2-[[4-[ethyl(2-hydroxyethyl)amino]phenyl]azo]-6-methoxy-3-methylbenzothiazolium acetate

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Reproductive toxicity study

Based on the data available from different studies, NOAEL for test material was considered to be in range of  300-500mg/kg bw/day for F0, F1 generation. When male and female CD rats were treated with test material orally. Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Experimental data from various test chemicals

Justification for type of information:

Weight of evidence approach based on the available information from various test chemicals.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

WoE report is based on reproductive toxicity studies on rats

GLP compliance:

not specified

Limit test:

no

Justification for study design:

No data available

Species:

rat

Strain:

other: 1. Crj: CD (SD)IGS 2.wistar

Details on species / strain selection:

No data available

Sex:

male/female

Details on test animals or test system and environmental conditions:

Details on test animals and env. conditions
TEST ANIMALS
- Source: Charles River Co., Ltd. Atsugi Breeding Center Co., Ltd.
- Age at study initiation: 5 weeks of age
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g
- Fasting period before study:
- Housing: The animals were placed in a bracket-type metallic wire mesh floor cage
- Use of restrainers for preventing ingestion (if dermal): yes/no
- Diet (e.g. ad libitum): gamma-irradiated solid feed CRF-1 (Oriental Yeast Co., Ltd.) ad libitum
- Water (e.g. ad libitum): drinking water was freely ingested by Sapporo city tap water
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 to 24 ° C
- Humidity (%):45 to 68%
- Air changes (per hr): 10 to 15 times / hour
- Photoperiod (hrs dark / hrs light): an illumination time of 8 to 20 hours

IN-LIFE DATES: From: To:

Route of administration:

oral: gavage

Vehicle:

other: 0.5% methyl cellulose aqueous solution

Details on exposure:

1.Details on exposure
PREPARATION OF DOSING SOLUTIONS:
The test substance was suspended using a 0.5% methyl cellulose aqueous solution.
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): corn oil
- Concentration in vehicle: 0, 250, 500, and 1000 mg / kg
- Amount of vehicle (if gavage): 5 mL / kg

- Lot/batch no. (if required): No data available
- Purity: No data available
2.Details on exposure
PREPARATION OF DOSING SOLUTIONS:
Test material dissolved in water (Milli-U)
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): water (Milli-U)
- Concentration in vehicle: 0, 100, 300 and 1000 mg/kg bw
- Amount of vehicle (if gavage): 10 ml/kg bw

- Lot/batch no. (if required): No data available
- Purity: No data available

Details on mating procedure:

Details of mating
- M/F ratio per cage: 1:1
- Length of cohabitation: No data available
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy the presence of vaginal plugs or sperm in the vaginal smear referred to as day 0 of pregnancy

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

Study 1
28 days
Study 2.
15 days (from day 6 to day 20 post-coitum)

Frequency of treatment:

daily

Details on study schedule:

No data available

Remarks:

Study 1.
0, 250, 500, and 1000 mg / kg
Study 2.
0, 100, 300 and 1000 mg/kg bw

No. of animals per sex per dose:

Study 1.
Total: 42 male and 42 female
0 mg / kg:7 male and 7 female
250 mg / kg:7 male and 7 female
500 mg / kg:7 male and 7 female
1000 mg / kg:7 male and 7 female
Recovery group
0 mg / kg:7 male and 7 female
1000 mg / kg:7 male and 7 female
Study 2.
Total:96
0 mg/kg bw/day:24female
100mg/kg bw/day:24female
300mg/kg bw/day:24female
1000mg/kg bw/day:24female

Control animals:

yes, concurrent vehicle

Details on study design:

No data available

Positive control:

No data available

Parental animals: Observations and examinations:

1.Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule: daily


BODY WEIGHT: Yes
Time schedule for examinations: Body weight of individual animals were measured on days 1, 7, and 14 before administration on days 1, 2, 7, 14, 21 and 28, and on the autopsy day of each day after the end of each period
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):Yes food intake of individual animals were measured on days 1, 7, and 14 before administration on days 1, 2, 7, 14, 21 and 28, and on the autopsy day of each day after the end of each period,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:
2.Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule: daily for the clinical signs

BODY WEIGHT: Yes
Time schedule for examinations: Body weights
was determined at periodically during pregnancy.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):Yes food consumption
was determined at periodically during pregnancy.
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):

