Nondestructive quantification of neutral lipids by thin-layer chromatography and laser-fluorescent scanning: Suitable methods for "lipidome" analysis

A simple method for separation and quantification of neutral lipids was dev eloped using thin-layer chromatography (TLC) and high-performance fluoresce nt scanning. Neutral Lipid classes were separated using the double-developi ng TLC method and detected by rhodamine 6G and a laser-excited fluorescent scanner. The amount of lipids applied correlated with scanned intensity vol ume in a dose-dependent manner. The mass of each neutral lipid band was det ermined by comparing band intensities of unknown samples to dilution curves of authentic standards. After scanning the dye-sprayed TLC, acyl chain spe cies of triglyceride (TG) extracted from TLC could be determined by gas chr omatography. Using this method, we quantified the amounts of TG in mouse li ver and found that the measured total mass of TG correlated with that obtai ned by enzymatic methods. Our method should provide the basic technique for "lipidome" analysis, designed to determine and compare total lipid classes and mass present in biological samples. (C) 2001 Academic Press.