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Qualitative Characterization of the Rat Liver Mitochondrial Lipidome Using All Ion Fragmentation on an Exactive Benchtop Orbitrap MS

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Book cover Mitochondrial Medicine

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1264))

Abstract

Untargeted lipidomics profiling by liquid chromatography-mass spectrometry (LC-MS) allows researchers to observe the occurrences of lipids in a biological sample without showing intentional bias to any specific class of lipids and allows retrospective reanalysis of data collected. Typically, and in the specific method described, a general extraction method followed by LC separation is used to achieve nonspecific class coverage of the lipidome prior to high-resolution accurate mass (HRAM) MS detection. Here we describe a workflow including the isolation of mitochondria from liver tissue, followed by mitochondrial lipid extraction and the LC-MS conditions used for data acquisition. We also highlight how, in this method, all-ion fragmentation can be used to identify species of lower abundances, often missed by data-dependent fragmentation techniques. Here we describe the isolation of mitochondria from liver tissue, followed by mitochondrial lipid extraction and the LC-MS conditions used for data acquisition.

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Acknowledgments

These studies were funded by U01-ES16048 (BSK, PI) and P30-DK040561 (W. Allan Walker, PI). The authors thank Thermo Fisher for the loan of an Exactive benchtop Orbitrap for demonstration testing (later purchased) and financial support for scientific meeting attendance.

Financial Disclosures

SSB currently works for Thermo Fisher.

IGS, RMG, and FT have no financial disclosures.

BSK is the inventor on general metabolomics-related IP that has been licensed to Metabolon via Weill Medical College of Cornell University and for which he receives royalty payments via Weill Medical College of Cornell University. He also consults for and has a small equity interest in the company. Metabolon offers biochemical profiling services and is developing molecular diagnostic assays detecting and monitoring disease. Metabolon has no rights or proprietary access to the research results presented and/or new IP generated under these grants/studies. BSK interests were reviewed by the Brigham and Women’s Hospital and Partners Healthcare in accordance with their institutional policy. Accordingly, upon review, the institution determined that BSK financial interest in Metabolon does not create a significant financial conflict of interest (FCOI) with this research. The addition of this statement where appropriate was explicitly requested and approved by BWH.

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    Authors and Affiliations

    1. Thermo Fisher Scientific, Cambridge, MA, 02139, USA

      Susan S. Bird

    2. Department of Neurosurgery, Harvard Medical School and Brigham and Women’s Hospital, Boston, MA, 02115, USA

      Irina G. Stavrovskaya, Rose M. Gathungu & Fateme Tousi

    3. Department of Neurosurgery, Brigham and Women’s Hospital, 221 Longwood Avenue, Boston, MA, 02115, USA

      Bruce S. Kristal

    Authors
    1. Susan S. Bird
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    2. Irina G. Stavrovskaya
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    3. Rose M. Gathungu
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    4. Fateme Tousi
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    5. Bruce S. Kristal
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    Corresponding author

    Correspondence to Bruce S. Kristal .

    Editor information

    Editors and Affiliations

    1. Midwestern University, Glendale, Arizona, USA

      Volkmar Weissig

    2. ISANH, Paris, France

      Marvin Edeas

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    Bird, S.S., Stavrovskaya, I.G., Gathungu, R.M., Tousi, F., Kristal, B.S. (2015). Qualitative Characterization of the Rat Liver Mitochondrial Lipidome Using All Ion Fragmentation on an Exactive Benchtop Orbitrap MS. In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine. Methods in Molecular Biology, vol 1264. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2257-4_36

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    • DOI: https://doi.org/10.1007/978-1-4939-2257-4_36

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    • Publisher Name: Humana Press, New York, NY

    • Print ISBN: 978-1-4939-2256-7

    • Online ISBN: 978-1-4939-2257-4

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