Tomatidine, a natural steroidal alkaloid from unripe green tomatoes has been shown to exhibit many health benefits. We recently provided in vitro evidence that tomatidine reduces the infectivity of Dengue virus (DENV) and Chikungunya virus (CHIKV), two medically important arthropod-borne human infections for which no treatment options are available. The objective of this mass spectrometry analysis was to determine possible proteomic changes in the human cell line Huh7 induced by tomatidine which may be indicative of the underlying antiviral activity of tomatidine.