Abstract N , N -dimethyltryptamine (DMT) is a psychoactive indole alkaloid present in beverages consumed in religious ceremonies and in neo-shamanic rituals all around the world. It is a substance banned in most countries, which makes its acquisition difficult. In Brazil, a beverage rich in DMT named ayahuasca is legally consumed in a religious context. On the other hand, DMT is a controlled drug, enforced by the Brazilian National Health Surveillance Agency ( Agencia Nacional de Vigilância Sanitaria ). The present study describes a simple and fast method to obtain N , N -dimethyltryptamine (DMT) from inner barks of Mimosa tenuiflora for the purpose of using it as a chromatographic analytical standard. Fourier transform infrared spectroscopy (FTIR), single and tandem stage mass spectrometry (MS), nuclear magnetic resonance spectroscopy ( 1 H and 13 C NMR) and melting point measurements were performed for the structural characterization of N , N -dimethyltryptamine. The results obtained were in agreement with previous literature reports. The purity of the compound (> 95%) was determined using ultraviolet (UV) absorption spectrometry with a tryptamine analytical standard.