Abstract: Developing a selective and sensitive tool for recognizing 2,4,6-trinitrotoluene (TNT) is of great importance from the both security and contamination viewpoints. Here, a novel molecularly imprinted polymer (MIP) based chemiluminescence (CL) assay was described for the determination of TNT in environmental samples. ZnO quantum dots (QDs) were synthesized using a simple and economical process and applied as an efficient support for MIP layer. The MIP layer was produced by (3-Aminopropyl) triethoxysilane (APTES) as a monomer and tetraethoxysilane (TEOS) as a cross linker, in the presence of TNT as template. The MIP-labeled ZnO QDs composite (MIP@QDs) was characterized using photoluminescence emission, X-ray powder diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and transmission electron microscopy (TEM). The prepared composite showed a remarkable promoting effect on the weak chemiluminescence emission of alkaline permanganate-rhodamine B (RB) system. This effect was described based on the catalytic activity of the composite. Moreover, in the presence of TNT, its molecules were adsorbed onto specific sites of MIP and placed near the QDs, resulted in a sensible decrease in the CL intensity of KMnO4-RB-MIP@QDs system. Based on this effect, a selective and high sensitive CL assay was introduced for TNT determination. Under the optimal conditions, a linear relationship was obtained between the decreased CL intensity and TNT concentration in the ranges of 0.02-50 and 50-200 ng mL-1 with a detection limit (3σ) of 6.8 pg mL-1. This method had a good selectivity toward TNT but not the similar species such as 2-nitrotoluene, 2,4-dinitrotoluene, etc