The invention discloses a method for separating syringin from ilex rotunda. The medicinal herb is crushed into fine powder and is reflowed and extracted through benzene. The extracted gruff is dried in air and is reflowed and extracted through acetone. Filtrates are mixed and extracting solution is condensed and stands for the isolation of white powder, and the extracted gruff is filtrated after the isolation is completed. Educt is washed by acetone and undergoes repeated recrystallization with mixed acetone-methanol solution, and finally methanol is used again for recrystallization to obtain the white raphide of syringin. In the first place, the method adopts ilex rotunda as the raw material with low cost and abundant resource. The average content of syringin in ilex rotunda is determined as 2.55 percent through an HPLC method. Thus, abundant raw material for preparing syringin and derivates thereof can be provided. In the next place, through the method, a plurality of experiments on the dry bark of holly ilex rotunda are carried out to obtain the syringin with high purity; in addition, the method which saves time and labor with good stability is an effective and simple method for extraction and separation.