An anti-cancer active substance fromAntrodia camphorata, produced from mycelium ofAntrodia camphoratawith following steps:(1) culturing mycelium ofAntrodia camphorataby liquid fermentation (2) extracting the thus-obtained mycelium with ethanol and concentrating the extracted solution to obtain an ethanol extract (3) dissolving said ethanol extract in water, and extracting with equal volume of ethyl acetate to obtain ethyl acetate extract (4) purifying said ethyl acetate extract by silica gel column chromatography using hexane/ethyl acetate gradient as the mobile phase to obtain an eluate fraction g and (5) purifying said eluate fraction g by silica gel column chromatography using hexane/ethyl acetate gradient as the mobile phase to obtain a specific eluate fraction which could be purified further to obtain said anti-cancer active substance fromAntrodia camphorata.一種牛樟芝抗癌活性物質,係將牛樟芝菌絲體經過下列步驟所產生者:(1)將牛樟芝菌絲體以液體醱酵培養;(2)將所得菌絲體經過乙醇萃取,得到乙醇萃取物;(3)將所得乙醇萃取物濃縮後溶於水中,並加入等體積的乙酸乙酯(ethyl acetate)萃取,得到乙酸乙酯萃取物;(4)將所得乙酸乙酯萃取物以膠體管柱層析純化,並以正己烷/乙酸乙酯(hexane/ethyl acetate)梯度流洗液為移動相,得到流洗液片段g;以及(5)將所得流洗液片段g再以膠體管柱層析純化,並以正己烷/乙酸乙酯梯度流洗液,取特定流洗液片段,即可製得牛樟芝抗癌活性物質。