Two novel polyacetylenic compounds were successfully separated from the dichloromethane extract of the aerial parts of Bidens parviflora Willd., known for its ethnomedicinal use in traditional Dai medicine. These compounds were structurally elucidated and identified as (2R)-trideca-3E,5Z,11E-triene-7,9-diyne-1,2,13-triol (1), and (2R)-trideca-11E-ene-5,7,9-triyne-1,2,13-triol (2). Their chemical structures were confirmed through a comprehensive analysis involving ultraviolet (UV) spectroscopy, infrared (IR) spectroscopy, high-resolution electrospray ionization mass spectrometry (HR-ESI-MS), as well as detailed one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) spectroscopic data, and measurements of specific optical rotation.