4-(phenylazo)benzene-1,3-diamine

Toxicity to reproduction

Administrative data

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Data is from peer-reviewed journal

Data source

Reference

Reference Type:

publication

Title:

Evaluation of the teratogenic potential of the Oxidative dyes 6-chloro-4-nitro-2-aminophenol And o-chloro-p-phenylenediamine

Author:

J. C. PICCIANO, W. E. MORRIS and B. A. WOLF

Year:

1984

Bibliographic source:

Fd. Chem. Toxic. Vol. 22, no. 2, pp. I47 149, 1984

Materials and methods

Test guideline

Qualifier:

no guideline available

Principles of method if other than guideline:

The present study was conducted to determine the reproductive toxicity potential of 2-chloro-p-phenylenediamine (o-chloro-p-PD) (CAS No.- 615-66 -7) when administered orally to rats.

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

2-chloro-p-phenylenediamine

EC Number:

210-441-2

EC Name:

2-chloro-p-phenylenediamine

Cas Number:

615-66-7

Molecular formula:

C6H7ClN2

IUPAC Name:

2-chlorobenzene-1,4-diamine

Details on test material:

- Name of test material (as cited in study report): o-chloro-p-phenylenediamine (o-chloro-p-PD)
- Molecular formula (if other than submission substance): C6H7N2Cl
- Molecular weight (if other than submission substance): 142.59 g/mole
- Substance type: Organic
- Physical state: Solid
- Impurities (identity and concentrations): No data available

Specific details on test material used for the study:

- Name of test material (as cited in study report): 2-chloro-p-phenylenediamine (o-chloro-p-PD)
- Molecular formula: C6H7N2Cl
- Molecular weight:142.59 g/mole
- Substance type: Organic
- Physical state: Solid
- Impurities (identity and concentrations):No data available

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

- Source: Charles River Breeding Laboratories, Wilmington,MA
- Age at study initiation: (P) x wks; (F1) x wks : No data available
- Weight at study initiation: P - 225-250 g
- Fasting period before study: No data available
- Housing: Individually housed
- Diet (e.g. ad libitum): Purina Laboratory Rodent Chow, ad libitum
- Water (e.g. ad libitum):Water, ad libitum
- Acclimation period: 2 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data available
- Humidity (%):No data available
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): No data available

Administration / exposure

Route of administration:

oral: gavage

Type of inhalation exposure (if applicable):

not specified

Vehicle:

propylene glycol

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: o-chloro-p-phenylenediamine was dissolved in propylene glycol before administration.

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Propylene glycol
- Concentration in vehicle: 0, 100, 200 and 400 mg/kg/day
- Amount of vehicle (if gavage): 10 ml/kg
- Lot/batch no. (if required): No data available
- Purity: No data available

Details on mating procedure:

- M/F ratio per cage:1 : 2 ratio
- Length of cohabitation:No data available
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy: Sperm plug or the presence of sperm in a vaginal smear referred to as day 0 of pregnancy.
- After … days of unsuccessful pairing replacement of first male by another male with proven fertility. No data available
- Further matings after two unsuccessful attempts: [no / yes (explain)]: No data available
- After successful mating each pregnant female was caged (how): Caged separately
- Any other deviations from standard protocol: No data available

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

10 days

Frequency of treatment:

Daily

Details on study schedule:

Details on study schedule
- F1 parental animals not mated until [...] weeks after selected from the F1 litters: No data available
- Selection of parents from F1 generation when pups were [...] days of age. No data available
- Age at mating of the mated animals in the study: [...] weeks
(Explain how study was performed on perents and offspring separately whatever information we have): No data available

Doses / concentrations

Remarks:

Doses / Concentrations:
0,100, 200 and 400 mg/kg/day
Basis:
actual ingested

No. of animals per sex per dose:

Total : 53
10 ml/kg : 20 female
100 mg/kg/day : 11 female
200 mg/kg/day : 11 female
400 mg/kg/day : 11 female

Control animals:

yes, concurrent vehicle

Examinations

Parental animals: Observations and examinations:

Mortality , general health and condition and body weight were observed.

