We administered 4 g of azlocillin sodium intravenously to 24 patients scheduled to undergo elective intraocular surgery and collected specimens of serum, tears, and aqueous humor from zero to six hours after infusion for assay by high-pressure liquid chromatography. We found that azlocillin is distributed into tears and penetrates into the aqueous humor of human volunteers with noninflamed eyes after a single intravenous dose. Levels of 4.17 micrograms/ml and 4.44 micrograms/ml were achieved in tears and aqueous humor, respectively.