A two-step calcination synthesis, considering the potential for mass production, of lithium titanium oxide powder was carried out to fabricate a single Li4Ti5O12 phase, which is useful for anode electrode material of Li-based rechargeable battery as well as an electrode for supercapacitor. The final composition is controlled by adding more TiO2 powder into powder gained at one calcination process during the two calcination process. We investigated the influence of excess TiO2 on the structural characteristics of lithium titanium oxide synthesized by the two-step calcination method. X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FT-IR) measurements showed that the as-synthesized powder had a spinel crystal structure as well as A composition of 4 : 5 : 12. In addition, a high resolution transmission electron microscopy (HRTEM) analysis revealed that the fabricated powder exhibited a single crystalline phase formation. These results indicated that the powder synthesized in the one-step calcination process showed coexistence crystalline phases, which are the Li4Ti5O12 and Li2.39Ti3.4O8 phase. However, in the two-step calcination process, the powder synthesized showed the single crystalline Li4Ti5O12 phase. A very uniform grain size of the as-synthesized powder was shown in a field emission scanning electron microscopy (FESEM). These results suggested that the two-step calcination process can be used for synthesis of single crystalline Li4Ti5O12 powder with uniform grain shape and provide motivation to pursue mass production of lithium titanium based oxide powder for bulk type batteries.