One of the most widely studied organisms responsible for Harmful Algal Blooms (HABs) is the marine dinoflagellate Karenia brevis. This organism produces neurotoxic compounds known as brevetoxins. A related dinoflagellate, Karenia papilionacea, has been reported to occasionally co-bloom with K. brevis but has received little attention as a possible toxin producing species. Therefore, our aim was to investigate the toxin profile for K. papilionacea. A toxic fraction was identified using a cell based cytotoxicity assay and the toxin was isolated and identified as the ladder frame polyether brevetoxin-2 (PbTx-2) using mass spectrometry (MS) and nuclear magnetic resonance (NMR). Toxin production in K. papilionacea increased in response to hypoosmotic stress, as previously observed in K. brevis.