Six adult mares were given a single dose of aqueous suspension procaine penicillin G (300,000 IU/ml) IM at a dosage of 22,000 IU/kg of body weight (15.4 mg of penicillin G/kg). Serum, synovial fluid, peritoneal fluid, and urine penicillin concentrations were measured serially over a 48-hour period. The mean peak serum penicillin concentration was 1.42 microgram/ml at 3 hours. Penicillin was detected in synovial fluid and peritoneal fluid, which obtained mean peak penicillin concentrations of 0.62 microgram/ml and 0.58 microgram/ml, at 4 hours and 3 hours, respectively. These concentrations steadily decreased in parallel with serum concentrations and were still measurable at 48 hours. Urine concentrations of penicillin were relatively high, with a mean peak concentration of 525.81 microgram/ml at 8 hours.