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Characterization of complex lipid mixtures in contaminant exposed JEG-3 cells using liquid chromatography and high-resolution mass spectrometry

  • 14th EuCheMS International Conference on Chemistry and the Environment (ICCE 2013, Barcelona, June 25 - 28, 2013)
  • Published:
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Abstract

The aim of this study was to develop a method based on ultra high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS) for lipid profiling in human placental choriocarcinoma (JEG-3) cells. Lipids were solid–liquid extracted from JEG-3 cells using a solution of chloroform/methanol (2:1, v/v) in a simple procedure requiring minimal sample alteration. Simultaneous separation of complex lipid mixtures in their major classes was achieved with a reversed-phase (C8) UHPLC column and a mobile phase containing methanol with 1 mM ammonium formate and 0.2 % formic acid (A)/water with 2 mM ammonium formate and 0.2 % formic acid (B). Lipids were characterized using time-of-flight (TOF) and Orbitrap under full scan and positive electrospray ionization mode with both analyzers. A total of 178 species of lipids, including 37 phosphatidylcholines (PC), 32 plasmalogen PC, 9 lyso PC, 4 lyso plasmalogen PC, 30 triacylglycerols, 22 diacylglycerols, 7 cholesterol esters, 25 phosphatidylethanolamines, and 12 sphingomyelins, were identified using TOF and Orbitrap. The identification of all lipid classes was based on exact mass characterization with an error < 5 ppm. The developed methodology was applied to study lipid alterations in human placental cells against the exposure to perfluorinated chemicals (PFCs) and tributyltin (TBT).

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Acknowledgments

This study was financed by the INNPACTO project (IPT-2011-0709-060000). Dr. R. Chaler, D. Fanjul, and Eva Dalmau are acknowledged for TOF-MS support and Dr. Alberto Adeva for Orbitrap-MS support.

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Authors and Affiliations

  1. Department of Environmental Chemistry, Institute of Environmental Assessment and Water Research (IDAEA), Consejo Superior de Investigaciones Científicas (CSIC), Jordi Girona 18-26, Barcelona, 08034, Catalonia, Spain

    Eva Gorrochategui, Elisabet Pérez-Albaladejo, Cinta Porte & Sílvia Lacorte

  2. Department of Biomedicinal Chemistry, Institute of Advanced Chemistry of Catalonia (IQAC), Jordi Girona 18-26, Barcelona, 08034, Catalonia, Spain

    Josefina Casas

  3. Scientific and Technological Centers, University of Barcelona, Baldiri Reixac, 10-12, Barcelona, 08028, Catalonia, Spain

    Olga Jáuregui

Authors
  1. Eva Gorrochategui
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  2. Josefina Casas
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  3. Elisabet Pérez-Albaladejo
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  4. Olga Jáuregui
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  5. Cinta Porte
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  6. Sílvia Lacorte
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Corresponding author

Correspondence to Sílvia Lacorte.

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Responsible editor: Philippe Garrigues

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Table S1

Elemental composition of glycerophospholipid, glycerolipid, sterol lipid, and sphingolipid species found in human placental choriocarcinoma JEG-3 cells, calculated by mass accuracy within error of 5 ppm, with atom constraints and with −0.5 ≤ DBE ≤ 15.0. DBE double-bond equivalent. Elemental composition of PC, Plasmalogen PC, Lyso PC, Lyso plasmalogen PC, PE, and SM refer to the [M + H]+ ions, whereas TAG, DAG, and CE refer to ammonium adducts [M + NH4]+. Lipid species were detected under ESI (+) using an UHPLC system coupled to a TOF analyzer with an Acquity UPLC BEH C8 column (1.7 μm particle size, 100 mm × 2.1 mm), with the exception of lipid species highlighted with symbol (*), which were identified using an UHPLC-Orbitrap system with the same column (DOCX 65 kb)

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Gorrochategui, E., Casas, J., Pérez-Albaladejo, E. et al. Characterization of complex lipid mixtures in contaminant exposed JEG-3 cells using liquid chromatography and high-resolution mass spectrometry. Environ Sci Pollut Res 21, 11907–11916 (2014). https://doi.org/10.1007/s11356-014-3172-5

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  • DOI: https://doi.org/10.1007/s11356-014-3172-5

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