A preparation method of a touch screen tempered glass film belongs to the technical field of tempered glass films of electronic products. The preparation method comprises the steps that a glass substrate and sticking glue are prepared, the lower surface of the glass substrate is stuck with the sticking glue to obtain the touch screen tempered glass film, and the preparation method further comprises the following steps that A, the prepared glass substrate is placed into a reactor filled with SOCl2 liquid after cleaned and dried, the reactor is stood for 3-5 h, the glass substrate is taken out and placed into a resistance furnace, the temperature of the glass substrate is raised to 450 DEG C to 550 DEG C with the furnace temperature, the temperature is kept for 1-3 h, and a pretreatment layer is formed on the surface of the glass substrate; B, the glass substrate treated by the step A is soaked in nitrate with the temperature of 330 DEG C to 550 DEG C, insulation treatment is performed for 2-10 h, and an ion exchange layer is obtained; C, the glass substrate treated by the step B and the sticking glue are combined so that the touch screen tempered glass film is obtained. The invention further discloses the touch screen tempered glass film prepared with the method, and the antibacterial effect of an antibacterial layer gathered on the surface of the tempered glass is better.