Abstract Breadfruit (Artocarpus altilis) is an underutilized staple crop that may be a good source of pro-vitamin A carotenoids (pVACs). The current study assessed the total carotenoid contents and carotenoid profiles of 94 cultivars of breadfruit and three related species to identify pVAC-rich germplasm and determine if total carotenoid content alone could be used for future screening efforts. Lutein, a non-pVAC, was the predominant carotenoid with an average of 45.0 μg/100 g fresh fruit (SE ± 4.80). The most abundant pVAC was β-carotene, with an average of 15.3 μg/100 g fresh fruit (SE ± 0.54). The ratio of lutein to pVACs varied considerably among cultivars, precluding the use of flesh color or total carotenoid measurements as valid screening methods to identify pVAC rich germplasm. The cultivar “Samoan 1” contained significantly more pVACs (61.9 ± 10.68 μg/100 g β-carotene, 22.9 ± 3.05 μg/100 g α-carotene, and 3.3 ± 1.66 μg/100 g β-cryptoxanthin) than other cultivars. Cooking the fruit resulted in higher measured pVAC contents. Consumption of a standard daily serving of boiled “Samoan 1” would provide 62–84% of the vitamin A requirement of an adult male or 43%-48% of the requirement of a pregnant woman. These data indicate the potential of breadfruit to improve food security and nutrition in the tropics.