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Multidimensional Mass Spectrometry-Based Shotgun Lipidomics

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Mass Spectrometry in Metabolomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1198))

Abstract

Multidimensional mass spectrometry-based shotgun lipidomics (MDMS-SL) has become a foundational analytical technology platform among current lipidomics practices due to its high efficiency, sensitivity, and reproducibility, as well as its broad coverage. This platform has been broadly used to determine the altered content and/or composition of lipid classes, subclasses, and individual molecular species induced by diseases, genetic manipulations, drug treatments, and aging, among others. Herein, we briefly discuss the principles underlying this technology and present a protocol for routine analysis of many of the lipid classes and subclasses covered by MDMS-SL directly from lipid extracts of biological samples. In particular, lipid sample preparation from a variety of biological materials, which is one of the key components of MDMS-SL, is described in detail. The protocol for mass spectrometric analysis can readily be expanded for analysis of other lipid classes not mentioned as long as appropriate sample preparation is conducted, and should aid researchers in the field to better understand and manage the technology for analysis of cellular lipidomes.

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References

  1. Fahy E, Subramaniam S, Brown HA, Glass CK, Merrill AH Jr, Murphy RC, Raetz CR, Russell DW, Seyama Y, Shaw W, Shimizu T, Spener F, van Meer G, VanNieuwenhze MS, White SH, Witztum JL, Dennis EA (2005) A comprehensive classification system for lipids. J Lipid Res 46:839–861

    Article  PubMed  CAS  Google Scholar 

  2. Han X, Gross RW (2003) Global analyses of cellular lipidomes directly from crude extracts of biological samples by ESI mass spectrometry: a bridge to lipidomics. J Lipid Res 44:1071–1079

    Article  PubMed  CAS  Google Scholar 

  3. Shevchenko A, Simons K (2010) Lipidomics: coming to grips with lipid diversity. Nat Rev Mol Cell Biol 11:593–598

    Article  PubMed  CAS  Google Scholar 

  4. Han X, Gross RW (2005) Shotgun lipidomics: Electrospray ionization mass spectrometric analysis and quantitation of the cellular lipidomes directly from crude extracts of biological samples. Mass Spectrom Rev 24:367–412

    Article  PubMed  CAS  Google Scholar 

  5. Han X, Gross RW (2005) Shotgun lipidomics: multi-dimensional mass spectrometric analysis of cellular lipidomes. Expert Rev Proteomics 2:253–264

    Article  PubMed  CAS  Google Scholar 

  6. Han X, Yang K, Gross RW (2012) Multi-dimensional mass spectrometry-based shotgun lipidomics and novel strategies for lipidomic analyses. Mass Spectrom Rev 31:134–178

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  7. Sun G, Yang K, Zhao Z, Guan S, Han X, Gross RW (2008) Matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis of cellular glycerophospholipids enabled by multiplexed solvent dependent analyte-matrix interactions. Anal Chem 80:7576–7585

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  8. Kiebish MA, Bell R, Yang K, Phan T, Zhao Z, Ames W, Seyfried TN, Gross RW, Chuang JH, Han X (2010) Dynamic simulation of cardiolipin remodeling: greasing the wheels for an interpretative approach to lipidomics. J Lipid Res 51:2153–2170

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  9. Yang K, Han X (2011) Accurate Quantification of Lipid Species by Electrospray Ionization Mass Spectrometry–Meets a Key Challenge in Lipidomics. Metabolites 1:21–40

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  10. Jiang X, Cheng H, Yang K, Gross RW, Han X (2007) Alkaline methanolysis of lipid extracts extends shotgun lipidomics analyses to the low abundance regime of cellular sphingolipids. Anal Biochem 371:135–145

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  11. Yang K, Zhao Z, Gross RW, Han X (2007) Shotgun lipidomics identifies a paired rule for the presence of isomeric ether phospholipid molecular species. PLoS ONE 2:e1368

    Article  PubMed  PubMed Central  Google Scholar 

  12. Han X, Yang K, Cheng H, Fikes KN, Gross RW (2005) Shotgun lipidomics of phosphoethanolamine-containing lipids in biological samples after one-step in situ derivatization. J Lipid Res 46:1548–1560

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  13. Wang M, Fang H, Han X (2012) Shotgun lipidomics analysis of 4-hydroxyalkenal species directly from lipid extracts after one-step in situ derivatization. Anal Chem 84:4580–4586

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  14. Han X, Yang J, Cheng H, Ye H, Gross RW (2004) Towards fingerprinting cellular lipidomes directly from biological samples by two-dimensional electrospray ionization mass spectrometry. Anal Biochem 330:317–331

    Article  PubMed  CAS  Google Scholar 

  15. Han X, Gross RW (1994) Electrospray ionization mass spectroscopic analysis of human erythrocyte plasma membrane phospholipids. Proc Natl Acad Sci U S A 91:10635–10639

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  16. Koivusalo M, Haimi P, Heikinheimo L, Kostiainen R, Somerharju P (2001) Quantitative determination of phospholipid compositions by ESI-MS: effects of acyl chain length, unsaturation, and lipid concentration on instrument response. J Lipid Res 42:663–672

