title: Bioassay of N-nitrosodiphenylamine for possible carcinogenicity.
abstract: A bioassay of N-nitrosodiphenylamine for possible carcinogenicity was conducted by administering the test chemical in feed to F344 rats and B6C3F1 mice. Groups of 50 rats of each sex were administered N-nitrosodiphenylamine at one of two doses, either 1,000 or 4,000 ppm, for 100 weeks. Matched controls consisted of 20 untreated rats of each sex. All surviving rats were killed at the end of administration of the test chemical. Groups of 50 male mice were administered N-nitrosodiphenylamine at one of two doses, either 10,000 or 20,000 ppm, for 101 weeks. Groups of 50 female mice were administered the test chemical at one of two doses, initially 5,000 or 10,000 ppm, for 38 weeks. Because of excessive depression in the amount of mean body weight gained in the dosed groups, the doses for the females were then reduced to 1,000 and 4,000 ppm, respectively, and administration at the lowered doses was continued for 60 weeks. The time-weighted average doses for the female mice were either 2,315 or 5,741 ppm. Matched controls consisted of 20 untreated mice of each sex. All surviving mice were killed at the end of administration of the test chemical. Mean body weights of dosed rats and mice of each sex were lower than those of corresponding controls, and were dose related throughout the bioassay, except for those of female rats during the first part of the bioassay. Mortality was dose related in the female rats, but was not affected when the test chemical was administered to the male rats or the male or female mice. Survival at the end of the bioassay was 64% or greater in the dosed and control groups of rats and mice of each sex, and sufficient numbers of animals were at risk in all groups for the development of late-appearing tumors. Transitional-cell carcinomas of the urinary bladder occurred at incidences that were dose related (P</= 0.001) in both male and female rats, and in direct comparisons the incidences of these tumors in the high-dose groups of each sex were significantly higher (P</= 0.001) than those in the corresponding controls (males: controls 0/19; low-dose 0/46; high-dose 16/45; females: controls 0/18; low-dose 0/48; high-dose 40/49). The possible mechanism by which these tumors were induced, such as calculi formation in the bladder or nitrosation of amines present in feed to a carcinogenic nitrosoamine, is unknown. Fibromas of the integumentary system occurred in male rats at incidences that were dose related (P=0.003), although in direct comparisons the incidences of these tumors in the individual dosed groups were not significantly higher than those in the control group (controls 1/20, or 5%; low-dose 1/50, or 2%; high-dose 10/50, or 20%). The incidence of fibromas of the integumentary system in historical-control male F344 rats at this laboratory is 6/285, or 2%. These results suggest an association of the fibromas in the male rats with the administration of the test chemical. No tumors occurred in the mice of either sex at incidences that were significantly higher in the dosed groups than in the corresponding control groups. The only changes related to compound administration were chronic inflammatory lesions in the urinary bladders of dosed mice. It is concluded that under the conditions of this bioassay, N-nitrosodiphenylamine was carcinogenic for both sexes of F344 rats, including transitional-cell carcinomas of the urinary bladder, but was not carcinogenic for B6C3F1 mice of either sex.