Design space exploration and multi-color analytical profiling of a BBD assisted RP-HPLC method for simultaneous...

Analytical Quality-by-Design principles were used to develop and validate a reverse-phase high-performance liquid chromatography method for the simultaneous quantification of butamirate citrate and chlorpheniramine maleate in pharmaceutical formulations. Box-Behnken Design systematically optimized three critical variables mobile phase pH, flow rate, and column temperature with response surface plots confirming robust chromatographic performance. Using an isocratic mobile phase of ethanol: water (40:60 v/v) containing 0.33% triethylamine and adjusted to pH 6.0 using 1% orthophosphoric acid, separation was accomplished on a Waters Spherisorb cyano column (250mm×4.6mm, 5μm) at 1.2mL/min with photodiode array detection at 225nm. Validation per ICH Q2(R2) demonstrated excellent linearity (r²=0.999) across 112-337μg/mL and 10-30μg/mL for both analytes, with percentage recoveries of 99% and 98%, respectively. Environmental sustainability was confirmed through Analytical Eco-scale (score: 80), Analytical Greenness metric (0.69), and Complex Modified Green Analytical Procedure Index (83). Analytical performance was evaluated using the Red Analytical Performance Index (70) and Multi-Color Assessment Tool (72.6%), collectively reflecting strong scope, sensitivity, accuracy, and precision. Reliable quantification of both compounds in pharmaceutical dosage forms is offered by a validated, environmentally friendly approach, supporting routine quality control and research applications.