Abstract: A method of determining skin tissue cholesterol includes preparing a section of skin surface, applying a dosed amount of an enzyme-containing mixture onto the skin, exposing, evaluating the concentration of cholesterol in a reaction solution and calculating the concentration of skin cholesterol, wherein the area of contact of the skin with the mixture is limited on the section of the skin surface by means of a sealing vessel without a bottom with a base fixed on the skin, a mixture comprising 2.0-2.5 units of cholesteroloxidase, 0.04-0.06% by weight of sodium dezoxycholate, 0.1-0.2.% by weight of 3-(dodecyl-dimethyl-ammonium)-propanesulfonate in a buffer solution with pH=6.8 is used as the mixture, evaluating the concentration of cholesterol in the reaction solution is carried out by measuring the level of hydrogen peroxide. Measurements of the level of hydrogen peroxide may be carried out by immersing an electrochemical sensor or a colorimetric indicator in the reaction solution.