Skip to main content
SpringerLink
Log in

Liquid Chromatography–High-Resolution Mass Spectrometry for Quantitative Analysis of Gangliosides

  • Methods
  • Published:
Lipids

Abstract

Gangliosides are a large family of glycosphingolipids that are abundant in the brain, and have been shown to affect neuronal plasticity during development, adulthood, and aging. We developed a fast, efficient, and sensitive liquid chromatography–mass spectrometry method to quantify eight different classes of gangliosides (GM1, GM2, GM3, GD3, GD1a, GD1b, GT1b, GQ1b) in the brains of 2-day-old and 80-day-old Wistar rats. The gangliosides were extracted from rat brain using a modified Svennerholm and Fredman method. After ganglioside class separation using a hydrophilic high performance liquid chromatography (HPLC) column, the resolving power of the LTQ-Orbitrap™ mass spectrometer was used to extract and sum the major species of each ganglioside class, generating fully resolved extracted ion current peaks for both standards and samples. The flexibility and the specificity of this method are such that it can be applied to the analysis of other ganglioside species/classes not discussed in this paper, provided appropriate standards are available. The method had good repeatability (coefficient of variation 4.8–12.3%) and mean recoveries in the range 92–107%.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

CV:

Coefficient of variation

LBSA:

Lipid-bound sialic acid

LOD:

Limit of detection

LOQ:

Limit of quantification

NANA:

N-acetylneuraminic acid

NGNA:

N-glycolylneuraminic acid

References

  1. Hungund BL, Ross DC, Gokhale VS (1994) Ganglioside gm1 reduces fetal alcohol effects in rat pups exposed to ethanol in utero. Alcohol Clin Exp Res 18:1248–1251

    Article  PubMed  CAS  Google Scholar 

  2. Laev H, Karpiak SE, Gokhale VS, Hungund BL (1995) In utero ethanol exposure retards growth and alters morphology of cortical cultures: gm1 reverses effects. Alcohol Clin Exp Res 19:1226–1233

    Article  PubMed  CAS  Google Scholar 

  3. Saito M, Mao RF, Wang R, Vadasz C, Saito M (2007) Effects of gangliosides on ethanol-induced neurodegeneration in the developing mouse brain. Alcohol Clin Exp Res 31:665–674

    PubMed  CAS  Google Scholar 

  4. Wang B, McVeagh P, Petocz P, Brand-Miller J (2003) Brain ganglioside and glycoprotein sialic acid in breast-fed compared with formula-fed infants. Am J Clin Nutr 78:1024–1029

    PubMed  CAS  Google Scholar 

  5. Ando S, Chang NC, Yu RK (1978) High-performance thin-layer chromatography and densitometric determination of brain ganglioside compositions of several species. Anal Biochem 89:437–450

    Article  PubMed  CAS  Google Scholar 

  6. Kasperzyk JL, El-Abbadi MM, Hauser EC, d’Azzo A, Platt FM, Seyfried TN (2004) N-butyldeoxygalactonojirimycin reduces neonatal brain ganglioside content in a mouse model of GM1 gangliosidosis. J Neurochem 89:645–653

    Article  PubMed  CAS  Google Scholar 

  7. Nakabayashi H, Iwamori M, Nagai Y (1984) Analysis and quantitation of gangliosides as para-bromophenacyl derivatives by high-performance liquid-chromatography. J Biochem 96:977–984

    PubMed  CAS  Google Scholar 

  8. Ullman MD, McCluer RH (1985) Quantitative-analysis of brain gangliosides by high-performance liquid-chromatography of their perbenzoyl derivatives. J Lipid Res 26:501–506

    PubMed  CAS  Google Scholar 

  9. Miyazaki K, Okamura N, Kishimoto Y, Lee YC (1986) Determination of gangliosides as 2,4-dinitrophenylhydrazides by high-performance liquid-chromatography. Biochem J 235:755–761

    PubMed  CAS  Google Scholar 

  10. Traylor TD, Koontz DA, Hogan EL (1983) High-performance liquid-chromatographic resolution of para-nitrobenzyloxyamine derivatives of brain gangliosides. J Chromatogr 272:9–20

    Article  PubMed  CAS  Google Scholar 

  11. Wagener R, Kobbe B, Stoffel W (1996) Quantification of gangliosides by microbore high performance liquid chromatography. J Lipid Res 37:1823–1829

    PubMed  CAS  Google Scholar 

  12. Gazzotti G, Sonnino S, Ghidoni R (1985) Normal-phase high-performance liquid chromatographic separation of non-derivatized ganglioside mixtures. J Chromatogr 348:371–378

