A simple, rapid procedure for the determination of organic mercury in sediments, plants and fish tissues has been developed and validated. Extraction and separation of organic mercury compounds from the sample matrix was achieved by an established procedure based on an acid leaching of the sample (H 2 SO 4 /KBr/CuSO 4 ), followed by extraction of the organic mercury halide with toluene and back-extraction with an aqueous solution of thiosulphate. Detection and quantification of mercury, in the liquid extracts, was made by atomic absorption spectrometry (AAS), following thermal decomposition of the sample. The method was evaluated using Certified Reference Material (CRM) BCR 463 (tuna fish), BCR 580 (estuarine sediment), IAEA-140TM (sea plant homogenate) and NRCC TORT-2 (lobster hepathopancreas). The recovery factors for organic mercury in all tested CRM were between 81-107%. The precision of the method has relative standard deviations of less than 10% for sediments and fish tissues and of less than 16% for plant material. The method was successfully applied to natural samples of sediments, plants, macroalgae and fish tissues collected from an estuarine ecosystem and could, therefore, be used for routine analyses.