Arylamidation and arylation by the carcinogen N-2-fluorenylacetamide: a sensitive and rapid radiochemical assay

Abstract

N-acetoxy-N-arylacetamides, which are generally considered as an ultimate carcinogenic form of the corresponding N-arylacetamides, react with the cellular macromolecules (nucleic acids, proteins, etc.) to give two types of adducts: (I) arylamidation and (II) arylation addition products. In this paper, we present a radiochemical determination of the amount of N-2-fluorenylacetamide bound to DNA via arylamidation or arylation, respectively. This assay is based upon the difference of stability under weak alkali hydrolysis conditions (0.1 N NaOH, 75 degrees C, 2 h) of the specifically 14C-labeled N-acetyl group of the N-2-fluorenylacetamide residue linked to the macromolecule either via arylamidation or arylation. Native DNA which has been reacted with N-acetoxy-N-2-[14C]acetylaminofluorene exhibits 16% of the fluorene adducts linked to the bases via arylation. On the other hand, denatured DNA reacts with the fluorene derivative to give almost only arylamidation addition products.