Oil crops have become increasingly farmed worldwide because of their numerous functions in foods and health. In particular, oil derived from the seeds of evening primrose (Oenothera biennis) (EPO) comprises essential fatty acids of the omega-6 (ω-6) series. It is well recognized to promote immune cells with a healthy balance and management of female ailments. The nutrients of interest in this oil are linoleic acid (LA, 70-74%) and γ-linolenic acid (GLA, 8-10%), which are polyunsaturated fatty acids (PUFA) that account for EPO's popularity as a dietary supplement. Various other chemicals in EPO function together to supply the body with PUFA, elevate normal ω-6 essential fatty acid levels, and support general health and well-being. The inclusive EPO biochemical analysis further succeeded in identifying several other components, i.e., triterpenes, phenolic acids, tocopherols, and phytosterols of potential health benefits. This comprehensive review capitalizes on EPO, the superior product of O. biennis, highlighting the interrelationship between various methods of cultivation, extraction, holistic chemical composition, sensory characters, and medicinal value. Besides the literature review, this study restates the numerous health advantages of primrose oil and possible drug-EPO interactions since a wide spectrum of drugs are administered concomitantly with EPO. Modern techniques to evaluate EPO chemical composition are addressed with emphasis on the missing gaps and future perspectives to ensure best oil quality and nutraceutical benefits.