Calix[4]arene based polymer supported chelating resin 8 has been synthesized by covalently linking calix[4]arene-o-vanillinthiosemicarbazone via its 'lower rim to Merrifield resin and was characterized by FT-IR, H-1-NMR and elemental analysis. Resin 8 was competently used to separate and preconcentrate toxic metal ions Cr(VI), As(III) and Tl(I) in a column prior to their determination by UV-Visible and GF-AAS with R.S.D. between 1.0-1.4%. Various physico-chemical parameters like pH, concentration of eluting agents, flow rate, total sorption capacity, metal-ligand stoichiometry, exchange kinetics, preconcentration factor, distribution coefficient, breakthrough capacity, resin stability, effect of electrolytes, and associated metal ions have been studied. Detection limits (3 sigma(B)) of 5.83, 8.48 and 5.56 mu g L-1 , along with preconcentration factors of 117, 90 and 105 for Cr(VI), As(III) and Tl(I) respectively were achieved. Resin 8 was successfully applied to the separation and trace determination of Cr(VI), As(III) and Tl(I) from natural water samples of Ahmedabad city and the standard environmental, biological and geological reference materials.