Phytochemical investigation of roots from Solanum asterophorum led to the identification of 37 compounds, including 9 phenolic acid derivatives, 2 alkamides, 7 hydroxy-carboxylic acid derivatives, and 19 steroidal alkaloids. The dereplication of compounds was performed by extensive analysis by ultra-performance liquid chromatography coupled with a diode array and quadrupole time-of-flight mass spectrometry. With the exception of alkaloid isojurubidine, all compounds were reported from Solanum asterophorum for the first time. The chemotaxonomic significance of the isolated compounds was discussed.