QBD Driven Analytical Method Development and Validation for Ganaxolone in Bulk and Pharmaceutical Dosage Form by RP-HPLC

Abstract

A robust and reliable Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) method was developed and validated for the estimation of Ganaxolone in bulk and pharmaceutical dosage forms in accordance with ICH Q2(R1) guidelines. Method optimization was carried out using a Box–Behnken design to evaluate the influence of flow rate, mobile phase ratio, and buffer pH on key responses such as tailing factor and retention time. The optimized chromatographic conditions included a SPURSIL C18-EP column (3.0×150mm, 3μm), a mobile phase of 40% formic acid and 60% acetonitrile, a flow rate of 1.0 mL/min, and detection at 220 nm. The developed method yielded sharp, symmetrical peaks with satisfactory system suitability parameters. Validation results confirmed excellent performance with assay (100.6%), linearity (10–50 µg/mL, R² = 0.9998), precision (%RSD<1), accuracy (mean recovery ~100.2%), LOD (0.03 ppm), and LOQ (0.11 ppm). Robustness studies demonstrated the stability of the method under small deliberate variations in chromatographic conditions. The results establish that the proposed RP-HPLC method is simple, precise, accurate, sensitive, and reproducible, making it highly suitable for routine quality control of Ganaxolone in bulk and dosage forms. Its reliability, cost-effectiveness, and compliance with regulatory requirements further support its applicability in stability studies, batch release testing, and industrial pharmaceutical analysis.