This chapter summarizes the application of polysaccharide-based chiral stationary phases (CSPs) for separation of enantiomers in high-performance liquid chromatography (HPLC). Since this book contains dedicated chapters on enantioseparations using supercritical fluid chromatography (SFC), or capillary electrochromatography (CEC), the application of polysaccharide-based materials in these modes of liquid-phase separation techniques is touched just superficially. Special emphasis is directed toward a discussion of the optimization of polysaccharide-based chiral selectors, their attachment onto the carrier, and the optimization of the support. The optimization of the separation of enantiomers based on various parameters such as mobile phase composition and temperature is discussed.