Untargeted metabolomic profiling of unripe and ripe Solanum nigrum Linn. berries using GC-MS and LC-HRMS/MS

: Solanum nigrum Linn. is a notable health-promoting plant and a rich source of bioactive compounds. Despite their health benefits, pharmaceutically active berries remain underutilised and yet to be explored. As the berries ripen, they undergo natural changes in their metabolic composition that have not been comprehensively reported. In this present study, an untargeted metabolomics approach integrating GC-MS and LC-HRMS/MS was employed to investigate metabolic differences among unripe and ripe berries. A total of 1154 features were initially detected, of which 445 metabolites were retained after curation. Principal Component Analysis (PCA) confirmed a clear separation between unripe and ripe berries. Univariate analysis identified 125 differentially abundant metabolites, including 70 enriched in unripe berries and 55 enriched in ripe berries. Unripe berries accumulated steroidal glycoalkaloids, terpenoids and other defence-related metabolites, indicating active plant defence during early development. In contrast, ripe berries showed increased levels of flavonoids, phenylpropanoids, sugars and amino acids, reflecting enhanced palatability and nutritional quality. Metabolite sets enrichment analysis disclosed that unripe berries were associated with pathways like biotin, pyrimidine, ascorbate, and aldarate metabolism. Whereas ripe berries exhibited enrichment of carbohydrate and amino acid metabolism. While these pathways demonstrated significance based on p- value ( p ≤ 0.05) but, following FDR correction ( q > 0.05) they are interpreted as exploratory trends. Overall, this study demonstrates a metabolic shift and provides comprehensive insights into the metabolic dynamics underlying fruit ripening in Solanum nigrum Linn. berries.