Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Effect on fertility:

No Observed Adverse Effect Level (NOAEL) was considered to be 10mg/kg/day and LOAEL was considered to be 50mg/kg bw . When female Sprague Dawley rats were treated with 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7) orally.

Link to relevant study records
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature
Justification for type of information:
Data from secondary source
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Principles of method if other than guideline:
Reproduction and developmental toxicity study 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloridewas performed on Sprague Dawley rats.
GLP compliance:
not specified
Limit test:
no
Justification for study design:
No data available
Specific details on test material used for the study:
- Name of test material: Basic Violet 2
- IUPAC name: 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride
- Molecular formula: C22H23N3ClH
- Molecular weight: 365.906 g/mol
- Substance type: Organic
- Physical state: No data
- Purity:No data
- Impurities (identity and concentrations): No data
Species:
rat
Strain:
Sprague-Dawley
Remarks:
Hsd: SD strain
Sex:
female
Details on test animals and environmental conditions:
No data available
Route of administration:
oral: unspecified
Vehicle:
water
Remarks:
Distilled water
Details on exposure:
PREPARATION OF DOSING SOLUTIONS:
Test material dissolved in distilled water
DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food )
- Storage temperature of food: No data available
VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 2,10 and 50 mg/kg bw/day
- Amount of vehicle (if gavage): 10ml/kg bw/day

- Lot/batch no. (if required): No data available
- Purity: No data available
Details on mating procedure:
No data available
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
9 days ( from day 6 through day 15 gestation)
Frequency of treatment:
daily
Details on study schedule:
No data available
Remarks:
2,10 and 50 mg/kg bw/day
No. of animals per sex per dose:
Total:72
2mg/kg bw/day:24
10mg/kg bw/day:24
50 mg/kg bw/day:24
Control animals:
not specified
Details on study design:
No data available
Positive control:
No data available
Parental animals: Observations and examinations:
Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule:


BODY WEIGHT: Yes
Time schedule for examinations:
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes Food consumption was determined weekly.

Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data: No data available


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
Time schedule for examinations:
Oestrous cyclicity (parental animals):
No data available
Sperm parameters (parental animals):
No data available
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: [yes/no]
- If yes, maximum of [...] pups/litter ([...]/sex/litter as nearly as possible); excess pups were killed and discarded.

PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 / F3] offspring:
[number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities, anogenital distance (AGD), presence of nipples/areolae in male pups, other:]foetuses were weighed, sexed and Number of implantation sites were observed.

GROSS EXAMINATION OF DEAD PUPS:: yes, subjected to external,soft tissue or skeletal examinations.

ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY:No data

ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY:No data
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]
- Maternal animals: All surviving animals [describe when, e.g. after the last litter of each generation was weaned.]yes

GROSS NECROPSY: yes
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.] final body weight, uterus weight and corrected body weight were Observed


HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.No data
Postmortem examinations (offspring):
SACRIFICE
- The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at [#?] days of age.
- These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows:

GROSS NECROPSY
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]: No data

HISTOPATHOLOGY / ORGAN WEIGTHS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.:The foetuses were subjected to external, soft tissue or skeletal examinations

Statistics:
No data available
Reproductive indices:
No data available
Offspring viability indices:
No data available
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Violet coloured faeces in 10mg/kg bw/day and 50mg/kg bw/day dose groups while Dyspnoea in 50mg/kg bw/day group were observed.
Dermal irritation (if dermal study):
not specified
Mortality:
not specified
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Reduced body weight in 50mg/kg bw/day group from day 8
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:
effects observed, treatment-related
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):
The number of implantations was decreased in 50mg/kg bw/day dose group.
Dose descriptor:
NOAEL
Effect level:
10 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
clinical signs
body weight and weight gain
organ weights and organ / body weight ratios
reproductive performance
Remarks on result:
other: No effect was observed at given dose
Dose descriptor:
LOAEL
Effect level:
50 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
organ weights and organ / body weight ratios
reproductive performance
Remarks on result:
other: toxic effects observed at given dose level
Critical effects observed:
not specified
System:
other: not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
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):
Foetal weight was decreased in the 50mg/kg bw/day dose group
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
Gross pathological findings:
no effects observed
Description (incidence and severity):
Foetal visceral exam and Foetal skeletal exam no treatment related effects was observed.
Histopathological findings:
not specified
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
10 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
body weight and weight gain
gross pathology
other: Foetal visceral and skeletal finding
Remarks on result:
other: No effect was observed
Dose descriptor:
LOAEL
Generation:
F1
Effect level:
50 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
body weight and weight gain
Remarks on result:
other: effect on body weight was observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Reproductive effects observed:
no
Treatment related:
not specified
Relation to other toxic effects:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Conclusions:
No Observed Adverse Effect Level (NOAEL) was considered to be 10mg/kg/day and LOAEL was considered to be 50 mg/kg bw.When female Sprague Dawley rats were treated with 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7) orally.
Executive summary:

The reproduction and development toxicity study of 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7)was performed on female Sprague Dawley  Hsd: SD strainrats. 72 rats were divided as 24 rats /dose group. The test materialdissolved in water were administers in dose concentration 0, 5, 10and 50 mg/kg bw/day from day 6 through day 15of gestationby oral gavage route. Animals were observed for clinical signs, Food consumption and body weight .On day 20 post-coitum, the dams were sacrificed and subjected to a macroscopic examination. The litter parameters like number of implantation sites. The foetuses were weighed and subjected to external, soft tissue or skeletal examinations.

Clinical sings like violet coloured faeces in 10mg/kg bw/day and 50mg/kg bw/day dose groups while dyspnoea in 50mg/kg bw/day group were observed.Body weightreduced in 50mg/kg bw/day group from day 8 and reductionin food consumptionwas observed during treatment. At necropsy final body weight, uterus weight and corrected body weight were decreased in 50mg/kg bw/day dose group in dams were noted. In litter the number of implantations was decreased in 50mg/kg bw/day dose group and foetal weight decrease in 50mg/kg bw/day dose group but no treatment related effects were observed in Foetal visceral and skeletal examination at any dose group. HenceNo Observed Adverse Effect Level (NOAEL) was considered to be 10mg/kg/day and LOAEL was considered to be 50mg/kg bw.When femaleSprague Dawley rats were treated with4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7)orally.

 

Effect on fertility: via oral route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LOAEL
50 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Data is Klimicsh 4 and from secondary source
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

The reproductive toxicity via oral route

In different studies, 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7) has been investigated for reproductive toxicity to a greater or lesser extent. Often are the studies based on in vivo experiments and estimated data in rodents, i.e. most commonly in rats for 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7)

This study published in a report by Scientific Committee on Consumer Safety (the Scientific Committee on Consumer Products, SCCS/1340/10,2011) The reproduction and development toxicity study of  4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7)was performed on female Sprague Dawley  Hsd: SD strain rats. 72 rats were divided as 24 rats /dose group. The test material dissolved in water were administers in dose concentration 0, 5, 10and 50 mg/kg bw/day from day 6 through day 15of gestation by oral gavage route. Animals were observed for clinical signs, Food consumption and body weight .On day 20 post-coitum, the dams were sacrificed and subjected to a macroscopic examination. The litter parameters like number of implantation sites. The foetuses were weighed and subjected to external, soft tissue or skeletal examinations.

Clinical sings like violet coloured faeces in 10mg/kg bw/day and 50mg/kg bw/day dose groups while dyspnoea in 50mg/kg bw/day group were observed. Body weight reduced in 50mg/kg bw/day group from day 8 and reduction in food consumption was observed during treatment. At necropsy final body weight, uterus weight and corrected body weight were decreased in 50mg/kg bw/day dose group in dams were noted. In litter the number of implantations was decreased in 50mg/kg bw/day dose group and foetal weight decrease in 50mg/kg bw/day dose group but no treatment related effects were observed in Foetal visceral and skeletal examination at any dose group. Hence No Observed Adverse Effect Level (NOAEL) was considered to be 10mg/kg/day and LOAEL was considered to be 50mg/kg bw .When female Sprague Dawley rats were treated with4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91 -7)orally.

This is further supported by experimental study conducted by Ryo Ohta, Atsuya Takagi, Hideo Ohmukai, Hideki Marumo, Atsushi Ono, Yuko Matsushima, Tohru Inoue, Hiroshi Ono, Jun Kanno (The Journal of Toxicological Sciences, Vol. 37, No. 5, 879-889, 2012) The reproductive toxicity study of New Fuchsin (3248-91-7)was performed on female  C57BL/6Jrats.18 mice were divided as 6 rats /dose group.6 week old C57BL/6J ovariectomized mice were checked for vaginal smears 4 days before the start of the test. The animals were acclimatized for a period of 2 weeks. At 8 weeks of age, non-estrus mice were selected. The test material dissolved in water were administers in dose concentration 0,300mg/kg bw/day by oral gavage route of exposure at 24 hrs interval for 7 consecutive days. The dosing volume was 5ml/Kg were used .17α-ethynyl estradiol was used as the reference control. Vehicle control animals were also observed. Based on the dose range finding study performed highest dose 300mg/kg bw were selected. The animals were observed daily for clinical signs with the body weight recorded.24 hrs after the last treatment; the animals were euthanized by cervical dislocation. The uterus was carefully dissected at the level of the vaginal fornix, trimmed of fascia and fat under a stereomicroscope and weighed including the luminal fluid (wet weight), then pierced, gently blotted on moistened filter and weighed again for blot weight. Significant change in uterine weight was noted. Test material showed weak but significant agonistic and antagonistic effect at dose concentration 300mg/kg bw by oral route. Hence the Low Observed Effect Level (LOEL) was considered to be 300 mg/Kg/day. When femaleC57BL/6Jmice were treated with New Fuchsin (3248-91-7)orally. 

So, based on the above mentioned studies for target substance 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7), reduction in body weight , food consumption , adverse effects on reproductive organ and decrease in number of implantations was observed at dose concentration 50mg/kg bw .Therefore, according to CLP criteria, the substance 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7) can be classified as“Category 2” for  reproductive toxicity.

Effects on developmental toxicity

Description of key information

Developmental toxicity

No Observed Adverse Effect Level (NOAEL) was considered to be 10mg/kg/day and LOAEL was considered to be 50 mg/kg bw .When female Sprague Dawley rats were treated with 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7) orally.

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
secondary literature
Justification for type of information:
Data from secondary source
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Principles of method if other than guideline:
Prenatal developmental toxicity study 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloridewas performed on Sprague Dawley rats.
GLP compliance:
not specified
Limit test:
no
Specific details on test material used for the study:
- Name of test material: Basic Violet 2
- IUPAC name: 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride
- Molecular formula: C22H23N3ClH
- Molecular weight: 365.906 g/mol
- Substance type: Organic
- Physical state: No data
- Purity:No data
- Impurities (identity and concentrations): No data
Species:
rat
Strain:
Sprague-Dawley
Remarks:
Hsd: SD strain
Details on test animals and environmental conditions:
No data available
Route of administration:
oral: unspecified
Vehicle:
water
Remarks:
Distilled water
Details on exposure:
Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test material dissolved in distilled water
DIET PREPARATION
- Rate of preparation of diet (frequency):No data available
- Mixing appropriate amounts with (Type of food )
- Storage temperature of food: No data available
VEHICLE
- Justification for use and choice of vehicle (if other than water):
- Concentration in vehicle: 2,10 and 50 mg/kg bw/day
- Amount of vehicle (if gavage): 10ml/kg bw/day

- Lot/batch no. (if required): No data available
- Purity: No data available
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
No data available
Duration of treatment / exposure:
9 days ( from day 6 through day 15 gestation)
Frequency of treatment:
Daily
Duration of test:
9 days ( from day 6 through day 15 gestation)
Remarks:
2,10 and 50 mg/kg bw/day
No. of animals per sex per dose:
Total:72
2mg/kg bw/day:24
10mg/kg bw/day:24
50 mg/kg bw/day:24
Control animals:
not specified
Details on study design:
No data available
Maternal examinations:
Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule:


BODY WEIGHT: Yes
Time schedule for examinations:
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes Food consumption was determined weekly.

Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes / No / No data: No data available


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

OTHER:
Ovaries and uterine content:
The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: No data
- Number of implantations: Yes
- Number of early resorptions: No data
- Number of late resorptions: No data
- Other:
Fetal examinations:
- External examinations: Yes:
- Soft tissue examinations: Yes:
- Skeletal examinations: Yes:
- Head examinations: Yes:
Statistics:
No data available
Indices:
No data available
Historical control data:
No data available
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Violet coloured faeces in 10mg/kg bw/day and 50mg/kg bw/day dose groups while Dyspnoea in 50mg/kg bw/day group were observed.
Dermal irritation (if dermal study):
not specified
Mortality:
not specified
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
body weight reduced in 50mg/kg bw/day group from day 8
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Reduction in food consumption was observed during treatment
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:
effects observed, treatment-related
Description (incidence and severity):
At necropsy final body weight, uterus weight and corrected body weight were decreased in 50mg/kg bw/day dose group
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Number of abortions:
not specified
Pre- and post-implantation loss:
no effects observed
Description (incidence and severity):
The number of implantations was decreased in 50mg/kg bw/day dose group.
Total litter losses by resorption:
not specified
Early or late resorptions:
not specified
Dead fetuses:
not specified
Changes in pregnancy duration:
not specified
Description (incidence and severity):
Migrated Data from removed field(s)
Field "Effects on pregnancy duration" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsMaternalAnimals.MaternalDevelopmentalToxicity.EffectsOnPregnancyDuration): not specified
Changes in number of pregnant:
not specified
Other effects:
not specified
Dose descriptor:
NOAEL
Effect level:
10 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
clinical signs
body weight and weight gain
food consumption and compound intake
gross pathology
pre and post implantation loss
Remarks on result:
other: No effect observed in given dose
Dose descriptor:
LOAEL
Effect level:
50 mg/kg bw/day (nominal)
Based on:
test mat.
Basis for effect level:
body weight and weight gain
organ weights and organ / body weight ratios
other: The number of implantations was decreased in 50mg/kg bw/day dose group.
Remarks on result:
other: Adverse effects was observed at given dose level
Abnormalities:
not specified
Fetal body weight changes:
effects observed, treatment-related
Description (incidence and severity):
Foetal weight was decreased in the 50mg/kg bw/day dose group
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:
not specified
Changes in litter size and weights:
not specified
Changes in postnatal survival:
not specified
External malformations:
no effects observed
Description (incidence and severity):
no treatment related effects was observed in any dose group.
Skeletal malformations:
no effects observed
Description (incidence and severity):
no treatment related effects was observed in any dose group.
Visceral malformations:
no effects observed
Description (incidence and severity):
no treatment related effects was observed in any dose group.
Other effects:
not specified
Dose descriptor:
NOAEL
Effect level:
10 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
fetal/pup body weight changes
external malformations
skeletal malformations
visceral malformations
Remarks on result:
other: No effect was observed
Dose descriptor:
LOAEL
Effect level:
50 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
not specified
Basis for effect level:
fetal/pup body weight changes
Remarks on result:
other: Decrease in body weight was observed at given dose level
Abnormalities:
not specified
Developmental effects observed:
no
Treatment related:
not specified
Relation to maternal toxicity:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified
Conclusions:
No Observed Adverse Effect Level (NOAEL) was considered to be 10mg/kg/day and LOAEL was considered to be 50 mg/kg bw .When female Sprague Dawley rats were treated with 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7) orally.
Executive summary:

