This experiment was conducted to evaluate the effects of l-carnitine on performance, egg quality and certain biochemical parameters in laying hens fed a diet containing high levels of copper proteinate. Forty-eight 42-week-old laying hens were divided into four groups with four replicates. The laying hens were fed with a basal diet (control) or the basal diet supplemented with either 400 mg carnitine (Car)/kg diet, 800 mg copper proteinate (CuP)/kg diet or 400 mg carnitine + 800 mg copper (Car+CuP)/kg diet, for 6 weeks. Supplemental CuP decreased feed consumption (p < 0.01), feed efficiency and egg production (p < 0.001), as compared to control. The combination of Car and CuP increased (p < 0.001) egg production and feed efficiency as compared to CuP. The activities of alanine aminotransferase (p < 0.05) and alkaline phosphatase (p < 0.01) were increased, while lactate dehydrogenase activity was decreased (p < 0.001) by supplemental CuP and Car+CuP. Supplemental CuP caused an increase in plasma malondialdehyde (p < 0.01) and nitric oxide levels (p < 0.05). In the Car+CuP group, this increase was observed to have been reduced significantly (p < 0.05). Furthermore, Car+CuP increased (p < 0.05) glucose level. These results indicate that the carnitine and copper combination may prevent the possible adverse effects of high dietary copper on performance and lipid peroxidation in hens.