The glucose level measurement in the blood of diabetic patients without significant variations is important. The level of glycated hemoglobin (HbA1c) in the blood provides an authentic tool for glucose level measurement. In our study, a biosensing system established on properly aligned single-crystal zinc oxide (ZnO) nanorods structures grown on indium-tin oxide coated glass plate (ITO) electrode. ZnO nanorods were immobilized with fructosyl amino-acid oxidase (FAO) enzyme through physical adsorption integrated with cross linking molecules N-5-azido-2-nitro-benzoyloxysuccinimide (ANB-NOS). Whole blood samples were first hemolyzed & then properly digested with protease prior to measuring the HbA1c through the sensor. This enzyme biosensor reported an optimum response at +0.2 V. This biosensor displayed a significant sensitivity and detection limit (0.1μM), fast response time (4s) and wide linear range (from 0.1 to 2000 μM). The enzyme/working electrode is stable for about 4 months, when kept at 4 oC. This recommended biosensor method may apply for detecting HbA1c in blood samples obtained from apparently healthy as well as diabetic patients.