Electrospun molybdenum disulfide (MoS2) nanoparticles composite carbon nanofibers (MoS2-CNF) were first applied for determining the amount of Vanillin. The synthesized MoS2-CNF was characterized by scanning electron microscopy (SEM), Raman, X-Ray diffraction (XRD) and Electrochemical impedance technique (EIS). The electrochemical detection of vanillin was successfully conducted in 0.1 M Phosphate Buffer (pH 10) and showed a good linear response in the range of 0.3 to 135 mu M and the detection limit reached 0.15 mu M. The sensor showed a good amperometric response signal toward vanillin and result in the determination of real sample.