Introduction: Metabolomics aims to measure the occurrence and concentration of as many metabolites as possible in a complex matrix [1]. Chromatographic and spectroscopic methods such as LC-MS, GC-MS or NMR are most commonly used. In this work we focus on tissue spray ionization mass spectrometry (TSI-MS), which is derived from the electrospray ionization (ESI) and allows metabolome analysis of in natura plant tissues [2]. In this technique, a high voltage source is connected to the fresh sample and a potential difference is generated between the sample and the mass spectrometer inlet. A spray is created by adding a suitable solvent on the sample surface. This spray contains solubilized metabolites, which are ionized through the same process that occurs in ESI process [3]. Amides are the most common secondary metabolites in species of the Piper genus [4]. This class of compounds shows high structural diversity and biological potential [5]. Thus, this work proposes the use of TSI-MS in the identification of amides in Piper tuberculatum leaves in order to prove the efficacy of this technique. Methodology: Leaves were washed, dried, chopped and connected to a mass spectrometer Bruker® Amazon (ESI-IT-MS) with modified ionization source. Methanol (10,0 μl) was used as solvent. Results and discussion: Analysis showed a clean full scan spectrum in which was possible to observe three signals in m/z that match with those of protonated molecules of piplartine, piperlonguminine and 4,5-dihydro-piperlonguminine. To confirm the presence of these compounds, collision induced dissociation (CID) was performed. Piplartine TSI-MS2 mass spectra showed the protonated molecular ion at m/z 318.07 and its predominant product ion at m/z 221.00; Piperlonguminine ([M+H]+ m/z 274.27) mass spectra exhibited its main fragment ion at m/z 200.98; and 4,5-dihydro-piperlonguminine ([M+H]+ m/z 276.05) mass spectra showed two important product ions at m/z 202.97and 135.08 (this last is regarding to the formation of methylene dioxy tropylium ion [6]). Conclusions: This analysis allowed the identification of three piperamides of P. tuberculatum in a rapid and simple way, using minimum sample preparation in a “green chemistry” way.