The paper presents application of a new resin dedicated to copper(II) flow-injection on-line preconcentration prior its flame atomic absorption spectrometric (FAAS) determination. The new sorbent, obtained by suspension polymerization technique, was styrene-divinylbenzene copolymer modified with 5-dodecylsalicylaldoxime-copper(II) complex. In flow mode leaching of initially imprinted Cu(II) ions from polymer beads was effective with 1% (v/v) nitric acid, however for elution of ions retained on the sorbent during the loading process sufficient efficiency was obtained for 0.5% (v/v) nitric acid. The most effective copper(II) sorption was observed within sample pH ca. 6.3 at flow rate 7.5 mL min−1. Furthermore, preconcentration studies of Cu(II) ions realized in the presence of popular foreign ions like Cd(II), Pb(II), Zn(II), Ni(II), Mn(II), Co(II) did not reveal significant interference. The expected effect of Cu(II)-imprinting was confirmed by higher tolerance level for interferents ions concentration for the new sorbent than for the control polymer. It was found that alkaline metals ions and humic acid had the most relevant influence on copper(II) uptake. Accuracy of the evaluated method was assessed for analysis of water samples (tap and mineral water, river water, artesian water) and certified water reference materials compare them to results obtained by inductively plasma mass spectrometry. The satisfactory relative error values obtained with use of standard addition calibration method, confirms the feasibility of this method for Cu(II) determination in water samples. Application of 120 s sorption time enabled to obtain 74-fold enrichment factor and limit of detection (3σ) equal to 0.4 μg L−1.