The effect of dehydrated Chlorella vulgaris and rapeseed oil on the cholesterol, vitamin and carotenoid content in eggs was evaluated in 40-wk old Hisex Brown hens, fed a wheat/soybean meal diet. Detailed analyses of excreta sterols were carried out. Diets were supplemented with rapeseed oil (0 or 5%) and Chlorella (0 or 0.5%). Chlorella significantly decreased the concentration of cholesterol in eggs and increased the concentrations of retinol, lutein, zeaxanthin and β-carotene, but not the concentration of α-tocopherol. Rapeseed oil doubled the α-tocopherol concentration in yolks and significantly increased the concentration of carotenoids in hens fed Chlorella. Cholesterol and coprostanol concentrations in the excreta of hens were similar in different groups. Sterols of plant origin dominated over other neutral sterols. The major plant sterols were β-sitosterol and campesterol. Chenodeoxycholic acid was the most important bile acid in excreta. Concentrations of bile acids in excreta were low. Supplementation of diets with Chlorella did not significantly influence the total concentrations of neutral sterols, bile acids and phytosterols. Taurocholic acid and taurochenodeoxycholic acid were identified in the small intestinal contents. No bile acids conjugated with glycine were found. Concentrations of conjugated bile acids in the small intestine were considerably higher than the concentrations of their non-conjugated counterparts in the excreta.