The formation of an inclusion complex of glibenclamide (GL) with β-cyclodextrin (β-CD) in an aqueous ethanolic buffer solution of pH 7.0 has been investigated by UV spectrophotometry and differential pulse voltammetry and its stability constant is determined to be 855 and 354.15 M−1, respectively. The phase solubility profile, based on the spectrophotometric absorbance’s variations, was classified as AL-type, indicating the formation of 1:1 stoichiometric inclusion complex of glibenclamide with β-CD with a stability constant value, KS, of 846 M−1.