A novel method for the small scale synthesis of N-acetoxy-2-acetylaminofluorene and its adducts

The carcinogen N-acetoxy-2-acetylaminofluorene (N-acetoxy-AAF) can be synthesized effectively in milligram amounts by reacting N-hydroxy-2-acetylaminofluorene (N-hydroxy-AAF) with acetic anhydride in a continuously mixed, closed 2-phase system consisting of ethyl ether/petroleum ether (2 : 1 v/v) and dilute sodium bicarbonate. The organic phase containing N-acetoxy-AAF can be added directly to 10 mm sodium citrate/20% ethanol to synthesize nucleoside or nucleotide adducts without the need to crystallize or resuspend N-acetoxy-AAF in a polar solvent. The advantage of the method is that it makes possible the synthesis of nucleic acid derivatives of N-acetoxy-AAF with high specific activity in small quantities.