A novel composite film which contains ordered mesoporous carbon (OMC) along with the incorporation of poly(neutral red)(PNR) has been synthesized on glassy carbon elect rode by potentiostatic method. This composite film was characterized by scanning electron microscope (SEM) and cyclic voltammetry (CV) Two pairs of the redox peaks appear at formal potential E-0' = +0.045 V (peak 1) and E-0' = 0.49 V (peak II) at the PNR/OMC/GC electrode And it is found that only the redox waves (peak I) exhibits good electrocatalytic activity towards nicotinamide adenine dinucleotide (NADH) and 2-mercaptoethanol (2-ME). Under a lower operation potential of +0.07 V. amperometry method was used to determine the concentration of NADH and 2-ME, respectively. In pH 70, sensors for two molecules under their corresponding optimized conditions were developed with acceptable sensitivity and low detection limits in large determination ranges. In addition, these sensors have good reproducibility and stability. (C) 2010 Elsevier Ltd. All rights reserved