Cloud point extraction (CPE) was carried out to extract Eosin B dye from aqueous solution using non-ionic surfactant Triton x-114. The enrichment of dye was determined by a spectrophotometric study. The effects of different operating parameters, e.g., concentrations of surfactant, salt, temperature, pH, the heating time, equilibration temperature and time of centrifuge have been studied. The extracted surfactant-rich phase was diluted with ethanol and its absorbance was measured at 533 nm. The calibration curve was linear in the range of 0.0067-0.7300 mgL -1 with r 2 =0.995. The detection limit, preconcentration and enrichment factor were obtained 1.08 ng mL -1 , 15.00 and 11.20, respectively. The effect of some foreign species including cations, anions and similar dye, fluorescein was also investigated. This method was applied successfully for determination of the amount of Eosin B in water samples.