Identification of fluorescent 2'-deoxyadenosine adducts formed in reactions of conjugates of malonaldehyde and acetaldehyde, and of malonaldehyde and formaldehyde.

2'-Deoxyadenosine was reacted with malonaldehyde in the presence of formaldehyde or acetaldehyde. The reactions were carried out at 37 degrees C in aqueous solution at acidic conditions. The reaction mixtures were analyzed by HPLC. In both reactions, two major products were formed. The reaction products were isolated and purified by C18 chromatography, and their structures were characterized by UV absorbance, fluorescence emission, (1)H and (13)C NMR spectroscopy, and mass spectrometry. The reaction products isolated from the mixture containing formaldehyde, malonaldehyde, and deoxyadenosine were identified as 3-(2'-deoxy-beta-D-ribofuranosyl)-7H-8-formyl[2,1-i]pyrimidopurine (M(1)FA-dA) and 9-(2'-deoxy-beta-D-ribofuranosyl)-6-(3,5-diformyl-1, 4-dihydro-1-pyridyl)purine (M(2)FA-dA). In the reaction mixture consisting of acetaldehyde, malonaldehyde, and deoxyadenosine, the identities of the products were determined to be 3-(2'-deoxy-beta-D-ribofuranosyl)-7-methyl-8-formyl[2, 1-i]pyrimidopurine (M(1)AA-dA) and 9-(2'-deoxy-beta-D-ribofuranosyl)-6-(3,5-diformyl-4-methyl-1, 4-dihydro-1-pyridyl)purine (M(2)AA-dA). The yields of the compounds were 1.8 and 0.7% for M(1)FA-dA and M(2)FA-dA, respectively, and 6.8 and 10% for M(1)AA-dA and M(2)AA-dA, respectively. All compounds exhibited marked fluorescent properties. These findings show that in addition to direct reaction of a specific aldehyde with 2'-deoxyadenosine, aldehyde conjugates also may react with the base. Although three of the adducts (M(1)FA-dA, M(2)FA-dA, and M(1)AA-dA) could not be detected in reactions carried out under neutral conditions, the possibility that trace amounts of the adducts may be formed under physiological conditions cannot be ruled out. Therefore, conjugate adducts must be considered in work that aims at clarifying the mechanism of aldehyde genotoxicity.