Objective To select the best storage temperature and accurate detection way of the blood alcohol content of drunk driving and provide technical support of judging objectively drunk driving behaviors for traffic management department. Methods This study selects EDTA-2 vacuum tubes, take human vein blood after drinking and respectively store at four temperature conditions -20℃、4℃~8 ℃、 25 ℃ normal temperature and high temperature of 35℃~42 ℃.GC method is used by testing 0,3d, 7d, 14d, 21d and 28d blood alcohol content, and the test result statistics and data are compared and analyzed. Results The blood alcohol contents within 0-3d stored at 35℃~42 ℃and 25 ℃temperature remain stable, decrease significantly after 3d (P<0.05); the blood alcohol contents stored at 4℃~8 ℃ temperature in 0~14d is basically stable and decrease significantly after 14d (P<0.05); at -20℃temperature there are no significant differences among the 28d testing results of blood alcohol contents. Conclusions It is suggested that the blood samples should be collected at low temperature and the best preservation temperature of blood samples is -20℃.