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Anthropogenic oil spills such as the 2010 Deepwater Horizon oil spill present a number of unique challenges in environmental remediation, detection, and monitoring of a wide range of toxicants in complex environments. We have previously reported the use of cyclodextrin derivatives to accomplish the selective extraction of polycyclic aromatic hydrocarbons (PAHs) and facilitate their detection via proximity-induced energy transfer. Reported herein is the ability of these cyclodextrins to operate in a real-world scenario: extracting PAHs from oil collected from an oil spill site and from tar ball extracts into crude seawater from the Narragansett Bay in Rhode Island. The ability of this system to operate in this complex environment highlights the practical significance of cyclodextrin-based systems, and a direct comparison of the results obtained in seawater with those obtained using a variety of aqueous solvent systems provides significant insights into the factors responsible for efficient performance.