Abstract 1. This study was conducted to determine the effects of hempseed (HS) on performance, egg traits, serum lipid profile and antioxidant activity in Japanese quail (Coturnix coturnix japonica). 2. A total of 120 eight-week old laying quail were divided into 4 experimental groups with 10 replicates. The treatments were as follows: 1) Control diet (C, no HS in the diet); 2) 5% HS in the diet (HS5); 3) 10% HS in the diet (HS10); and 4) 20% HS in the diet (HS20). The quail were fed the experimental diets from 8 to 14 weeks of age. 3. There was no significant difference in body weight (BW), feed intake (FI) and feed conversion ratio (FCR) of the birds overall in the experiment. The egg production was not influenced by the HS contents in the diet; however the HS10 diet increased egg weight and egg specific gravity. The carcass traits were not affected by the HS contents. 4. The serum triglyceride, cholesterol and the high density lipoprotein (HDL) were not significantly altered; however, low density lipoprotein (LDL) concentration in HS supplemented groups were lower than that of the C group. 5. The MDA, SOD, CAT and NO concentrations were not significantly affected but in the HS10 and HS20 GSH-Px concentration was higher than in the C and HS5 groups. 6. The omega-3 fatty acid content of eggs increased linearly with increasing dietary HS content in the diet. 7. In conclusion, HS could be a potential feed and health benefit as a natural antioxidant in relation to decreasing serum LDL and increasing GSH-Px concentration in the liver of laying quail.