Quantitative analysis of N-(guanin-8-yl)-N-acetyl-2-aminofluorene and N-(guanin-8-yl)-2-aminofluorene in modified DNA by hydrolysis in trifluoroacetic acid and high pressure liquid chromatography.

N-acetyl-2-aminofluorene (AAF)- and 2-aminofluorene (AF)-modified DNA was hydrolyzed in dry trifluoroacetic acid (TFA). The hydrolysate was analyzed by high-pressure liquid chromatography. Using this procedure N-(guanin-8-yl)-[G-3H]2-aminofluorene was released quantitatively from DNA, modified by the reaction with [G-3H]N-hydroxy-2-aminofluorene. From DNA that had been reacted with [G-3H]N-acetoxy-N-acetyl-2-aminofluorene, 70% of the total bound radioactivity was isolated as N-guanin-8-yl)-[G-3H]-N-acetyl-2-aminofluorene. Thirty eight percent of the DNA bound radioactivity after i.p. injection of [G-3H]N-acetyl-2-aminofluorene followed by isolation of the rat liver DNA and subsequent hydrolysis in trifluoroacetic acid was identified as N-(guanin-8-yl)-[G-3H]-N-acetyl-2-aminofluorene. In this DNA the corresponding deacetylated compound N-(guanin-8-yl)-[G-3H]2-aminofluorene could not be detected.