The invention relates to the field of medicinal chemistry, in particular to a crystal form preparation process and application of isotretinoin. A crystal form A takes positions 2theta of diffraction peaks as spectrogram feature parameters, wherein the positions 2theta sequentially comprise 5.8+/-0.2, 10.52+/-0.2, 11.46+/-0.2, 11.87+/-0.2, 12.56+/-0.2, 15.83+/-0.2, 16.49+/-0.2, 18.11+/-0.2, 19.70+/-0.2, 20.24+/-0.2, 21.68+/-0.2, 24.83+/-0.2, 25.40+/-0.2, 26.94+/-0.2, 28.62+/-0.2, and 30.87+/-0.2. A specific preparation method comprises the steps that an isotretinoin raw material is dissolved with organic alcohol to obtain an isotretinoin organic alcohol solution, crystallization is conducted with a temperature differential method, solid-liquid separation is conducted, a solid is dried and then the crystal form A is obtained; the crystal form A is put under light in air and the stability of the crystal form A is obviously better than that of a raw material medicine.