Enhancing Pharmaceutical Analysis with Analytical Quality by Design in UHPLC: A Review of Methodological Innovations (2014–2025)

The integration of analytical quality by design (AQbD) in ultrahigh-performance liquid chromatography (UHPLC) has enhanced pharmaceutical method development by improving robustness, efficiency, and regulatory compliance. Despite its growing adoption, a dedicated evaluation remains limited. Among 26 review articles on AQbD, none exclusively focuses on UHPLC, while 52 research papers detail its pharmaceutical applications. This review bridges this gap by consolidating reported methods and advancements in risk assessment, experimental design, and optimization strategies for defining the method operable design region (MODR). Screening designs like Plackett-Burman design (PBD) and risk assessment tools like Ishikawa diagrams identify critical method parameters (CMPs), while response surface methodology (RSM)-based models optimize performance. The impact of stationary phases, mobile phase composition, and flow rate adjustments is examined, emphasizing ethylene-bridged hybrid octadecylsilane columns and mobile phase trends favoring water-acetonitrile (CAN) combinations with formic acid (FA) or ammonium acetate. Gradient elution is frequently applied for complex separations, with optimized flow rates (0.2-0.5 mL/min) enhancing efficiency and sustainability. Unlike previous reviews, this study systematically evaluates AQbD-driven UHPLC, offering critical insights for regulatory compliance, lifecycle management, and future pharmaceutical analysis advancements.