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Vancomycin concentrations in serum, tissues, and sternum, administered as prophylaxis to patients during coronary artery bypass surgery, were measured. Vancomycin (15 mg/kg) was administered to 15 patients 1 hour before skin incision. Blood, tissue, and sternum samples were collected before, during, and after bypass. The concentration in serum at the end of infusion was 55.1 ± 22.8 μg/mL, the mean elimination half-life was 9 ± 4 hours, the areas under the concentration–time curve (AUC) from 0 to 12 hours and from 0 to infinity were 90.6 ± 25.1 and 289.7 ± 86.5 μg/h per mL, respectively, the mean residence time (MRT) was 11.9 ± 5.0 hours, the mean volume of distribution was 51.1 ± 12.2 L, and the total clearance was 78.3 ± 32.6 mL/min. Vancomycin concentrations in serum, tissues, and sternum during the operation were greater than the MIC90 for most staphylococci and ranged from 16 to 55 μg/mL in serum and from 4 to 39 μg/g in sternum and tissues.