The origin and authenticity of feed for laying hens is an important and fraud-susceptible aspect in the production of organic eggs. Chemical fingerprinting in combination with chemometric methods is increasingly used in conjunction with administrative controls to verify and safeguard the authenticity of food commodities. On the basis of fatty acid fingerprinting data of 36 organic and 60 conventional feeds, we have developed a chemometric classification model to discriminate between organic and conventional chicken feed. A two-factor partial least squares-discriminant analysis (PLS-DA) model was developed using 70% of the original data. External validation of the model with the remaining 30% of the data showed that all of the organic feeds and 90% of the conventional feeds (18 of 20) were correctly identified by the model. These results indicate that the PLS-DA model developed in this study could be routinely used to verify the identity of unknown or suspicious feed for laying hens.