Application of H-point standard additions method to spectrophotometric and spectrofluorimetric determinations of glafenine and glafenic acid in mixtures

H-point standard additions method (HPSAM), based on spectrophotometric and spectrofluorimetric measurements, was proposed for simultaneous determination of glafenine (G) and glafenic acid (GA). A study of the absorption spectra of G and GA in various pH media has been carried out. Reasonably resolved UV-absorption spectra were obtained with a solution adjusted at pH 4.5 with citric acid–phosphate buffer. Additionally, the fluorescence properties in aqueous micellar systems of anionic, cationic and non-ionic surfactants were investigated. Well resolved fluorescence spectra were established in aqueous Triton X-100 solution at pH 7.8 (citric acid–phosphate buffer). As a comparative method, UV-derivative spectrophotometry (based on zero-crossing technique) was suggested. First-derivative value at 352 nm (1D352) and second-derivative value at 366 nm (2D366) were selected for the quantification of G and GA, respectively. The relative standard deviations of the proposed methods approximate 2%. The proposed methods were evaluated through the analysis of commercial tablets. The results were accurate and precise.