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 ThermoFisher for the loan of an Exactive Benchtop orbitrap for demonstration testing (later purchased) and financial support for scientific meeting attendance.
Financial Disclosures
IGS and RMG have no financial disclosures.
SSB currently works for ThermoFisher.
BSK is the inventor of general metabolomics-related IP that has been licensed to Metabolon via Weill Medical College of Cornell University, for which he receives royalty payments via Weill Medical College of Cornell University. He also 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’s 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’s 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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Stavrovskaya, I.G., Gathungu, R.M., Bird, S.S., Kristal, B.S. (2021). 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 2275. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1262-0_24
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DOI: https://doi.org/10.1007/978-1-0716-1262-0_24
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