A second-derivative spectrophotometric method based on zero-crossing over technique is developed in simultaneous determination of copper(II) and nickel(II) ions. Methylthymol blue (MTB) as a chromogenic reagent and cetyltrimethylammonium bromide as a surfactant were used, and measurements were carried out in buffered solution at pH 6 and at a temperature of 25°C. The amplitude of derivative spectra was measured at wavelengths of 631.9 and 587.7 nm for the simultaneous determination of Ni 2+ and Cu 2+ , respectively. Linearity was obtained in the range of 0.5-5.0 mg mL -1 for both ions in the presence of 0.0-5.0 mg mL -1 of the other ion as an interfering ion. IUPAC detection limits for Cu 2+ and Ni 2+ ions were obtained at 0.48 and 0.43 mg mL -1 , respectively. The proposed procedure has been applied successfully for the simultaneous determination of copper and nickel in synthetic binary mixtures and real samples.