A biosensor enabling accurate measurement of the concentration of a substrate contained in a trace amount of a sample solution is disclosed. The electrochemical analysis element in accordance with the present invention which should be included in the biosensor comprises a sensor body having a hollow space which space is open at its both ends, an electrode system having a working electrode and a counter electrode, a reagent segment containing an enzyme, wherein the electrode system and the reagent segment are disposed on an inner wall of the hollow space.