RP-HPLC Method Development and Stability-Indicating Assay of Vericiguat: An Analytical Quality by Design and Green Chemistry Approach

Abstract

Objectives: The present studies describe the systematic development and validation of a stability-indicating and environmentally sustainable RP-HPLC method for the quantification of Vericiguat in bulk form by applying AQbD and green chemistry principles. Materials and Methods: Method optimization was performed by employing custom design, where critical method parameters and critical analytical attributes were evaluated statistically. Optimal chromatographic separation was achieved on a Grace C18 column (250 × 4.6 mm, 5 µm) under isocratic conditions using potassium dihydrogen phosphate buffer and methanol (60:40, v/v, pH 4.5) at a flow rate of 0.75 mL/min, with detection at 254 nm. Method validation and degradation studies conducted according to ICH guidelines. Environmental sustainability was assessed using green analytical assessment tools. Results and Discussion: The optimized method showed a global desirability of 0.75. Vericiguat exhibited a retention time of 3.713 min, plate count of 8253, and tailing factor of 1.53. The method demonstrated excellent linearity over 50-150 µg/mL with an R² value of 0.999. The limit of detection and quantification were 0.082 and 0.20 µg/mL, respectively. Significant degradation was observed under acidic and thermal stress conditions, with degradation of 11.07% and 11.00% within 20% limit. AGREE and MoGAPI scores of 0.79 and 86 confirmed sustainability. The Custom Design approach enabled optimization through contour plots and desirability studies, while validation, stability-indicating, and greenness were established according to ICH guidelines. Conclusion: The developed method is novel, reliable, accurate, environmentally sustainable, and suitable for routine quality control analysis of Vericiguat in bulk drug.