High-performance liquid chromatographic analysis of p-nitrophenol and its conjugates in biological samples.

M E Morris, S B Hansel

Index: J. Chromatogr. A. 532(2) , 285-93, (1990)

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Abstract

p-Nitrophenol (pNP) and its conjugated metabolites, generated in a perfused rat liver preparation, are readily separated and quantitated in serum perfusate and bile samples using a reverse-phase high-performance liquid chromatographic method. Serum perfusate samples can be analyzed following protein precipitation with acetonitrile: following protein precipitation with 1.5 M perchloric acid (1 part to 2 parts serum) there was degradation of pNP sulfate to pNP when samples were stored at room temperature. pNP can also be analyzed in blood perfusate samples following extraction with a number of organic solvents including ethyl acetate or isobutanol-methylene chloride (4:1, v/v). Rat liver perfusions at a constant input concentration of 40 microM demonstrated a high hepatic extraction ratio of pNP (mean of 0.90) due to the formation of the sulfate and glucuronide conjugates; no pNP glucoside was detected in perfusate or bile samples.