Abstract: The present invention provides an analytical element which is free from evaporation of a sample during measurement and therefore capable of quantifying a substrate using a very small amount of sample with high accuracy and which is free from scattering of the sample during and after the measurement and therefore hygienically excellent; and a measuring device and a substrate quantification method using the same. The analytical element comprises a cavity for accommodating a sample, a working electrode and a counter electrode exposed to an inside of the cavity, a reagent layer which comprises at least an oxidoreductase and is formed inside or in the vicinity of the cavity, an opening communicating with the cavity and a member covering the opening.