DESENVOLVIMENTO E VALIDAÇÃO DE METODOLOGIAS ANALÍTICAS PARA QUANTIFICAÇÃO E DISSOLUÇÃO IN VITRO DE DAPAGLIFLOZINA

Abstract

The commercial formulation containing dapagliflozin (Forxiga®) was recently approved by regulatory agencies in Europe, Brazil and United States (EMA, ANVISA and FDA, respectively). The drug is not described in the pharmacopoeia and only two analytical methods are found in the literature. Thus, it is necessary the development and validation of analytical methodology for the stability indicating assay of Dapagliflozin in tablets as well as in vitro dissolution studies. The analytical method for the determination of dapagliflozin tablets and their degradation products by HPLC using Phenomenex® Luna C18 reverse phase chromatographic column (150 mm x 4.60 mm, 5 μm) and mobile phase composed of acetonitrile and formic acid 0.1% (42:58, v/v) with flow 1 mL min-1 was validated, showing to be specific and linear in the range of 1 to 100 μg mL-1 (y= 7019.x + 1535 r2 = 0.999). The limits of detection and quantification were 0.09 and 0.28 μg mL-1, respectively, indicating a more sensitive method compared to that found in the literature. The methodology was confirmed precise and accurate, with recovery within the specified values. The robustness was determined by Factorial experimental design. The method was successfully applied to the quantification of dapagliflozin tablets. A degradation product was found in the photolytic (UV light) degradation study and was evaluated by UPLC-MS/MS and characterized as a possible dimer of dapagliflozin. So far, this degradation product wasn’t described in the literature and its safety should be further evaluated. For the dissolution studies, the analytical methodology was validated using apparatus II and simulated gastric fluid without enzymes (pH 1.2) as the dissolution medium, with assays being performed at 50 rpm. The method was specific and linear in the interval from 0.5 to 15 μg mL-1 (y= 0.052.x + 0.008, r2= 0.998), with limit detection and quantitation of 0.05 and 0.15 μg mL-1, respectively. It proved to be precise and accurate, with values within the requirement made by the International Conference Harmonization (ICH), and also robust, through a systematic approach (Factor 23). The applicability of the method was confirmed by the study of the dissolution profile of differents batches of Forxiga®. With the duly validated methodologies, it is important to emphasize the importance of this procedure both from the regulatory point of view and in relation to the quality of the pharmaceutical product.