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Diaporthe phaseolorum, an endophyte isolated from Combretum lanceolatum, has exhibited diverse biological activities including antimicrobial, leishmanicidal, antitumoral, and herbicidal properties. Regardless of these activities, there remains limited research on its chemical constituents. Therefore, this study aimed to investigate the metabolome of D. phaseolorum employing mass spectrometry data dereplication through molecular networking and MolNetEnhancer tools. Twenty two molecular families were annotated, including prenol lipids, benzenoids, organoheterocyclic compounds, and nucleosides. Among the 41 nodes annotated, 36 metabolites were confirmed via spectral references, including alismol, farnesol, linoleic acid, tyramine, N-acetyl-tyramine, 5-methoxy-1H-indole-3-carbaldehyde, 6-pentyl-2H-pyran-2-one, and adenosine. Further node annotation was achieved through MolNetEnhancer, resulting in the classification of an additional 68 nodes into various molecular families. Consequently, this study represents a significant contribution to the identification of the D. phaseolorum metabolome, highlighting its secondary metabolites and providing valuable chemical insights to elucidate its demonstrated biological activities. lanceolatum antimicrobial leishmanicidal antitumoral properties constituents Therefore D tools annotated lipids benzenoids compounds nucleosides 4 3 references alismol farnesol acid tyramine Nacetyltyramine, Nacetyltyramine N acetyl N-acetyl-tyramine 5methoxy1Hindole3carbaldehyde, 5methoxy1Hindole3carbaldehyde methoxyHindolecarbaldehyde 5 methoxy 1H indole carbaldehyde, carbaldehyde H 5-methoxy-1H-indole-3-carbaldehyde 6pentyl2Hpyran2one, 6pentyl2Hpyran2one pentylHpyranone 6 pentyl 2H pyran 2 one, one 6-pentyl-2H-pyran-2-one adenosine Consequently Hindole Hpyran