QUANTITATIVE ANALYSIS OF SIMPLE LIPID CLASSES BY THIN-LAYER CHROMATOGRAPHY

1. 1. A quantitative thin-layer Chromatographie procedure for the analysis of simple lipid classes ('neutral lipids') is described. They were separated by two-step development on silica gel thin-layer chromatoplates. In the first step the solvent diisopropyl ether-acetic acid (96:4, v/v) was used and in the second step, petroleum ether-diethyl ether-acetic acid (90:10:1, v/v/v). 2. 2. After separation, lipid bands were detected by Rhodamine 6G spray, the silica gel with adhered lipids was scraped off and the lipids were eluted. Triglycerides, diglycerides, free fatty acids and hydrocarbons were eluted with diethyl ether, whereas monoglycerides, free cholesterol and cholesteryl esters were eluted by chloroform-methanol (4:1, v/v). The eluates were filtered through sintered glass ('fine' porosity) funnels. 3. 3. The eluted lipids were analyzed in the following ways: quantity of triglycerides, diglycerides, monoglycerides and hydrocarbons were determined by quantitative infrared spectrophotometry; free fatty acids by titration; and cholesteryl esters and cholesterol by a standard chemical procedure. 4. 4. The average recovery of individual lipid classes was in the range 98.9-102.0%. 5. 5. Applicability of the procedure for quantitative analyses of tissue lipids is demonstrated on lipid extracted from rat livers. © 1968.