To study the alkaloids of Solanum lyratum Thunb. ODS and silica gel column chromatography combined with HPLC were used to separate and purify the alkaloids. Seven alkaloids were identified via UHPLC-ESI-Q-TOF-MS, 1D and 2D NMR, respectively 15β-ethoxy-(3β,5α,25R)-16,23-epoxy-23,24-imino-cholestan-16,20,23(N)-triene-3β-ol-3-O-β-D-glucopyranosyl-(1→2)- β-D-glucopyranosyl-(1→4)-β-D-galactopyran-oside (1), 15β-hydroxyl-(3β,25R)-16,23-epoxy-23,24-imino-cholestan-5,16,20, 23(N)-tetraene-3β-ol-3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl-(1→4)-β-D-galactop-yranoside (2), Solalyraine A (3), Solalyraine B (4), 16,23-epoxy-22,26-imino-cholest-22(N),23,25(26)-trien-3β-ol-3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl- (1→4)-β-D-galactopyranoside (5), N-(4-amino-butyl)-3-(3-hydroxy-4-methoxy-phenyl)-E-acrylamide (6) and magnoflorine (7). Among them, compounds 1 and 2 were two new steroidal glycoalkaloids that has not been reported in the literature, and compound 7 was isolated from S. lyratum for the first time. Copyright © 2021 Journal of Chinese Pharmaceutical Sciences, School of Pharmaceutical Sciences, Peking University