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This study investigates the antioxidant components [lycopene, total phenolics, total flavonoids, ascorbic acid (AsA) and dehydroascorbic acid (DHA)] as well as antioxidant activities of the hydrophilic and lipophilic fractions (AAHF and AALF) of peel, pulp and seed fractions isolated from red-ripe berries of the ordinary tomato cultivar Rio Grande and the two high-lycopene tomato breeding lines HLT-F61 and HLT-F62 simultaneously grown in an open-field of Northern Tunisia. Significant differences (p < 0.05) were found among cultivars for each trait studied. All fractions isolated from the red-ripe berries of HLT lines showed higher lycopene, total phenolics and total flavonoid contents, as well as higher AAHF and AALF, than those isolated from Rio Grande. Regardless of the fraction, HLT-F61 had the highest lycopene content (893.0 mg per kg fw, 280.0 mg per kg fw, and 47.5 mg per kg fw in peel, pulp and seed fractions, respectively) and total phenolics at least 2-fold and 3-fold higher than HLT-F62 and Rio Grande, respectively. Peel and seed fractions from HLT-F61 red-ripe tomato berries had the highest AsA content (345 mg per kg fw and 115 mg per kg fw, respectively), while no significant difference was found in the seed fraction between HLT-F62 and Rio Grande. The HLT-F62 pulp fraction showed the highest content of AsA (186 mg per kg fw) and DHA (151 mg per kg fw) among all the assayed cultivars. Except for the peel fraction, where HLT-F61 had similar AAHF values to HLT-F62, the high-lycopene line HLT-F61 showed higher AAHF values than HLT-F62 and Rio Grande. Regardless of the fraction, the highest AALF values were recorded in HLT-F61 berries. Thus, both HLT tomato lines are promising for the introduction, as advanced hybrids, in either fresh market or processing industry.