Registration Dossier

Toxicological information

Toxicity to reproduction

Currently viewing:

Administrative data

Endpoint:
one-generation reproductive toxicity
Remarks:
based on generations indicated in Effect levels (migrated information)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)

Data source

Reference
Reference Type:
publication
Title:
Chronic Toxicity, Teratologic, and Reproduction Studies with Hair Dyes
Author:
THEODORE WERNICK, BEN MARR LANMAN AND JEAN LOUIS FIUUX
Year:
1975
Bibliographic source:
TOXICOLOGY AND APPLIED PHARMACOLOGY 32,450-460 (1975)

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: as mentioned below
Principles of method if other than guideline:
To assess the reproductive toxicity of the test chemical in rats
GLP compliance:
no
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report):2-nitro-p-phenylenediamine- Substance type:Organic- Physical state:Solid

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALSSource: Charles River breeding laboratoriesAge at study initiation: (P) x wks; (F1) x wks : No dataWeight at study initiation: (P) Males: x-x g: 240 to 280 g Females: x-x g: 180 to 220 g.Fasting period before study: during mating periodHousing: Cages Use of restrainers for preventing ingestion (if dermal): yes/no : No Diet (e.g. ad libitum): basal diet of Purinalaboratory chow, ad libitumWater (e.g. ad libitum): Tap water, ad libitumAcclimation period: No dataENVIRONMENTAL CONDITIONSTemperature (°C): No dataHumidity (%): No dataAir changes (per hr):No dataPhotoperiod (hrs dark / hrs light):No dataIN-LIFE DATES: From: To: No data

Administration / exposure

Route of administration:
oral: feed
Type of inhalation exposure (if applicable):
not specified
Vehicle:
other: basal diet of Purina laboratory chow
Details on exposure:
DIET PREPARATIONRate of preparation of diet (frequency): Diets were prepared twice weekly.Mixing appropriate amounts with (Type of food): The composite was mixed into the basal diet of Purina laboratory chow at concentrations of 0, 97.50 and 390 mg/kgbw
Details on mating procedure:
- M/F ratio per cage:One male was placed in a cage with two females from 4 PM to 8 AM the following day.- Length of cohabitation:from 4 PM to 8 AM. This procedure was continued until copulation was confirmed.- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancyappearance of sperm in a vaginal smear (day 0 of pregnancy- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.Males were rotated within their dietary groups at I0-day intervals until conception wasconfirmed or until each female had been mated with a maximum of two males. If oneof the two females caged with the male became pregnant, the male was consideredfertile. A female was considered infertile if she failed to become pregnant after matingwith two different males for 10 days each.- Further matings after two unsuccessful attempts: [no / yes (explain)]Males were rotated within their dietary groups at I0-day intervals until conception wasconfirmed or until each female had been mated with a maximum of two males. If one of the two females caged with the male became pregnant, the male was considered fertile. A female was considered infertile if she failed to become pregnant after mating with two different males for 10 days each.- After successful mating each pregnant female was caged (how):Individual cages- Any other deviations from standard protocol:No data
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No data
Duration of treatment / exposure:
The study was divided into two parts. In Part I, the females received the basal diet from 8 wk prior to mating through the weaning of their litters. The males siring these litters were fed the test diets for 8 wk prior to mating and during the mating period. In Part II, males received the basal diet for 8 wk prior to and during mating,while the females received the test diets 8 wk prior to mating and during gestation and21 days of lactation.
Frequency of treatment:
Daily
Details on study schedule:
No data
Doses / concentrations
Remarks:
Doses / Concentrations:0, 195 and 780 mg/kg bw/day Basis:no data
No. of animals per sex per dose:
six groups of 10 males and 20 females each.
Control animals:
not specified
Details on study design:
No data
Positive control:
No data

