This communication presents the isolation and detailed structural elucidation of a novel sesquiterpene lactone derived from the leaves of Artemisia afra, a medicinal plant extensively used in traditional African medicine. The structure of the compound was determined through comprehensive spectroscopic analyses, including one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) spectroscopy, alongside high-resolution mass spectrometry (HRMS). These complementary techniques enabled the clear identification of the compound's molecular architecture and stereochemistry, thereby enriching the phytochemical profile of A. afra and highlighting its potential pharmacological relevance. The HRMS data further supported the proposed molecular formula, with an observed mass of 285.1098 [M+Na⁺], closely matching the calculated value for C₁₅H₁₈O₄Na (285.1103), thereby confirming the compound’s structural integrity.