The study focused on Melampodium divaricatum, with above ground organs collected from plants grown in the New York Botanical Garden. Dried, powdered plant material was extracted with dichloromethane and methanol. The methanolic extract was purified, and the alkaloid extract was analyzed by GLC-MS and TLC. Pyrrolidine-2-acetic acid methyl ester, tussilagin, isotussilagin, neo-tussilagin, and neo-isotussilagin were identified. Chemotaxonomically, these atypical pyrrolizidine alkaloids (characterized by a C-3 methyl group and C-1 oxidation) co-occur with pyrrolidine-2-acetic acid in Melampodium, the third genus in Heliantheae known for this co-occurrence, suggesting these compounds are more common than previously expected.