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An optimized electrochemistry-liquid chromatography-mass spectrometry method for studying guanosine oxidation

Oxidative stress can disrupt the integrity of genetic material. Due to its importance in the pathogenesis of different kinds of disease, including neurodegenerative disease, cardiovascular disease and cancer, major efforts are put into the elucidation of mechanisms involved. Herein, the combination...

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Autores principales: Erb, Robert, Plattner, Sabine, Pitterl, Florian, Brouwer, Hendrik-Jan, Oberacher, Herbert
Formato: Online Artículo Texto
Lenguaje:English
Publicado: WILEY-VCH Verlag 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3474898/
https://www.ncbi.nlm.nih.gov/pubmed/22451054
http://dx.doi.org/10.1002/elps.201100406
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author Erb, Robert
Plattner, Sabine
Pitterl, Florian
Brouwer, Hendrik-Jan
Oberacher, Herbert
author_facet Erb, Robert
Plattner, Sabine
Pitterl, Florian
Brouwer, Hendrik-Jan
Oberacher, Herbert
author_sort Erb, Robert
collection PubMed
description Oxidative stress can disrupt the integrity of genetic material. Due to its importance in the pathogenesis of different kinds of disease, including neurodegenerative disease, cardiovascular disease and cancer, major efforts are put into the elucidation of mechanisms involved. Herein, the combination of electrochemistry/liquid chromatography/mass spectrometry (EC/LC/MS) is presented as convenient, fast and simple method to study nucleic acids oxidation. Guanosine was selected as test compound. 8-Hydroxyguanosine and (guanosine-H)(2) were identified as primary oxidation products. Oxidation was accomplished in an electrochemical thin-layer cell integrated in the flow path of the autosampler of the chromatographic system. The reaction mixture was separated and mass analyzed by LC/MS. The use of LC was found to be particularly beneficial to resolve isobaric oxidation products. Another advantage of the setup used was the ability to decouple the electrochemical cell and the electrospray ionization source from each other eliminating any kind of cell potential interaction. Separation of EC from LC/MS, furthermore, facilitates method optimization. Experimental parameters were optimized for both techniques independently. Highest yields and best detectability of oxidation products were obtained with 10 mM ammonium formate at physiological pH delivered at a flow rate of 2.5-5 μL/min through the electrochemical cell.
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spelling pubmed-34748982012-10-18 An optimized electrochemistry-liquid chromatography-mass spectrometry method for studying guanosine oxidation Erb, Robert Plattner, Sabine Pitterl, Florian Brouwer, Hendrik-Jan Oberacher, Herbert Electrophoresis Part I: Instrumental and Methodical Developments Oxidative stress can disrupt the integrity of genetic material. Due to its importance in the pathogenesis of different kinds of disease, including neurodegenerative disease, cardiovascular disease and cancer, major efforts are put into the elucidation of mechanisms involved. Herein, the combination of electrochemistry/liquid chromatography/mass spectrometry (EC/LC/MS) is presented as convenient, fast and simple method to study nucleic acids oxidation. Guanosine was selected as test compound. 8-Hydroxyguanosine and (guanosine-H)(2) were identified as primary oxidation products. Oxidation was accomplished in an electrochemical thin-layer cell integrated in the flow path of the autosampler of the chromatographic system. The reaction mixture was separated and mass analyzed by LC/MS. The use of LC was found to be particularly beneficial to resolve isobaric oxidation products. Another advantage of the setup used was the ability to decouple the electrochemical cell and the electrospray ionization source from each other eliminating any kind of cell potential interaction. Separation of EC from LC/MS, furthermore, facilitates method optimization. Experimental parameters were optimized for both techniques independently. Highest yields and best detectability of oxidation products were obtained with 10 mM ammonium formate at physiological pH delivered at a flow rate of 2.5-5 μL/min through the electrochemical cell. WILEY-VCH Verlag 2012-02 2012-03-27 /pmc/articles/PMC3474898/ /pubmed/22451054 http://dx.doi.org/10.1002/elps.201100406 Text en Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim http://creativecommons.org/licenses/by/2.5/ Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation.
spellingShingle Part I: Instrumental and Methodical Developments
Erb, Robert
Plattner, Sabine
Pitterl, Florian
Brouwer, Hendrik-Jan
Oberacher, Herbert
An optimized electrochemistry-liquid chromatography-mass spectrometry method for studying guanosine oxidation
title An optimized electrochemistry-liquid chromatography-mass spectrometry method for studying guanosine oxidation
title_full An optimized electrochemistry-liquid chromatography-mass spectrometry method for studying guanosine oxidation
title_fullStr An optimized electrochemistry-liquid chromatography-mass spectrometry method for studying guanosine oxidation
title_full_unstemmed An optimized electrochemistry-liquid chromatography-mass spectrometry method for studying guanosine oxidation
title_short An optimized electrochemistry-liquid chromatography-mass spectrometry method for studying guanosine oxidation
title_sort optimized electrochemistry-liquid chromatography-mass spectrometry method for studying guanosine oxidation
topic Part I: Instrumental and Methodical Developments
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3474898/
https://www.ncbi.nlm.nih.gov/pubmed/22451054
http://dx.doi.org/10.1002/elps.201100406
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