Simultaneous determination of psychosine and cerebrosides

Takao Ichioka, Yasuo Kishimoto, Andrew M. Yeager

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

A new method was developed for the simultaneous determination of psychosine and cerebrosides in tissues. Total lipids extracted from the tissues were treated with [3H]acetic anhydride in toluene-methanol. Known amounts of nonradioactive N-acetylpsychosine were added to the reaction product and then subjected to mild alkaline methanolysis. After the product was washed, it was fractionated by silica gel column chromatography and the fraction containing glycolipids was benzoylated. The benzoylated product was finally fractionated on TLC. The amounts of benzoylated derivatives of nonhydroxy- and hydroxycerebrosides and N-acetylpsychosine were determined using a scanning densitometer. The amounts of psychosine in tissues were calculated from the radioactivity in the spot of N-acetylpsychosine and the recovery of added carrier N-acetylpsychosine. This method allowed us to determine 5 to 1000 pmol of psychosine and 1 to 20 nmol of cerebrosides in peripheral nerves and other tissues of the twitcher mouse as well as transfected Schwann cells derived from the sciatic nerves.

Original languageEnglish (US)
Pages (from-to)178-182
Number of pages5
JournalAnalytical Biochemistry
Volume166
Issue number1
DOIs
StatePublished - Oct 1987

Keywords

  • benzoylation
  • cerebroside
  • lipids
  • psychosine
  • radioactivity measurement
  • sphingolipidosis
  • thin-layer chromatography

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Simultaneous determination of psychosine and cerebrosides'. Together they form a unique fingerprint.

  • Cite this