The invention discloses polypeptide separated from centipede toxin, and determination on anti-tumour activity thereof is carried out. Ultrafilteration and reversed-phase chromatography separation methods are adopted to separate and purify one polypeptide composed of twelve amino acids from centipede toxin, the sequence thereof detected by applying edman degradation method is Phe-Thr-Gly-Gly-Asp-Glu-Ser-Arg-Ile-Gln-Glu-Gly, and the molecular weight thereof measured by applying mass-spectrometric method is 1296.05Da. The anticancer cell viability of the polypeptide is determined by applying MTT method. The polypeptide in the invention has anti-tumour activity, has inhibition action on propagation and growth of human umbilical vein endothelial cell, is in dose-response relationship and can be applied to treatment and auxiliary treatment of various cancers.