A dispersive liquid-liquid microextraction procedure coupled to spectrophotometry is described for the determination of the trace levels of Sudan Blue II. Analytical parameters, such as pH, volume of extraction solvent (carbon tetrachloride), volume of dispersant (ethanol), volume of sample, and extraction time, were optimized. Matrix effects were also investigated. Preconcentration factor was found to be 200. Detection limit and relative standard deviation (RSD) were 0.55 mu g L-1 and 3.9%, respectively. The procedure was successfully used for the determination of trace levels of Sudan Blue II in food, ink, antifreeze, and industrial waste-water samples.