QBD-Driven RP-HPLC Method Development and Validation for Ipragliflozin and Saxagliptin in Pharmaceutical Dosage Forms Following ICH Guidelines

Abstract

A robust and validated Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) method was developed for the simultaneous estimation of Ipragliflozin and Saxagliptin in pharmaceutical dosage forms. Method optimization was achieved using a Box-Behnken Design, evaluating the influence of key chromatographic parameters such as buffer pH and flow rate. Optimal separation was accomplished using an Inspire C18-EP column (4.6 × 250 mm, 5 µm) with a mobile phase of methanol and phosphate buffer (pH 6.0) in a 70:30 ratio, at a flow rate of 1 mL/min and UV detection at 254 nm. The method demonstrated excellent resolution (3.52), acceptable tailing factors (<1.2), and high theoretical plate counts, confirming efficient peak performance. ANOVA results validated the statistical significance of the model, and all system suitability parameters met regulatory standards. The method was thoroughly validated in accordance with ICH guidelines, showing excellent linearity for Ipragliflozin (30–150 µg/mL, R² = 0.9993) and Saxagliptin (3–15 µg/mL, R² = 0.9994). Assay results confirmed high accuracy with recoveries of 99.7% and 99.6% respectively. Precision studies yielded %RSD values below 2%, indicating strong repeatability and intermediate precision. Sensitivity was established through low LOD and LOQ values, while robustness was confirmed under deliberate variations in chromatographic conditions. Overall, the developed RP-HPLC method is simple, precise, accurate, and reliable, making it highly suitable for routine quality control of Ipragliflozin and Saxagliptin in bulk and formulated pharmaceutical products.