Amaryllidaceae alkaloids from the Amaryllidoideae subfamily exhibit broad pharmacological activities, including neuroprotection and anticancer effects. Galanthamine is a key compound for Alzheimer's therapy. The genus, particularly in Bolivia, offers significant potential for novel drug discovery, emphasising the need for conservation and further phytochemical research. Twenty-seven samples from Bolivian species were investigated in terms of their alkaloid profile and anticholinesterase activity. The phytochemical analysis of Bolivian species via GC-MS identified 48 Amaryllidaceae alkaloids, displaying diverse structural groups with potential pharmacological significance. Lycorine- and Homolycorine-type alkaloids were predominant, particularly in and , with high concentrations of lycorine, a promising anticancer compound. The species and contained notable quantities of Galanthamine type alkaloids, relevant for Alzheimer's treatment. This study also highlights variability in acetylcholinesterase and butyrylcholinesterase inhibitory activities, with and demonstrating strong inhibition. These findings suggest that species are a valuable source of bioactive compounds, warranting further research into their therapeutic applications.