Immunochemical detection of rodent hepatic and urinary metabolites of cooking-induced food mutagens.
Monoclonal antibodies, previously selected to bind either 2-amino-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-PhIP antibodies bound N-hydroxy-PhIP, N-sulfinamide-glutathione-PhIP and N-hydroxy-glucuronide-PhIP. Immunoaffinity 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 conjugate metabolites, e.g. N-sulfamate-MeIQx and N(OH)-gluc-PhIP, while others are as yet unidentified. Immunoaffinity chromatography is demonstrated as a promising means of selectively concentrating metabolites of PhIP and MeIQx from urine.
Vanderlaan
M
M
Biomedical Sciences Division, Lawrence Livermore National Laboratory, Livermore, CA 94550.
Alexander
J
J
Thomas
C
C
Djanegara
T
T
Hwang
M
M
Watkins
B E
BE
Wallin
H
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D002846:Chromatography, Affinity; D005502:Food; D006358:Hot Temperature; D007093:Imidazoles / Q000276:immunology / Q000378:metabolism* / Q000652:urine; D007150:Immunohistochemistry; D008099:Liver / Q000378:metabolism*; D008460:Meat / Q000633:toxicity*; D009153:Mutagens / Q000378:metabolism* / Q000652:urine; D011804:Quinolines / Q000276:immunology / Q000378:metabolism* / Q000652:urine; D051381:Rats; D011916:Rats, Inbred F344
D000911:Antibodies, Monoclonal; D007093:Imidazoles; D009153:Mutagens; D011804:Quinolines; C049584:2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine; C036989:2-amino-3,4-dimethylimidazo(4,5-f)quinoline