The increasing demand for enantiopure drugs has led to the development of a variety of stereoselective separation technologies. Among them, high performance liquid chromatography (HPLC) is well recognized as a powerful, fast and efficient technique, which has been successfully employed for analysis and preparation of enantiomers of drugs. Nowadays, liquid chromatographic techniques are the focus of intensive research, which lead to the rational design and production of highly selective and efficient chromatographic materials. This review focuses on various HPLC methods and related hyphenated techniques (including LC-MS, LC-OR detector, LC-CD detector) for chiral separation of pharmaceuticals, many new developments and applications are introduced in chiral HPLC separations in recent years. In this paper, the usage of new materials as chiral stationary phases (CSPs) in liquid chromatography for enantiomeric discrimination is investigated in detail. Moreover, hyphenated techniques are very useful for chiral separation of HPLC, especially for separation of new pharmaceuticals.