The Last 10 Years of Research in the Enantioseparation of Pharmacologically Relevant Phytocannabinoids: An Updated Review

ABSTRACT As research on Cannabis sativa L. and its derivatives expands, an increasing number of novel phytocannabinoids are being identified. Many of these compounds exhibit molecular chirality due to the presence of one or more stereogenic centers in their structures. Given the pharmacological and toxicological relevance of chiral molecules, regulatory authorities are expected to demand more stringent analytical characterization of the enantiomeric composition of phytocannabinoids in C. sativa L. extracts. The growing availability and enhanced performance of advanced analytical technologies, particularly chiral chromatographic systems, have renewed interest in the stereochemical profiling of these natural products. Chiral separation techniques have become central to research efforts aimed at determining enantiomeric purity and understanding stereoisomeric distributions in complex plant matrices. Among the limited studies addressing chiral chromatographic methods for separating phytocannabinoid enantiomers and diastereomers, two techniques are predominant: high‐performance liquid chromatography (HPLC) and supercritical fluid chromatography (SFC). Method development in both approaches typically relies on two major classes of chiral stationary phases (CSPs). The first comprises Pirkle‐type CSPs, which utilize the “Inverted Chirality Columns Approach” (ICCA) and are based on rational design principles for enantioselective interactions. The second group includes polysaccharide‐based CSPs employing cellulose or amylose derivatives, recognized for their broad applicability in resolving diverse phytocannabinoid enantiomers. This work will review a selection of recent, representative studies, highlighting key analytical strategies for chiral separation, methodological challenges, and implications for both scientific research and regulatory frameworks.