The invention relates to the technical field of thin film materials, in particular to a preparation method of periodic multi-directional thickness gradient thin films. The method comprises the following steps that a mask plate is fixed above a substrate in a suspended mode, the mask plate is putted into a vacuum cavity of a sputtering instrument, a target material is fixed on a cathode, the substrate is putted on an anode opposite to a target surface, and after vacuumizing is carried out, inert gas is introduced, and the periodic multi-directional thickness gradient thin films are prepared through a sputtering method, wherein the distance between the mask plate and the substrate is h, the mask plate is provided with meshes, the mesh size is recorded as w, the rib width is recorded as d, h,w and d are adjustable, and h is greater than 0. According to the method, the multi-directional thickness gradient thin films can be prepared at a time and are periodically and densely arranged, theperiodic multi-directional thickness gradient thin films have the characteristic of high flux, the large-scale integration research and application are facilitated, and the application prospects in the field of flexible electronic materials and devices is wide.