Multistep electrochemical anodization is an electrochemical experiment orderly conducting in different electrolytes. In this paper, TiO2 nanotube arrays have been firstly anodic grown in aqueous electrolyte (H3PO4/HF) and later anodic grown in organic electrolyte (glycerol/NH4F). Compared with separately anodizing in aqueous and organic electrolyte, the morphology of the resulting nanotube arrays can be optimized. SEM images showed that the obtained nanotubes have a length of more than 2 μm and single-pore diameter ranging from 120 to 150 nm. The current work indicates that the multistep electrochemical anodization has a contribution to the optimization of the morphology of TiO2 nanotube arrays.