Integrated orbitrap-HRMS and chemometric profiling reveals metabolite diversity and antioxidant signatures in cerrado fruits

Abstract

Fruits from the Brazilian Cerrado, such as pitanga (Eugenia uniflora), jambolao (E. jambolana) and murta (E. punicifolia), are recognized for their nutritional and therapeutic potential, largely due to their phenolic-rich profiles. However, systematic studies integrating high-resolution mass spectrometry (HRMS) with chemometric tools to correlate metabolite diversity with antioxidant activity remain scarce. Hydroalcoholic fruit extracts were analysed using Orbitrap-HRMS with data-dependent MS/MS for metabolite annotation. Spectrophotometric and voltammetric assays were performed to assess antioxidant activity, while total phenolic content was quantified. Chemometric analyses, including Fold change analysis, PCA, HCA, PLS-DA and OPLS-DA, were applied to classify samples and identify discriminant metabolites. Visualization strategies such as Van Krevelen diagrams, heatmaps and chord plots were used to explore compositional similarities and metabolite classes. Jambolao extract showed high antioxidant activity (DPPH IC50: 3.10 ± 0.10 µg/mL, ABTS: 189.0 ± 3.2 µg/mL) and total phenolic content (41.60 ± 1.05 mg GAE/100 g), confirming a strong correlation between phenolic content and antioxidant potential. HRMS profiling revealed both shared and species-specific metabolites, including anthocyanins, flavonoids, tannins and phenolic acids. Chemometric models allowed the identification of cyanidin and delphinidin derivatives as key discriminant markers. Fold change analysis revealed distinct metabolic signatures among the species, with jambolao enriched in low-mass aglycone polyphenols, while pitanga and murta were characterized by higher-mass glycosylated polyphenols. Multivariate clustering grouped jambolao and pitanga together due to their similar metabolic and antioxidant profiles, whereas murta formed a distinct cluster. The findings highlight jambolao and pitanga as rich sources of bioactive polyphenols, supporting their potential for nutraceutical and functional food applications.