Prenatal development toxicity study of  with4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7)was performed on female Sprague Dawley  Hsd: SD strainrats. 72 rats were divided as 24 rats /dose group. The test materialdissolved in water were administers in dose concentration 0, 5, 10and 50 mg/kg bw/day from day 6 through day 15of gestationby oral gavage route. Animals were observed for clinical signs, Food consumption and body weight .On day 20 post-coitum, the dams were sacrificed and subjected to a macroscopic examination. The litter parameters like number of implantation sites. The foetuses were weighed and subjected to external, soft tissue or skeletal examinations.Clinical sings like violet coloured faeces in 10mg/kg bw/day and 50mg/kg bw/day dose groups while dyspnoea in 50mg/kg bw/day group were observed.Body weightreduced in 50mg/kg bw/day group from day 8 and reductionin food consumptionwas observed during treatment. At necropsy final body weight, uterus weight and corrected body weight were decreased in 50mg/kg bw/day dose group in dams were noted. In litter the number of implantations was decreased in 50mg/kg bw/day dose group and foetal weight decrease in 50mg/kg bw/day dose group but no treatment related effects were observed in Foetal visceral and skeletal examination at any dose group. Hence No Observed Adverse Effect Level (NOAEL) was considered to be 10mg/kg/day and LOAEL was considered to be 50mg/kg bw.When female Sprague Dawley rats were treated with 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7) orally.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LOAEL
50 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Data is Klimicsh 4 and from secondary source
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

Developmental toxicity

In different studies, 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7) has been investigated for developmental toxicity to a greater or lesser extent. Often are the studies based on in vivo experiments and estimated data in rodents, i.e. most commonly in rats for 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7)

This study published in a report by Scientific Committee on Consumer Safety (the Scientific Committee on Consumer Products, SCCS/1340/10,2011) The reproduction and development toxicity study of  4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7)was performed on female Sprague Dawley  Hsd: SD strain rats. 72 rats were divided as 24 rats /dose group. The test material dissolved in water were administers in dose concentration 0, 5, 10and 50 mg/kg bw/day from day 6 through day 15of gestation by oral gavage route. Animals were observed for clinical signs, Food consumption and body weight .On day 20 post-coitum, the dams were sacrificed and subjected to a macroscopic examination. The litter parameters like number of implantation sites. The foetuses were weighed and subjected to external, soft tissue or skeletal examinations.

Clinical sings like violet coloured faeces in 10mg/kg bw/day and 50mg/kg bw/day dose groups while dyspnoea in 50mg/kg bw/day group were observed. Body weight reduced in 50mg/kg bw/day group from day 8 and reduction in food consumption was observed during treatment. At necropsy final body weight, uterus weight and corrected body weight were decreased in 50mg/kg bw/day dose group in dams were noted. In litter the number of implantations was decreased in 50mg/kg bw/day dose group and foetal weight decrease in 50mg/kg bw/day dose group but no treatment related effects were observed in Foetal visceral and skeletal examination at any dose group. Hence No Observed Adverse Effect Level (NOAEL) was considered to be 10mg/kg/day and LOAEL was considered to be 50mg/kg bw. When female Sprague Dawley rats were treated with4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7)orally.

So, based on the above mentioned study for target substance 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7),Foetal visceral and skeletal examination did not reveal teratogenic effects. Maternal toxicity and a delay of foetal development were observed at 50 mg/kg bw/day.Therefore, according to CLP criteria, the substance 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7) can be classified as“Category 2” for developmental toxicity.

Justification for classification or non-classification

Therefore, according to CLP criteria, the substance 4,4'-[(4-imino-3-methylcyclohexa-2,5-dien-1-ylidene)methylene]bis(2-methylaniline) hydrochloride(3248-91-7) can be classified as“Category 2” for reproductive and developmental toxicity.