Parameters examined in [all/P/F1/F2] male parental generations:
testis weight and epididymis weight were observed

Litter observations:

number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain were observed

Postmortem examinations (parental animals):

1.SACRIFICE
- Male animals: All surviving animals
- Maternal animals: All surviving animals

GROSS NECROPSY:yes
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]
HISTOPATHOLOGY / ORGAN WEIGHTS: yes
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.
Study2.
Postmortem examinations (Parent Animal)
SACRIFICE :
On GD 21, all females were subjected to an examination post-mortem and external, thoracic and abdominal macroscopic findings were recorded. The ovaries and uterine horns were dissected and examined for the number of corpora lutea, the weight of the gravid uterus, the number of implantation site scars, the number and distribution of live/dead foetuses and embryo-foetal deaths

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 NECROPSYfoetal sex and externally visible foetal macroscopic abnormalities were observed
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]

HISTOPATHOLOGY / ORGAN WEIGTHS:lAternate foetuses of each litter were preserved in 96% aqueous ethanol or
Bouin's fluid, and subjected to skeletal or visceral examinations respectively

The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.

Statistics:

Quantitative items of urinalysis (excluding urine specific gravity), hematological test (excluding percentage of leukocytes), blood chemistry test, results of absolute and relative weight of organs, body weight, weight gain and rate of increase, We analyzed equi dispersibility by Bartlett's test method, and analyzed by one-way analysis of variance in the case of equal variance and by Kruskal-Wallis's test in case of unequal variance. If a significant difference is found as a result of one-way ANOVA, Dunnett's test method is used, and if there is a significant difference as a result of analysis by the Kruskal-Wallis method, Mann-Whitney's U- , And compared with the control group, respectively.
For the qualitative items of urinalysis, the results of urine specific gravity and white blood cell percentage, the tendency of each group is analyzed by the Kruskal-Wallis test method, and when a significant difference is
observed, the Mann-Whitney U-test method is used And compared with controls. For each percentage of
leukocytes, average values and standard deviations were also calculated. Regarding the comparative test with the control group, statistically significant less than the risk ratio of 5% was taken.

Reproductive indices:

No data available

Offspring viability indices:

No data available

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

1.No abnormality was observed in both sexes in each administration group in either administration period or recovery period.
2.Orange to red urine was noted in all animals in the 300mg/kg and 1000mg/kg dose groups and in some of the 100mg/kg dose group

Dermal irritation (if dermal study):

not specified

Mortality:

mortality observed, treatment-related

Description (incidence):

2.There was one death in the 1000mg/kg dose group on GD 13, but there were no relevant toxicologically clinical or pathological signs. Since the cause of death is unclear, a treatment-related effect can not be excluded

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

1.Only the males in the 250 mg / kg administration group showed significant increase in body weight gain and body weight gain during 28 days of administration, but not a dose-dependent change.
2.Body weight and body weight gain were comparable to the control in the 100mg/kg and 300mg/kg dose group, but there was a statistically significant decrease in the 1000mg/kg dose group

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

1.Only the males in the 250 mg / kg administration group showed a significant increase compared with the control group on the 28th administration but not the dose-dependent change.
2.The decrease was mirrored in the food consumption in the 1000mg/kg dose group, In the 300mg/kg dose group, food consumption decreased from GD12.

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

A significant lower value was found in males of the group receiving 250 mg / kg or more in the γ-globulin fraction ratio as compared with the control group, and at the end of the recovery period, females with a dose of 1000 mg / kg showed significant High level was recognized.

Clinical biochemistry findings:

no effects observed

Description (incidence and severity):

There was no significant change in each treatment group of males and females as compared with the control group in either the administration period or the recovery period.

