Liver, kidney, and lung homogenates from 45- and 630-d-old rats were incubated with 0.5 muCi of 14C-labeled di(2-ethylhexyl) phthalate (DEHP) for 40 min at 37 degrees C. Radiochromatogram scans of the ether extracts of the incubated mixtures showed the presence of a peak corresponding to the hydrolytic product monoethylhexyl phthalate (MEHP), in addition to the parent DEHP. Preparations from old animals revealed another radioactive zone along with MEHP. A dramatic reduction in the formation of MEHP was observed only with homogenates of livers from the old animals. It is shown that this difference is probably attributable to differences in Km values of the enzymes from adult and old rats respectively. Protein content in the three tissues did not differ between young and old animals. Formation of MEHP is a major step in the metabolic pathway of the plasticizer DEHP. Impairment of this conversion could possibly alter the rate of its excretion and hence its toxicologic significance.