QbD assisted green and white analytical UPLC method quantification of pharmacopeia impurities of Pravastatin in bulk drug and pharmaceutical formulations

Abstract

Background: A QbD-assisted green and white UPLC method was optimized for the simultaneous quantification of pravastatin and its EP impurities B, C, and D. Methods: A Design of Experiments (DoE) approach was used to evaluate the influence of critical method parameters (CMPs) on key chromatographic responses. Results and discussion: The statistical modeling using quadratic response surface methodology yields excellent regression coefficients (R² = 0.9399 for Pravastatin–impurity C and R² = 0.9758 for impurities C-D) and significant F-values (p < 0.0001), confirming robust model performance. The optimized chromatographic conditions were ethanol and 0.1% formic acid (65:35, v/v) at 0.3 mL/min as the mobile phase, with detection at 239 nm and a Waters ACQUITY BEH C18 column (100 × 2.1 mm, 1.7 µm). These conditions provide sharp, symmetrical peaks at retention times of 0.31, 0.89, 1.16, and 1.43 min for impurity B, pravastatin, impurity C, and D, respectively. Method was validated in accordance with ICH Q2(R2) guidelines, and results demonstrate excellent linearity (r² &gt; 0.999), precision (%RSD &lt; 1%), recovery (98.17–101.09%), and sensitivity (LOD = 0.015 µg/mL; LOQ = 0.05 µg/mL). The AGREE metric yields the greenness score of 0.81, and the GAPI pictogram confirms the minimum environmental impact (E-factor = 7.0 × 10⁻²). Furthermore, RGB 12 whiteness assessment provides a whiteness brilliance (MB) score of 80.6%, indicating an optimal balance among analytical performance, eco-friendliness, and operational efficiency. Conclusion: The proposed method was robust, rapid, environmentally sustainable, and practically efficient for routine impurity profiling and quality control of pravastatin formulations.