A Schiff base compound E-2-(((4-aminophenyl)imino)methyl)-5-(difluoromethoxy)phenol was synthesized and characterized by FT-IR, 1H and 13C NMR, ESI-mass spectroscopy. The synthesized compound also selectively detects Al3+, Fe2+ and Cu2+ without any interference of other metal ions. Fluorescence titrations carried out to find the selectivity of Al3+, Fe2+ and Cu2+ in turn-on system, with binding modes of 2:1 complex, confirmed by Job′s plot. The presence of metal ions Al3+, Fe2+ and Cu2+ with receptor conformed by ESI-MS spectrum, which changed the base value at 298.00 m/z. Moreover, among the binding constant of three metals calculated (20 μM), Al3+ showed a high value of 5.7 × 104 M-1 compared to Fe2+ and Cu2+ metal ions. Prominently, the cytotoxicity activities of probe with HeLa cells were also calculated