A micelle-mediated procedure was presented for enrichment of Cu2+ using 4-(2-chloroacetamido)-salicylic acid and Triton X-114 as complexing and extracting agents, respectively. The experimental parameters that affect the extraction were investigated and optimized. Good analytical features were achieved including limit of detection (0.25 µg L-1), limit of quantification (0.83 µg L-1), linearity (1.0-500 µg L-1) and reproducibility (2.2%). For 50.0 mL sample, the enrichment factor was 49.5. Most concomitant ions not interfere with Cu2+ determination. The procedure was employed for preconcentration and determination of Cu2+ in different samples (water, blood and urine) by flame atomic absorption spectrometry with satisfying results.