Pro-vitamin A(beta-carotene) and the predominant forms of vitamin A in human skin (retinol and dehydroretinol) were analyzed in hydrolyzed specimens from 4 cadavers and 22 healthy subjects. Beta-carotene was identified in extracts of epidermis by its specific absorption spectrum. Vitamin A was determined by high-pressure liquid chromatography. The concentrations were related to the wet weight or protein content of the sample. The analysis of different skin compartments showed that the concentrations of the 3 constituents were usually related as follows: carotene greater than retinol greater than dehydroretinol. The concentrations were always higher in the epidermis than in the upper dermis. Skin surface lipids contained carotene and retinol but not in amounts sufficient to contribute to the higher epidermal values. Analysis of epidermal autopsies from 5 different skin areas (back, breast, arm, leg and foot) and of epidermal biopsies from the back of the healthy subjects showed that the interindividual differences were larger for carotene and dehydroretinol than for retinol, whereas the intraindividual variations were larger for retinol. The mean (+/- SD) concentrations of carotene, retinol and dehydroretinol in back skin epidermis of healthy subjects were 13 +/- 5, 1.7 +/- 0.4 and 0.4 +/- 0.2 microgram/g protein, respectively. No significant variations with age and sex were found.