This article first reported that a simple synthesis of carbon dots (UCDs) by pyrolysis of uric acid was proposed. The excitation wavelength was 350 nm and the emission wavelength was 402 nm for the synthesized UCDs. And the corresponding fluorescence quantum yield was 52.06%. The obtained UCDs could be served as a fluorescence probe to recognize Ag+ and glutathione (GSH), respectively. The fluorescence of UCDs was quenched after the addition of Ag+. The obtained UCDs had a linear relationship with Ag+ in the detection range of 0.1 μM to 2.0 μM, and the detection limit was 39 nM. The quenching fluorescence of this system could be restored after adding GSH. The fluorescence intensity increased linearly on increasing the concentration of glutathione in the range of 4 to 9 μM, and the detection limit was 66 nM. Based upon these phenomena, we proposed a novel fluorescence probe to detect Ag+ and detect GSH, respectively. In addition, the prepared UCDs were successfully applied to analyze Ag+ in mineral waters and GSH in blood serums.