Cottonseed oil fatty acid composition was determined for several cotton genotypes included in the 2006 and 2007 years of the National Cotton Variety Trials. Seed was collected from a total of nine environments that included six locations, and 35 genotypes were included in the analysis. Oil was extracted from seed with hexane, and the glycerides were converted to fatty acid methyl esters and analyzed by gas chromatography. Results indicated that commercially acceptable cotton genotypes vary modestly in their distribution of fatty acids, covering a range slightly greater than the range specified in the Codex trading standard for cottonseed oil. Analysis of variance, based on a random effects model, indicated the relative level of most fatty acids was influenced by environment and genetics, but the interaction of these effects was relatively small. Correlations were found between the levels of several major and minor fatty acids. Many of the observed associations appeared to have some foundation with known and proposed biosynthesis pathways. Although the results indicate that breeding cotton for modified oil composition should be feasible, the range of variation observed within the genotypes studied was insufficient to provide useful traits for breeding. A more extensive survey of cottonseed genotypes will be needed for this purpose.