The enzyme indoleamine 2,3-dioxygenase (IDO), which uses free oxygen radicals to cleave the pyrrole ring of indoleamines and give kynurenamines, has previously been found in most tissues, but not in the eye. In this study, IDO activity was measured in post-mortem bovine eyes using Yamazaki's method with L-tryptophan as substrate. Because of the physiological importance of IDO in the protection against free oxygen radical damage, a search was conducted to find this enzyme in the eye. Products of tryptophan degradation by IDO, the kynurenine and 3-hydroxyanthranilic acid were detected and measured in the aqueous humor, iris/ciliary body, and the retina by high-performance liquid chromatography (HPLC) coupled with electrochemical detection. IDO activity was 3.2, 9.0 and 10 nmol/mg protein per h for the aqueous humor, iris/ciliary body, and retina, respectively. These findings suggest that, because of its scavenger properties, IDO is involved in the protection of the eye where, because of its transparency, free radicals are formed not only in the normal oxidation process, but also photochemically.