An integral multilayer analytical element for analysis of cholesterol which comprises a water-impermeable light-transmissive support, a hydrophilic layer and a spreading layer superposed in this order, and contains cholesterol esterase, cholesterol oxidase, peroxidase and a coloring reagent composition, which is characterized, in that the above coloring reagent composition is a combination of 4-aminoantipyrine or its derivative and 2-hydroxy-3,5-dichlorobenzene sulfonic acid, and in that pH of the layer where coloring reaction proceeds is kept in the range of 7.5 to 9.This analytical element is stable, and capable of preserving more than 8 months without fogging.