A two year, comprehensive, quantitative investigation was conducted to analyze and identify the spatial distribution of petrogenic and biogenic hydrocarbons in sediments, surface waters, fish and shellfish of Biscayne Bay, Florida. The goal for the first year of the project was to establish baseline information to support oil spill impact assessment and clean-up. One hundred fifty-five sediment and eleven biota samples were collected. The areas sampled included the Miami River, Intracoastal Waterway, tidal flats, access canals and environmentally sensitive shorelines. The second year of the study centered on areas exhibiting petroleum contamination. These areas included the Miami River, Little River, Goulds Canal, Black Creek and Military Canal. Surface and subsurface sediment, biota and surface water were collected. Sample collection, analyses, and data handling for the two year project were conducted so that all information was court-competent and scientifically accurate. Chain of custody was maintained for all samples. Total hydrocarbon content of surface sediments ranged from below detection limits to a high of 2663.44 pg/g. Several sample stations contained petroleum contamination. The majority of biota samples exhibited hydrocarbon concentrations and characteristics that indicated little, if any, petroleum contamination. Surface water samples ranged from 0.78 to 64.47 ?g/L and several samples contained petroleum hydrocarbons. Our results indicate several areas of petroleum contamination. These areas are characterized by industrial complexes, port facilities, marinas, major boating routes and many of the major tributaries emptying into Biscayne Bay.