Ocular clearance of amphotericin B after direct intravitreal injection was studied in a rabbit model. Unmodified phakic eyes, Candida-infected eyes, aphakic eyes, and aphakic vitrectomized eyes were employed. Using high pressure liquid chromatography to assess drug level, the half-lives of drug disappearance after single 10-microgram (microgram) intravitreal injections were 9.1, 8.6, 4.7, and 1.4 days, respectively. The disappearance slope for vitrectomized eyes was significantly different from all nonvitrectomized eyes with P less than 0.001. The rapid disappearance of amphotericin from vitrectomized eyes must be considered in clinical management of patients with fungal endophthalmitis.