This paper presents the synthesis and physico-chemical characterization of iron(II) gluconate as a potential antianemic drug for oral administration. Iron(II) gluconate was synthesized from sodium gluconate and iron(II) sulphate as precursors. The reaction product was purified by treating the crude reaction mixture with the acidic and basic ion exchange resins for the removal of Na+ i SO42- ions. Iron(II) gluconate with the purity greater than 90% was obtained after evaporation in a vacuum evaporator, the treatment with aprotic solvent and drying in a desiccator over a dehydration agent. Molar ratio of 1:2 of iron and gluconate remains was determined by applying the Job's method. Structural properties of the obtained iron(II) gluconate were determined by using UV-VIS spectrophotometry, infrared spectroscopy (FT-IR), proton nuclear magnetic resonance (1H-NMR) and X-ray diffraction (XRD) methods. Thermal degradation of iron(II) gluconate was monitored by differential scanning calorimetry (DSC) and the results showed that synthetized and purified Iron(II)-gluconate was most probably in the form of dihydrate. The qualitative and quantitative analysis of iron(II) gluconate was performed by high performance liquid chromatography (HPLC).