The present study estimates biological half-life (BHL) of tritium by analysing routine bioassay samples of radiation workers. During 2007-2009 year, 72,100 urine bioassay samples of the workers were analysed by liquid scintillation counting technique for internal dose monitoring for tritium. Two hundred and two subjects were taken for study with minimum 3 μCiL(-1) tritium uptake in their body fluid. The BHL of tritium of subjects ranges from 1 to 16 d with an average of 8.19 d. Human data indicate that the biological retention time ranges from 4 to 18 d with an average of 10 d. The seasonal variations of the BHL of tritium are 3.09 ± 1.48, 6.87 ± 0.58 and 5.73 ± 0.76 d (mean ± SD) for summer, winter and rainy seasons, respectively, for free water tritium in the coastal region of Karnataka, India, which shows that the BHL in summer is twice that of the winter season. Also three subjects showed the BHL of 101.73-121.09 d, which reveals that organically bound tritium is present with low tritium uptake also. The BHL of tritium for all age group of workers is observed independent of age and is shorter during April to May. The distribution of cumulative probability vs. BHL of tritium shows lognormal distribution with a geometric mean of 9.11 d and geometric standard deviation of 1.77 d. The study of the subjects is fit for two-compartment model and also an average BHL of tritium is found similar to earlier studies.