The molecular species of dihydrosphingosines and phytosphingosines and their 1-phosphates with carbon chain lengths from 16 to 20 have been tagged with the fluorescent amino group reagent, 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. All these derivatives could be resolved by reversed phase HPLC on a C18 column. A convenient one-pot method is described whereby lipid extracts from strains of Saccharomyces cerevisiae containing carbon chain length homologs of sphingolipid long chain bases and their phosphorylated derivatives were directly reacted with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate, ester lipids were deacylated, and the reaction mixtures were subjected to liquid chromatography. Five molecular species of both sphingolipid long chain bases and their phosphorylated derivatives are for the first time separated and analyzed. The procedure is quite sensitive, requiring only ∼108 wild-type cells.
|Number of pages||10|
|State||Published - Dec 15 2001|
Bibliographical noteFunding Information:
This work was supported by National Institutes of Health Grant GM 41302.
- 6-aminoquinolyl-N-hydroxyuccinimidyl carbamate
- Dihydrosphingosine phosphate
- Phytosphingosine phosphate
- Saccharomyces cerevisiae
- Sphinganine phosphate
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology