Aspidosperma pyrifolium Mart., Apocynaceae, is a species popularly known with the popular Brazilian names of “pereiro” and “pau-pereiro.” It is a native tree from the Caatinga biome, and the flowers frequently attract bees. The species is known for the presence of plumeran indole alkaloids. In this study, the alkaloidal fraction of A. pyrifolium flowers resulted in the characterization of eighteen plumeran indolic alkaloids via ultra-performance liquid chromatography coupled with diode array and quadrupole time-of-flight mass spectrometry. Four of these alkaloids were isolated, and their structures were established by different spectroscopic methods. The novel indole alkaloid N-formyl-15-methoxy-aspidofractinin, in addition to the known alkaloids pyrifoline, aspidospermine, and 15-methoxy-aspidospermine, was identified from the analyzed plant. [Figure not available: see fulltext.] © 2020, Sociedade Brasileira de Farmacognosia.