Quantitative analysis of human serum cholesterol by thin-layer chromatographic spot test

A fast, inexpensive, simple method is described for the determination of the total cholesterol concentration in human serum. The cholesterols (free and esterified) are extracted from serum by a double extraction procedure, using isopropanol-water-10 N sodium hydroxide (250:125:10) and then n-octane. An aliquot of the octane extract is spotted on a thin-layer plate and the cholesterol in the spot is rendered visible using an aqueous phosphomolybdic acid staining solution and a heat treatment for the color development. The colored spot is scanned with a densitometer.