An electrochemical sensor modified with tetra-(N-phenylpyrazole)-porphyrin/b- cyclodextrin/carbon nanotube (TPPyPh/b-CD/CNT) film was fabricated for determination of hemoglobin (Hb) in this paper. The electrochemical behavior of the proposed sensor was investigated by cyclic voltammetry (CV). TPPyPh has been shown preferably for the fabrication of electrochemical sensor. The peak currents decreased significantly with the increase of the Hb concentration in solution. The response mechanism of TPPyPh/b-CD/CNT film for Hb was suggested to be based on the coordinated interaction between Hb and TPPyPh. It showed excellent selectivity for the determination of Hb in PBS.