The invention provides a tildipirosin 1,4-dioxane solvate and a preparation method thereof. Characteristic peaks occur in an X-ray powder diffraction pattern represented by 2theta when 2theta is equal to 6.06+/-0.2, 8.42+/-0.2, 9.34+/-0.2, 9.94+/-0.2, 10.28+/-0.2, 12.34+/-0.2, 13.44+/-0.2, 13.74+/-0.2, 14.66+/-0.2, 15.58+/-0.2, 15.78+/-0.2, 16.40+/-0.2, 17.00+/-0.2, 17.90+/-0.2, 18.60+/-0.2, 19.32+/-0.2, 20.10+/-0.2, 21.40+/-0.2, 22.34+/-0.2, 22.68+/-0.2, 23.34+/-0.2 and 24.18+/-0.2. The preparation method employs a constant-temperature suspended crystal transformation process and is simple in process and low in energy consumption. The tildipirosin 1,4-dioxane solvate is in a rod shape and crystal is of a regular shape.