Quantitation of free sphingosine in liver by high-performance liquid chromatography.


Journal Article

Conditions were established for the extraction of free sphingosine from liver and the separation and quantitation of this and other long-chain (sphingoid) bases (e.g., sphingosine, sphinganine, phytosphingosine, and homologs) by reverse-phase high-performance liquid chromatography (HPLC). The long-chain bases were extracted with chloroform and methanol and then treated with base to remove interfering lipids. After preparation of the o-phthalaldehyde derivatives, the long-chain bases could be separated using C18 columns eluted isocratically with methanol:5 mM potassium phosphate, pH 7.0 (90:10). The HPLC analyses took 15 to 20 min per sample and had lower limits of detection in the picomole range. Quantitation was facilitated by using a 20-carbon long-chain base homolog as an internal standard. The utility of the method was demonstrated with rat liver, providing the first quantitation of free sphingosine in this tissue of approximately 7 nmol/g wet wt.

Full Text

Duke Authors

Cited Authors

  • Merrill, AH; Wang, E; Mullins, RE; Jamison, WC; Nimkar, S; Liotta, DC

Published Date

  • June 1988

Published In

Volume / Issue

  • 171 / 2

Start / End Page

  • 373 - 381

PubMed ID

  • 3407935

Pubmed Central ID

  • 3407935

International Standard Serial Number (ISSN)

  • 0003-2697

Digital Object Identifier (DOI)

  • 10.1016/0003-2697(88)90500-3


  • eng

Conference Location

  • United States