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Chestnut bee pollen is a high antioxidative natural bee product. In this study the phenolic compounds in chestnut bee pollen were determined by HPLC-DAD system. Ethanol, water and methanol were used for the extraction of antioxidant compounds from chestnut bee pollen. The total phenolic contents and antioxidant capacities of extracts were determined by Folin-Ciocalteu, CHROMAC (Cr (VI) reduction antioxidant capacity) and FRAP (ferric reducing antioxidant power) methods, respectively. When compared the amounts of phenolic compounds and spectroscopic results (total phenolic contents and antioxidant capacities), the most suitable solvent was found ethanol for the extraction of antioxidant compounds from bee pollen. The major phenolic compounds in the chestnut bee pollen were determined as pinocembrin, chrysin, galangin and hyperoside with the contents of 1.246, 0.332, 0.122 and 0.516 mg/g of bee pollen, respectively. And also it was determined that the chestnut bee pollen was inhibited 11% of DNA oxidation in Fenton medium.