Applications of hydrophilic interaction and mixed-mode liquid chromatography in pharmaceutical analysis

Hydrophilic Interaction Liquid Chromatography (HILIC) and Mixed-Mode Chromatography (MMC) excel in separating polar, hydrophilic, and charged analytes due to unique hydrophilic or mixed-mode retention mechanisms. They represent a complementary approach to the widely used Reversed Phase Liquid Chromatography (RPLC). Often, where RPLC struggles, HILIC and MMC thrive. The applications of HILIC and MMC in pharmaceutical analysis are expanding rapidly across a variety of drug modalities. This article reviews advances in the applications of HILIC and MMC in seven major areas of pharmaceutical analysis: synthetic small molecules, counterions and salts, lipids and surfactants, carbohydrates, amino acids and peptides, proteins, and nucleic acids in the past two decades. We aim to provide comprehensive information and strategic guidance to facilitate future research, development and applications in these areas.