A method for the quantitative separation and preconcentration of trace amount Cr(III) in real samples has been developed. It is based on the sorption of Cr(III) ions onto a column of Amberlite XAD-2 resin functionalized with 5-palmitoyl-8-hydroxyquin oline reagent. The resin was used for speciation and preconcentration of chromium because this functionalized resin is selective for Cr(III) ions. The effect of pH, flow-rate, sorption capacity, reusability and effect of various metal cations and salt anions on the sorption on to the resin were investigated. The sorption is quantitative in the pH range of 4.5–7.0, and Cr(III) ion was desorbed by using hydrochloric acid. The amount of metal ion in the eluate was determined by a flame atomic absorption spectrometer. Total chromium was obtained by an efficient reduction of Cr(VI) to Cr(III) by hydroxylamine. The procedure has been applied to the determination and speciation of chromium in lake water, and in the chromium-plating baths.