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Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Under the condition of the study, the 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).

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
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
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
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
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals 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
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
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
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
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: 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.
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
Critical effects observed:
no
Remarks on result:
not measured/tested
Critical effects observed:
not specified
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
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Reproductive performance:
No significant differences in the male:female sex ratio were observed as compare to control.
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
Critical effects observed:
no
Remarks on result:
not measured/tested
Critical effects observed:
not specified
Reproductive effects observed:
no
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.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
100 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The data is Klimicsh 2 and from peer-reviewed journal.
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Various studies has been investigated for reproductive toxicity to a greater or lesser extent for the test chemical 2-chloro-p-phenylenediamine (CAS No.- 615-66-7) along with its structurally similar read across substances 2,4,6-trichloroaniline (CAS No.- 634-93-5) and 3,4-dichloroaniline (CAS No : 95-76-1). The studies are summarized as below:

 

The present study was undertaken by J. C. Picciano et al. (Food and Chemical Toxicology. Vol. 22, no. 2, pp. I47 149, 1984)to determine the reproductive toxicity potential of 2-chloro-p-phenylenediamine(CAS No.- 615-66-7), female Sprague-Dawley rat were exposed 2-chloro-p-phenyle nediamine (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-phenylenediamine (o-chloro-p-PD) orally for 10 days.

 

In addition, reproductive toxicity study published by SRC Inc (2010), male and female rats were exposed to 2,4,6-trichloroaniline (CAS No.- 634-93-5) in the concentrations of 0, 0.3, 3 and 29 mg/kg/day. The results showed that 2,4,6 trichloroaniline is toxic at higher doses. Toxic changes were observed as decreased in weight gain, increased numbers of hypochromic RBCs and doubled levels of methemoglobin, anisocytosis, poikilocytosis, reticulocytosis, hypochromia and presence of Heinz bodies in the RBCs and decreased in SDH and LDH activities in liver and the kidney of 29 mg/kg/day treated rat. Increased relative weights of brain and decreased liver weight. In addition, Degenerative changes and evidence of hemorrhage in the myocardium, kidneys, liver, spleen and brain in 29 mg/kg/day treated rats. Decreased numbers of fetuses/dam and massive hematomas were observed in abdominal cavities of 3 mg/kg/day treated rat of F1 generation. In F1 generation, Pre and post implantation fetal mortality were also observed in 3 mg/kg/day treated rat. Therefore, the NOAEL was considered to be 0.3 mg/kg/day for both F0 and F1 generation when rats were treated with 2, 4, 6 trichloroaniline orally for 6 months.

 

Moreover, the study was designed and published by U.S. Environmental Protection Agency (U.S. Environmental Protection Agency TSCA, NTIS Technical Report : OTS0600391, 2008) to investigate the reproductive effects of 3,4-Dichloroaniline in Charles River CrI:CD BR rats. The substance3,4-dichloroaniline (CAS No : 95-76-1) was administered orally by gavage at three doses 0, 5, 25 and 125 mg/kg - to groups of pregnant Charles River CrI:CD BR rats. No statistically significant or toxicologically relevant adverse effects on any of the maternal reproductive parameters studied were observed in the 5 mg/kg dose groups. The test article did, however, produce a slight increase in absorptions and consequently in post-implantation loss at 125 mg/kg. No effects observed on fetuses. Hence, NOAEL for maternal toxicity study was considered to be 5 mg/kg bw/day whereas NOAEL for fetotoxicity study was considered to be 25 mg/kg bw/day in Charles River CrI:CD BR rats when 3,4-dichloroaniline was administered orally by gavage.

 

Based on the above mentioned studies for target substance and to its read across substance and by comparing these with the CLP criteria, it can be concluded that no adverse effects was observed, therefore the substance 2-chloro-p-phenylenediamine (CAS No.- 615-66-7) cannot be classified as reproductive toxicant.

Effects on developmental toxicity

Description of key information

Under the condition of the study, the NOAEL was considered to be 100 mg/kg/day for maternal study and 200 mg/kg/day for teratogenicity study when female rat were exposed to 2-chloro-p-phenylenediamine (o-chloro-p-PD).

