Disclosed are a crystallization water-free calcium dibutyryladenosine cyclophosphate crystal form, and a preparation method and a use thereof. In an X-ray powder diffraction pattern using Cu-Kα as a source of radiation, the crystallization water-free calcium dibutyryladenosine cyclophosphate crystal form has characteristic peaks at positions where diffraction angles 2θ are equal to 12.3°±0.2°, 17.6°±0.2°, 21.4°±0.2°, 24.7°±0.2°, 25.3°±0.2° and 27.8°±0.2°. The crystallization water-free calcium dibutyryladenosine cyclophosphate crystal form of the present invention has a high purity, and good stability; the preparation method is simple and convenient, has good reproducibility, and is easy to industrially popularize and apply.