<p>Piper guineense is a West African spice traditionally used for culinary and medicinal purposes. This study evaluated the phytochemical composition and antimicrobial activity of <i>P. guineense</i> seed extracts. Soxhlet extraction using n-hexane was employed, followed by phytochemical screening, antimicrobial assays, gas chromatography analysis, and spectroscopic characterization of the major compound. The extract contained several bioactive phytochemicals and exhibited measurable antibacterial activity against selected Gram-positive and Gram-negative clinical pathogens. The antimicrobial action was concentration-dependent but lower than that of the reference antibiotic. Spectroscopic analysis of the purified fraction suggested the presence of β-amyrin, a pentacyclic triterpenoid previously reported in medicinal plants. These findings provide scientific support for the ethnomedicinal use of <i>P. guineense</i> and indicate its potential as a source of bioactive compounds for further pharmacological investigation. This study is among the few to integrate non-polar compound isolation with antimicrobial evaluation and nutritional profiling of Piper guineense seeds, providing both phytochemical and ethnonutritional justification for its traditional use. These findings provide scientific support for its ethnomedicinal application and highlight its potential as a source of bioactive natural products.</p>