Atipamezole is a specific α2-adrenergic receptor antagonist, and there exists insufficient information on its use during pregnancy. The aim of this study was to determine the embryotoxic activity of Atipamezole through an in ovo method. During the first stage of the study, 210 fertile eggs were divided into seven groups of 30 fertile eggs and placed in an incubator. On the seventh day of the first stage, no application was made to the control group. The second group was administered 50 microliters (μL) of saline solution, while the other groups were given Atipamezole at doses of 250, 125, 62.5, 31.25 and 15.62 micrograms·egg-1 (μg·egg-1) in 50 μL saline solution. In the second stage, according to the embryotoxic dose range determined from the first stage, 150 fertile eggs were divided into five groups of 30 fertile eggs and placed in an incubator. On the seventh day of the second stage, no application was made to the control group. Fifty μL of saline solution was administered to the second group. The other groups were given Atipamezole at doses of 220, 190 and 160 μg·egg-1 in 50 μL saline solution. After the incubation period, the eggs hatched, and the embryonic mortality rates were calculated. The mortality rate was determined to be 39.3% at the highest dose (250 μg·egg-1 = 5 miligrams·kilograms-1 –mg·kg-1–) (P<0.05), while the mortality rate at other doses was determined to be the same as the control group (P>0.05). In conclusion, it can be stated that the dose determined for Atipamezole in this study was very high compared to the recommended doses and it can be used in pregnancy as a benefit-loss calculation when necessary. However molecular or histopathological studies regarding the development of organ drafts are necessary to determine the safety of its use during pregnancy.