Uranium concentration in groundwater reflect both redox conditions and uranium content in host rock. In the present study an attempt has been made to study the uranium concentration and activity ratios of uranium isotopes to present the geochemical conditions of the groundwater in Malwa region of Punjab state, India and the reason for high uranium levels and variation of activity ratios from secular equilibrium conditions. Uranium concentration in groundwater samples was found to be in the range of 13.9 ± 1.2 to 172.8 ± 12.3 μg/l with an average value of 72.9 μg/l which is higher than the national and international guideline values. On the basis of uranium concentration, the groundwater of the study region may be classified as oxidized aquifer on normal uranium content strata (20 %) or oxidized aquifer on enhanced uranium content strata (80 %). The U, U and U isotopic concentration in groundwater samples was found to be in the range of 89.2-1534.5, 4.4-68.5, and 76.4-1386.2 mBq/l, respectively. Activity ratios of U/U varies from 0.94 to 1.85 with a mean value of 1.11 which is close to unity that shows secular equilibrium condition. High value of U isotope than U may be due to alpha recoil phenomenon. The plot of AR of U/U against the total uranium content in log scale reveals that the groundwaters of the study region either belongs to stable accumulation or normal oxidized aquifer.