In contrast to more polar media, G-quadruplex assembly reveals remarkably high thermodynamic and kinetic stability in toluene solutions. Depending on the substituents installed at the lipophilic guanosine, either a dodecamer or an octamer complex is formed in the presence of K+ or Na+ salts that resist conditions of high dilution and elevated temperatures without exhibiting significant dissociation. Moreover, kinetic exchange between complexed and uncomplexed G is slow enough in NMR to monitor G-quadruplex formation along a day time scale.