Ultra-trace detection of Copper ions by complexation with Neocuproin as an organic chelating agentusing dispersive liquid-liquid microextraction combined with UV/VisSpectrophotometerywas investigated as a quite simple and rapid method in water samples.Some factors influencing on the extraction efficiency of copper were studied and optimized, such as the extraction and dispersive solvent type and volume, pH, chelating concentration, extraction time and salting effect. The proposed method showed a linear calibration curve in the concentration range of 20- 200 ppb(R2 = 0.9996) with a limit of detection of 0.1ppb (3Sb/m) following a very niceenrichment factor of >136 for the analyte.The relative standard deviation for seven replicate determination was ≤3.5%. The proposed protocol has been applied for determination of copper in standard and water samples with satisfactory results.