Rapid micromethod for flourometric determination of neutral lipids on thin-layer chromatograms

A rapid fluorometric procedure for in situ evaluation of neutral lipids on thin-layer chromatograms is described. The lipids are rendered fluorescent by treatment with perchloric acid, and the intensity of the emitted light is recorded by flying spot scanning. Detection limits (2.8 × SD) are 0.10, 0.11, and 0.17 nmol for triglycerides, cholesterol, and cholesterol esters, respectively. Range of linearity is 0-3.4 nmol for triglycerides, 0-2.0 nmol for cholesterol, and 0-4.0 nmol for cholesterol esters. When standard in triplicate and 2 samples of each unknown are applied to the chromatogram, standard error of the mean is 2.7% for triglycerides, 1.7% for cholesterol, and 1.0% for cholesterol esters. The sensitivity of the method allows simultaneous determination of the three lipid classes in 5 μl lipid extract representing 0.238 μl human plasma.