Lipidomics pp 57-63 | Cite as

High-Performance Chromatographic Separation of Cerebrosides

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1609)

Abstract

High-performance thin-layer chromatography (HPTLC) is a very robust, fast, and inexpensive technique that enables separation of complex mixtures. Here, we describe the analytical separation of glucosylceramide and galactosylceramide by HPTLC. This technique can be used for quantitation purposes but also with small modification for subsequent mass spectrum analyses for structural determination.

Key words

High-performance thin-layer chromatography Cerebrosides Galactosylceramide Glucosylceramide Orcinol reagent 
This is a preview of subscription content, log in to check access.

References

  1. 1.
    Fuchs B, Suss R, Teuber K, Eibisch M, Schiller J (2011) Lipid analysis by thin-layer chromatography--a review of the current state. J Chromatogr A 1218(19):2754–2774. doi: 10.1016/j.chroma.2010.11.066 CrossRefPubMedGoogle Scholar
  2. 2.
    Meisen I, Mormann M, Muthing J (2011) Thin-layer chromatography, overlay technique and mass spectrometry: a versatile triad advancing glycosphingolipidomics. Biochim Biophys Acta 1811(11):875–896CrossRefPubMedGoogle Scholar
  3. 3.
    Nakamura K, Suzuki Y, Goto-Inoue N, Yoshida-Noro C, Suzuki A (2006) Structural characterization of neutral glycosphingolipids by thin-layer chromatography coupled to matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight MS/MS. Anal Chem 78(16):5736–5743. doi: 10.1021/ac0605501 CrossRefPubMedGoogle Scholar
  4. 4.
    Levery SB, Momany M, Lindsey R, Toledo MS, Shayman JA, Fuller M, Brooks K, Doong RL, Straus AH, Takahashi HK (2002) Disruption of the glucosylceramide biosynthetic pathway in Aspergillus nidulans and Aspergillus fumigatus by inhibitors of UDP-Glc:ceramide glucosyltransferase strongly affects spore germination, cell cycle, and hyphal growth. FEBS Lett 525(1-3):59–64CrossRefPubMedGoogle Scholar
  5. 5.
    Rollin-Pinheiro R, Liporagi-Lopes LC, de Meirelles JV, LMd S, Barreto-Bergter E (2014) Characterization of Scedosporium apiospermum glucosylceramides and their involvement in fungal development and macrophage functions. PLoS One 9(5):e98149CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Folch J, Lees M, Sloane Stanley GH (1957) A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem 226(1):497–509PubMedGoogle Scholar
  7. 7.
    Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37(8):911–917CrossRefPubMedGoogle Scholar
  8. 8.
    Schnaar RL (1994) Isolation of glycosphingolipids. Methods Enzymol 230:348–370CrossRefPubMedGoogle Scholar
  9. 9.
    CWH X (2010) Lipid analysis-isolation, separation, identification and lipidomic analysis. Oily Press, Bridgwater, UKGoogle Scholar
  10. 10.
    Fewou SN, Bussow H, Schaeren-Wiemers N, Vanier MT, Macklin WB, Gieselmann V, Eckhardt M (2005) Reversal of non-hydroxy:alpha-hydroxy galactosylceramide ratio and unstable myelin in transgenic mice overexpressing UDP-galactose:ceramide galactosyltransferase. J Neurochem 94(2):469–481CrossRefPubMedGoogle Scholar
  11. 11.
    Pinto MR, Rodrigues ML, Travassos LR, Haido RMT, Wait R, Barreto-Bergter E (2002) Characterization of glucosylceramides in Pseudallescheria boydii and their involvement in fungal differentiation. Glycobiology 12(4):251–260CrossRefPubMedGoogle Scholar
  12. 12.
    Sakaki T, Zahringer U, Warnecke DC, Fahl A, Knogge W, Heinz E (2001) Sterol glycosides and cerebrosides accumulate in Pichia Pastoris, Rhynchosporium secalis and other fungi under normal conditions or under heat shock and ethanol stress. Yeast 18(8):679–695. doi: 10.1002/yea.720 CrossRefPubMedGoogle Scholar
  13. 13.
    Saito K, Maekawa K, Ishikawa M, Senoo Y, Urata M, Murayama M, Nakatsu N, Yamada H, Saito Y (2014) Glucosylceramide and lysophosphatidylcholines as potential blood biomarkers for drug-induced hepatic phospholipidosis. Toxicol Sci 141(2):377–386. doi: 10.1093/toxsci/kfu132 CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Brade L, Vielhaber G, Heinz E, Brade H (2000) In vitro characterization of anti-glucosylceramide rabbit antisera. Glycobiology 10(6):629–636CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Vivex powered by UMTBMiamiUSA
  2. 2.Department of Biochemistry and Molecular Biology, Sylvester Comprehensive Cancer Center, Miller School of MedicineUniversity of MiamiMiamiUSA

Personalised recommendations