Phospholipid and neutral lipid separation by one-dimensional thin-layer chromatography.

A simple and rapid method for separation of six major phospholipids and four major neutral lipids from cell extracts by one-dimensional preadsorbent thin-layer chromatography was developed. Due to the inert characteristics of the preadsorbent layer (Celite) no separation occurs until the sample reaches the preadsorbent (Celite)-silica gel junction. The compounds were applied to the preadsorbent (Celite) area (625 mm2) in 10-microliters aliquots (total volume of 0.15 ml). By this method, samples can be applied rapidly in large volumes without respotting any area of the preadsorbent layer. The time required to apply one sample was reduced considerably (2 min) compared to conventional methods (10 min). Since all the compounds move with the solvent front as a sharp, narrow band to the preadsorbent (Celite)-silica gel boundary, excellent separation was achieved when up to 650 micrograms of lipid material was applied on each lane (25 mm wide). Thus, this method is suitable for the separation of relatively large amounts of radiolabeled and non-radiolabeled lipids and free fatty acids from extracts of biological fluids, tissues, or cells maintained in monolayer culture.