This paper presents a method of optical fluorescence analysis for the evaluation of homogeneity of multicomponent grain mixtures. This method is based on the evaluation of the content of fluorescent marker. Maize with two degrees of fineness d1 = 1:25 mm and d2 = 2:00 mm was used as a tracer. Maize was covered with Rhodamine B, which emits red light under the influence of ultraviolet radiation. The tracer was introduced into the mixture before the mixing process began. Nine multicomponent grain mixtures were used. The proportion of fluorescent maize was evaluated on the basis of computer image analysis. Additionally, the fraction of the tracer was evaluated using a control method (validation of the accuracy of the proposed method). The results indicate that the degree of the tracer’s fineness influences the results obtained. The use of fluorescent maize with particle size d2 = 2:00 mm allowed to obtain results which differed less from the control method. The average size of the difference in results ranged from 0.20–0.38 for the 2.00 mm tracer and 0.38–1.34 for the 1.25 mm tracer.