The dynamic light scattering method was successfully applied to determine the molar mass of nitrocellulose. The methodology of nitrocellulose fractionation in acetonic solutions is described in detail; six polymer fractions with monomodal distribution were obtained. It was shown that the unfractionated colloxylin with polymodal molar mass distribution had mass average molecular mass values of 87.3 ± 14.1, 28.3 ± 7.3, and 0.54 ± 0.17 kDa when investigated by the dynamic light scattering method. The viscometric method only provided integral viscosity average molar mass equal to 56.7 ± 5.8 kDa.