DEVELOPMENT AND ANALYTICAL QUALITY BY DESIGN-BASED OPTIMIZATION OF REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC METHOD FOR QUANTIFICATION OF TRIMETAZIDINE DIHYDROCHLORIDE IN PHARMACEUTICAL DOSAGE FORMS
Abstract
Objectives: Analytical method development through quality by design (QbD) framework includes a systematic approach to establish robust analytical procedures in compliance with the International Council for Harmonisation (ICH). The present investigation describes the structured development and comprehensive validation of a reversed-phase high-performance liquid chromatographic technique for determination of trimetazidine dihydrochloride in bulk and modified-release tablet formulations. Methods: Central composite design incorporating circumscribed configuration was implemented for evaluating critical method parameters (CMPs) including mobile phase composition (methanol percentage) and volumetric flow rate. Separation was accomplished utilizing an Agilent C18 column (4.6×250 mm, 2.5 μm particle size) with methanol and 0.1% orthophosphoric acid aqueous solution (65:35, v/v) as mobile phase, maintained at a flow rate of 1.00 mL/min with spectrophotometric detection at 231 nm. Results: The established methodology demonstrated exceptional analytical characteristics: Linearity across 20–100 μg/mL concentration interval (r2=0.9996), intermediate precision with relative standard deviation (RSD) ≤0.309%, accuracy within 98–102% recovery bounds, limit of detection (0.678456 µg/mL), limit of quantification (2.055926 µg/mL) and assay performance of 99.15% label claim in pharmaceutical tablets. The analytical target profile specifications were consistently satisfied across all validation parameters, conforming to ICH Q2(R2) standards. Conclusion: Statistical analysis revealed mobile phase composition and flow rate as predominant CMPs influencing retention time, peak area, and chromatographic efficiency. Robustness evaluation confirmed method stability with %RSD values ≤1.734%, substantiating its applicability for routine quality control. The structured analytical QbD methodology facilitated establishment of a method-operable design region delineating operational boundaries for assured performance.