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Objective To extract and purify juglone from Cortex Juglandis Mandshuricae, and then study it s anti-tumor activity as well as mechanism.Methods Cortex Juglandis Mandshuricae were extracted with 75% ethanol and concentrated with chloroform.The chloroform portion was eluted gradiently on silica gel to get juglone.By the MTT assay method to study the inhibition of cell proliferation activity and by detecting the mitochondrial membrane potential to explore the juglone anti-tumor mechanism.Results Juglone was extracted from the Cortex Juglandis Mandshuricae successfully.MTT experiment results showed that juglone can inhibit cell proliferation effectively.And mitochondrial membrane potential experiment results prove that juglone can change the mitochondrial membrane potential in a dose-dependent manner.Conclusion Juglone is an important component of Cortex Juglandis Mandshuricae which has a strong inhibition of cell proliferation activity, and it s mechanism may be associated with lower mitochondrial membrane potential.