In this work, a novel non-enzymatic acetylcholine (ACh) sensor was constructed based on electrodepoition of Ni-Al layered double hydroxides (Ni-Al LDHs) on ordered mesoporous carbon (OMC) modified glassy carbon (GC) electrode. The obtained Ni-Al LDHs/OMC nanocomposite was characterized by scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD) and electrochemical methods. A novel non-enzymatic acetylcholine (ACh) sensor based on Ni-Al LDHs/OMC was developed. Compared with the pristine OMC and Ni-Al LDHs modified electrodes, Ni-Al LDHs/OMC displayed higher electrocatalytic activity toward ACh and gave a wide concentration interval of 2-49221 mu M and a low limit of detection (42 nM, R=0.999), which is better than some reports in the literatures. The sensor also exhibited long-term stability and remarkable antifouling property. The excellent electrocatalysis for ACh can be attributed to the combination of the unique properties of Ni-Al LDHs nanoparticles and the ordered mesostructure of OMCs matrix. (C) 2012 Elsevier Ltd. All rights reserved.