Background: Momordica charantia L. (Cucurbitaceae), is an herbaceous plant used for food and traditional medicine. It presents a proven antidiabetic activity in the literature, being a promising species for the development of phytotherapics. Objective: The objective was performing an anatomical study and characterizing the metabolites in leaves of M. charantia. Materials and Methods: Semipermanent histological slides were prepared for analysis of petiole and leaf blade in optical, polarization and scanning electron microscopy coupled with energy-dispersive X-ray spectrometry. Maceration and histochemical tests were also performed in the leaf blade. Results: The anatomical characterization revealed information about the type of trichomes, cuticle, vascular bundles and arrangement of the idioblasts and tissues that determine the botanical identity of this species. The histochemistry allowed determining the location of the metabolites and, along with the chemical microanalyses, to identify the type of crystal in the leaf blade. Conclusion: The study described new characters for M. charantia and the results provide support to quality control of the species.