The binding interactions of bovine serum albumin (BSA) with the unbranched fatty acids (FA) pentanoate (five-carbon chain length: C5) up to nonanoate (C9), and the carbamates n-methyl carbamate (equivalent to C3) up to n-hexyl carbamate (equivalent to C8) were examined using an ultrafiltration technique. A single, high-affinity site was observed for each of the FA, with an increasing number of secondary sites with increasing chain length. From binding affinity and competition data, there appear to be distinct albumin sites for the short-chain (less than or equal to C7) and the medium-chain (greater than or equal to C8) FA. Published data suggest that the medium-chain FA site is one of the major drug-binding sites on human serum albumin (HSA) or BSA, the indole/benzodiazepine site. Competition between the FA and warfarin for BSA or HSA binding was studied by ultrafiltration and fluorescence methods and suggests that the short-chain FA site may lie in the same region as a second major drug-binding site, the large warfarin-binding area. Thermodynamic parameters of the FA-BSA interactions are suggestive of primary binding being a combination of electrostatic and hydrophobic binding and secondary binding being purely hydrophobic in nature. Carbamate interactions with BSA show several primary sites and also suggest a disparity between the binding of ligands of less than or equal to 7 and greater than or equal to 8 in total length, but there was no evidence of competition between FA and carbamates. A model is proposed to explain these observations, which includes the suggestion that several classes of hydrophobic binding areas exist, each of which is specific for ligands of a restricted range of chain lengths.