QbD-Driven Analytical Method Development and Validation for Raloxifene Hydrochloride in Pure Drug and Solid Oral Dosage Form

Abstract

The present research work encompasses systematic development of a simple, fast, sensi- tive, reproducible and cost-effective reversed-phase high-performance liquid chromatographic (RP-HPLC) method, applying the principles of Quality by design (QbD) for quantification of raloxifene hydrochloride (RLX). Reversed-phase chromatography was carried out using Waters Acquity high-performance liquid chro- matographic system (HPLC), equipped with photodiode array (PDA) detector controlled by Empower 2^® software, using a Hibar Purospher^® STAR C₁₈ (250 x 4.6 mm; 5 μm) HPLC column. Initial risk assessment followed by screening studies was carried out using a four-factor-eight-run fractional factorial design (FFD). Subsequently, optimization studies were carried out employing a central composite design (CCD) with flow rate and mobile phase ratio as the critical method parameters (CMPs), and tailing factor (TF), % assay and theoretical plate count (TPC) as the critical analytical attributes (CAAs). The optimal chromatographic sepa- ration was conducted using graphical and numerical optimization with the mobile phase composition of methanol and sodium acetate buffer (pH 4) in the ratio of 50:50 (% v/v) at a flow rate of 1mL/min, an oven temperature of 40°C at a λ_max of 287 nm. Linearity was observed in the concentration range of 0.5-100 μg/mL, with a correlation coefficient of 0.999. Further, the LOD and LOQ were found to be 0.5432 ng/mL and 1.6461 ng/mL respectively. The developed method was validated as per the ICH Q2 (R1) guidelines, and the principles and science of analytical quality by design (AQbD) for establishing a high degree of linearity, accuracy, precision, sensitivity and robustness, for estimating RLX in bulk drugs and marketed formulations.

Key word::

• Raloxifene HCl
• RP-HPLC
• design of experiments (DoE)
• risk assessment