Our previous investigation on Litsea glutinosa (Lour.) C. B. Rob. (Lauraceae) resulted in the isolation of new aporphine alkaloids and flavone glycoside. In this study, we report the isolation and identification of ten known megastigmane O-glucopyranosides 1–10 for the first time from the n-BuOH-soluble fraction of an EtOH extract of L. glutinosa. The leaves and twigs of L. glutinosa were collected from Xishuangbanna County of Yunnan Province, P. R. China, extracted with EtOH, and the extract was partitioned into petroleum ether, EtOAc, and n-BuOH fractions. The n-BuOH-soluble fraction was subjected to silica gel column chromatography, Sephadex LH-20, and RP-18 to yield compounds 1–10. The structures of megastigmane O-glucopyranosides 1–10 were elucidated on the basis of their MS, 1H NMR, and 13C NMR spectra, with experimental data in agreement with literature IR, UV, 1D, and 2D NMR data and some compared with authentic samples. These megastigmane glycosides are reported for the first time from the genus Litsea. Megastigmane derivatives have never been reported from the genus Litsea before. The result of the present study suggests that L. glutinosa was a source of megastigmane glycosides, and the presence of this class of naturally occurring compounds in the genus Litsea could stimulate future phytochemical genomics studies.