SOLID-MATRIX, ROOM-TEMPERATURE PHOSPHORESCENCE IDENTIFICATION AND QUANTITATION OF THE TETRAHYDROTETROLS DERIVED FROM THE ACID-HYDROLYSIS OFBENZO[A]PYRENE-DNA ADDUCTS FROM HUMAN LUNG

A new method, suitable for human biomonitoring, that uses room tempera ture phosphorescence for the detection of DNA damage by carcinogenic m etabolites of polycyclic aromatic hydrocarbons is described. Samples o f human lung DNA (1 mg) that had been subjected to immunoaffinity chro matography (anti-benzo[a]pyrene-diol-epoxide deoxyguanosine monoclonal antibodies) were acid hydrolyzed (0.1 N HCl, 90 degrees C, 3 h) and t he resulting DNA lung hydrolyzates separated by high performance liqui d chromatography. Relevant fractions were combined with a solid matrix support which consisted of a mixture of alpha-cyclodextrin (alpha-CD) :NaCl (1:9) or alpha-CD:TINO3: aNO(3) (1:1:8). The dried and powdered sample-matrix material was analyzed by phosphorescence spectroscopy at room temperature. Certain fractions of human lung samples were found to contain materials that yielded phosphorescence spectra that were in distinguishable from those produced when an authentic r-7, t-8, t-9, c -10-tetrahydroxy-7,8,9, 10-tetrahydrobenzo[a]pyrene reference standard was analyzed. The data confirm previous studies that have reported th e presence of r-7, t-8 t-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene- DNA adducts in human tissues at levels of 1 adduct/l0(7)-10(8) nucleot ides. The alpha-cyclodextrin solid matrix, room temperature phosphores cence technique was performed with a commercially available instrument , but is 50 times more sensitive than the synchronous fluorescence spe ctroscopic technique previously used.