• Interactions between Ru with TX114 and Q with T80 are strong. • Interactions is mostly based on hydrogen bonds and hydrophobic forces. • For Ru with TX114 there are the highest partition coefficient. The behaviour of quercetin (Q) and rutin (Ru) in the aqueous solution of two nonionic surfactants, Triton X-114 (TX114) and Tween 80 (T80) was investigated using the spectroscopic techniques. The object of investigations were the interactions between the given flavonoid and the surfactant in a wide range of surfactant concentrations and the temperature range from 293 to 313 K. The obtained UV–Visible and emission spectra were the basis for determination of the binding constant, partition coefficient, Gibbs free energy of binding as well as partition, enthalpy and entropy of binding and partition. It was proved that the interactions between Ru with TX114 and Q with T80 are strong and become weaker with the increasing temperature. The nature of interactions between the molecules is mostly based on the hydrogen bonds and hydrophobic forces.