We present results of a study of chromatographic separation based on surface area exclusion induced by the irreversible adsorption of the diblock copolymer poly(vinylpyridine)-polystyrene on silica from toluene solutions using a radiolabeled polymer. This characteristic of the copolymer enabled the number of adsorbed copolymers to be determined as a function of the plate number inside the chromatographic column. We find that the initial step of the surface coverage may be simulated by the random sequential adsorption of a mixture of unimers (isolated copolymer) and micelles (structured form). The influences of the relative length of the two PV2P blocks and the concentration of the copolymer solution on the chromatogram were investigated by simulation and experimentally. The chromatograms showed that the surface area availability conditioned the copolymer distribution in the successive plates of the column, and a direct correlation between simulation and experiment could be found for a copolymer of size asymmetry ratio equal to 1. Deviation from this simple behavior was observed in other copolymers. Notably, the influence of the interfacial reconformation was investigated, and we found that the rate at which the process developed depended on the structure (unimer or micelle) of the adsorbed copolymer. Unimer and micelle adsorption was attributed to the existence of strong interactions between the pyridine group of the polymer and the silanol group of the silica.