The invention discloses a preparation method and application of a Bi2Ti4O11:Yb, Er up-conversion luminescence anti-counterfeiting film. HAC, DMF and TBOT solutions are uniformly mixed, bismuth nitrate, ytterbium nitrate and erbium nitrate are sequentially added, after the bismuth nitrate, the ytterbium nitrate and the erbium nitrate are fully dissolved, a mixture is evaporated to dryness to obtain gray black powder, the powder is annealed, TPU, THF and DMF are dissolved to obtain a uniform transparent solution with viscosity, and Bi2Ti4O11:Yb, Er powder is added under a stirring condition and is fully dissolved to obtain an electrostatic spinning precursor solution. The precursor solution is transferred into an electrostatic field, an electrostatic spinning process is performed, and a spinning product is obtained on a collecting plate. The collecting plate is put into a drying oven to be dried. Finally, the film is white and transparent under a fluorescent lamp, and no light emitting phenomenon exists. However, under the excitation of a 980 nm laser, the film shows excellent up-conversion luminescence performance. By adjusting the doping proportion of the ytterbium nitrate and the erbium nitrate, the anti-counterfeiting film with red, yellow and green up-conversion luminescence properties can be obtained.