In the present study, the nutritive composition and mineral content of the leaves of eight species of Acacia trees were evaluated to assess their potential for different purposes, such as food resources for ruminants and mineral sources in soils. Caco-2 cells were employed to assess cytotoxicity, revealing that the extracts exhibited no cytotoxic effects after cellular incubation, suggesting their suitability as an alternative animal feed. The leaves proved to be a promising source of protein and fiber, offering an alternative to meet the needs of ruminants. The protein content differed among species, ranging from 18.96% in A. cyclops to 14.04% in A. melanoxylon. Similarly, fiber content varied from 35.52% in A. melanoxylon to 16.43% in A. cyclops. The species displayed moderate to high levels of minerals, particularly concentrations of Ca, P, and K. These values varied among species, ranging from 8452.90 to 28,465.31 mg/kg DM for Ca, 309.61 to 1293.82 mg/kg DM for P, and 5557.24 to 11,475.85 mg/kg DM for K, respectively. Each species displayed a distinct profile regarding the analyzed parameters, suggesting varied potential for their respective use. Additionally, vibrational spectroscopy analysis proved to be a highly valuable and dependable method for distinguishing between species.