We published recently an integrated procedure for applying chemical marker analysis to characterize the microbiology of indoor environments comprising a scheme for extraction and analysis of markers of endotoxin, peptidoglycan/bacterial biomass, and fungal biomass. In the present paper, we report some significant improvements and also new possibilities of the described approach. We found that while 3-hydroxy fatty acids (3-OH FAs) of 10-14 carbon chain lengths are useful endotoxin markers, longer 3-OH FAs (i.e. with 16 carbon atoms and more) may rather serve as markers of Actinobacteria. We introduced 13C-labeled 3-hydroxytridecanoic acid, from labeled Pectinatus cerevisiiphilus, as an internal standard to improve quantification of the 3-OH FAs in the gas chromatography-mass spectrometry analysis. Finally, in experiments aiming to identify a suitable method for collection of house dust for chemical marker analysis, we found that the marker compositions of dusts sedimented on plexiglass plates that were spatially well-distributed in a studied room at different heights above floor level, were undistinguishable. This type of sampling thus appears to be well suited for use, e.g. in epidemiological studies. In summary, the presented work describes important new capabilities of chemical marker analysis in defining human exposure to microorganisms in indoor environments. We are developing an integrated methodology for characterizing the microbiology of indoor environments where specific microbial monomeric constituents in building materials and inhalable house dust particles are determined by using mass spectrometry-based methods. The methodology should represent a firm basis in research aiming to relate microbial exposure indoors to well-being and health.
