Zolpidem is an imidazopyridine agent for short term treatment of insomnia with a favorable adverse effect profile. A novel chemiluminescence (CL) method has been proposed for simple determination of zolpidem. The method is based on the fact that zolpidem can produce CL emission in the system of acidic KMnO4 (permanganate) and Na2SO3 (sulfite). On the optimized conditions of chemical variables, CL intensity was proportional to concentration of zolpidem in the range 2.5-295.1 ng mL-1. Limit of detection was 1.6 ng mL-1 (S/N=3). The percent of relative standard deviation was 4.5% for 98.4 ng mL-1 zolpidem. The proposed method was satisfactorily applied for the determination of zolpidem in pharmaceutical formulations and human plasma samples. Sampling rate of the method was calculated about 30 samples h-1. CL mechanism has been proposed using UV-Vis and CL spectra.