The aim of the present study was to investigate the phytochemical composition and the antioxidant potential of the Date Palm pollen grains (DPP) collected from Abu-Hamad, Sharkia, Egypt. Investigation of the lipid constituents of pollen grains of Phoenix dactylifera by GLC analyses led to identification of the fatty acids mixture which consists of nine fatty acids with palmitic acidas a major and the unsaponifiable fraction that contains a series of hydrocarbons, sterols, with pentadecane as a major. Estimation of total phenolic content (TPC) and total flavonoid content (TFC) of the methanolic extract (ME) were performed by Folin-Ciocalteau reagent and aluminium chloride methods respectively. Twenty three phenolic compounds and twenty flavonoid compounds were identified and quantified by HPLC to provide the scientific basis for its biological activity. The results revealed that the methanolic extract had total phenolic content value of 62.1 ± 1.8 mg GAE/g and total flavonoid content value of 90.2 ± 2.3mg QE/g. Antioxidant activity was screened by DPPH assay. The methanolic and ethyl acetate extracts of the DPP grains showed potent antioxidant activity and high cytotoxic activities at a concentration of 25 µM against RPE-1 and MCF-7 human cell lines using MTT assay, compared to doxorubicin as a reference drug.