The electrochemiluminescence (ECL) of doped silica nanoparticles (DSNPs), prepared by a reverse microemulsion method that leads to covalent incorporation of the Ru(bpy)(3)(2+), was investigated in acetonitrile and aqueous buffers. The emission was produced for the first time by cation-anion direct annihilation, and the position of ECL maxima indirectly allowed estimation of the E(1/2,IOx) and E(1/2,IRed) potentials for Ru(bpy)(3) inside DSNPs. The weak ECL emission is most likely generated by an intrananoparticle ruthenium unit annihilation rather than by the electron transfer between a reduced and oxidized DSNP due to the very low diffusivities of the nanoparticles. Thiol-terminated DSNPs were self-assembled on gold substrates, forming compact and stable monolayers which mimic probe-target assays with DSNPs as labels. The ECL intensity obtained by such functionalized substrates in aqueous media, using tripropylamine (TPrA) as coreactant, was surprisingly increased with respect to direct electrochemical oxidation because of the ability of oxidized TPrA to diffuse within the DSNPs structure and reach a higher number of emitting units with respect to direct electron tunneling. The experimental results have been explained by proposing a basic physical-chemical model which supports evaluation of the number of redox-active centers per nanoparticle. In the model the contrasting effects of increased luminescence quantum yield and decreased diffusion coefficient with respect to free (i.e., not bound within the silica structure) emitting molecules were taken into account. This allows, in principle, optimizing the ECL emission intensity as a function of DSNP size, doping material, charge, doping level, supporting electrolyte, electrode material, and solvent. Finally, it is worth noting that this study has provided a more than 1000-fold increase of the ECL signal of a chemically and electrochemically stable DSNP compared to that of a single dye, suggesting that use of this kind of nanostructures as luminescent labels represents a very promising system for ultrasensitive bioanalysis.