Metabolism of the food-derived carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine by lactating Fischer 344 rats and their nursing pups.
BACKGROUND
An important class of dietary mutagens and carcinogens are the heterocyclic arylamine compounds that have been identified in a variety of cooked, protein-containing foods. Among these heterocyclic amines, 2-amino-1-methyl-6-phenylimidazo[4,5-b]-pyridine (PhIP) is potentially the most important carcinogen for human cancer risk. We have recently observed that PhIP-derived radioactivity is excreted into the breast milk of lactating rats administered [3H]PhIP.


OBJECTIVE
To better assess the significance of breast milk as a route of exposure of the newborn to dietary heterocyclic amines, we examined the metabolites of PhIP in breast milk and in urine of nursing pups.


METHODS
Lactating Fischer 344 rats with 5-day-old pups were given a single oral dose of 10 mg/kg of [3H]PhIP. We collected milk from the dams and urine from the pups and then analyzed the samples for metabolites of PhIP, using high-pressure liquid chromatography (HPLC). PhIP-DNA adduct levels in the tissues of the pups were determined by 32P-postlabeling analysis.


RESULTS
Three radioactive peaks were observed by HPLC separation of milk samples: an unidentified early eluting peak, 4'-hydroxy-PhIP, and PhIP. Four metabolites and the parent compound were found in urine of the pups nursed by dams given radiolabeled PhIP: PhIP-4'-O-glucuronide, PhIP-4'-sulfate, 4'-hydroxy-PhIP, and N2-hydroxy-PhIP-N3-glucuronide. 4'-Hydroxy-PhIP and its conjugates contributed approximately 60% of the radioactivity found in the urine. By 32P-postlabeling analysis, PhIP-DNA adducts were detected in spleen, lung, heart, kidney, liver, and stomach of pups at mean levels ranging from 0.06 to 0.55 adducts/10(7) nucleotides.


CONCLUSIONS
The large percentage of 4-hydroxy-PhIP and its conjugates in the urine indicates that 5-day-old pups detoxify PhIP and further metabolize 4'-hydroxy-PhIP obtained from the breast milk. The presence of the glucuronide conjugate of N-hydroxy-PhIP in the urine of pups and the lack of detectable conjugate or N-hydroxylamine itself in breast milk suggest that PhIP from breast milk undergoes metabolic activation via N-hydroxylation in 5-day-old rat pups. This conclusion was further supported by the observation that hepatic S9 fractions from the pups activated PhIP to a mutagen in the Ames Salmonella mutagenicity assay and by the presence of PhIP-DNA adducts in the tissues of the pups.


CONCLUSIONS
The findings reported here may have carcinogenic and toxicologic implications for the offspring of women who breast-feed and consume a diet rich in cooked meat.
Davis
C D
CD
Laboratory of Experimental Carcinogenesis, National Cancer Institute, Bethesda, Md 20892.
Ghoshal
A
A
Schut
H A
HA
Snyderwine
E G
EG
D000818:Animals; D000833:Animals, Suckling / Q000652:urine*; D002273:Carcinogens / Q000378:metabolism*; D005260:Female; D007093:Imidazoles / Q000378:metabolism* / Q000652:urine; D007774:Lactation / Q000378:metabolism*; D008892:Milk / Q000378:metabolism*; D051381:Rats; D011916:Rats, Inbred F344; D013997:Time Factors
D002273:Carcinogens; D007093:Imidazoles; C049584:2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine