Electron capture/gas chromatography (EC/GC) and high pressure liquid chromatography (HPLC) were used to detect the metabolites of Direct Black 38 (1937377) and Pigment Yellow 12 (6358856) in hamster and human urine. Analytic procedures included: extraction of the free aromatic amines and neutral compounds; alkaline hydrolysis of the aqueous phase and extraction of any hydrolyzed conjugates as free amines; and analysis of the free amines and acetylated metabolites directly by high pressure liquid chromatography or electron capture gas chromatography after conversion of the amines to heptafluorobutyryl derivatives. Metabolites in hamster and human urine were detected at concentrations as low as 1 part per billion. The authors recommend this approach for monitoring exposed workers since it will also detect any other derivatizable aromatic amines.
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