The paper elaborates various methods with merits and drawbacks of acute oral toxicity test. Traditional methods of acute toxicity test such as Horne’s method, Karber’s method and others all require the accurate value of LD₅₀, which need a higher amount of tested animals and disobey the humanitarianism and animal protection. Thus, the new methods including up-down procedure, fixed does procedure and acute toxic classification method are developed and replaced by international organization. The new methods without an accurate value of LD₅₀ are worth of promotion and application because they can minimize the mortality of laboratory animals and save the resources of human and material.