AbstractThe attention to chiral drugs has been raised to an unprecedented level as drug discovery and development strategies grow rapidly. However, separation of enantiomers is still a huge task, which leads to an increasing significance to equip a wider range of expertise in chiral separation science to meet the current and future challenges. In the last few decades, remarkable progress of chiral resolution has been achieved. This review summarizes and classifies chiral resolution methods in analytical scale and preparative scale systematically and comprehensively, including crystallization‐based method, inclusion complexation, chromatographic separation, capillary electrophoresis, kinetic resolution, liquid–liquid extraction, membrane‐based separation, and especially one bold new progress based on chiral‐induced spin selectivity theory. The advances and recent applications will be presented in detail, in which the contents may bring more thinking to wide‐ranging readers in various professional fields, from analytical chemistry, pharmaceutical chemistry, natural medicinal chemistry, to manufacturing of drug production.