An enzyme sensor specific for hypoxanthine (Hx) was developed using immobilized xanthine oxidase-membrane and an oxygen probe. Xanthine oxidase (E.C. 184.108.40.206.) was covalently immobilized on a membrane prepared from cellulose triacetate, 1,8-diamino-4-aminomethyloctane, and glutaraldehyde. Hx is oxidized to uric acid by the immobilized enzyme, the output current of the oxygen probe decreasing due to oxygen consumption. A linear relationship was obtained between current decrease and Hx concentration in the range 0.06–1.5 mM. The enzyme sensor could be used for more than 100 assays without decrease of output current After 30-day-storage at 5°C, no remarkable decrease of output current was observed. The enzyme sensor system was applicable to the simple, rapid, and economical determination of Hx in several fish meats including sea bass, saurel, mackerel, yellowfish, and flounder.