Abstract
Conditions have been determined for the benzoylation of ceramides containing nonhydroxy and hydroxy fatty acids, and a high performance liquid chromatography system for the separation and measurement of these derivatives has been devised that is capable of good resolution and high sensitivity. These methods have been used to determine quantitatively the levels of ceramides in human tissues, and in serum and urine, and to demonstrate elevated amounts of ceramide in Farber's disease urine and tissues.
Highlights
Ceramides play a central role in sphingolipid metabolism
Using a sample of serum Nonhydroxy fatty acid (NFA) ceramide, benzoylated as described above, it was shown that elution of the sample from the H P L C column followed by mild alkaline hydrolysis and T L C in chloroform-methanol-acetic acid 94: 1:5 (v/v/v) resulted in recovery of NFA ceramide with an RF identical with that of standard NFA ceramide
The theoretical relative response of Hydroxy fatty acid (HFA) ceramide to NFA ceramide is 3 to 2 (3 vs. 2 hydroxyl groups); the observed deviation from this ratio is unexplained but may result from differences in the fatty acid compositions of the standards, since for convenience the results are expressed on a weight rather than molar basis
Summary
Ceramides play a central role in sphingolipid metabolism. They are the basic lipid moiety of the sphingolipids and may be the precursors of sphingomyelin, cerebrosides, sulfatides, and gangliosides [1]. T w o laboratories [2, 5, 7, 8] have reported finding increased amounts of both hydroxy and nonhydroxy fatty acid ceramides in several tissues of patients with Farber's lipogranulomatosis, a lipid storage disease.
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