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A single run LC-MS/MS method for phospholipidomics

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Abstract

Liquid chromatography coupled to tandem mass spectrometry has been compared to shotgun analysis with the objective of finding the best compromise for a single run analysis of whole cell phospholipids. Hydrophilic interaction liquid chromatography (HILIC), normal phase (NP), and reversed phase (RP) liquid chromatography were evaluated with reference phospholipids belonging to phosphatidic acid (PA), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylinositol (PI), and phosphatidylserine (PS) classes. NP-HPLC- and RP-HPLC-ESI-MS/MS were applied to yeast phospholipidome analysis, using a wild-type strain and two strains defective for acyltransferases that are known to be involved in de novo phospholipid synthesis or phospholipid remodeling. The MRM mode was used for relative quantitation of individual compounds based on reference phospholipids bearing fatty acid chains with an odd number of carbon atoms. Combined LC-MS/MS was found superior to shotgun analysis, leading to a larger number of quantified species than shotgun analysis. Finally, RP-HPLC-MS/MS was the preferred method for its higher selectivity, robustness, and better repeatability.

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Abbreviations

ESI-MS:

Electrospray ionization mass spectrometry

FA:

Fatty acid

MRM:

Multiple reaction monitoring

NP:

Normal phase

PA:

Phosphatidic acid

PC:

Phosphatidylcholine

PE:

Phosphatidylethanolamine

PG:

Phosphatidylglycerol

PI:

Phosphatidylinositol

PL:

Phospholipid

RP:

Reversed phase

PS:

Phosphatidylserine

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Acknowledgments

We acknowledge funding of the French Agence Nationale de la Recherche (programme blanc “PANACEA” NT09_517917 to CB, ET and JMS), of the Région Aquitaine and of platforms Protéome (http://www.pgfb.u-bordeaux2.fr/proteome/index.html), and Métabolome-Lipidome-Fluxome of Bordeaux (https://www.bordeaux.inra.fr/umr619/RMN_index.htm) for contribution to mass spectrometry equipment. The PhD position of S.A. was funded by the Centre National de la Recherche Scientifique and the Conseil Régional d'Aquitaine. We also gratefully acknowledge S. Mongrand for the critical reading of the manuscript.

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Author notes

  1. Sophie Ayciriex

    Present address: Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307, Dresden, Germany

Authors and Affiliations

  1. Chimie Biologie des Membranes et Nanoobjets CBMN—UMR 5248 Centre de Génomique Fonctionnelle Université Bordeaux 2, Université de Bordeaux, 146, rue Léo Saignat, 33076, Bordeaux cedex, France

    Corinne Buré & Jean-Marie Schmitter

  2. Laboratoire de Biogenèse Membranaire, UMR 5200 CNRS-Université Bordeaux Segalen, INRA Bordeaux Aquitaine BP81, Université de Bordeaux, 33 883, Villenave d’Ornon Cedex, France

    Sophie Ayciriex & Eric Testet

Authors
  1. Corinne Buré
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  2. Sophie Ayciriex
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  3. Eric Testet
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  4. Jean-Marie Schmitter
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Correspondence to Corinne Buré.

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Buré, C., Ayciriex, S., Testet, E. et al. A single run LC-MS/MS method for phospholipidomics. Anal Bioanal Chem 405, 203–213 (2013). https://doi.org/10.1007/s00216-012-6466-9

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  • DOI: https://doi.org/10.1007/s00216-012-6466-9

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