Enantioselective analytical approaches are essential for monitoring pharmacokinetics and acquiring accurate data to better understand the role of stereochemistry in pharmacokinetics. Enantioselectivity significantly impacts the pharmacokinetics of chiral drugs, especially in metabolic profile, leading to toxicity of enantiomer. Consequently, there is a need to study the pharmacokinetics of enantiomerically pure drugs and racemates as they differ in affinity with enzymes and proteins. Combining the best enantioseparation conditions with the specified biological matrix and the intended purpose of the analysis is a challenging task. This review discusses the importance of chirality in stereoselective pharmacokinetics with more relevant examples, various enantioselective analytical techniques, and stationary phases employed. Challenges such as lack of universal chiral columns, biological inversion of the isomers, and others have been discussed. Further presented the recent advances in the screening of chiral drugs and innovative improvements in the analytical approaches for chiral molecule analysis such as supercritical fluid chromatography, simulated moving bed chromatography, and other techniques are discussed.

Chirality resides in diverse fields such as pharmaceuticals, food, bio‐pharmaceuticals, and cosmetics. Drugs used in treating various disease conditions, the majority of the plant secondary metabolites, various chemicals used in cosmetics, and distinct flavoring agents used in food industries exhibit chirality. In this context, there is an obligation to extend the analysis to chiral aspects. Attempts have been made to summarize the systematic, successful approaches for enantiomeric production via both enantio‐selective synthesis and the resolution of enantiomers using various analytical techniques including crystallization, chiral chromatography, capillary electrophoresis, kinetic resolution mediated by enzymes. Efforts have also been made to describe the regulatory guidance regarding bridging studies in the milieu of chiral switching. The majority of the monoclonal antibodies, growth factors, blood components, deoxyribonucleic acids, enzymes, and even vaccines for various diseases like cancer, polio, malaria, tetanus, hepatitis B, cholera, diphtheria bind differently to the receptors because the majority of drug binding and antibody binding receptors are also exhibiting chirality. Recent advancements in the various analytical techniques involved in the chiral analysis of food to preserve the authenticity of distinct flavors and cosmetics to identify safer enantiomers and reduce side effects both to humans and the environment are described.