Dansyl-derivatized, triazole-linked, glucopyranosyl conjugates, 5FLOH, 2FLOH, 1FLOH, and 0FLOH were synthesized and characterized. While the 5FLOH acts as a molecular probe for CN-, 2FLOH, 1FLOH, and 0FLOH acts as control molecules. The reactivity of CN- toward 5FLOH has been elicited through the changes observed in NMR, ESI MS, emission, and absorption spectroscopy. The conjugate 5FLOH releases a fluorescent product upon reaction by CN- in aqueous acetonitrile medium by exhibiting an ∼125-fold fluorescence enhancement even in the presence of other anions. Fluorescence switch-on behavior has been clearly demonstrated on the basis of the nucleophilic substitution reaction of CN- on 5FLOH. A minimum detection limit of (2.3 ± 0.3) × 10-7 M (6 ± 1 ppb) was shown by 5FLOH for CN- in solution. All the other anions studied showed no change in the fluorescence emission. The utility of 5FLOH has been demonstrated by showing its reactivity toward CN- on a thin layer of silica gel as well as on Whatman No. 1 cellulose filter paper strips. The role of glucose moiety and the penta-fluorobenzenesulfonyl reactive center present in 5FLOH in the selectivity of CN- over other anions has been demonstrated by fluorescence, absorption and thermodynamics study. Similar studies carried out with the control molecules showed no selectivity for CN-. The mechanistic aspects of the reactivity of CN- toward 5FLOH were supported by DFT computational study.