Determination of vamorolone in pharmaceutical formulation in the presence of degradation products by liquid chromatography method based on design-based quality approach

Pharmaceutical products must have the required identity and purity characteristics. If a pharmaceutical product does not meet the established quality standards, it may lack the desired therapeutic effect and cause toxic and adverse reactions. The quality of the drug product should be monitored at every stage, including the initial Active Pharmaceutical Ingredients manufacturing process, packaging, transportation, and storage conditions. Vamorolone is an oral corticosteroid agent with proven efficacy and reliability and a low side effect profile developed for the treatment of duchenne muscular dystrophy. There is no method developed and validated in pharmaceutical formulations and biological fluids for the agent in question in the literature. In this study, a liquid chromatography technique was established for the analysis of VAM in pharmaceutical formulations. The analytical technique has stability indicators, and optimization studies were conducted in the presence of degradation products. All stages of the optimization studies of the analytical method were carried out by performing risk analysis in line with the quality approach in design and using screening designs that determine the parameters that will significantly affect the optimization. A three-level Box-Behnken response surface design was used for the final stage of optimization and for determining the optimized conditions. The conditions of the optimized method were determined as water and acetonitrile (60:40; v/v) containing 0.1% acetic acid as the mobile phase, 0,5 mL/min flow rate, 3 µL injection volume, column oven temperature 30 °C ± 0,1, and detection wavelength 241 nm. In addition, the validity of the developed method was verified according to the ICH Q2(R2) guideline. The sustainability and practicality of the developed method were evaluated. Greenness assessment was evaluated using NEMI, Analytical Eco-scale (87), and AGREE (0,62) The BAGI (77,52) tool was used in the practicality assessment. Finally, the method was evaluated with the RGB12 (92,9) metric in order to provide a common scaling of greenness and practicality assessments. The proposed method has the practicality to be applied in every analytical laboratory