The chemical properties are related to the secondary hydroxyl group and the double bond at C₅ – C₆.

Cholesterol, when oxidised in suitable conditions, undergoes rapid oxidation to corresponding ketone and cholestenone. The hydroxyl group readily forms esters with acids. The fatty acid esters, namely cholesterol acetate palmitate and propionate, are found in blood and tissues. Cholesterol palmitate, because of the double bond, gives addition and hydrogenation reactions of unsaturated compounds. The iodine value of cholesterol is 65.8.