Journal of Lipid Research, Vol 20, 879-907, Copyright © 1979 by Lipid Research, Inc.
Free ceramide, sphingomyelin, and glucosylceramide of isolated rat intestinal cells
JF Bouhours and H Guignard
Free ceramide, glucosylceramide, and sphingomyelin were isolated from
mature cells of adult rat small intestine. Free ceramide and ceramide
cleaved from sphingomyelin by enzymatic hydrolysis were fractionated by
thin-layer chromatography on borate-impregnated silica gel plates.
Sphingoid bases were characterized by gas-liquid chromatography of
aldehydes formed upon periodate oxidation. Fatty acids were quantified as
methyl esters. Ceramide structures were confirmed by direct-inlet mass
spectrometry. Free ceramide was found to contain two major long- chain
bases in nearly equal quantity: sphingosine, mainly linked to palmitic
acid, and 4D-hydroxysphinganine associated with C20 to C24 fatty acids, 22%
being hydroxylated. Sphinganine occurred as a minor component linked to
nonhydroxy fatty acids. Sphingomyelin contained the three long-chain bases
and 63% of its ceramide was N-palmitoyl- sphingosine. Mass spectrometry of
glucosylceramide confirmed 4D- hydroxyshingamine as the major sphingoid
base associated preferentially with longer chain hydroxy fatty acids.
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