Simultaneous quantitation of ceramides and 1,2-diacylglycerol in tissues by latroscan thin-layer chromatography-flame-ionization detection

Kenji Okumura, Kazunori Hayashi, Itsuro Morishima, Kichiro Murase, Hideo Matsui, Yukio Toki, Takayuki Ito

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)

Abstract

Ceramides and 1,2-diacylglycerol have been demonstrated in intracellular signaling pathways. A method of simultaneous mass determination of ceramides and 1,2-diacylglycerol in tissues was developed using the latroscan which combines thin layer chromatography and flame ionization detection (TLC/FID) techniques. Because of relatively low amounts of these components in tissues, the fraction of nonpolar lipids, which included ceramides and glycerides, was eluted with chloroform/acetone mixture (3:1, vol/vol) through a silicic acid column to eliminate the polar phospholipids. Development of Chromarods was carried out using three solvent systems in a four-step development technique. The relationship of the peak area ratio to weight ratio compared with cholesteryl acetate added as an internal standard was linear. The amount of ceramides increased with incubation of rat heart homogenate and human erythrocyte membranes in the presence of sphingomyelinase (E.C. 3.1.4.12). The location TLC/FID system provided a quick and reliable assessment of ceramides and 1,2-diacylglycerol.

Original languageEnglish
Pages (from-to)529-532
Number of pages4
JournalLipids
Volume33
Issue number5
DOIs
Publication statusPublished - 1998
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Organic Chemistry
  • Cell Biology

Fingerprint Dive into the research topics of 'Simultaneous quantitation of ceramides and 1,2-diacylglycerol in tissues by latroscan thin-layer chromatography-flame-ionization detection'. Together they form a unique fingerprint.

Cite this