    Article  PubMed  CAS  Google Scholar 

  13. Sorensen LK (2006) A liquid chromatography/tandem mass spectrometric approach for the determination of gangliosides GD3 and GM3 in bovine milk and infant formulae. Rapid Commum Mass Spectrom 20:3625–3633

    Article  Google Scholar 

  14. Gu JH, Tifft CJ, Soldin SJ (2008) Simultaneous quantification of G(M1) and G(M2) gangliosides by isotope dilution tandem mass spectrometry. Clin Biochem 41:413–417

    Article  PubMed  CAS  Google Scholar 

  15. Sommer U, Herscovitz H, Welty FK, Costello CE (2006) LC–MS-based method for the qualitative and quantitative analysis of complex lipid mixtures. J Lipid Res 47:804–814

    Article  PubMed  CAS  Google Scholar 

  16. Avrova NF, Zabelins Sa (1971) Fatty acids and long chain bases of vertebrate brain gangliosides. J Neurochem 18:675

    Article  PubMed  CAS  Google Scholar 

  17. Rosenber A, Stern N (1966) Changes in sphingosine and fatty acid components of gangliosides in developing rat and human brain. J Lipid Res 7:122

    Google Scholar 

  18. Yu RK, Ledeen RW (1970) Gas–liquid chromatographic assay of lipid-bound sialic acids: measurement of gangliosides in brain of several species. J Lipid Res 11:506–516

    PubMed  CAS  Google Scholar 

  19. Svennerholm L, Fredman P (1980) A procedure for the quantitative isolation of brain gangliosides. Biochim Biophys Acta 617:97–109

    PubMed  CAS  Google Scholar 

  20. Valsecchi M, Palestini P, Chigorno V, Sonnino S (1996) Age-related changes of the ganglioside long-chain base composition in rat cerebellum. Neurochem Int 28:183–187

    Article  PubMed  CAS  Google Scholar 

  21. Ikeda K, Shimizu T, Taguchi R (2008) Targeted analysis of ganglioside and sulfatide molecular species by LC/ESI–MS/MS with theoretically expanded multiple reaction monitoring. J Lipid Res 49:2678–2689

    Article  PubMed  CAS  Google Scholar 

  22. 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 

  23. Brugger B, Erben G, Sandhoff R, Wieland FT, Lehmann WD (1997) Quantitative analysis of biological membrane lipids at the low picomole level by nano-electrospray ionization tandem mass spectrometry. Pro Natl Acad Sci U S A 94:2339–2344

    Article  CAS  Google Scholar 

  24. Ahn EJ, Kim H, Chung BC, Moon MH (2007) Quantitative analysis of phosphatidylcholine in rat liver tissue by nanoflow liquid chromatography/tandem mass spectrometry. J Sep Sci 30:2598–2604

    Article  PubMed  CAS  Google Scholar 

  25. Vanier MT, Holm M, Ohman R, Svennerh L (1971) Developmental profiles of gangliosides in human and rat brain. J Neurochem 18:581–592

    Article  PubMed  CAS  Google Scholar 

  26. Rosner H, Alaqtum M, Rahmann H (1992) Gangliosides and neuronal differentiation. Neurochem Int 20:339–351

    Article  PubMed  CAS  Google Scholar 

  27. Yu RK, Macala LJ, Taki T, Weinfeld HM, Yu FS (1988) Developmental-changes in ganglioside composition and synthesis in embryonic rat-brain. J Neurochem 50:1825–1829

    Article  PubMed  CAS  Google Scholar 

  28. Rosner H (2003) Developmental expression and possible roles of gangliosides in brain development. Prog Mol Subcell Biol 32:49–73

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors would like to thank Kathleen Schoeman for technical support, Charles Yang and Alastair MacGibbon for LC–MS discussions, Claire Woodhall for editing this manuscript, and Mark Vickers (Liggins Institute, Auckland University) for providing the rat brains.

Author information

Authors and Affiliations

  1. Fonterra Research Centre, Dairy Farm Road, Private Bag 11029, Palmerston North, 4412, New Zealand

    Bertram Fong, Carmen Norris, Edwin Lowe & Paul McJarrow

Authors
  1. Bertram Fong
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Carmen Norris
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Edwin Lowe
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Paul McJarrow
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bertram Fong.

About this article

Cite this article

Fong, B., Norris, C., Lowe, E. et al. Liquid Chromatography–High-Resolution Mass Spectrometry for Quantitative Analysis of Gangliosides. Lipids 44, 867–874 (2009). https://doi.org/10.1007/s11745-009-3327-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11745-009-3327-1

Keywords

Search

Navigation