Spinach landraces cultivated from temprate regions to subtropicals in Iran. However, iron content in the different parts of leaf in Iranian spinach landraces have not been investigated in detail. This study has been conducted to evaluate the effect of different levels of iron (3, 6, 12 and 15 mg. L^-1 Fe form Fe-EDTA) on iron content in blade and petiol, SPAD index, total phenolics, total amino acids and soluble carbohydrates  in leaves of three spinach land races (‘Hamedan’, ‘Shirvan’, ‘Varamin-88’). Spinach plants were grown on cocopeat+ perlite (4:1) and Copper nutrient solution was use. Results showed that the content of Fe in petiol, blade and whole of leaf were significantly affected by concentration of Iron in nutrient solution. ‘Shirvan’ showed higher Fe content in petiol than the Fe content in petiols of ‘Varamin-88’ and ‘Hamedan’. The content of total amino acids, total phenolics, pentoses, hexoses and total soluble carbohydrates in leaf were reduced significantly by the lowered iron treatments. With respect to the contents of Fe in petiol and/or blade and biochemical characteristics, it was found that the quality of spinach was significantly affected by the iron treatments.