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
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.
Qualifier:
no guideline available
Principles of method if other than guideline:
The present study was conducted to determine the teratogenic potential of 2-chloro-p-phenylenediamine (o-chloro-p-PD) (CAS No.- 615-66 -7).
GLP compliance:
not specified
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
Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
Sex: Female
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
Analytical verification of doses or concentrations:
not specified
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
Duration of treatment / exposure:
10 days
Frequency of treatment:
Daily
Duration of test:
10 days
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
Maternal examinations:
Mortality , general health and condition and body weight and gross abnormalities were examined
Ovaries and uterine content:
Number of implantation sites and number of resorptions were also examined.
Fetal examinations:
Litter was observed for survival rate, body weight, number of litter, sex ratio, general appearance, 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.
Indices:
Fertility index, gestation index, delivery index and implantation index were examined.
Historical control data:
No data available
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):
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:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
Reproductive function:
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

Reproductive performance:
No effect were observed on reproductive performance of treated animals as compare to control.
Details on maternal toxic effects:
Maternal toxic effects:no effects

Details on maternal toxic effects:
Mortality :
No mortality were observed in treated female rats

Clinical signs:
No abmormalities were observed in treated female rats

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.

Reproductive function:
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

Reproductive performance:
No effect were observed on reproductive performance of treated animals as compare to control.


Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: No adverse effect on survival, clinical sign, body weight, reproductive performance.
Abnormalities:
not specified
Fetal body weight changes:
effects observed, treatment-related
Description (incidence and severity):
Weight of both male and female fetuses were significantly decreased as compare to control
Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not specified
Reduction in number of live offspring:
not specified
Changes in sex ratio:
no effects observed
Description (incidence and severity):
No significant differences in the male and female sex ratio of fetuses were observed as compare to control
Changes in litter size and weights:
not specified
Changes in postnatal survival:
not specified
External malformations:
no effects observed
Description (incidence and severity):
No significant differences were observed in number of abnormal foetuses and the types of gross anomalies in treated female rat as compare to contol.
Skeletal malformations:
no effects observed
Description (incidence and severity):
No significant differences were observed in number of abnormal foetuses and the types of skeletal anomalies in treated female rat as compare to contol.
Visceral malformations:
no effects observed
Description (incidence and severity):
No significant differences were observed in number of abnormal foetuses and the types of visceral anomalies in treated female rat as compare to contol.
Other effects:
not specified
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects: no effects
Dose descriptor:
NOAEL
Effect level:
200 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: teratogenicity
Abnormalities:
not specified
Developmental effects observed:
no
Conclusions:
NOAEL was considered to be 100 mg/kg/day for maternal study and 200 mg/kg/day for teratogenicity study when female rat were exposed to 2-chloro-p-phenylenediamine (o-chloro-p-PD).
Executive summary:

In a teratogenicity study, female Sprague-Dawley rat were exposed to 2-chloro-p-phenylenedia mine (o-chloro-p-PD) orally in the concentration of 0, 100, 200 and 400 mg/kg/day. In the parental generation, decreased in mean body weight were observed 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 male and female fetal weight was observed when treated with 400 mg/kg/day. Therefore, NOAEL was considered to be 100 mg/kg/day for maternal study and 200 mg/kg/day for teratogenicity study when rats are exposed to 2-chloro-p-phenylenediamine (o-chloro-p-PD) orally for 10 days.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
200 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The data is Klimicsh 2 and from peer-reviewed journal.
Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

In a teratogenicity study conducted by J. C. Picciano et al. (Food and Chemical Toxicology. Vol. 22, no. 2, pp. I47 149, 1984), female Sprague-Dawley rat were exposed to 2-chloro-p-phenylenediamine (o-chloro-p-PD) orally in the concentration of 0, 100, 200 and 400 mg/kg/day. In the parental generation, decreased in mean body weight were observed 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 male and female fetal weight was observed when treated with 400 mg/kg/day. Therefore, NOAEL was considered to be 100 mg/kg/day for maternal study and 200 mg/kg/day for teratogenicity study when rats are exposed to 2-chloro-p-phenylenediamine (o-chloro-p-PD) orally for 10 days.

Thus, on the basis of above available study and by comparing these with the CLP criteria, it can be concluded that no adverse effects on development of the offspring was observed. Therefore the substance 2-chloro-p-phenylenediamine(CAS No.- 615-66-7) can be regarded as not owing a significant potential for developmental toxicity.

Justification for classification or non-classification

Based on the available data for the assessment of reproductive toxicity and developmental toxicity and following CLP Regulation EC No. 1272/2008 no classifi cation of 2-chloro-p-phenylenediamine(CAS No.- 615-66-7) as reproductive toxicant is warranted.