Highly polar compounds ? such as small molecule pharmaceuticals, metabolites, and biomarkers ? can be challenging to separate using traditional reversed-phase separation methods. Several alternatives are available, including hydrophilic interaction liquid chromatography (HILIC), aqueous normal phase (ANP) chromatography, and HPLC using ion-exchange, polar-embedded, or polar-endcapped columns. So which option should you choose? Joseph Pesek of San Jos? State University explains the mechanisms, advantages, and disadvantages of these approaches.