Lutein, zeaxanthin and β-carotene are compounds showing health-promoting properties. The objective of the research study was to assess the selected cultivars of Cucurbita maxima D: ‘Amazonka’, ‘Justynka’, ‘Amber’, and ‘Otylia F1’ from the point of view of the content of the above named compounds in the lyophilized samples. The content of those carotenoids was determined using HPLC-DAD. It was found that the lyophilized fruits of the pumpkin cultivars analyzed were a valuable source of lutein, zeaxanthin, and β-carotene. Large amounts of lutein were determined in 1 g of dried fruits of the ‘Justynka’ cv. (0.186 mg) and ‘Amazonka’ cv. (0.118 mg ), whereas the smallest amounts of lutein were determined in the lyophilized fruits of ‘Amber’ cultivar, which were characterized by the highest level of β-carotene (0.411 mg). The lyophilized fruits of ‘Otylia F1’ cultivar were characterized by the smallest amount of β-carotene. Furthermore, it was reported that the lyophilized fruits of ‘Justynka’, ‘Amazonka’ and ‘Ambar’ cultivars were rich in natural carotenoids and might be widely used in food industry as a source of bioactive, health promoting phytochemical compounds.