The invention relates to a preparation method of epimedium aglycone, which comprises the steps that dried epimedium is pulverized or cut into segments, 30-95% ethanol or 30-100% methanol is added for reflux extraction, the extract solution is decompressed and concentrated to obtain paste, 10-100 ml of acetate-sodium acetate buffer solution with the pH of 3.5-9.0 is added into each gram of paste, snailase is added for hydrolysis, the mass ratio of enzyme to paste is 0.05-5, the reaction temperature is 15 to 80 DEG C, the reaction time is 2 to 72h, the reaction solution is centrifuged, the precipitate is dissolved with an organic solvent, filtration is carried out to obtain filtrate, the solvent is steamed away to obtain crude epimedium aglycone, and pure epimedium aglycone is obtained after recrystallization. The invention has the characteristic that the epimedium extract is directly converted into high-purity aglycone in one step by utilizing the strong enzymic hydrolysis of the snailase so as to simplify the operation steps with strong purposiveness and high conversion rate, and has the advantages of no pollution and large-scale production. The invention overcomes the shortcomings of large destruction on the aglycone, serious pollution, poor purposiveness, low conversion rate, difficult separation and purification and the like of the prior art.