Results from tests aimed at optimizing an instrumental procedure for the direct and fast determination of uranium in solution by laser fluorescence are presented. A comparison of sample fluorescence measured using different fluorescence enhancing reagents was performed: sodium pyrophosphate, orthophosphoric acid, sulphuric acid and a commercially available fluorescence enhancer were tested for the determination of uranium. From the experimental results, 0.01 M Na4P2O7·10H2O showed the best performance. Effects of reagent pH, different matrices, different concentrations of dissolved Th, and sample volume were investigated. Applications of the improved procedure for the determination of uranium in samples arising from UO2-based high level nuclear waste dissolution studies are described.