Dampness in buildings has been linked to adverse health effects, but the specific causative agents are unknown. Mycotoxins are secondary metabolites produced by molds and toxic to higher vertebrates. In this study, mass spectrometry was used to demonstrate the presence of mycotoxins predominantly produced by Aspergillus spp. and Stachybotrys spp. in buildings with either ongoing dampness or a history of water damage. Verrucarol and trichodermol, hydrolysis products of macrocyclic trichothecenes (including satratoxins), and trichodermin, predominately produced by Stachybotrys chartarum, were analyzed by gas chromatographytandem mass spectrometry, whereas sterigmatocystin (mainly produced by Aspergillus versicolor), satratoxin G, and satratoxin H were analyzed by high-performance liquid chromatography-tandem mass spectrometry. These mycotoxin analytes were demonstrated in 45 of 62 building material samples studied, in three of eight settled dust samples, and in five of eight cultures of airborne dust samples. This is the first report on the use of tandem mass spectrometry for demonstrating mycotoxins in dust settled on surfaces above floor level in damp buildings. The direct detection of the highly toxic sterigmatocystin and macrocyclic trichothecene mycotoxins in indoor environments is important due to their potential health impacts.Microorganisms are thought to be involved in health problems connected to damp buildings. However, the causative microbiological agents are unknown (22). Many molds that thrive in damp indoor environments are potent mycotoxin producers and may play a role in the reported adverse health effects ( 1,5,17,23,24,26,30). Mycotoxins are secondary metabolites, e.g., produced to give molds strategic advantages over encroaching organisms. Examples are sterigmatocystin (STRG), a carcinogenic mycotoxin produced mainly by Aspergillus versicolor; satratoxin G (SATG) and satratoxin H (SATH), which are cytotoxic mycotoxins produced by Stachybotrys chartarum; and citrinin, gliotoxin, and patulin, produced by, e.g., Aspergillus spp. and Penicillium spp. The latter three mycotoxins have been shown to be immunomodulatory, causing a polarization in cytokine production towards a Th2 phenotype (36), and citrinin caused depletion of intracellular glutathione at nontoxic concentrations (18). Based on spore counts, the airborne mycotoxin concentrations found in damp buildings have been estimated to be insufficient for causing adverse health effects (20). However, indoor molds may fragment into very small airborne mycotoxin-containing particles, resulting in up to a 500-fold larger exposure than assumed previously (4,11,21,32). In addition, Cho et al. (7) showed that the respiratory deposition of S. chartarum fragments was over 200-fold higher than that of spores in adults and an additional 4 to 5 times higher in infants. These aerosolized fragments could potentially also be the source of allergens (13).S. chartarum and A. versicolor are two commonly encountered molds in buildings with moisture problems (9,12,...