A multilayer analytical element for measuring enzyme activity in a liquid sample such as a body fluid which is less affected by intrinsic pyruvic acid contained in the liquid sample is disclosed. The analytical element comprises a support, a reagent layer, an optionally provided other layer, and a porous spreading layer. The reagent layer and/or the optionally provided other layer contain pyruvic acid oxidase. The characteristic feature of the analytical element lies in that the spreading layer has a specific void volume in the range of 3-15 .mu.l/cm.sup.2.