Abstract: Two rare olive cultivars, Dhokar and Gemri-Dhokar, growing in the south of Tunisia were investigated to identify their phenolic profile and evaluate their sugar content and antioxidant activity. The highest value of phenolic content was detected in Gemri-Dhokar cultivar extract (6.97 g gallic acid equivalents/kg of fresh olive). In addition, sugar content was quantified; glucose (45.17 g/kg of fresh olive) was the predominant sugar in Dhokar cultivar, followed by fructose (40.83 g/kg of fresh olive). The identification of phenolic compounds was based on separation by high-performance liquid chromatography equipped with a diode array detector followed by liquid chromatography-mass spectrometry analysis. In both cultivars, Oleuropein aglycon derivatives and elenolic acid were the main phenolic components. Oleuropeins were the major compounds quantified in the Gemri-Dhokar drupes olives (61.04 mg/100 g of fresh olive), while 0.25 mg/100 g were found in fresh Dhokar. A new iridoid compound, isolated as pure compound, was not previously reported in the literature. Its structure was established by spectroscopic analyses (NMR, UV, MS, and IR). DPPH, ABTS, and FRAP assays showed that the most important antioxidant capacity of olive extracts was found in with Gemri-Dhokar cultivar.