An experimental study was designed using male Sprague-Dawley rats treated with a single dose of 1800 rads to an area of skin and soft tissue of the back measuring 2 X 3 cm. This dose was estimated to produce changes equivalent to 6000 rads in divided doses over 6 weeks. At intervals of 5, 10, and 15 weeks after irradiation, punch biopsies were taken from both irradiation, and nonirradiated skin areas of each animal 30 minutes after the intraperitoneal administration of gentamicin. Skin homogenates were prepared, and the antibiotic levels in these samples were determined by a bacterial growth inhibition assay. The antibiotic levels were found to be equal (16.1 +/- 6 micrograms/ml vs. 16.0 +/- 5 micrograms/ml) in both irradiated and nonirradiated skin at 5 weeks after radiation. However, at 10 and 15 weeks after radiation, the antibiotic levels had dropped to 9.9 +/- 3 micrograms/ml in irradiated skin compared with 14.1 +/- 4 micrograms/ml in normal skin (p less than 0.001) and with 5.4 micrograms/ml in irradiated skin vs. 11.8 +/- 5 micrograms/ml in nonirradiated skin (p less than 0.001), respectively. Results demonstrate that in spite of adequate gentamicin levels in the circulation and nonirradiated tissue in rats, gentamicin has a decreasing ability to diffuse into irradiated tissues with increasing intervals after therapeutic doses of radiation.