The invention provides an extraction method and application of tobacco anti-tumor active components, and belongs to the technical field of processing and extraction of tobacco products. According to the method, tobacco raw materials are pretreated, operations such as normal-temperature extraction, high-temperature extraction, methanol extraction, acetone extraction and liquid chromatography separation are sequentially carried out on the pretreated tobacco raw materials, high-polarity solvent purified water, methanol and acetone are used as extraction solvents, the tobacco active components inthe tobacco raw materials can be fully extracted by adopting different extraction solvents and different extraction conditions, and the extraction content of the tobacco active components is increased. Experiments show that the target component has an anti-tumor function and can be developed into drugs for preventing and treating tumors.