Objective To evaluate the optimal anesthesia in animal experiments. Methods Several animal species ware used in this study: mice, rats, guinea pigs, rabbits, dogs, cats, pigs and gouts. Combined anesthesia was tested on several animal species, in different ways and with different anesthetics, and the results were evaluated. Results The combination of ketamine hydrocholoride, sumianxin and diazepam in a ratio of 4:2:1 was found to be an optimal intravenous anesthetic agent. For canine, feline and swine, combined anesthesia of both intravenous and intramuscular was most satisfactory. Combined use of atropine, which is an anticholinergic agent, can effectively avoid overanesthesia. Conclusion Satisfactory results of anesthesia in experimental animals can be obtained with an appropriate combined anesthesia.