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The detection of metal ions is of particular importance for monitoring environmental pollution and life metabolic activities. However, it is still a challenge to achieve Fe3+ detection with specific sensitivity and rapid response, especially in the presence of chelating agents for Fe3+ ions. Herein, a novel fluorescence probe for Fe3+, i.e., amide derivative of [1,2,4]triazolo[1,5-a] pyrimidine (TP, Id), was synthesized, featuring specific Fe3+ selectivity, rapid quenching (5 s), low limit of detection (0.82 μM), good permeability and low cytotoxicity. More importantly, Id can be used to identify and detect Fe3+ in the presence of existing strong chelating agents (e.g., EDTA) for Fe3+ ions. The results show that the as-synthesized fluorescence probe is particularly suitable as a bioimaging reagent to monitor intracellular Fe3+ in living HeLa cells. Furthermore, we proposed the binding mode for Id with Fe3+ ions and the light-emitting mechanism through high-resolution mass spectra and density function theory calculations, respectively. An Id -based test paper can be used to rapidly identify Fe3+. These results are expected to improve the development of new sensitive and specific fluorescent sensors for Fe3+. [Display omitted] • A novel amide derivative of [1,2,4]triazolo[1,5-a]pyrimidine fluorescence probe Id for Fe3+ ions was first reported with specific, rapid response to Fe3+ ions and low cytotoxicity. • The probe Id could detect Fe3+ ions in the presence of strong chelating agents (e.g., EDTA) of Fe3+ ions and the Id -based test paper could identify Fe3+ ions rapidly. [ABSTRACT FROM AUTHOR]