Examinations

Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes / No / No data: No dataTime schedule: No dataCage side observations checked in table [No.?] were included.: No dataDETAILED CLINICAL OBSERVATIONS: Yes / No / No data: YesTime schedule: No dataBODY WEIGHT: Yes / No / No data: YesTime schedule for examinations:No dataFOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): YesFood consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes / No / No data: No dataCompound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data: No dataWATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes / No / No data: No dataTime schedule for examinations: No data
Oestrous cyclicity (parental animals):
No data
Sperm parameters (parental animals):
No data
Litter observations:
Yes
Postmortem examinations (parental animals):
SACRIFICEMale animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]: No data Maternal animals: All surviving animals [describe when, e.g. after the last litter of each generation was weaned.]: One female pregnant by each male was killed by chloroform inhalation on day 13 of her pregnancy to obtain information regarding the early stages of gestation.The uterus was examined for the number and distribution of embryos, the presence of empty implantation sites, and the number of embryos undergoing resorption. Each embryo was examined under a dissecting microscope. The remaining dams were allowed to deliver normally. A necropsy was performed on all females that did not deliver a litter to determine whether pregnancy had occurred. The duration of gestationwas noted and the litters’ were examined for numbers of live and stillborn pups andgross abnormalities.
Postmortem examinations (offspring):
SACRIFICEThe F1 offspring not selected as parental animals and all F2 offspring were sacrificed at [#?] days of age.: F1 : The litters’ were examined for numbers of live and stillborn pups and gross abnormalities.These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:The litters were examined for numbers of live and stillborn pups and gross abnormalities. The pups were weighed at birth, and at 4 and 21 days. At 21 days all surviving pups were killed by chloroform inhalation and examined grossly for abnormalities.
Statistics:
The data were subjected to statistical analysis, using the 95% confidence level. The methods used included chi square test, analysis of variance and t test, and the Fisher exact probability testFemale Fertility Index: (No of pregnant females/No. of females mated ) as percentMale Fertility Index: (No of males siring the litter/ No. mated to fertile females) as percentGestation Indices =%age of pregnancies resulting in litters cast liveViability Indices: %age of pups cast alive that survived at 4 daysLactation Indices: %age of pups alive at 4 days that survived to weaning at 21 days
Reproductive indices:
Part I: Female fertility indices: 85,95,65 ;Male fertility indices: 90, 100, 80Gestation Indices: 100,100,100Part II: Female fertility index: 95,90,95Gestation Index: 100,100,100
Offspring viability indices:
Part I:Viability Indices: 98,99,96Lactation Indices: 98,100,98Part IIViability Indices: 92,86,92Lactation Indices: 99,98,99

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
no effects observed
Description (incidence and severity):
No signs of toxicity were seen throughout the study.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
At the dietary concentrations fed, there were no effects on food consumption and body weight gains of either males or females.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
At the dietary concentrations fed, there were no effects on food consumption and body weight gains of either males or females.
Organ weight findings including organ / body weight ratios:
not specified
Histopathological findings: non-neoplastic:
not specified
Other effects:
not specified

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
effects observed, treatment-related
Description (incidence and severity):
The female fertility index in the high dosage group in Part I and the average pup weight in the high dosage group in Part II were lower than the control values, but the differences were not statistically significant at the 95 % confidence level.

Effect levels (P0)

Dose descriptor:
NOAEL
Effect level:
616 mg/kg diet
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: NO effect on body weight gains, food consumption and fertility

Results: F1 generation

General toxicity (F1)

Clinical signs:
not specified
Mortality / viability:
not specified
Body weight and weight changes:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
The female fertility index in the high dosage group in Part I and the average pup weight in the high dosage group in Part II were lower than the control values, but the differences were not statistically significant at the 95 % confidence level.
Histopathological findings:
not specified

Effect levels (F1)

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
616 mg/kg diet
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effect on live pups , survival, pup body weights and gross pathology

Overall reproductive toxicity

Reproductive effects observed:
not specified

Applicant's summary and conclusion

Conclusions:
NOAEL for F1 and F2 generation was considered to be 616 mg/kg when Sprague- Dawley CD male and female rats treated with 2-nitro-p-phenylenediamine orally in feed for 13 weeks.
Executive summary:
in a Fertility and reproductive study, Sprague- Dawley CD male and female rats were treated with 2-nitro-p-phenylenediamine in the concentration of 0, 195 and 616 mg/kg bw/day orally in feed. No significant effect food consumption and body weight gains of male and female were observed as compared to control. Similarly, no effect on male and female fertility, length of gestation, numbers of females with resorption sites, live pups per litter, pup body weights, and pup survival were observed as compared to control. Decrease in female fertility index and average pup weight was observed at 616 mg/kg bw/day as compared to control, but the differences were not statistically significant at the 95 % confidence level. In addition, no effects on abnormal pups were seen upon dissection of embryos after 13 days of gestation or upon gross examination at weaning after 21 days. Therefore, NOAEL for F1 and F2 generation was considered to be 616 mg/kg when Sprague- Dawley CD male and female rats treated with 2-nitro-p-phenylenediamine orally in feed for 13 weeks.