High-performance liquid chromatography (HPLC) remains the most widely adopted analytical tool for the estimation, purity assessment, and stability profiling of anti-viral agents. Significant advancements in column technologies, mobile phase optimization, detectors, and analytical quality-by-design (AQbD) approaches have enhanced the robustness and reliability of HPLC methods. Anti-viral drugs-including nucleoside analogues, protease inhibitors, integrase inhibitors, polymerase inhibitors, and fixed-dose combinations-require highly selective and stability-indicating methods due to their complex structures, degradation pathways, and combination formulations. This review highlights the fundamental principles of HPLC method development, recent advancements in stationary phases, ultra-high-performance liquid chromatography (UHPLC), AQbD-driven method optimization, and green analytical approaches. Detailed discussion on validation parameters based on ICH Q2 (R2), emerging trends, and comparative evaluation of published methods is provided. The review summarizes more than 20 years of literature to present a comprehensive understanding of how HPLC continues to evolve for the estimation of anti-viral agents.
Gali Haritha1*, Madhulapally Swathi2, Arunabha Mallik3 (Sun,) studied this question.