Abstract A new, simple and low cost spectrophotometric method for the determination of Glibenclamide in pharmaceutical preparations was developed. The method was based on the interaction of Glibenclamide as n-electron donor with Picric acid as the π – acceptor. The absorbance maximum (λmax) of the resultant complex was at 419 nm. Different variables affecting the reaction were carefully studied and optimized. Of the different solvents tried, Acetonitrile was found to be the most suitable solvent. Beer’s law was obeyed in concentration range of 20 – 100 μg/ml Glibenclamide. The correlation coefficient was found to be (r = 0.9920). The limit of detection and limit of quantification were 1.40 µg/ml and 4.12 µg/ml, respectively. The reaction ratio between Glibenclamide and Picric acid was studied and found to be 1:1. The method was validated and results obtained for the assay of three different brands of Glibenclamide tablets were compared with the BP method (titrimetric). The repeatability and reproducibility of the developed method were evaluated and the obtained results quoted. The proposed method was successfully applied to the determination of Glibenclamide with good accuracy and precision.