A new, simple UV-Vis spectrophotometric method for the separation-preconcentration and determination of rhodamine B based on its adsorption onto multi-walled carbon nanotubes has been described. The effects of parameters for the quantitative recoveries of rhodamine B, including pH, flow, sample volumes, etc., were optimized. Matrix effects of concomitant ions or other dyes were also examined. The preconcentration factor and LOD were calculated as 125 and 0.80 mu g/L, respectively. The procedure was applied to the spectrophotometric detection of rhodamine B in a soft drink, dialysis water, textile industry wastewater, and nail polish samples.