Methods for Testing Compounds for DNA Adduct Formation

Abstract

This article, based on a presentation on DNA adduct detection given at a Genetic Toxicology Association workshop, is an overview of methods used for testing compounds for DNA adduct formation. A DNA adduct study may be initiated on a case by case basis when there are conflicting results within the standard battery of genetic toxicology tests or when tumors are detected in the animal bioassay for nongenotoxic compounds. Methods for adduct detection include the 32P-postlabeling assay, the use of radioactive test chemicals, physicochemical methods, and immunoassays. Of these, the 32P-postlabeling assay and the use of radiochemicals are discussed in greater detail, since only these two methods are readily applicable to test a compound for the formation of uncharacterized DNA adducts. The other methods are applicable to those adducts that have been chemically characterized or that contain a fluorophore or electrochemically active groups. Evaluation of mutagenic and carcinogenic risk from DNA adducts would require the understanding of various parameters, including the chemical nature, quantity and stability of adducts, proliferation rates for target cells to fix adducts into mutations, mutagenic and repair efficiencies of adducts, and the extent of modifications in critical genes. Since such data cannot be readily obtainable, the toxicological risk from uncharacterized adducts is difficult to assess.