Holistic Approach to Investigate Pyrrolizidine Alkaloids in Honeys from Diverse Botanical Origin: From Target to Suspect and Nontarget Screening Analysis

This study investigates the presence of pyrrolizidine alkaloids (PAs) in honey from diverse botanical origins using a combination of target analysis, through ultrahigh-performance liquid chromatography coupled with tandem mass spectrometry and suspect and nontarget screening via ultrahigh-performance liquid chromatography coupled with quadrupole-Orbitrap high-resolution mass spectrometry. A total of 80 honey samples were analyzed, with 61% were found to contain at least one PA. Eucalyptus and wildflower honeys showed the highest contamination levels, reaching 27 μg/kg when expressed as the sum of regulated PAs. Indicine and lycopsamine, commonly associated with <i>Echium</i> plant species, were the most frequently detected compounds. Statistical analysis revealed variability in PA profiles across different honey types, though classification based on botanical origin remained limited. Additional strategies, including molecular networking and the computationally generated spectral library, expanded the detection capabilities of these compounds. In particular, molecular networking can be a powerful tool to reveal structural relationships among PAs.