The interaction of 38 nonionic surfactants containing various hydrophobic moieties with dimethyl-β-cyclodextrin (DIMEB) was studied by charge transfer chromatography. DIMED modified the retention behaviour of each surfactant, however, the strength of interaction varied considerably. Both the lipophilicity and specific hydrophobic surface area of surfactants significantly influenced the strength of interaction. The presence of ester bond and double bond in the hydrophobic moiety of surfactants has impact on the complex formation.