The maximum dietary protein intake that does not cause adverse effects in a healthy population is uncertain. We tested whether a high protein intake enhances oxidative stress. Adult rats were adapted to different casein-based diets containing either an adequate (13.8%; AP), medium (25.7%; MP), or high (51.3%; HP) level of crude protein; a fourth group received a HP diet but no RRR-alpha-tocopherol acetate (HP-toc). After 15 wk of feeding, plasma protein carbonyl concentration, liver lipid peroxide levels [thiobarbituric acid-reacting substances (TBARS)], reduced glutathione (GSH) status and leucine kinetics ([1-(13)C]leucine) were measured. Higher concentrations of protein carbonyls and TBARS were found in rats fed the AP and the HP-toc diets compared with those fed the MP and HP diets (P: < 0.05). GSH concentrations in plasma did not differ but total blood GSH concentrations were significantly (P: < 0.05) lower in rats fed the HP-toc diet compared with those fed the AP, MP and HP diets. Liver GSH concentrations were significantly (P: < 0.01) lower in rats fed the AP diet compared with the other groups. Rates of postabsorptive leucine oxidation (LeuOX) and flux (Q(Leu)) were positively correlated with the dietary protein level (for AP, MP, and HP, respectively: LeuOX, 74.9 +/- 28.5, 109 +/- 35.2, 142.3 +/- 38.4 micromol/(kg. h); Q(Leu), 425 +/- 102, 483 +/- 82, 505 +/- 80 micromol/(kg. h). Only HP-toc resulted in a significantly greater protein breakdown (PB(Leu)) and Q(Leu). No difference was seen in nonoxidative leucine disposal. Long-term intake of high protein diets did not increase variables of oxidative stress, in contrast to our initial hypothesis. An unexpected finding was that adequate protein feeding (AP) may in fact induce oxidative stress.