Quantitative thermodynamic analyses of spectroscopic titration curves

•Spectroscopic methods rely on spectroscopic property responses to complex formations.•In general, spectroscopic titration curves are not thermodynamic binding isotherms.•The examined spectroscopic property should be related to the true degree of binding.•Methods to obtain true degree of bindings in spectroscopic titrations are discussed.•Described methodologies are particularly useful for multiple-ligand binding processes.