The usefulness of caffeine (CF) as a chelating agent to increase the luminescence intensity of terbium (Tb3+) was studied. The efficiency of the developed Tb3+/CF coordination nanoparticles was evaluated as a sensing platform for the detection of fluoroquinolone antibiotics using norfloxacin (NF) as a model analyte. The luminescence conditions of Tb3+/CF-NF were investigated and optimized. Under the optimized experimental conditions, the linearity of the method was established with a limit of detection (LOD) of 5.8 nM, and the effect of some expected interferes was examined. The developed method was successfully applied for the detection of norfloxacin in tablet and urine samples with recoveries between 88.9–96.0% and 90.7–96.3%, respectively.