While numerous Fabaceae seeds are a good nutritional source of high-quality protein, the use of some species is hampered by toxic effects caused by exposure to metabolites that accumulate in the seeds. One such species is the faba or broad bean (Vicia faba L.), which accumulates vicine and convicine. These two glycoalkaloids cause favism, the breakdown of red blood cells in persons with a glucose-6-phosphate dehydrogenase deficiency. Because this is the most common enzyme deficiency worldwide, faba bean breeding efforts have focused on developing cultivars with low levels of these alkaloids. Consequently, quantification methods have been developed; however, they quantify vicine and convicine only and not the derivatives of these compounds that potentially generate the same bio-active molecules. Based on the recognition of previously unknown (con)vicine-containing compounds, we screened the fragmentation spectra of LC-MS/MS data from five faba bean cultivars using the characteristic fragments generated by (con)vicine. This resulted in the recognition of more than a hundred derivatives, of which 89 were tentatively identified. (Con)vicine was mainly derivatized through the addition of sugars, hydroxycinnamic acids, and dicarboxylic acids, with a group of compounds composed of two (con)vicine residues linked by dicarboxyl fatty acids. In general, the abundance profiles of the different derivatives in the five cultivars mimicked that of vicine and convicine, but some showed a derivative-specific profile. The description of the (con)vicine diversity will impact the interpretation of future studies on the biosynthesis of (con)vicine, and the content in potentially bio-active alkaloids in faba beans may be higher than that represented by the quantification of vicine and convicine alone.

Callus cultures derived from three species of Fabaceae were compared under identical hormonal conditions using untargeted UHPLC–MS/MS metabolomics together with phenotypic and antioxidant capacity analyses. Principal component analysis with 290 metabolites revealed species identity as the dominant determinant of the metabolic profile, indicating that species-associated metabolic signatures persist in dedifferentiated tissues. Soybean calli accumulated saponins, primarily soyasapogenol derivatives. Lupin calli were characterized by diverse isoflavones detected as aglycones, glycosylated and malonylated conjugates. Calli from the pea cultivar Karacter were dominated by hydroxycinnamate derivatives: coumaroyl methylhexose, feruloyl-coumaroyl glycoside derivatives, and caffeoyl amino acid conjugates. Within each species, the calli metabolomes were furthermore influenced by genotype, explant origin, and independent callus line establishment. Antioxidant capacity correlated with metabolite subclass composition rather than total metabolite abundance, with polyphenol-rich profiles displaying higher reducing potential than saponin-dominated metabolomes. Together, these findings provide the first systematic comparative evidence that species-specific metabolic signatures are maintained in Fabaceae callus cultures, while remaining quantitatively modulated by genotype, explant origin, and somaclonal variation. Soybean, lupin, and pea callus cultures thus provide tractable model systems for the species-specific study of respectively triterpenoid saponin, isoflavonoids, and hydroxycinnamate metabolism, offering a foundation for future biotechnological development.