Papaya (C. papaya) is a rich source of bioactive compounds. However, fruit bioactive content varies greatly depending on factors such as the variety and growing location. In this study, three yellow-fleshed papaya cultivars (Lāʻie Gold, Rainbow, Kapoho Solo) and two red-fleshed cultivars (Sunset and Sunrise) were harvested from different locations throughout the Hawaiian Islands and analyzed for their mineral and carotenoid content using ICP-MS and HPLC, respectively. Bioaccessibility of carotenoids across papaya cultivars were compared using an in-vitro digestion model. Yellow-fleshed papayas contained two major carotenoids: β-carotene and β-cryptoxanthin. In addition to these two carotenoids, red-fleshed papayas also contained high lycopene levels. Varietal and geographical differences were evident in both carotenoid content and their bioaccessibility. β-cryptoxanthin was the main carotenoid among yellow-fleshed cultivars, Laie Gold, Rainbow and Kapoho Solo (242.9–739.5 μg/100 g), followed by β-carotene (152.4–331.0 μg/100 g). The red-fleshed varieties, Sunset and Sunrise, contained 1089.6–1570.4 μg lycopene /100 g. Papayas (100 g) contained 6% and 8% of the dietary reference intake (DRI) for Cu and Mg, respectively, but less than 3% of the DRI for other minerals. Among yellow-fleshed papayas, total carotenoid bioaccessibility was highest in the Rainbow variety from Keaʻau and Kapoho farms on the Island of Hawaii. Bioaccessibility of lycopene from red-fleshed papayas ranged from 1.5–11.4%. Altogether, these findings suggest that not only variety, but also different growing location alter the content and bioaccessibility of carotenoids in papaya.