Poly[1-(2-aminophenyl) pyrrole] film was electropolymerized on a gold electrode and used for the first time as the matrix for the electrochemical determination of glucose. The working electrode where glucose oxidase was chemically immobilized by amine groups of the poly[1-(2-aminophenyl) pyrrole] film was used for online glucose determination with flow-injection operation. The optimum conditions for glucose were a flow rate of 4mL/min at +0.7V in 100mM using pH 6.5 phosphate buffer as the carrier. The electrode provided a long linear dynamic range between 0.2mM and 30mM, a response time of 9s, a detection limit of 63 mu M, a relative standard deviation of 0.7%, and an accuracy of 99.95%. The working electrode preserved 64% of its initial current value following six weeks. The optimized device was employed to determine glucose in wastewater from the Zonguldak Biological Treatment Plant, Turkey.