This experiment was conducted at the poultry farm of the college of Agriculture, University of Al-Qasim Green for the period from 26/9/2019 to 8/11/2019. The study was aimed to the effect of adding different levels of Biotin to the diet in some biochemical characteristics of the blood serum of Japanese quail exposed to oxidative stress . in the experiment, 300 unsexed Japanese quail birds were used. The eggs were obtained from the Agricultural Research Department in Abu Ghraib and the eggs hatched in Al-Nasr hatchery in Babylon province. They were raised together until the age of 8 days and then were randomly distributed to 5 treatments with 3 replicates each treatment ( 20 birds / replicate). The oxidative stress was induced by adding 0.5% H2O2 to drinking water and the experimental treatments were as follows: The first treatment: Negative control treatment is free of any addition. The second treatment: a positive control treatment adding water with H2O2 at just by only 0.5%. The third, fourth and fifth treatment: adding biotin at a concentration of 500, 600 and 700 micrograms / kg feed + water with H2O2 added at a concentration of 0.5%, respectively. The treatment lasted from 8 to 42 days. The results are as follows:The results of the experiment showed a significant improvement (p≤0.05) for the adding of Biotin (third, fourth and fifth) in the total protein and albumin concentration, and a significant decrease in the concentration of glucose compared to the first treatment and the second treatment, as for the concentration of liver enzymes ALT and AST, the results showed a significant decrease in the first treatment and adding treatment (Biotin ) compared to the second treatment (positive control), while the results showed a significant increase (p≤0.05) in the enzyme Glutathione for the adding treatment Biotin (third, fourth and fifth treatment) Compared to the first treatment and the second treatment .Biotin adding treatment (third, fourth and fifth) showed a significant decrease (p≤0.05) in the level of cholesterol and triglycerides concentration as well as low-density lipoproteins and the level of MDA and an increase in the level of high-density lipoproteins in the Serum blood of bird compared to the first treatment and the second treatment.