Chromatographic separation of the urinary metabolites of the carcinogen N-2-fluorenylacetamide into free compounds, sulfuric acid, and glucuronic acid esters.

The urinary metabolites of N-2-fluorenylacetamide were separated by chromatography on an alumina column into three classes: (a) unconjugated materials, amounting to 5-10 per cent of the urinary activity; (b) sulfuric acid conjugates (20-25 per cent); (c) glucuronic acid conjugates (60-70 per cent). Evidence for the structure of these materials was adduced by solvent partition, paper chromatography, and hydrolysis by acid and by appropriate enzymes. Taka-diastase, often used to split sulfuric acid esters, did not quantitatively hydrolyze the sulfates of metabolites of N-2-fluorenylacetamide under our experimental conditions.