Enzalutamide is a potent second‐generation androgen receptor inhibitor that is used to treat metastatic castration‐resistant prostate cancer. It was developed by Medivation and Astellas and was approved by the Food and Drug Administration in 2012 under the brand name Xtandi. Enzalutamide has three major anticancer mechanisms, it inhibits the binding of androgens to the ligand‐binding domain of androgen receptors; inhibits nuclear translocation of androgen receptors; inhibits binding of androgen receptors to deoxyribonucleic acid. It demonstrates reduced expression of androgen receptor‐dependent genes, decreased prostate cancer cell proliferation, and induction of cancer cell death and tumor regression. As a result, patients with metastatic castration‐resistant prostate cancer have a higher chance of survival. Enzalutamide is very important in anticancer therapy, hence, it's necessary to compile all the analytical and bio‐analytical methods to monitor the bioequivalence, bioavailability, and therapeutic monitoring of a drug substance during the course of patient follow‐ups. Thus, this study presents a comprehensive review of the literature on characteristics, properties, and analytical and bio‐analytical methods in various matrices, including formulations, biological fluids, and drug delivery systems.