Oil was extracted from silkworm (Bombyx mori) pupae using supercritical carbon dioxide extraction.Principal component analysis (PCA) was used to establish a mathematical model with cluster analysis based on 90 silkworm varieties using monomer fatty acids (MUFAs) and polyunsaturated fatty acids (PUFAs) as indices.The influence of the variety of the silkworm pupae on the crude fat and MUFAs was remarkably significant.Single-factor correlation analysis indicated that the PUFAs content was significantly positively correlated with the α-linolenic acid (ALA) and negatively correlated with the oleic acid (OA) contents.The amount of ALA and PUFAs were the key factors determining the nutritional value of the pupae fat, because their loading coefficients in the mathematical model were higher than those of the other factors.Cluster analysis on the composite scores of the principal component divided the 90 varieties into three groups.A cluster of 5 varieties with the best nutritional value was characterized by high contents of ALA and PUFAs and a low content of oleic acid.