Separation of acidic phospholipids by one-dimensional thin-layer chromatography

Abstract

1. 1. Three one-dimensional thin-layer Chromatographie systems for separation of acidic phospholipids such as cardiolipin, phosphatidic acid and phosphatidyl-glycerol, are described. System I: adsorbent is silica gel without CaSO 4 binder. Two step developing system is used. The first solvent is acetone-light petroleum (1:3, by vol.); the second solvent is a mixture of chloroform-methanol-acetic acid-water (80:13:8:0.3, by vol.). The separated compounds from the top of the chromatogram are: monoglyceride, cardiolipin, phosphatidic acid, ceramidemonohexosides (cerebrosides), phosphatidylglycerol and phosphatidylethanolamine. System II: silica gel thin-layer plates are prepared with 0.1 M Na 2CO 3. The first solvent is pyridine-light petroleum (3:1, by vol.); the second solvent is chloroform-methanol-pyridine-2 M NH 4OH (35:12:65:1, by vol.). The following compounds are separated by the two-step developing system (from the top of chromatogram): ceramidemonohexosides, sulfatides and ceramidedihexosides (overlapping), phosphatidic acid, phosphatidylglycerol and cardiolipin. System III: one-step development modification of System II. This system is suitable only for quantitative determination of phospholipids if the presence of glycolipids is not deemed detrimental. 2. 2. Application of the Systems I and II for separation of acidic phospholipids along with some glycolipids is demonstrated on total lipid extracts from animal tissues. 3. 3. The systems described for the separation of acidic phospholipids are complementary to the previously reported system for separation of phospholipids.