A novel multifunctional biosensor was prepared by dropping graphene on the 2-Amino-thiazol (AT) film at glassy carbon electrode (GCE). The surface morphology of graphene, AT and graphene-AT film modified GCE has been examined by using SEM and AFM. The advantages of proposed film were demonstrated for the determination of H2O2 reduction using amperometric, DA and UA oxidation using differential pulse voltammetry. The graphene-AT/GCE as a multifunctional biosensor exhibited a linear response range for H2O2 determine (from 1.010-5 to 3.110-4 M), low detection limit (10-6 M), high sensitivity (1.73 mA mM-1 cm-2) and the relative standard deviation (RSD %) less than 2% (n = 5). Similarly, the multifunctional biosensor can simultaneously determine the DA and UA with the interference of AA. For the linear response range of DA and UA were from 510-6 to 2.510-5 M and 2.510-6 to 1.2510-4 M. In addition, the multifunctional biosensor by graphene-AT film modified GCE was advantageous in terms of its simple preparation, specificity, stability and selectivity.