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 developed using thin-layer chromatography (TLC) and high-performance fluorescent scanning. Neutral lipid classes were separated using the double-developing TLC method and detected by rhodamine 6G and a laser-excited fluorescent scanner. The amount of lipids applied correlated with scanned intensity volume in a dose-dependent manner. The mass of each neutral lipid band was determined by comparing band intensities of unknown samples to dilution curves of authentic standards. After scanning the dye-sprayed TLC, acyl chain species of triglyceride (TG) extracted from TLC could be determined by gas chromatography. Using this method, we quantified the amounts of TG in mouse liver and found that the measured total mass of TG correlated with that obtained 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.
Kishimoto
K
K
Research Institute for Food Science, Kyoto University, Kyoto, Gokasho, Uji, 611-0011, Japan.
Urade
R
R
Ogawa
T
T
Moriyama
T
T
D000818:Animals; D002138:Calibration; D002855:Chromatography, Thin Layer / Q000379:methods*; D005456:Fluorescent Dyes; D007834:Lasers; D008055:Lipids / Q000032:analysis*; D008099:Liver / Q000737:chemistry; D051379:Mice; D012235:Rhodamines; D013050:Spectrometry, Fluorescence / Q000379:methods*
D005456:Fluorescent Dyes; D008055:Lipids; D012235:Rhodamines; C026188:rhodamine 6G