This study described the relationship between the concentration of the branched-chain fatty acid (BCFA) in rat skin surface lipid and the serum level of the branched-chain amino acid (BCAA) and branched-chain alpha-keto acid (BCKA). The concentrations of the BCFAs in the monoester fraction of the skin surface lipid, and BCAAs and BCKAs in the serum were analyzed in rats fed varying amounts of protein (0 to 40%) and different types of BCAAs (3%) for 2 weeks. The serum concentrations of BCAA were proportional to the protein level by day 10 of the feeding period. This dose response was not sustained by day 14 at the end of the feeding. Protein level dependence was not so evident in the concentration of BCKA. The concentrations of BCFA, even carbon number iso-acid in particular, increased in linear proportion to protein intake in the skin surface lipid. Supplementation of valine and isoleucine to the diet at a 3% level specifically raised the concentration of the respective BCAA and corresponding BCKA in serum, and related BCFA in the skin surface lipid. Addition of leucine, however, did not affect the related BCFA concentration in spite of elevated concentration of leucine and its alpha-keto acid. A good linear correlation between the average concentration of the substrates in the circulation and the concentrations of the product BCFA on the skin surface was thus obtained for valine, isoleucine and their respective alpha-keto acid. This relationship did not appear to hold up for leucine and its alpha-keto acid.