Separation and Quantitation of Acetaminophen and Its Metabolites in the Bile of Mice

Abstract

After a single dose of ~4C-acetaminophen (250 mg/kg, orally) was administered to bile duct cannulated mice, 18.1 + 2.4% of the radioactivity was recovered in the bile in the first five hours following administration. The biliary metabolites were separated and quantltated by a combination of paper and Sephadex gel G-10 column chromatography and reversed-phase high-performance liquid chromatography. The metabolites of acetaminophen identified were: glutathione (61.17 _+ 2.94%), glucuronide {6.89  2.04%), sulfate (13.00 +_ 1.51%), unchanged acetaminophen (7.55  0.39%), and unknown (11.66_+ 2.83%). The identification of acetaminophen glutathione in the bile represents the first time that this metabolite has been detected In biological fluid after acetaminophen administration.