Simultaneous determination of glucose and sucrose by a dual-working electrode multienzyme sensor flow-injection system

A dual‐working electrode (K1 and K2) multienzyme sensor, made of carbon paste, glucose oxidase (GOD) membrane (on K1) and invertase (INV)/mutarotase (MUT)/GOD membrane (on K2), is proposed for simultaneous determination of glucose and sucrose. Interference to the K2 caused by H2O2 produced in the enzyme reaction on K1 can be eliminated by depositing a catalase (CAT) layer between K1 and K2. Acceleration of glucose mutarotation by MUT was found to be three times faster than by phosphate buffer. Glucose was determined directly from the response of K1, while sucrose was computed based on a formula derived from a two‐point calibration method. The maximum concentration of sucrose that could be determined depended strongly on the concentration of coexisting glucose. The activities of immobilized GOD, INV, MUT, and CAT were examined individually and found stable at room temperature for at least 1 month. The final loss of sucrose response activity was found to be due to denaturation of INV.