IMPROVEMENT OF REVERSE-PHASE HIGH-PRESSURE LIQUID-CHROMATOGRAPHIC RESOLUTION OF BENZO[A]PYRENE METABOLITES USING ORGANIC AMINES - APPLICATION TO METABOLITES PRODUCED BY FISH

Addition of primary organic amines, such as n-butylamine, to the mobile phase altered the capacity factors and selectivity of benzo[a]pyrene metabolites obtained with reverse-phase high pressure liquid chromatography on an ODS column. Separation of benzo[a]pyrene phenols in particular was improved with 8 of the 10 available metabolites resolved, including those known to be biologically produced. The method offers sufficiently improved resolution or convenience that it should prove useful in comparative studies of metabolism of benzo[a]-pyrene and other polynuclear aromatic hydrocarbons. Applying the method to analysis of benzo[a]pyrene metabolites produced in vitro by hepatic microsomes from the marine fish Stenotomus versicolor indicated the principal phenolic derivatives produced by this fish were 1-hydroxy-, 3-hydroxy-, 7-hydroxy-, and 9-hydroxybenzo[a]pyrene. © 1979.