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.