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A Robust Lipidomics Workflow for Mammalian Cells, Plasma, and Tissue Using Liquid-Chromatography High-Resolution Tandem Mass Spectrometry

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Lipidomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1609))

Abstract

Lipids have been analyzed in applications including drug discovery, disease etiology elucidation, and natural products. The chemical and structural diversity of lipids requires a tailored lipidomics workflow for each sample type. Therefore, every protocol in the lipidomics workflow, especially those involving sample preparation, should be optimized to avoid the introduction of bias. The coupling of ultra-high-performance liquid chromatography (UHPLC) with high-resolution mass spectrometry (HRMS) allows for the separation and identification of lipids based on class and fatty acid acyl chain. This work provides a comprehensive untargeted lipidomics workflow that was optimized for various sample types (mammalian cells, plasma, and tissue) to balance extensive lipid coverage and specificity with high sample throughput. For identification purposes, both data-dependent and data-independent tandem mass spectrometric approaches were incorporated, providing more extensive lipid coverage. Popular open-source feature detection, data processing, and identification software are also outlined.

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Acknowledgments

This work was supported by the Southeast Center for Integrated Metabolomics (SECIM)—NIH Grant #U24 DK097209.

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Authors and Affiliations

  1. Department of Chemistry, University of Florida, Chemistry Laboratory Bldg., 210C, P.O. Box 117200, 125 Buckman Drive, Gainesville, 32611, FL, USA

    Candice Z. Ulmer, Rainey E. Patterson, Jeremy P. Koelmel & Richard A. Yost

  2. Department of Pathology, Immunology and Laboratory Medicine, College of Medicine, University of Florida, Gainesville, 32603, FL, USA

    Timothy J. Garrett & Richard A. Yost

Authors
  1. Candice Z. Ulmer
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  2. Rainey E. Patterson
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  3. Jeremy P. Koelmel
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  4. Timothy J. Garrett
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  5. Richard A. Yost
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Corresponding author

Correspondence to Richard A. Yost .

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Editors and Affiliations

  1. University of Miami, Miami, Florida, USA

    Sanjoy K. Bhattacharya

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Ulmer, C.Z., Patterson, R.E., Koelmel, J.P., Garrett, T.J., Yost, R.A. (2017). A Robust Lipidomics Workflow for Mammalian Cells, Plasma, and Tissue Using Liquid-Chromatography High-Resolution Tandem Mass Spectrometry. In: Bhattacharya, S. (eds) Lipidomics. Methods in Molecular Biology, vol 1609. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6996-8_10

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  • DOI: https://doi.org/10.1007/978-1-4939-6996-8_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6995-1

  • Online ISBN: 978-1-4939-6996-8

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