High-resolution melting analysis of chloroplast markers for species authentication and mislabeling detection in commercial açaí and juçara products

Abstract Authentication of açaí products is increasingly important due to the risk of species substitution among morphologically similar Euterpe taxa, with implications for food quality, labeling accuracy, and consumer trust. Despite advances in molecular methods, rapid and cost-effective tools for species authentication of closely related Euterpe taxa in processed commercial matrices remain limited. This study evaluated High-Resolution Melting (HRM) analysis targeting two complementary chloroplast markers—psbK-I and ycf1b—as a practical approach for species-level authentication of açaí ( Euterpe oleracea and E. precatoria ) and juçara ( E. edulis ) products. In silico analysis predicted amplification of the ycf1b region to be confined to the Arecaceae family, with no amplification of non-palm species commonly present in commercial formulations; species discrimination in both markers relies on differential melting profiles rather than primer exclusivity. Applied sequentially, the two markers enabled reliable differentiation of all target species, including closely related taxa, with a detection limit of at least 10% in binary DNA admixtures. When applied to 50 commercial products, HRM successfully authenticated 46 samples, providing a higher authentication success rate for this particular set of processed samples than DNA sequencing, which was limited by amplification failure and mixed chromatograms. Mislabeling was detected in one açaí sorbet and three frozen açaí pulps marketed as açaí but molecularly identified as E. edulis . These findings indicate that HRM analysis represents a promising approach for species authentication in processed plant-based matrices, with potential future integration into food quality control workflows pending broader validation.