Identification, Determination, and Potency Quantification of Roxithromycin by a Novel Validated RP-HPLC Method for Bulk and Pharmaceutical Formulations

Introduction: The project visualizes a robust HPLC method for the separation and quantification of roxithromycin (ROX) using Stat-Ease software and Design of Experiments (DoE) access. Methods: The UV method quantified ROX in ex vivo studies, confirmed excipient compatibility, and modelled how solvent concentration, flow rate, and column temperature affect chromatographic performance. Results: The retention time of pure drug roxithromycin is 4.606 have sharp peak, and the retention time of drug roxithromycin with excipients is 4.610. A central composite design was applied to assess factor interactions and quadratic effects on selected responses, which were subsequently validated. ROX was eluted using a Quasar C18 SS column (250mm × 4.6mm id, 5μm) and detected with UV at 207 nm in a quick procedure. The mobile phase was 80% methanol and 20% acetic acid by volume. Discussion: This technique showed linearity above 20-120μg/mL concentration scale with a correlation coefficient of 0.9994. Roxithromycin dosages of 60μg/mL, 120μg/mL, and 180μg/mL resulted in recovery rates of 99.89%, 99.88%, and 99.78%. The LLOQ & LLOD values showed that ROX can be estimated at 3.04μg/mL and precisely at 9.2μg/mL. In method applicability of % drug loading (NEF-1, NEF-2 & NEF-3) have % claim of drug (101%, 98% & 102%). The analytical robustness of the design was confirmed by variations in mobile/solvent phase flow rate (± 0.5 mL/min), column temperature (± 5°C), and solvent/mobile phase concentration (± 5.0 mL), which affected chromatographic system suitability parameters, although maintaining overall system suitability. Conclusion: DoE optimization established a robust, specific HPLC method for accurate quantification of roxithromycin in pure form and formulations