Abstract
This protocol describes the analysis, specifically the identification, of blood plasma lipids. Plasma lipids are extracted using methyl tert-butyl ether (MTBE), methanol, and water followed by separation and data acquisition of isolated lipids using reversed-phase liquid chromatography coupled to quadrupole/time-of-flight mass spectrometry (RPLC–QTOFMS) operated in MS/MS mode. For lipid identification, acquired MS/MS spectra are converted to the mascot generic format (MGF) followed by library search using the in-silico MS/MS library LipidBlast. Using this approach, lipid classes, carbon-chain lengths, and degree of unsaturation of fatty-acid components are annotated.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Cajka T, Fiehn O (2014) Comprehensive analysis of lipids in biological systems by liquid chromatography-mass spectrometry. Trac-Trend Anal Chem 61:192–206. doi:10.1016/j.trac.2014.04.017
Hyotylainen T, Oresic M (2015) Optimizing the lipidomics workflow for clinical studies--practical considerations. Anal Bioanal Chem 407(17):4973–4993. doi:10.1007/s00216-015-8633-2
Cajka T, Fiehn O (2016) Toward merging untargeted and targeted methods in mass spectrometry-based metabolomics and lipidomics. Anal Chem 88(1):524–545. doi:10.1021/acs.analchem.5b04491
Blanksby SJ, Mitchell TW (2010) Advances in mass spectrometry for lipidomics. Annu Rev Anal Chem 3:433–465. doi:10.1146/annurev.anchem.111808.073705
Kind T, Liu KH, Lee do Y, DeFelice B, Meissen JK, Fiehn O (2013) LipidBlast in silico tandem mass spectrometry database for lipid identification. Nat Methods 10 (8):755-758. doi:10.1038/nmeth.2551
Herzog R, Schuhmann K, Schwudke D, Sampaio JL, Bornstein SR, Schroeder M, Shevchenko A (2012) LipidXplorer: A software for consensual cross-platform lipidomics. Plos One 7(1.) ARTN:e29851. doi:10.1371/journal.pone.0029851
Song H, Hsu FF, Ladenson J, Turk J (2007) Algorithm for processing raw mass spectrometric data to identify and quantitate complex lipid molecular species in mixtures by data-dependent scanning and fragment ion database searching. J Am Soc Mass Spectr 18(10):1848–1858. doi:10.1016/j.jasms.2007.07.023
Matyash V, Liebisch G, Kurzchalia TV, Shevchenko A, Schwudke D (2008) Lipid extraction by methyl-tert-butyl ether for high-throughput lipidomics. J Lipid Res 49(5):1137–1146. doi:10.1194/jlr.D700041-JLR200
Cajka T, Fiehn O (2016) Increasing lipidomic coverage by selecting optimal mobile-phase modifiers in LC–MS of blood plasma. Metabolomics 12(2):article 34. doi:10.1007/s11306-015-0929-x
Acknowledgments
This study was supported by the U.S. National Institutes of Health (NIH) Grants P20 HL113452 and U24 DK097154.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Cajka, T., Fiehn, O. (2017). LC–MS-Based Lipidomics and Automated Identification of Lipids Using the LipidBlast In-Silico MS/MS Library. 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_14
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6996-8_14
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6995-1
Online ISBN: 978-1-4939-6996-8
eBook Packages: Springer Protocols