Abstract: An analytical element is designed for the colorimetric determination of total cholesterol in aqueous liquids, such as biological fluids. This element comprises an absorbent material and critical amounts of cholesterol ester hydrolase (from about 1500 to about 12,000 I.U./m.sup.2) and a nonionic surfactant (from about 5 and up to, but less than 11 g/m.sup.2). The use of these ranges of reagents reduces the potential for interference by triglycerides present in a test sample. Total cholesterol is determined with this element by detection of a color change resulting from a series of enzymatic reactions which produce hydrogen peroxide.