<p>The rhizomes of <i>Atractylodes macrocephala</i> are used as both a food source and traditional Chinese medicine in China. A phytochemical investigation was carried out on wild <i>A. macrocephala</i> grown in Qimen County in eastern China, and yielded a novel bisesquiterpenoid lactone, namely, biatractylenolide II (<b>1</b>), along with two known compounds, atractylenolide II (<b>2</b>) and taraxeryl acetate (<b>3</b>). The structure and relative configuration of the new compound were elucidated mainly by 1D and 2D NMR spectroscopic methods in combination with HRESIMS experiments. This paper describes the isolation and structural elucidation of the new bisesquiterpenoid lactone (<b>1</b>).</p