K. Kishimoto et al., Nondestructive quantification of neutral lipids by thin-layer chromatography and laser-fluorescent scanning: Suitable methods for "lipidome" analysis, BIOC BIOP R, 281(3), 2001, pp. 657-662
Citations number
11
Categorie Soggetti
Biochemistry & Biophysics
Journal title
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
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.