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.
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References
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
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):875–896
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
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):59–64
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):e98149
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):497–509
Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37(8):911–917
Schnaar RL (1994) Isolation of glycosphingolipids. Methods Enzymol 230:348–370
CWH X (2010) Lipid analysis-isolation, separation, identification and lipidomic analysis. Oily Press, Bridgwater, UK
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):469–481
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):251–260
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
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
Brade L, Vielhaber G, Heinz E, Brade H (2000) In vitro characterization of anti-glucosylceramide rabbit antisera. Glycobiology 10(6):629–636
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Sicard, R., Landgraf, R. (2017). High-Performance Chromatographic Separation of Cerebrosides. 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_7
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DOI: https://doi.org/10.1007/978-1-4939-6996-8_7
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