Highly fluorescent gold nanodots have been synthesized through a novel rapid, facile and one-pot room temperature route using trithiocyanuric acid as mild reducing agent and surface ligand. The proposed synthesis overcomes limitations of other synthetic routes in terms of cost, time, complexity and environmental risks, and gives rise to highly fluorescent gold nanodots within 10 min at room temperature, with a maximum emission wavelength at 623 nm and a large Stokes shift (213 nm). Moreover, the synthesized gold nanodots showed a large emission QY (9.62 × 10-2) and excellent photostability and colloidal properties during long periods. Increasing concentrations of CN- in aqueous solution progressively quenched the fluorescence emission and produced a slight blue shift of the synthesized gold nanodots. A good linear relationship was observed for CN- concentrations between 0.29 and 8.87 μM, obtaining a detection limit estimated according to the 3s IUPAC criteria of 150 nM. Besides, the influence on the fluorescence signal of potential interferents at high concentrations (1000 μM) was studied, including I-, F-, citrate, [Formula: see text] [Formula: see text] [Formula: see text] CH3COO-, EDTA, Br-, [Formula: see text] Cl- and S2- K+, Na+, Li+, Mg2+, Ca2+, Ba2+, Cu2+, Zn2+, Ni2+, Al3+, Fe2+, Fe3+, Pb2+, Cd2+, Hg2+ and Co2+. Results showed a high selectivity towards all the investigated ions, except for Pb2+, Cd2+ and Hg2+, although the use of glutathione and BSA as masking agents drastically minimized the effect of such cations at high concentrations. The synthesized gold nanodots were successfully evaluated as highly sensitive and selective probes for cyanide determination in environmental water samples, including tap, river, lake and sea water, indicating the validity of TCA-AuNDs for analytical CN- contamination control.