This paper proposes a non-parametric method for the classification of thin-layer chromatographic (TLC) images from patterns represented in a dissimilarity space. Each pattern corresponds to a mixture of Gaussian approximation of the intensity profile. The methodology comprises various phases, including image processing and analysis steps to extract the chromatographic profiles and a classification phase to discriminate among two groups, one corresponding to normal cases and the other to three pathological classes. We present an extensive study of several dissimilarity-based approaches analysing the influence of the dissimilarity measure and the prototype selection method on the classification performance. The main conclusions of this paper are that, Match and Profile-difference dissimilarity measures present better results, and a new prototype selection methodology achieves a performance similar or even better than conventional methods. Furthermore, we also concluded that simplest classifiers, such as k-NN and linear discriminant classifiers (LDCs), present good performance being the overall classification error less than 10% for the four-class problem.