In nature, uranium is composed of three isotopes, U-238, U-235, and U-234. Emitting alpha particles leads to radionuclides decay. The aim of this work is to set up a database for uranium concentrations in groundwater samples collected from Kufa city, Al-Najaf governorate, Iraq. Twenty four samples have been examined for detecting the presence of uranium levels using a CR-39 detector. The measured uranium concentrations were used to determine uranium isotopes with their ingested radiological toxicity risk (annual effective dose of uranium isotopes and excess cancer risk) and chemical toxicity risk (lifetime average daily dose and hazard quotient) from consumption of the water samples in the present study. The results show that the average value of uranium concentrations, uranium isotopes U-238, U-235, and U-234 (1.20 +/- 0.04) mu g/L, (1.48 +/- 0.06) Bq/L, (0.069 +/- 0.002) Bq/L and (1.49 +/- 0.06) Bq/L respectively. Also, it is concluded that the total average annual effective dose and excess cancer risk in all regions under study were (0.10 +/- 0.004) mSv/y and (0.31 +/- 0.013) x 10(-3) respectively. Moreover, the average value lifetime average daily dose, and hazard quotient was 0.0389 +/- 0.0016 mu g/kg.day and 0.0649 +/- 0.0026 respectively. All results of radiological and chemical risk assessment from uranium concentrations in groundwater samples collected from the Al-Kufa area when used as drinking water were within the global limitations of the organization International Commission on Radiological Protection (1.9 mu g/L; 1 mSv/y and 0.18 mSv/y) and World Health Organization recommended (0.6 mu g/kg/day). Finally, it can be concluded that uranium concentrations do not affect human health according to radiation contents.