Mushroom, as a kind of higher fungus, is a precious homology resource of medicine and foods. In this study, total lipids were extracted from eight wild edible mushrooms and subsequently characterized by ultra-high-performance liquid chromatography-Quadrupole-Exactive Orbitrap mass spectrometry. 20 lipid classes and 173 molecular species were identified and quantified. Lipid molecules and their concentrations in Boletus speciosus, Boletus bainiugan, and Tricholoma matsutake exhibited significantly different behaviors compared with the remaining mushrooms. Hierarchical cluster analysis revealed that lipid profiles of B. bainiugan were most similar to B. speciosus followed by T. matsutake, Canthar-ellus cibarius, Sarcodon aspratu, Termitomyces eurrhizus, Laccaria laccata, and Thelephora ganbajun. In addition, several differential lipids can be considered as potential biomarkers to distinguish different mushroom species, for instance, lysophosphatidylethanolamine (16:1) and ceramide non-hydroxy fatty acid-dihydrosphingosine (d23:0-10:0). This study provided a new perspective to discriminate the mushroom species from the perspective of lipidomics.