Basella alba L.is a frequently utilised plant that belongs to the Baselaceae family. The purpose of the present research is to isolate and characterize the bioactive flavonoid quercetin from the hexane, ethyl acetate, and ethanol extracts of the whole plant of B. alba. Conventional qualitative techniques were employed for the purpose of conducting a phytochemical examination. The analysis of TFCs (total flavonoid contents) was conducted on the extracts of the entire plant. Following its isolationvia Preparative HPLC (High-performance liquid chromatography), quercetin underwent evaluation through various standard analytical techniques such as mass spectrometry (MS), HPLC, 1H NMR (proton NMR), 13C NMR (carbon NMR), among others. It was revealed that the ethyl acetate extract exhibited a higher concentration of flavonoids in comparison to the other extracts. The present investigation documented the very first extraction of quercetin from a comprehensive B. alba plant using ethyl acetate. Furthermore, it was observed that no potential toxicological effects were detected in the B. alba extract at doses of 300 milligrammes per kilogramme of body weight and 2000 milligrammes per kilogramme of body mass in female Swiss albino mice.