Dietary vitamin D may compensate for inadequate sun exposure; however, there have been few investigations into the vitamin D content of Australian foods. We measured vitamin D₃ and 25-hydroxyvitamin D₃ (25(OH)D₃) in four species of white fish (barramundi, basa, hoki and king dory), and chicken eggs (cage and free-range), purchased from five Australian cities. Samples included local, imported and wild-caught fish, and eggs of varying size from producers with a range of hen stocking densities. Raw and cooked samples were analysed using high performance liquid chromatography with photodiode array. Limits of reporting were 0.2 and 0.1 μg/100 g for vitamin D₃ and 25(OH)D₃, respectively. The vitamin D₃ content of cooked white fish ranged from <0.1 to 2.3 μg/100 g, and the 25(OH)D₃ content ranged from 0.3 to 0.7 μg/100 g. The vitamin D₃ content of cooked cage eggs ranged from 0.4 to 0.8 μg/100 g, and the 25(OH)D₃ content ranged from 0.4 to 1.2 μg/100 g. The vitamin D₃ content of cooked free-range eggs ranged from 0.3 to 2.2 μg/100 g, and the 25(OH)D₃ content ranged from 0.5 to 0.8 μg/100 g. If, as has been suggested, 25(OH)D₃ has five times greater bioactivity than vitamin D₃, one cooked serve (100 g) of white fish, and one cooked serve of cage or free-range eggs (120 g) may provide 50% or 100%, respectively, of the current guidelines for the adequate intake of vitamin D (5 µg) for Australians aged 1–50 years. Full article