[49] Separation of Sphingosine Bases by Chromatography on Columns of Silica Gel

Yechezkel Barenholz, Shimon Gatt

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The three main sphingosine bases of animal tissues are sphingosine (CH3-(CH2)12-CH=CH-CHOH-CHNH2-CH2OH) and dihydrosphingosine (CH3-(CH2)14-CHOH-CHNH2-CH2OH), while in plants phytosphingosine (CH3-(CH2)13-CHOH-CHOH-CHNH2-CH2OH) predominates. Similar bases with smaller or greater chain length have been described. These bases are released from the phospho- or glycosphingolipids by acid or alkaline hydrolysis. Such hydrolyses yield mixtures of the bases as well as several derivatives which are formed as a result of the chemical reaction. The reaction medium must therefore be subjected to a purification step in which the individual bases are separated from the above mixture. In this procedure silica gel chromatography is used to separate, on a preparative scale, the pure bases from acid hydrolysis reaction mixtures. This is here exemplified by two experiments. In the first, pure sphingosine and dihydrosphingosine were obtained from the hydrolyzate of bovine spinal cord sphingolipids. In the second, phytosphingosine and dihydrosphingosine were isolated from the hydrolysate of the acetylated bases secreted into the growth medium of the yeast Hansenula ciferri.

Original languageEnglish
Pages (from-to)529-533
Number of pages5
JournalMethods in Enzymology
Issue numberC
StatePublished - 1 Jan 1975


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