Metal ions are essential for the life of plants, animals and humans. Among several metal ions of importance in human life, Zn2+ is involved in a variety of biological processes and hence its detection is an important aspect of research. In this regard, we synthesised a 1,3-bis(pyridine-2-ylmethyl) amido derivative of calix[4]arene (L) and studied its interaction towards five different biologically relevant metal ions and three different environmentally important ones. Among these, the L exhibits notable selectivity and sensitivity towards Zn2+ as demonstrated based on fluorescence spectral intensity. The Zn2+ forms a 1:1 complex with L and exhibits a binding constant (Ka) of 48255 ± 264 M–1 as derived based on the fluorescence data. The 1:1 complex formed was further confirmed by mass spectrometry. Based on all these data, a six coordinated structure having [ZnN6]2+ coordination core was proposed. The proposed structure was supported by DFT computations from which the geometry-based features of the complex have been derived, thereby obtaining a better understanding of the Zn2+ coordination features with the conjugate.