A cation-exchange column (TSK IC-Cation, 4.6 mm i.d., 10 mm) was used as a reaction/concentration/separation medium in an HPLC system. Cr(VI) and 1,5-diphenylcarbazide (DPC) were on-line mixed and reacted in a flow tube. When the reagents flowed into the column, both DPC and the complex were concentrated on the column, and the higher DPC concentration accelerated the remaining Cr(VI) to quickly complete the reaction with the DPC on the column. After the complexation and preconcentration, the complex and the extra DPC were eluted by a mixed solution containing lanthanum chloride and 1-propanol. The absorbance of the Cr(III)-DPCO complex at 540 nm was continuously monitored. In addition, the dissolved Cr(III) could be oxidized to Cr(VI) by 185-nm irradiation using a low pressure ultraviolet (UV) lamp for 8 min, and then analyzed by the present method as total chromium, thus the quantitative speciation analysis of Cr(III) and Cr(VI) was realized. In the range of 1 ng dm -3 -100 μg dm -3 Cr(VI) concentration, the calibration curve was linear. The analytical time was 8 min for Cr(VI) and 24 min for total Cr. The detection limit (3σ) of the method was 0.6 ng dm -3 for Cr(VI) and 0.8 ng dm -3 for total chromium when using a 3.9-cm 3 water sample. The present method could be successfully applied to the speciation analysis of dissolved chromium in natural water, and the leaching test of Cr(III) and Cr(VI) from stainless steel.