A new cerebroside (1), together with three known ones (2–4) have been isolated from the whole plants of Euphorbia platyphyllos L. The structures were established by FT-IR spectroscopy, FAB-MS, advanced two-dimensional NMR, including 1 H-NMR, 1 H,1 H-COSY, HMQC and HMBC experiments and chemical reactions. The structures of the cerebrosides were characterized as 1-O-β-Dglycosides of phytosphingosines, which comprised a common long-chain base, (2S,3S,4R,8Z)-2-aminooctadec-8-ene-1,3,4-triol (1–3) and (2S,3S,4E,8E)-2 aminooctadeca-4,8-diene-1,3-diol (4) with 2-hydroxy fatty acids of varying chain lengths (C16, C24, C26:1, C28:1) linked to the amino group. The isolated compounds have been evaluated for their antifungal and antitubercular activities.