    PubMed  Google Scholar 

  17. DeLong CJ, Baker PRS, Samuel M, Cui Z, Thomas MJ (2001) Molecular species composition of rat liver phospholipids by ESI-MS/MS: the effect of chromatography. J Lipid Res 42:1959–1968

    PubMed  CAS  Google Scholar 

  18. Yang K, Cheng H, Gross RW, Han X (2009) Automated lipid identification and quantification by multi-dimensional mass spectrometry-based shotgun lipidomics. Anal Chem 81:4356–4368

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  19. Han X, Yang K, Gross RW (2008) Microfluidics-based electrospray ionization enhances intrasource separation of lipid classes and extends identification of individual molecular species through multi-dimensional mass spectrometry: Development of an automated high throughput platform for shotgun lipidomics. Rapid Commun Mass Spectrom 22:2115–2124

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  20. Yang K, Fang X, Gross RW, Han X (2011) A practical approach for determination of mass spectral baselines. J Am Soc Mass Spectrom 22:2090–2099

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  21. Han X, Gross RW (2001) Quantitative analysis and molecular species fingerprinting of triacylglyceride molecular species directly from lipid extracts of biological samples by electrospray ionization tandem mass spectrometry. Anal Biochem 295:88–100

    Article  PubMed  CAS  Google Scholar 

  22. Christie WW, Han X (2010) Lipid Analysis: Isolation, Separation, Identification and Lipidomic Analysis, 4th edn. The Oily Press, Bridgwater, England

    Book  Google Scholar 

  23. Han X, Yang K, Yang J, Cheng H, Gross RW (2006) Shotgun lipidomics of cardiolipin molecular species in lipid extracts of biological samples. J Lipid Res 47:864–879

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  24. Vance DE, Vance JE (2008) Biochemistry of Lipids, Lipoproteins and Membranes, 5th edn. Elsevier Science B.V, Amsterdam

    Google Scholar 

  25. Yang K, Zhao Z, Gross RW, Han X (2009) Systematic analysis of choline-containing phospholipids using multi-dimensional mass spectrometry-based shotgun lipidomics. J Chromatogr B 877:2924–2936

    Article  CAS  Google Scholar 

  26. Han X (2002) Characterization and direct quantitation of ceramide molecular species from lipid extracts of biological samples by electrospray ionization tandem mass spectrometry. Anal Biochem 302:199–212

    Article  PubMed  CAS  Google Scholar 

  27. Han X, Cheng H (2005) Characterization and direct quantitation of cerebroside molecular species from lipid extracts by shotgun lipidomics. J Lipid Res 46:163–175

    Article  PubMed  CAS  Google Scholar 

  28. Hsu FF, Turk J (2001) Structural determination of glycosphingolipids as lithiated adducts by electrospray ionization mass spectrometry using low-energy collisional-activated dissociation on a triple stage quadrupole instrument. J Am Soc Mass Spectrom 12:61–79

    Article  PubMed  CAS  Google Scholar 

  29. Hsu F-F, Bohrer A, Turk J (1998) Electrospray ionization tandem mass spectrometric analysis of sulfatide. Determination of fragmentation patterns and characterization of molecular species expressed in brain and in pancreatic islets. Biochim Biophys Acta 1392:202–216

    Article  PubMed  CAS  Google Scholar 

  30. Jiang X, Han X (2006) Characterization and direct quantitation of sphingoid base-1-phosphates from lipid extracts: A shotgun lipidomics approach. J Lipid Res 47:1865–1873

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  31. Jiang X, Yang K, Han X (2009) Direct quantitation of psychosine from alkaline-treated lipid extracts with a semi-synthetic internal standard. J Lipid Res 50:162–172

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  32. Cheng H, Jiang X, Han X (2007) Alterations in lipid homeostasis of mouse dorsal root ganglia induced by apolipoprotein E deficiency: A shotgun lipidomics study. J Neurochem 101:57–76

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  33. Su X, Han X, Mancuso DJ, Abendschein DR, Gross RW (2005) Accumulation of long-chain acylcarnitine and 3-hydroxy acylcarnitine molecular species in diabetic myocardium: identification of alterations in mitochondrial fatty acid processing in diabetic myocardium by shotgun lipidomics. Biochemistry 44:5234–5245

    Article  PubMed  CAS  Google Scholar 

  34. Kalderon B, Sheena V, Shachrur S, Hertz R, Bar-Tana J (2002) Modulation by nutrients and drugs of liver acyl-CoAs analyzed by mass spectrometry. J Lipid Res 43:1125–1132

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

This work was supported by National Institute on Aging Grants R01 AG31675 and intramural institutional research funds. Special thanks are expressed to Ms. Imee Tiu for her editorial assistance.

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Correspondence to Xianlin Han .

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Wang, M., Han, X. (2014). Multidimensional Mass Spectrometry-Based Shotgun Lipidomics. In: Raftery, D. (eds) Mass Spectrometry in Metabolomics. Methods in Molecular Biology, vol 1198. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1258-2_13

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  • DOI: https://doi.org/10.1007/978-1-4939-1258-2_13

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1257-5

  • Online ISBN: 978-1-4939-1258-2

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