In recent years, several countries have implemented new regulations regarding the limitation or labeling of the trans fatty acid (TFA) content of foods and dietary supplements. GC methods for fatty acid (FA) analysis have been updated by improving the separation of TFAs from other FAs, especially trans- and cis-18:1, and by focusing more attention on the FAs contained in fats and oils in lower amounts. FA analysis is affected by the limited availability of reference materials. Identifications are frequently made simply by comparison with separations reported in the literature. This report describes the preparation of mixtures containing fatty acid methyl esters (FAMEs) that are not available as reference materials. These mixtures can be used for FAME identifications. The prepared mixtures are analyzed under the experimental conditions of the American Oil Chemists' Society (AOCS) Official Method Ce 1h-05 and AOCS Recommended Practice Ce 1j-07.