4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and its analogues substituted with deuterium at the methylene carbon, 4,4-dideutero-4-(methylnitrosamino)-1-(3-pyndyl)-1-butanone [4,4-D2)NNK], and the methyl carbon, 4-(trideuteromethylnitrosamino)-1-(3-pyridyl)-1-butanone [(CD3NNK) adjacent to the N-nitroso group were tested for tumorigenlcity in F344 rats. Each compound was administered by 60 s.c. injectioiis over a 20-week period such that the total doses were either 1.0 or 0.33 mmol/kg. The experiment was terminated after 104 weeks. Survival of the rats treated with the higher dose of (4,4-D2)NNK was significantly less than survival the groups treated with the same doses of NNK or (CD3)NNK Target tissues were liver, lung and nasal cavity for all three compounds. The higher dose of (4,4-D2)NNK induced higher numbers of nasal tumors and malignant nasal tumors than did NNK. The lower dose of (4,4-D2)NNK induced a higher number of nasal tumors than did NNK. No other significant differences in tumor incidence were observed. The results suggest that 4-(3-pyridyl)-4-oxobutylation DNA might be important in induction of nasal cavity tumors by NNK.