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A sensitive and robust electrochemical aptasensor for determining dopamine (DA) was developed using a grass carp skin collagen-graphene oxide (GCSC-GO) composite as a transducer and a label-free aptamer as a biological recognition element for the first time. In order to fabricate this sensor, the GCSC-GO composite was firstly prepared by ultra-sonication method and characterized by atomic force microscope, infrared spectroscopy, Raman spectroscopy, and electrochemical impedance spectroscopy. Subsequently, a label-free DA-binding aptamer was immobilized through strong interaction between collagen and aptamer. The fabricated electrochemical aptasensor was used to determine DA by differential pulse voltammetry. The results indicated that the peak current changes of the developed aptasensor was linear relationship with the DA concentrations from 1 to 1000 nM, and the detection limit was 0.75 nM (S/N = 3). Moreover, the fabricated aptasensor showed high selectivity for DA. More importantly, the obtained aptasensor exhibited satisfactory recovery toward DA in human serum specimens with excellent stability.