Oestrous cyclicity (parental animals):

Any irregularities in the estrous cycle were investigated.

Sperm parameters (parental animals):

No data available

Litter observations:

Litter was observed for survival rate, body weight, number of litter, sex ratio and general appearance were examined.

Postmortem examinations (parental animals):

Gross abnormalities were examined.

Number of implantation sites and number of resorptions were also examined.

Postmortem examinations (offspring):

Gross, visceral and skeletal anomalies in fetuses were observed.

Statistics:

Weight changes, the number of fetal implantations, fetal weights and the fetal sex ratio were analyzed by using Student's t test, The numbers of foetal resorptions and abnormal foetuses were compared by chi square analysis and vehicle control data were combined after an analysis of variance.

Reproductive indices:

Fertility index, gestation index, delivery index and implantation index were examined.

Offspring viability indices:

No data available

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:

no effects observed

Description (incidence and severity):

No abmormalities were observed in treated female rats.

Dermal irritation (if dermal study):

not specified

Mortality:

no mortality observed

Description (incidence):

No mortality were observed in treated female rats.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Body weight and weight gain: Mean weight gain was decreased on 6-16 day in 200 mg/kg/day and on 6-20 day in 400 mg/kg/day treated female rats.

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: 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):

No effect were observed on reproductive performance of treated animals as compare to control.
No change were observed in fertility index, gestation index and implantation index of treated female rat as compare to control. Statistically significant increase in the number of resorptions were observed in 400 mg/kg/day treated rats.

Effect levels (P0)

Dose descriptor:

NOAEL

Effect level:

100 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No effect on survival, clinical sign, body weight, Reproductive function and Reproductive performance

Target system / organ toxicity (P0)

Critical effects observed:

no

Results: P1 (second parental generation)

Effect levels (P1)

Remarks on result:

not measured/tested

Target system / organ toxicity (P1)

Critical effects observed:

not specified

Results: F1 generation

General toxicity (F1)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

not specified

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Body weight: Weight of both male and female fetuses were significantly decreased as compare to control

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

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):

No significant differences were observed in number of abnormal foetuses and the types of gross and skeletal anomalies as compare to contol.

Histopathological findings:

no effects observed

Description (incidence and severity):

No visceral anomalies were ovbserved in fetuses of treated female rats.

Other effects:

not specified

Developmental neurotoxicity (F1)

Behaviour (functional findings):

not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:

not specified

Details on results (F1)

Reproductive performance:
No significant differences in the male:female sex ratio were observed as compare to control.

Effect levels (F1)

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

200 mg/kg bw/day

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No effect on survival, body weight, Gross pathology and Histopathology

Target system / organ toxicity (F1)

Critical effects observed:

no

Results: F2 generation

Effect levels (F2)

Remarks on result:

not measured/tested

Target system / organ toxicity (F2)

Critical effects observed:

not specified

Overall reproductive toxicity

Reproductive effects observed:

no

Applicant's summary and conclusion

Conclusions:

NOAEL was considered to be 100 mg/kg/day for F0 generation and 200 mg/kg/day for F1 generation when female rat were exposed to 2-chloro-p -phenylenediamine (o-chloro-p-PD).

Executive summary:

In a reproductive toxicity study, female Sprague-Dawley rat were exposed 2-chloro-p-phenylenediamine (o-chloro-p-PD) orally in the concentration 0, 100, 200 and 400 mg/kg/day. In the parental generation, decreased in mean body weight were obsrved in 200 and 400 mg/kg/day treated rat as compared to control . In addition,significant increase in the number of resorptions were observed in female rat. Effect on fetal weight was observed when treated with 400 mg/kg/day. Therefore, NOAEL is considered to be 100 mg/kg/day for F0 generation and 200 mg/kg/day for F1 generation when rats are exposed to 2-chloro-p-phenylene diamine (o-chloro-p-PD) orally for 10 days.