This study aimed to investigate the influence of sequential extraction with solvents of increasing polarity on the antifungal and tyrosinase inhibitory activities of root extracts from three Dalea species (D. elegans Gillies ex Hook., D. boliviana Britton, and D. leporina (Aiton) Bullock), Fabaceae, considering the chemical profiles of the obtained extracts through UPLC-MS/MS analysis, with the goal of identifying specific compounds responsible for the observed activities.The roots were extracted, in sequential form, with solvents of increasing polarity, and the extracts obtained were evaluated for their antifungal activity against Candida albicans and their ability to inhibit tyrosinase. Chloroform extract of D. boliviana demonstrated the most potent antifungal activity with a minimum inhibitory concentration value of 250 μg/ml. All ethyl acetate extracts exhibited superior tyrosinase inhibition across all species, with the D. elegans extract being the most active (IC50 1.44 ± 0.012 µg/ml). UPLC-MS/MS analysis of these active extracts revealed the presence of prenylated flavanones, with varying relative abundances. In ethyl acetate extracts, flavanone (2S)-5,7,2′,4′-tetrahydroxy-5′-(1′′′,1′′′-dimethylallyl)-8-prenylflavanone was identified, suggesting its potential contribution to tyrosinase inhibition, particularly in D. elegans. While in the chloroform extracts, (2S)-5,7,2′-trihydroxy-5′-(1′′′,1′′′-dimethylallyl)-8-prenylflavanone was predominant in D. boliviana, correlating with its enhanced antifungal activity. Also, the presence of prenylated flavanones in all the evaluated Dalea species confirms them as marker compounds for this genus. This study highlights the chemical diversity of Dalea species and the potential of their extracts as sources of antifungal and tyrosinase inhibitory compounds. The identified flavanones represent promising leads for the development of novel therapeutic agents against fungal infections and hyperpigmentation disorders.