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

Abstract

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.

Key Words:

• cholesterol
• fluorometry
• thin-layer chromatography
• triglycerides