A novel rigid anion receptor 1 containing thiosemicarbazone groups were synthesized. Receptor 1 displayed competitive turn-off fluorescence and colorimetric response for hydrogen pyrophosphate. This recognition process was confirmed by UV–vis, fluorescence titrations. A detailed analysis of the TD-density functional theory calculations was consistent with the observed colorimetric experiments. These experiments revealed that the colorimetric and fluorescent response is triggered by thiourea/ HP2O7 3− hydrogen bonding.