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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http://hdl.handle.net/10138/335776

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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

Titel: Multiplexed analysis of amino acids in mice brain microdialysis samples using isobaric labeling and liquid chromatography-high resolution tandem mass spectrometry
Författare: Heininen, Juho; Julku, Ulrika; Myöhänen, Timo; Kotiaho, Tapio; Kostiainen, Risto
Upphovmannens organisation: Division of Pharmaceutical Chemistry and Technology
Drug Research Program
Division of Pharmacology and Pharmacotherapy
Regenerative pharmacology group
PREP in neurodegenerative disorders
Faculty of Pharmacy
Divisions of Faculty of Pharmacy
Department of Chemistry
Tapio Kotiaho / Principal Investigator
University Management
Risto Kostiainen / Principal Investigator
Datum: 2021-10-25
Språk: eng
Sidantal: 10
Tillhör serie: Journal of Chromatography. A
ISSN: 0021-9673
DOI: https://doi.org/10.1016/j.chroma.2021.462537
Permanenta länken (URI): http://hdl.handle.net/10138/335776
Abstrakt: 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
Referentgranskad: Ja
Licens: cc_by
Användningsbegränsning: openAccess
Parallelpublicerad version: publishedVersion


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