The invention discloses a fluorescent probe Eu-dtpa-bis(adenine) and application thereof to detection of orotic acid in urine. A preparation method of the fluorescent probe comprises the followingsteps: making diethylenetriamine pentaacetic acid, acetic anhydride and pyridine reflow with stirring at the temperature of 65 DEG C for 24 hours; cooling, performing suction filtration under reducedpressure, washing and drying; making prepared diethylenetriamine pentaacetic acid dianhydride, triethylamine, dimethyl formamide (DMF) and adenine reflow with stirring at the temperature of 100 DEG Cfor 24 hours; performing cooling, rotational evaporation, washing and drying; heating prepared diethylenetriamine pentaacetic acid-di(adenine) and Eu(NO3)3.6H2O with stirring at the temperature of 60DEG C for 2 hours to obtain a target product. The Eu-dtpa-bis(adenine) is taken as a probe for detecting the orotic acid in conjunction with a fluorescence method. The method disclosed by the invention is simple and novel, is low-cost, achieves high efficiency, and can be applied to an actual urine sample.