Terpenes and terpenoids are the primary bioactive substances present in essential volatile oils, condensed liquids extracted from diverse plant parts. These substances demonstrate remarkable biological activity and are frequently used as medicines, food additives, and scent molecules. Terpenoids have a wide range of pharmacological effects on the human body, including the treatment, prevent, and reduce the discomfort associated with a number of chronic illnesses. Therefore, these bioactive substances are crucial to our everyday existence. As most terpenoids are present in complex form, coupled with many other raw plant elements, it is important to identify and characterize these molecules. This article addresses various classes of terpenoids, their biochemical processes, and their biological functions. Additionally, it includes a comprehensive description of several hyphenated procedures and recently popular analytical approaches used for isolation, identification, and absolute characterization. It also includes a discussion of the various advantages, drawbacks, and challenges encountered during the collection of the sample and throughout the entire research process.