In this research, the toxicity of nano-[Cu-(DIP)-EA], a metal nano-complex consisting of ellagic acid and bathophenanthroline ligands, on human serum albumin (HSA) at a protein level was investigated. Molecular docking simulations and spectral analyses were conducted in a simulated physiological environment at pH 7.4 to explore the interaction of nano-[Cu-(DIP)-EA] with HSA. The results represented an increase in albumin absorption upon exposure to nano-[Cu-(DIP)-EA], demonstrating significant interaction between the two compounds. Steady-state and time-resolved fluorescence measurements pointed out that nano-[Cu-(DIP)-EA] induced static quenching of the albumin's intrinsic fluorescence with a high binding affinity of approximately 10 mol/L in a 1:1 interaction ratio. The thermodynamic variables clarified that binding of nano-[Cu-(DIP)-EA] to albumin occurs spontaneously and primarily driven by van der Waals interactions and H-bonds. The results of the computer simulations and the binding displacement experiments utilizing the site markers warfarin and ibuprofen revealed that nano-[Cu-(DIP)-EA] binds to site I within the subdomain IIA of albumin. Circular dichroism analysis elaborated that nano-[Cu-(DIP)-EA] slightly perturbed the microenvironment around of tryptophan residues and diminished the α-helix structure stability to a negligible amount.