Abstract.This work proposes a new extraction method for chromium based on the two-phase aqueous system isopropyl alcohol-ammonium sulfate-ammonium thiocyanate (i-PrOH-(NH4)2SO4-NH4SCN), and the related experimental conditions are optimized. The results show that chromium (III) can be quantitatively extracted under the selected conditions: 4mL of i-PrOH, 200L of 2mol/L sulfuric acid, 1mL of 4mol/L NH4SCN and 3mL of saturated (NH4)2SO4 solution (Vtotal=10mL). Application of the proposed method to speciation analysis of plasma chromium was also investigated and satisfactory results were obtained.