Fe(III) ion doped Ni L-Histidine Hydrochloride monohydrate crystals (Fe(III)-NiLHICL) are grown at room temperature using slow evaporation technique. The Fe(III) doped NiLHICL crystals are characterized by spectroscopic techniques such as X-Ray diffraction studies, Electron Paramagnetic Resonance(EPR), Optical Absorption and FTIR studies. Thermal stabilities were studied by TGA/DTA analysis. The powder diffraction patterns of prepared crystals have been recorded and lattice cell parameters are evaluated as a = 1.5286, b = 0.8933, c = 0.6852 nm. From EPR studies, g and hyperfine splitting parameters for Fe(III) ion in the host crystals are determined as g = 2.0301 indicating octahedral symmetry. Crystal field and inter-electronic parameters are evaluated from the optical absorption studies in addition to obtaining the confirmation for octahedral symmetry for the ions in the host lattice. The FT-IR spectrum exhibited characteristic vibrations of the groups present in the crystal indicating bond formation between the metal ion and the amino acid.