A couple of new fluorescent probes, based on diethoxycarbonyl glycoluril, were synthesized and characterized by 1H NMR, X-ray crystal structure. The probes were found to be relatively highly fluorescence with quantum yields of 0.20 and 0.18, respectively, at room temperature in THF solution. Both of the proposed probes were tested using different phenolic compounds, showing very high selectivity and sensitivity for 2-nitrophenol by forming a 1:1 complex in THF-MeOH (9:1, v/v). Additionally, the sensors could be used for the qualitative analysis of 2-nitrophenol in real water samples.