It is difficult to measure the particle size distribution for multi-modal particle system in dynamic light scattering technique, because of the ill posed problem in the data inversion. In this paper, the smooth constraint penalty term is added to the objective function of Tikhonov regularization, to improve the performance of inversion by enhancing the constraint to the solution. Three groups of simulated bimodal particle system (190/443 nm, 282/953 nm, 457/553 nm), one group of simulated unimodal particle system (564 nm), and one group of simulated trimodal particle system (292/591/889 nm), as well as two groups of experimental particle system (306/974 nm, 300/502 nm), were tested in this paper. The results show that, in the regularization inversion, adding the smooth constraint penalty term, can effectively eliminate the spike and spurious peaks in reversed particle size distribution, improve the resolution of particle size distribution and increase the ability of noise reduction, which, sequentially can make better use of the advantage that Multi-angle Dynamic Light Scattering (MDLS) technique could be able to provide more ultrafine particle size distribution information in medium and large size range, and then realize the accurate bimodal and multi-modal particle measurement from nanometer to submicron size range.