Zephyranthes irwiniana (Ravenna) Nic. García is an endemic, red list threatened species from the Brazilian savanna (Cerrado) with pharmacological potential to treat the symptoms of Alzheimer’s Disease (AD). This work analyzed the vegetative growth and phytochemistry of its potential compounds, in response to variations in sucrose concentration and activated carbon (AC). Seeds were germinated in vitro and in the greenhouse. The in vitro bulbs were separated in six treatments with different sucrose concentrations (30, 45 and 60 gL−1) and/or AC (1 gL−1). Biomass increases in individuals grown in the greenhouse were higher than those cultivated in vitro. Sucrose concentration significantly increased biomass and root number. AC had a positive influence on leaf and root size, and a negative influence on root number. GC–MS analyses indicated great variation in the abundance of α-terpenyl-acetate, ethyl linoleate, clionasterol and lycorine between treatments, with maximum concentrations of 53.06%, 38.68, 14.34% and 2.57%, respectively. Histolocalization tests indicated the presence of alkaloids in the leaf chlorenchyma and bulb cataphylls. Finally, the present study provided new evidence that the constitution of the culture medium directly influences the vegetative growth and phytochemistry of this species, providing a good medium condition for propagating the species under threat.