A micelle mediated separation and preconcentration method for flame atomic absorption spectrophotometric (FAAS) determination of Cu was developed. The method is based on complex formation between Cu and 1-(2-pyridylazo)-2-naphthol and cloud point extraction with Tergitol NP-7 surfactant prior to FAAS determination. The parameters affecting quantitative extraction of copper, including pH, surfactant and ligand concentrations, incubation time and temperature, were examined and optimized. Under optimum conditions, interference effects of other ions were studied, and tolerance ratios of interference ions were given. Under the optimal conditions, detection and quantitation limits were found as 3 and 8 μg/L, respectively. Linear dynamic range for Cu²⁺ was determined to be 0.13–1.5 μg/mL. Relative standard deviations were below 6% throughout the experiments. Enhancement factor was determined as 25. Applicability of the method was proved by analysis of standard reference materials. The developed method was applied to determine trace copper contents in different mushroom species.