Unmasking the Aliphatic Repertoire: New Polyunsaturated Metabolites in <i>Bupleurum falcatum sensu lato</i> Provide Chemotaxonomic Insights

<i>Bupleurum falcatum</i> (Apiaceae) <i>sensu lato</i> includes multiple infraspecific taxa with longstanding taxonomic ambiguities, often resulting from incomplete morphological and chemical characterizations. Herein, diethyl ether extracts were analyzed from four Balkan populations that were tentatively identified as <i>B. falcatum</i> subsp. <i>falcatum</i> (<i>syn</i>. <i>B. falcatum</i>) and <i>B. falcatum</i> subsp. <i>cernuum</i> (<i>syn</i>. <i>B. sibthorpianum</i>). Comprehensive chromatography and spectroscopic techniques (GC-MS and 1D/2D NMR) enabled the isolation of several newly identified aliphatic polyunsaturated esters, including compounds bearing uncommon conjugated tetraene and triyne backbones. These novel structures differ from canonical falcarinol derivatives by lacking the usual 3-hydroxylation, suggesting a divergent branch in the crepenynate pathway. The chemical profiles of each sample correlated closely with leaf morphology and infraspecific designations: for example, the Galičica Mt. population of <i>B. falcatum</i> featured a unique newly detected heptadecadientriyne, while the populations from Šar Planina and Suva Planina displayed distinct polyunsaturated repertoires. Extracts from <i>B. sibthorpianum</i> likewise contained stereoisomeric compounds that highlight metabolic divergence. Collectively, these findings demonstrate significant chemotypic variation within the <i>Bupleurum falcatum</i> complex and provide the first account of less-polar secondary metabolites, including newly discovered polyunsaturated metabolites. Future research integrating molecular markers and bioactivity assays may elucidate how these specialized metabolites contribute to both the taxonomy and pharmacological potential of these understudied taxa.