In order to determine cholesterol content in eggs, the several methods were compared. Then the cholesterol content in avian eggs were determined by enzymatic method. The results obtained were as follows; 1) After saponification of the lipids in egg yolk, acetone was used for the extraction of cholesterol before the extraction by n-hexane to prevent emulsion. The recovery by this method was 95±5%. 2) The cholesterol content in eggs was determined by the colorimetric (Liebermann-Bur-chard, Zak-Henry, o-phthalaldehyde), enzymatic and gas chromatographic methods, and the experimental results of those assays were nearly equal. All the methods could be applied for the determination of cholesterol in eggs. 3) The cholesterol content in egg yolk laid by layers was significantly lower than those of egg and meat type hens. 4) The cholesterol content of the egg yolk from Osaka Dack, Japanese Quail, Green Pheasant, Korian-Ring Necked Pheasant ant Guinea Fowl were 15.5, 13.8, 15.1, 12.9 and 14.0mg/g yolk, respectively.