In this study, the profile of tocopherols and tocotrienols in chestnut ( Castanea sativa Mill.) kernel oil was evaluated. Four Portuguese chestnut varieties were selected: Aveleira, Boaventura, Judia, and Longal. The vitamin E determination had already been applied to similar matrices, but, to the authors' knowledge, it is the first time that chestnut kernel oil has been evaluated. The prevalent vitamer was gamma-tocopherol, often present in trace amounts in other natural products. Due to the high commercial value of chestnut, a statistical analysis of the obtained results was also conducted to define the tocopherol and tocotrienol profile as a reliable indicator of a specific chestnut variety. To achieve this objective, an analysis of variance was performed to evaluate the accuracy of the method as well as the uniformity of results for each variety. A discriminant analysis was also carried out revealing quite satisfactory results. Four varieties were clustered in four individual groups through the definition of two discriminant analysis dimensions.