The influence of heat treatment on the anodic oxide film of TA1 pure titanium substrate was investigated. By comparing the macroscopic photographs and microscopic morphologies of titanium substrate samples before and after anodic oxidation under different heat treatment conditions, the effects of substrate heat treatment on the performance and coloration of the anodic oxide films were determined. The research results show that as the number of heat treatments increases, the substrate grain size increases and the roughness becomes larger. During anodic oxidation, it is easier to generate a continuous oxide film, and the coating thickness is greater. The grain size and roughness of the substrate significantly affect the brightness and uniformity of the anodic oxide coating coloration, and the color distribution follows the substrate morphology, grain size and orientation. Furthermore, substrates with large grain sizes and high roughness formed during heat treatment are prone to developing anatase-phase crystal structures on the surface after high-voltage anodic oxidation.