Preruminant calves are regarded as a model for studying beta-carotene bioavailability in humans. The objectives of this trial were to determine the relationship between multiple beta-carotene doses and plasma steady-state concentration, accumulation in selected tissues, and vitamin A balance in liver. Seventy newborn Holstein calves in six treatments (n = 10/treatment) were fed a complete milk replacer diet low in vitamin A and supplemented with beta-carotene doses of 0, 0.23, 0.46, 0.92, 1.84 or 3.68 mumol/(kg body wt.d) for 28 d. Ten calves were killed on d 1. Plasma beta-carotene increased in relation to log transformations of dose and time (P < 0.05) in all supplemented calves and steady state was attained after 4 wk. For doses up to 0.92 mumol/(kg body wt.d), the dose-response relationship was linear. A dose-dependent accumulation of beta-carotene was found for liver, heart, lungs, adrenals and adipose tissue. All-trans-beta-carotene was the only isomer in plasma and adrenals and the predominant isomer in the remaining tissues. In liver, vitamin A increased with beta-carotene uptake. Hepatic balance between vitamin A accumulation and loss was achieved at beta-carotene intake of 0.36 mumol/(kg body wt.d) for a calf of 45 kg. It is concluded that preruminant calves within 1 mo of age utilize beta-carotene as a source of vitamin A, and that for testing bioavailability of beta-carotene sources, doses up to 0.92 mumol beta-carotene/(kg body wt.d) are most appropriate.