Principles for Stereoselective Separation of Chiral Drug Compounds Enantiomers and Diastereomers in Pharmaceuticals and Biopharmaceuticals Using Liquid Chromatography

ABSTRACT In the fields of pharmaceuticals and biopharmaceuticals, chiral liquid chromatography techniques including high‐performance liquid chromatography and ultra‐performance liquid chromatography are frequently used to isolate, identify, separate, and quantify chiral isomers, including enantiomers and diastereomers (stereoisomers), due to the significant differences in biological activity and therapeutic effects of stereoisomers. The authors have provided a comprehensive overview of the fundamental principles necessary for using liquid chromatography to separate and accurately estimate chiral compounds that exhibit stereoisomerism (both enantiomers and diastereomers). The development strategies outlined include the selection of chromatographic conditions, optimization of sample preparation, evaluation of degradation pathways, establishment of system suitability criteria, and execution of method validation studies. Additionally, this article supports the development of robust and stability‐indicating methods by applying one factor at a time and design of experiments concepts for chiral drugs and their chiral impurities in pharmaceuticals and biopharmaceuticals. The method validation attributes essential to evaluate the characteristics of the developed method were discussed in this write‐up. The validation parameters include specificity, linearity, detection limit, quantitation limit, accuracy, precision, solution stability.