATE
METABOLITES
INCREASES
STRIATUM
GABA
1182 Biochemistry, cell and molecular biology
116 Chemical sciences
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