Journal of Lipid Research, Vol. 13, 525-530, July 1972
Copyright © 1972 by Lipid Research, Inc.

Identification and quantitation of free ceramides in human platelets

William Krivit and Sven Hammarström

Department of Pediatrics, University of Minnesota, Minneapolis, Minnesota 55455, and Department of Medical Chemistry, Royal Veterinary College, Fack, S-10405, Stockholm 50, Sweden

Free ceramides were isolated from human platelets. Their structures were unequivocally determined by gas-liquid chromatography-mass spectrometry of the trimethylsilyl ether derivatives. The major components were N-(palmitoyl) sphingosine, N-(stearoyl) sphingosine, N-(eicosanoyl) sphingosine, N-(docosanoyl) sphingosine, N-(tetracosanoyl) sphingosine, and N-(tetracosenoyl) sphingosine. Sphinganine-and sphingadienine-containing ceramides as well as ceramides containing other unsaturated acids were also present. The amount of ceramides was determined by quantitative gas-liquid chromatography, using radioactive ceramide as internal standard and synthetic crystalline ceramides for comparison of peak areas. The concentration of ceramides was found to be 1.31 µg/10⁹ platelets or 0.47 µg/mg of platelet protein.

Supplementary key words gas-liquid chromatography-mass spectrometry • trimethylsilyl ethers

Submitted on August 23, 1971
Accepted on March 10, 1972

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