Relationship of rat urinary metabolites of N-nitrosomethyl-N-alkylamine to bladder carcinogenesis.

Nitrosomethylalkylamines with chain lengths from C4 (n-butyl-) to C14 (n-tetradecyl-) were each administered in three rats at doses equimolar with 12 mg of the butyl compound. All of the compounds administered to rats at this dose, twice a week for 30 weeks, induced tumors in 100% of the animals. Some of the compounds with even-numbered alkyl chains induced bladder tumors, and a connection was sought with the metabolites of these excreted in urine. The pooled 24-hr urine was extracted with ethyl acetate before and after acidification to provide a neutral fraction and a fraction containing nitrosoamino acids. The fraction containing the acids was analyzed by capillary gas chromatography and by gas chromatography-mass spectrometry after esterification with diazomethane; the neutral fraction was analyzed similarly. The principal metabolite of the nitrosoamines with odd-numbered chains was found in the acidic fraction and was identified as nitrosomethyl-2-carboxy-ethylamine. There were several acids in the mixtures derived from the nitrosamines with even-numbered chains, nitrososarcosine and nitrosomethyl-3-carboxypropylamine being the major components. There was no trend in the yields of the nitrosamino acids that could be correlated with the differences in carcinogenic potency between the nitrosamines; the maximum yield of acids was more than 30% (from the tetradecyl compound). The principal component of the neutral fraction (less than or equal to 1% of the nitrosomethylalkylamine administered) was nitrosomethyl-2-oxopropylamine. The yield of this compound increased with length of the even-numbered chain nitrosamines.