Chestnut bee pollen has potential nutritional and medicinal effects and is an important natural bee product. This study focused on the investigation of the antioxidant capacity and DNA damage inhibition ability of chestnut bee pollen (CBP) from Bursa (Turkey). The phenolic compounds (rosmarinic acid, vitexin, hyperoside, pinocembrin, trans-chalcone, apigenin, protocatechuic, and galangin) and carotenoids in CBPE were determined by HPLC-DAD (high-performance liquid chromatography-diode array detection). Additionally, the protective ability of CBPE against DNA damage by oxidation was investigated. In this study, it was determined that CBPE has a high total phenolic compound content, and the antioxidant capacity of CBPE inhibits DNA oxidation (34% reduction of DNA damage in Fenton reaction media). This study could reveal new information regarding the use of CBPE as a protective agent for DNA in the future. Practical applications Phenolic compounds and carotenoids prevent some diseases because of their important biological activities. One of the potential food sources chestnut bee pollen contains sugar, carbohydrates, amino acids, proteins, lipids, vitamins, hormones, enzymes, and flavonoids. Chestnut bee pollen, which has protective activity against DNA oxidation, could be an excellent potential source of a protective agent against some degenerative diseases through future applications.