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Complementary precursor ion and neutral loss scan mode tandem mass spectrometry for the analysis of glycerophosphatidylethanolamine lipids from whole rat retina

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Abstract

A “shotgun” tandem mass spectrometry (MS) approach involving the use of multiple lipid-class-specific precursor ion and neutral loss scan mode experiments has been employed to identify and characterize the glycerophosphatidylethanolamine (GPEtn) lipids that were present within a crude lipid extract of a normal rat retina, obtained with minimal sample handling prior to analysis. Characterization of these lipids was performed by complementary analysis of their protonated and deprotonated precursor ions, as well as their various ionic adducts (e.g., Na+, Cl-), using a triple-quadrupole mass spectrometer. Notably, the application of novel precursor ion and neutral loss scans of m/z 164 and m/z 43, respectively, for the specific identification of sodiated GPEtn precursor ions following the addition of 500 μM NaCl to the crude lipid extracts was demonstrated. The use of these novel MS/MS scans in parallel provided simplified MS/MS spectra and enhanced the detection of 1-alkenyl, 2-acyl (plasmenyl) GPEtn lipids relative to the positive ion mode neutral loss m/z 141 commonly used for GPEtn analysis. Furthermore, the novel use of a “low energy” neutral loss scan mode experiment to monitor for the exclusive loss of 36m/z (HCl) from [M+Cl]- GPEtn adducts was demonstrated to provide a more than 25-fold enhancement for the detection of GPEtn lipids in negative ion mode analysis. Subsequent “high-energy” pseudo MS3 product ion scans on the precursor ions identified from this experiment were then employed to rapidly characterize the fatty acyl chain substituents of the GPEtn lipids.

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Authors and Affiliations

  1. Department of Physiology, Michigan State University, East Lansing, MI, 48824, USA

    Todd A. Lydic & Julia V. Busik

  2. Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI, 48824, USA

    Walter J. Esselman

  3. Department of Chemistry, Michigan State University, East Lansing, MI, 48824, USA

    Gavin E. Reid

  4. Department of Biochemistry and Molecular Biology, Michigan State University, 229 Chemistry Building, East Lansing, MI, 48824, USA

    Gavin E. Reid

Authors
  1. Todd A. Lydic
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  2. Julia V. Busik
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  3. Walter J. Esselman
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  4. Gavin E. Reid
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Corresponding author

Correspondence to Gavin E. Reid.

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Supplementary Table 1

Summary of glycerophosphatidylethanolamine (GPEtn) lipids identified from a whole rat retina crude lipid extract by complementary neutral loss and precursor ion scan mode tandem mass spectrometry. Fatty acyl species were determined by negative ion mode product ion CID-MS/MS or by PI scans of m/z of specific fatty acids. Lipid nomenclature used is outlined in [26] (DOC 41 kb)

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Lydic, T.A., Busik, J.V., Esselman, W.J. et al. Complementary precursor ion and neutral loss scan mode tandem mass spectrometry for the analysis of glycerophosphatidylethanolamine lipids from whole rat retina. Anal Bioanal Chem 394, 267–275 (2009). https://doi.org/10.1007/s00216-009-2717-9

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  • DOI: https://doi.org/10.1007/s00216-009-2717-9

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