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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
short-term repeated dose toxicity: oral
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 subacute repeated dose toxicity study of 2-chloro-p-phenylenediamine (CAS No. 615-66-7) in rats was conducted to evaluate adverse effects by oral route.
GLP compliance:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
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

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories, Wilmington,MA
- Age at study initiation: No data available
- Weight at study initiation: 225-250 g
- Fasting period before study: No data available
- Housing: Animals were housed separately
- Diet (e.g. ad libitum): Purina Laboratory Rodent Chow, ad libitum
- Water (e.g. ad libitum): 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

IN-LIFE DATES: From: To: No data available

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on oral 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: 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
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
10 days
Frequency of treatment:
Daily
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 100, 200 and 400 mg/kg/day

Basis:
nominal in diet
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
Details on study design:
- Dose selection rationale: The dose was determined from previously estimated maximum tolerated doses.
- Rationale for animal assignment (if not random): Pregnant females were assigned on a random basis to 100, 200 and 400 mg/kg/day
- Rationale for selecting satellite groups: No data available
- Post-exposure recovery period in satellite groups: No data available
- Section schedule rationale (if not random): No data available

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS:
- Time schedule: Daily
- Cage side observations checked in table [No.?] were included: Mortality was observed.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily general health and condition were observed.

BODY WEIGHT: Yes
- Time schedule for examinations: On days 0, 6, 16 and 20

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data available
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data available

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data available

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data available
- Time schedule for examinations: No data available

OPHTHALMOSCOPIC EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available

HAEMATOLOGY: No data available
- Time schedule for collection of blood: No data available
- Anaesthetic used for blood collection: No data available
- Animals fasted: No data available
- How many animals: No data available
- Parameters checked in table [No.?] were examined. No data available

CLINICAL CHEMISTRY: No data available
- Time schedule for collection of blood: No data available
- Animals fasted: No data available
- How many animals: No data available
- Parameters checked in table [No.?] were examined. No data available

URINALYSIS: No data available
- Time schedule for collection of urine: No data available
- Metabolism cages used for collection of urine: No data available
- Animals fasted: No data available
- Parameters checked in table [No.?] were examined. No data available

NEUROBEHAVIOURAL EXAMINATION: No data available - Time schedule for examinations: No data available
- Dose groups that were examined: No data available
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data available


Sacrifice and pathology:
GROSS PATHOLOGY:
Implantation sites, external fetal malformations, head, trunk and limbs were examined.

One half of the fetuses randomly selected from each litter were placed in Bouin's fixative for subsequent visceral examination according to Wilson's procedure (Wilson & Warkany, 1965). The remaining half of the fetuses were eviscerated, fixed in 95 % isopropyl alcohol, macerated in potassium hydroxide and stained with Alizarin Red S by a method similar to that of Dawson (1926), for subsequent skeletal evaluation.

HISTOPATHOLOGY: No data available

Other examinations:
Number of fetus, foetal malformations, and skeletal abnormalities of fetus 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, vehicle control data were combined after an analysis of variance.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
No abmormalities were observed in treated female rats.
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):
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
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
No gross pathological effect was observed in treated female rats as compared to control.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
Statistically significant increase in the number of resorptions was observed in 400 mg/kg/day treated female rats as compared to control.

Effect levels

Dose descriptor:
NOAEL
Effect level:
200 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
gross pathology
other: No adverse effect on survival, clinical sign, body weight, gross pathology and fetal development was observed.

Target system / organ toxicity

Critical effects observed:
no

Applicant's summary and conclusion

Conclusions:
Under the condition of the study, the no-observed adverse effect level (NOAEL) of 2-chloro-p-phenylenediamine (o-chloro-p-PD) (CAS No. 615-66-7) in rats was considered to be 200 mg/kg/day.
Executive summary:

The subacute repeated dose toxicity study of 2-chloro-p-phenylenediamine (CAS No. 615-66-7) in female Sprague-Dawley rats was conducted to evaluate the adverse effects by oral route. The test chemical was administered by oral gavage in the concentration of 100, 200 and 400 mg/kg/day. The results showed reduction in mean body weight gain of 200 and 400 mg/kg/day treated rats. No mortality were observed. Clinical signs was not observed. In addition, statistically significant increase in the number of resorptions was observed in 400 mg/kg/day treated female rats as compared to control.  No changes were observed in gross pathology of treated rats as compared to control. Thus, the no-observed adverse effect level (NOAEL) of 2-chloro-p-phenylenedi amine (o-chloro-p-PD) (CAS No. 615-66-7) in rats was considered to be 200 mg/kg/day.