Abstract A method, using two different systems, is described for the high-performance liquid chromatographic analysis of retinol, retinal, retinoic acid, retinyl acetate, retinyl palmitate, α-, β- and γ-carotene, β-apo-6′-,β-apo-8′-, β-apo-10′- and β-apo-12′-carotenal, ethyl β-apo-8′-carotenoate, α-tocopherol and α-tocopheryl acetate. The first system consists of a laboratory-packed Hypersil-ODS 3-μm column and a mobile phase of acetonitrile—methylene chloride—methanol—water (70:10:15:5, v/v). The second system consists of a laboratory-packed Nucleosil C 18 3-μm column and a mobile phase of acetonitrile—0.1 M ammonium acetate (80:20, v/v). The detection limits in standard solutions were 10 ng/ml for retinoids and carotenoids and 60 ng/ml for the E vitamers. Analysis of the tissues and plasma of rats, after 2 weeks on a diet supplemented with either β-carotene or canthaxanthin (both 2 mg/g), led to the conclusion that the rats were able both to transport and store β-carotene and canthaxanthin and to convert β-carotene to retinol. Incubation of cytosol preparations from the mucosa of the small intestine of rat with 1 μg of β-carotene resulted in the formation of 10–20 ng of retinal within 1 h.