A facility to measure total body nitrogen (TBN) by photoactivation analysis in rats, using the 14N(gamma, n)13N reaction, was designed, calibrated and evaluated. TBN provides a quantitative estimate of nutritional status. TBN in humans can be measured by neutron activation, however the small absolute amount of nitrogen in rats causes difficulties with this technique. Photoactivation analysis involved irradiating the specimen with high energy photons from an 18 MV Medical Linear Accelerator to a dose of approximately 2 Gy. The technique was calibrated using urea solution rat phantoms with 2.0-3.0% nitrogen content. By analysing the measured decay curve, the initial nitrogen count rate of 0.51 MeV photons was determined and the initial oxygen count rate, being an interfering source of 0.51 MeV photons, was shown to be constant. Within the range of nitrogen concentrations studied, the response is linear (r2 = 0.9) with a CV of 3.5%. Thus, photoactivation provides a relatively simple, in vivo technique for determining total body nitrogen in rats.