In order to reduce the stress response of animals in animal transportation, it is often used to feed the animals with sedative drugs combined with compound vitamin B. In this article, we will randomly divide 16 amur tiger into the control group and the experimental group. The drug intervention is carried out in the experimental group. The sedative is mainly dexmedetomidine, combined with compound vitamin B, while control group did not feed the drug. The urine of two groups of individuals was collected before and after transportation and the cortisol content in urine was determined by radioimmunoassay as an indication of stress. Finally, referring to the changes of cortisol level in amur tiger, we analyzed the effect of vitamin B6 combined with sedative drugs on the anti stress effect of amur tiger. The results showed that the cortisol level in the control group was significantly different (P < 0.05), but there was no significant difference in the cortisol level in the experimental group (P > 0.05). This kind of veterinary medicine which is mainly composed of sedative dexmedetomidine and vitamin B6 is obviously helpful for reducing the stress of the Amur tiger in transferred transportation.