An Analytical Approach to Metabolic Profiling of Aromatic Compounds Using Liquid Chromatography

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Abstract
An analytical approach to metabolic profiling of aromatic compounds is described for both conjugated and “free” metabolites in biological systems. Initially, an ethyl acetate extraction removes the less polar metabolites. A salting-out procedure using Sephadex G-10 is combined with reverse-phase high-performance liquid chromatography (HPLC) to analyze the water-soluble conjugates directly using sequential uv and fluorescence detection. The “free” metabolites are also derived from individual conjugate peaks by enzymatic or hydrolytic procedures and then re-chromatographed by HPLC. Metabolites in the ethyl acetate fraction are similarly analyzed by reverse-phase HPLC. The utility of this method is demonstrated for 2-acetylaminofluorene (2-AAF and 4-ethylsulfonyl-naphthalene-1-sulfonamide (ENS), two compounds of interest in the study of murine bladder carcinogenesis.