Reports by several investigators describing the biological activity of coal fly ash have presented a variety of results which in some cases are conflicting. The biological activity of coal fly ash may differ because of one or more of the following factors: (1) the samples studied were from different sources; (2) the samples were prepared for bioassay differently; (3) the sampling method differed, and, therefore, collected samples were different in chemical or physical properties which affect the biological activity. Several variables involved in coal fly ash studies -- source, sample collection land preparation methods, bioassay method -- are undoubtedly responsible for the diversity of biological effects observed. The objective of this study were to examine the sample preparation and collection factors which may affect the observed biological activity caused by coal fly ash and to evaluate the mutagenicity and cytotoxicity of fluidized-bed combustion (FBC) fly ash from experimental and commercial units. The bioassays used in this study were the Ames Salmonella plate incorporation test for mutagenicity and the rabbit alveolar macrophage (RAM) system for cytotoxicity.