A QBD BASED RP-HPLC METHOD DEVELOPMENT AND VALIDATION OF KETOCONAZOLE

Abstract

A stability-indicating reverse-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for the quantitative estimation of ketoconazole in bulk drug and pharmaceutical dosage forms using the Analytical Quality by Design (AQbD) framework. Critical method parameters were identified through risk assessment and optimized via Design of Experiments (DoE), ensuring robustness, reliability, and regulatory compliance. Chromatographic separation was achieved on an Agilent Zorbax SB-Aq column (250 × 4.6 mm, 5 µm) employing a mobile phase of phosphate buffer and acetonitrile (35:65, v/v) at a flow rate of 0.8 mL/min, with detection at 225 nm. Ketoconazole eluted at approximately 5.47 min within a total runtime of 15 min. Validation, performed in accordance with ICH Q2(R1) guidelines, confirmed excellent specificity, linearity, precision, accuracy, robustness, sensitivity, and reproducibility. The calibration curve was linear across 80–120 µg/mL with a correlation coefficient (R²) of 1.000. Precision studies yielded %RSD values below 2%, while recovery ranged between 99.75% and 100.07%, indicating high accuracy. Limits of detection and quantification were 0.31 µg/mL and 0.94 µg/mL, respectively. Robustness evaluation revealed negligible effects of minor variations in pH and detection wavelength. Forced degradation under acidic, alkaline, oxidative, thermal, and photolytic conditions demonstrated the method’s stability-indicating capability. Maximum degradation was observed under oxidative stress (16.14%), followed by thermal (9.34%), photolytic (8.97%), alkaline (6.71%), and acidic (6.09%) conditions, with degradation products well resolved from the analyte peak. The AQbD-based RP-HPLC method is rapid, accurate, economical, and highly robust, making it suitable for routine quality control, stability testing, and regulatory applications in ketoconazole analysis