Concentrations of heavy metals (Cu, Zn, Cd, Pb and As) were measured in water, sediment and two fish species, Sparus aurata and Solea senegalensis, from the estuary of Tinto and Odiel rivers in Huelva (Spain), one of the most metallic polluted estuaries in Europe. As a forward step to understand metal bioavailability and assess the potential impact on aquatic biota, a study of heavy metal speciation of sediments and water were achieved. High levels of total and dissolved Zn and Cu were found in water and high pollution of Zn, Pb, As and Cu were found in sediments. Availability of metals was established as following ranking: Cd > Zn > Cu > Pb in both water and sediment. In addition, the effect of this pollution was studied by evaluation of metal bioaccumulations and correlations obtained between metal levels in fractions of water and sediment and metal levels in fish tissues. High Cu and Zn levels were observed in liver tissue of both species, in according with higher total content and more available metals in water and sediment. Correlations among metal content in tissues and different fractions of metal in water for S. aurata and sediment for S. senegalensis were found.