Quantitation of 1,2-diacylglycerol in rat heart by iatroscan TLC/FID

Abstract

1,2-Diacylglycerol, which has been recognized as one of the intracellular second messengers, was measured quantitatively in the lipid extract from rat hearts using the thin layer chromatography and flame ionization detection (TLC/FID) method. Cholesterol acetate was added to the tissue as an internal standard, and the crude lipids from the tissue were purified with silicic acid column to eliminate phospholipids. Development of Chromarods was carried out using two solvent systems and a three-step development technique. The relationship of the peak area ratio detected by flame ionization detector to weight ratio was linear compared with cholesteryl acetate. The 1,2-diacylglycerol content in the rat heart in the unstimulated condition was 72.5 +/- 15.3 ng/mg wet wt (mean +/- SD).