Polycyclic aromatic hydrocarbon (PAH)-diol-epoxide-DNA and -protein adducts have been prepared synthetically and subjected to analysis using spectrophotometric, spectrophotofluorimetric and immunological methods. In addition some of the PAH-DNA adducts have been used as both solid phase and fluid phase antigens in competitive and noncompetitive enzyme linked immunosorbant assays (ELISA) to identify human anti-PAH-DNA antibodies in normal healthy individuals. A series of validation studies for the detection of adducts by synchronous fluorescence spectroscopy (SFS) and an ultrasensitive enzyme-linked radioimmunoassay (USERIA) have shown that these methods are corroborative for model adducts studied in controlled systems. However, when applied to the detection of PAH-DNA adducts in human peripheral blood, discrepancies are observed that may be related to the presence in these samples of complex mixtures of adducts. The presence of multiple adduct types in biological macromolecules presents a major problem for both qualitative and quantitative analysis of human biological samples. Analysis of a synthetic mixture containing 5-PAH-DNA adducts has been shown to be possible using SFS, HPLC and computer-based spectral analysis. A method for the extraction of PAH-hemoglobin adduct hydrolysis products has also been developed in order to improve the sensitivity of this method by boosting the total adduct pool.
