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Determination of Oxidized Phosphatidylcholines by Hydrophilic Interaction Liquid Chromatography Coupled to Fourier Transform Mass Spectrometry

A novel liquid chromatography-mass spectrometry (LC-MS) approach for analysis of oxidized phosphatidylcholines by an Orbitrap Fourier Transform mass spectrometer in positive electrospray ionization (ESI) coupled to hydrophilic interaction liquid chromatography (HILIC) was developed. This method depe...

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Autores principales: Sala, Pia, Pötz, Sandra, Brunner, Martina, Trötzmüller, Martin, Fauland, Alexander, Triebl, Alexander, Hartler, Jürgen, Lankmayr, Ernst, Köfeler, Harald C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4425085/
https://www.ncbi.nlm.nih.gov/pubmed/25874761
http://dx.doi.org/10.3390/ijms16048351
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author Sala, Pia
Pötz, Sandra
Brunner, Martina
Trötzmüller, Martin
Fauland, Alexander
Triebl, Alexander
Hartler, Jürgen
Lankmayr, Ernst
Köfeler, Harald C.
author_facet Sala, Pia
Pötz, Sandra
Brunner, Martina
Trötzmüller, Martin
Fauland, Alexander
Triebl, Alexander
Hartler, Jürgen
Lankmayr, Ernst
Köfeler, Harald C.
author_sort Sala, Pia
collection PubMed
description A novel liquid chromatography-mass spectrometry (LC-MS) approach for analysis of oxidized phosphatidylcholines by an Orbitrap Fourier Transform mass spectrometer in positive electrospray ionization (ESI) coupled to hydrophilic interaction liquid chromatography (HILIC) was developed. This method depends on three selectivity criteria for separation and identification: retention time, exact mass at a resolution of 100,000 and collision induced dissociation (CID) fragment spectra in a linear ion trap. The process of chromatography development showed the best separation properties with a silica-based Kinetex column. This type of chromatography was able to separate all major lipid classes expected in mammalian samples, yielding increased sensitivity of oxidized phosphatidylcholines over reversed phase chromatography. Identification of molecular species was achieved by exact mass on intact molecular ions and CID tandem mass spectra containing characteristic fragments. Due to a lack of commercially available standards, method development was performed with copper induced oxidation products of palmitoyl-arachidonoyl-phosphatidylcholine, which resulted in a plethora of lipid species oxidized at the arachidonoyl moiety. Validation of the method was done with copper oxidized human low-density lipoprotein (LDL) prepared by ultracentrifugation. In these LDL samples we could identify 46 oxidized molecular phosphatidylcholine species out of 99 possible candidates.
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spelling pubmed-44250852015-05-20 Determination of Oxidized Phosphatidylcholines by Hydrophilic Interaction Liquid Chromatography Coupled to Fourier Transform Mass Spectrometry Sala, Pia Pötz, Sandra Brunner, Martina Trötzmüller, Martin Fauland, Alexander Triebl, Alexander Hartler, Jürgen Lankmayr, Ernst Köfeler, Harald C. Int J Mol Sci Article A novel liquid chromatography-mass spectrometry (LC-MS) approach for analysis of oxidized phosphatidylcholines by an Orbitrap Fourier Transform mass spectrometer in positive electrospray ionization (ESI) coupled to hydrophilic interaction liquid chromatography (HILIC) was developed. This method depends on three selectivity criteria for separation and identification: retention time, exact mass at a resolution of 100,000 and collision induced dissociation (CID) fragment spectra in a linear ion trap. The process of chromatography development showed the best separation properties with a silica-based Kinetex column. This type of chromatography was able to separate all major lipid classes expected in mammalian samples, yielding increased sensitivity of oxidized phosphatidylcholines over reversed phase chromatography. Identification of molecular species was achieved by exact mass on intact molecular ions and CID tandem mass spectra containing characteristic fragments. Due to a lack of commercially available standards, method development was performed with copper induced oxidation products of palmitoyl-arachidonoyl-phosphatidylcholine, which resulted in a plethora of lipid species oxidized at the arachidonoyl moiety. Validation of the method was done with copper oxidized human low-density lipoprotein (LDL) prepared by ultracentrifugation. In these LDL samples we could identify 46 oxidized molecular phosphatidylcholine species out of 99 possible candidates. MDPI 2015-04-14 /pmc/articles/PMC4425085/ /pubmed/25874761 http://dx.doi.org/10.3390/ijms16048351 Text en © 2015 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Sala, Pia
Pötz, Sandra
Brunner, Martina
Trötzmüller, Martin
Fauland, Alexander
Triebl, Alexander
Hartler, Jürgen
Lankmayr, Ernst
Köfeler, Harald C.
Determination of Oxidized Phosphatidylcholines by Hydrophilic Interaction Liquid Chromatography Coupled to Fourier Transform Mass Spectrometry
title Determination of Oxidized Phosphatidylcholines by Hydrophilic Interaction Liquid Chromatography Coupled to Fourier Transform Mass Spectrometry
title_full Determination of Oxidized Phosphatidylcholines by Hydrophilic Interaction Liquid Chromatography Coupled to Fourier Transform Mass Spectrometry
title_fullStr Determination of Oxidized Phosphatidylcholines by Hydrophilic Interaction Liquid Chromatography Coupled to Fourier Transform Mass Spectrometry
title_full_unstemmed Determination of Oxidized Phosphatidylcholines by Hydrophilic Interaction Liquid Chromatography Coupled to Fourier Transform Mass Spectrometry
title_short Determination of Oxidized Phosphatidylcholines by Hydrophilic Interaction Liquid Chromatography Coupled to Fourier Transform Mass Spectrometry
title_sort determination of oxidized phosphatidylcholines by hydrophilic interaction liquid chromatography coupled to fourier transform mass spectrometry
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4425085/
https://www.ncbi.nlm.nih.gov/pubmed/25874761
http://dx.doi.org/10.3390/ijms16048351
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