Integrated suspect screening and non-targeted metabolomic profiling of extractable compounds in Southern Italian and Spanish beans by UHPLC-Q-Orbitrap-HRMS combined with chemometric analysis

Common beans are chemically complex food matrices whose composition may reflect both endogenous metabolic features and exposure to agricultural or environmental residues. This study applied an integrated ultra-high-performance liquid chromatography coupled with quadrupole-Orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap-HRMS) workflow combining a QuEChERS-based extraction, suspect screening, non- targeted metabolomic profiling, and chemometric analysis to investigate extractable compounds in bean samples from Southern Italy and Spain (n = 44). Overall, 53 tentatively identified compounds, mainly associated with pesticides and pharmaceuticals, were detected. Among these, 11 compounds, including boscalid, penconazole, and azoxystrobin, were confirmed and their concentrations were semi-quantitatively estimated. Non-targeted profiling further provided molecular information potentially related to sample variability. Chemometric modeling indicated sample-group-related trends, with the orthogonal partial least squares discriminant analysis (OPLS-DA) model showing strong performance (R²Y > 0.8; Q² > 0.7) and a 91.7% correct classification in external validation. These findings highlight the value of integrated HRMS-based workflows for residue profiling and for exploring chemical variability across legume samples from different geographical contexts.