Journal of Lipid Research
Volume 55, Issue 8, August 2014, Pages 1797-1809
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Methods
A monophasic extraction strategy for the simultaneous lipidome analysis of polar and nonpolar retina lipids[S]

https://doi.org/10.1194/jlr.D050302Get rights and content
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Lipid extraction using a monophasic chloroform/methanol/water mixture, coupled with functional group selective derivatization and direct infusion nano-ESI-high-resolution/accurate MS, is shown to facilitate the simultaneous analysis of both highly polar and nonpolar lipids from a single retina lipid extract, including low abundance highly polar ganglioside lipids, nonpolar sphingolipids, and abundant glycerophospholipids. Quantitative comparison showed that the monophasic lipid extraction method yielded similar lipid distributions to those obtained from established “gold standard” biphasic lipid extraction methods known to enrich for either highly polar gangliosides or nonpolar lipids, respectively, with only modest relative ion suppression effects. This improved lipid extraction and analysis strategy therefore enables detailed lipidome analyses of lipid species across a broad range of polarities and abundances, from minimal amounts of biological samples and without need for multiple lipid class-specific extractions or chromatographic separation prior to analysis.

lipidomics
lipid extraction
mass spectrometry
ganglioside
glycerolipid
sphingolipid

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    Abbreviations:

    Cer

    ceramide

    DMBNHS

    S,S′-dimethylthiobutanoylhydroxysuccinimide ester

    HCD

    higher-energy collision-induced dissociation

    nESI

    nano-ESI

    PA

    phosphatidic acid

    PC

    phosphatidylcholine

    PE

    phosphatidylethanolamine

    PG

    phosphatidylglycerol

    PI

    phosphatidylinositol

    PS

    phosphatidylserine

This work was supported by funding from the National Institutes of Health to G.E.R. and J.V.B. (GM103508 and EY016077). The authors have no conflicts of interest to report.

[S]

The online version of this article (available at http://www.jlr.org) contains supplementary data in the form of three figures.