Thallium (Tl) is a typical toxic element, whose biological effects and geochemical behavior are closely related with its chemical speciation in the environment. In this context, the objective of the present study was to develope an effective method for separation of Tl (I) and Tl (III) based on solid-phase extraction (SPE) using anion exchange resin AG1-X8 as a sorbent and ICP-MS measurement. In this proposed method, Tl (I) and Tl (III) could be separated by selective adsorption of Tl (III)-DTPA in the resin, while Tl (III) was eluted by the solution mixed with HCl and SO2. The validity of this method was confirmed by assays of standard solutions of Tl (I) and Tl (III), as well as with spike of contaminated samples. The present study results revealed that higher concentration of Tl (I) (245.48 mu g/l) and Tl (III) (20.92 mu g/l) had been found near the acid mine drainage (AMD) sample of sediment pore water. The results revealed that Tl (I) of 61.47 mu g/l and Tl (III) of 9.73 mu g/l were present in the river water contaminated by acid mine drainage. This thallium speciation analysis implied that the dominant Tl (I) species in the river water studied might be due to the weathering of sulfide mineral-bearing rocks, mining, and smelting activities in the studied area.