Tissue drug residue research often involves the killing of an animal every time tissue concentrations are determined. To decrease the number of animals required to perform tissue depletion studies and to circumvent the statistical problems associated with determining tissue depletion kinetic properties, using multiple animals, the renal depletion profile of gentamicin from individual sheep was studied, using a bilateral renal translocation technique. Seven ewes were surgically altered, allowed to stabilize, and then allocated into 2 groups; groups-1 sheep (n = 4) were given 3 mg of gentamicin/kg, IM, q 12 h for 10 days, and group-2 sheep (n = 3) were not given gentamicin. The kidneys from all ewes were biopsied 9 times over 74 days after the termination of gentamicin treatment. The renal concentrations of gentamicin were measured by use of a validated tissue digestion procedure coupled with a liquid-phase fluorescence polarization immunoassay. On days 75 and 77 after the end of gentamicin treatment, all ewes were euthanatized and necropsied. The concentrations of gentamicin in the biopsy specimens ranged from 71.9 to 183 microgram/g on days 1 and 2 after dosing, and decreased to concentrations ranging from 3.99 to 7.35 microgram/g on days 73 and 74 after the end of dosing. The decrease in renal gentamicin concentrations was best described by a biexponential equation, The early phase half-life was 2.8 days, whereas the terminal phase half-life was 59 days (harmonic means). There was no difference in the appearance or histologic features of the kidneys from groups 1 and 2. The only lesions noticed were linear fibroses that were attributed to the biopsy procedure.