Cerebrosides are important high-value dietary molecules with unique biological functions which are found extensively in mushrooms. However, tedious detection/extraction methods and lack of adequate reports led mushrooms to be an under-utilized cerebroside source. In this study, two of glycosphingolipids, cerebrosides B and E were identified and further confirmed from seven edible mushrooms (Pleurotus eryngii, Agaricus bisporus, Hypsizygus marmoreus, Pleurotus ostreatus, Agrocybe aegerita, Flammulina velutipes and Lentinus edodes) by high-resolution tandem mass spectrometry and hydrogen-1/carbon-13 nuclear magnetic resonance. A rapid method using high-performance liquid chromatography coupled with an evaporative light-scattering detector (HPLC-ELSD) was explicitly developed to detect and isolate cerebrosides from edible fungal matrices. The developed method accurately quantified cerebrosides B (0.15-1.05 mg/g, dry matter) and E (0.18-1.80 mg/g, dry matter), thus verified its applicability. Furthermore, silica solid-phase extraction was established to obtain cerebroside-rich extracts from shiitake mushroom (>46%, peak area%) and was later...