Immunochemical detection of rodent hepatic and urinary metabolites of cooking-induced food mutagens

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Abstract
Monoclonal antibodies, previously selected to bind either 2-ainino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) or 2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine (PhIP) were evaluated to determine their binding properties with several known metabolites of these compounds purified from rat hepatocyte cultures. Both 2-N- and 5-substituted MeIQx metabolites were recognized by antibodies AIA-2 and AIA-11, while antibodies AIA-1 and AIA-12 bound N-substituted metabolites only. Anti-PuIP antibodies bound N-hydroxy PhIP, N-sulfinamide-glutathione-PhIP and N-hydroxy glucuronide-PhIP. imniunoaffinity columns incorporating these antibodies were used to concentrate and purify both the unmetabolized parent compounds and several relatively non-polar metabolites from the urine of rats exposed either to MeIQx or PhIP. Several of these metabolites corresponded with available standards of previously identified polar con jugate metabolites, e.g. N-sulfamate-MeIQx and N(OH)-gluc PhIP, while others are as yet unidentified. Inununoaffinity chromatography is demonstrated as a promising means of selectively concentrating metabolites of PhIP and MeIQx from urine.

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