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:

effects observed, treatment-related

Description (incidence and severity):

1.Only mild changes such as kidney, liver, pituitary gland and the like were observed in one males and two females in the 1000 mg / kg administration group. Even at the end of the recovery period, only mild changes were observed in the lungs, liver, kidneys, etc. of 3 males in the 1000 mg / kg administration group

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):

2.Eight animals were not pregnant (3 control; 2 in 100mg/kg dose group, 2 in 300mg/kg dose group and 1 in 1000mg/kg dose group ) and in 300 mg/kg dose group one animal only showed implantation sites. Post-implantation losses increased at the 1000mg/kg dose resulting in decreased foetal numbers and increased foetal deaths. Embryonic resorption was not affected. No treatment related effects were seen at the other doses

No effects on reproductive organ weight was observed

Dose descriptor:

NOAEL

Effect level:

> 300 - <= 500 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

clinical signs

body weight and weight gain

food consumption and compound intake

haematology

clinical biochemistry

organ weights and organ / body weight ratios

gross pathology

histopathology: non-neoplastic

reproductive performance

Remarks on result:

other: No effects on reproductive organ weight and performance 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

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

not specified

Body weight and weight changes:

not specified

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):

2.At the 1000mg/kg dose, there was a high incidence of foetuses with major visceral abnormalities, i.e. severe umbilical hernia and associated visceral changes, including displacement of organs and absence of diaphragm. A number of the abnormal foetuses also exhibited malrotated hind limbs, atypical skeletal ossification. Some ossification parameters showed slight retardation that could be explained by the reduction in mean foetal weight recorded, whilst other parameters were slightly better ossified. The toxicological significance of the latter finding is unclear.

Histopathological findings:

not specified

Other effects:

not specified

Behaviour (functional findings):

not specified

Developmental immunotoxicity:

not specified

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

300 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

gross pathology

Remarks on result:

other: No developmental toxic effects were 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:

not specified

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 in range of 300-500mg/kg/day, Whenmale and female Crj:CD(SD)IGS rats were treated with test material orally.

Executive summary:

Data available from different studies were reviewed to determine the reproductive toxicity of testchemical.The studies are as mentioned below:

 Study 1

The reproductive organ toxicity study of test material was performed using twenty-eight-day Repeat Dose Oral Toxicity study of test material in Crj: CD (SD)IGS male and female rats. The test material was suspended using a 0.5% methyl cellulose aqueous solutionand administeried in dose concentration 0, 250, 500, and 1000 mg / kg orally for 28 days. Each dose group and recovery group contain 7 male and 7 female. Doses were selected on preliminary study in which male and female SD rats were administered 0, 250, 500 and 1000 mg / kg for 14 days, Body weight and food intake of individual animals were measured on days 1, 7, and 14 before administration on days 1, 2, 7, 14, 21 and 28, and on the autopsy day of each day after the end of each period, All the cases were autopsied on day 28 of administration or the next day after recovery 14 days.

No abnormality was observed in both sexes in each administration group in either administration period or recovery period. Only the males in the 250 mg / kg administration group showed significant increase in body weight gain and body weight gain during 28 days of administration, but not a dose-dependent change. Only the males in the 250 mg / kg administration group showed a significant increase in food intake compared with the control group on the 28th administration but not the dose-dependent change. There was no significant change in Urinalysis each treatment group of males and females as compared with the control group in either the administration period or the recovery period. Male of 250 mg / kg or higher administration group and female of 500 mg / kg or more administration group were reddish purple jejunum ileum contents, dark greening of contents of cecum in males and females of 250 mg/ kg or more administered group , Pale reddening of jejunum ileum and intraabdominal fat tissue were observed. These changes showed an increase in the number of cases as the dose increased and it was judged that the change in color tone itself had no toxicological significance.

Significant higher values of absolute and relative weights of liver in male of 250 mg / kg or more in comparison with control group, significant lower value of absolute and relative weight of thymus in males and females of 500 mg / kg administered group Significantly high values of relative weight of liver, high value trend of absolute weight, significant low value of ovarian relative weight, low value trend of absolute weight were observed in females at low trend, 1000 mg / kg administration group, recovery At the end of the period, a significant high value of the absolute weight of the liver, a high value tendency of the relative weight, a low tendency of the absolute weight of the spleen, and a significant low value of the relative weight were observed in the male of 1000 mg / kg administration group. Besides, a significant high value of the absolute weight of the adrenal glands was seen in the males of the 250 mg / kg administration group, but it was not a dose-dependent change, and at the end of the recovery period female of 1000 mg / kg administration group received pituitary Significant lower values of absolute and relative weights were observed, but not changed at the end of the administration. Only mild changes such as kidney, liver, pituitary gland and the like were observed in one males and two females in the 1000 mg / kg administration group. Even at the end of the recovery period, only mild changes were observed in the lungs, liver, kidneys, etc. of 3 males in the 1000 mg / kg administration group. As no effects on reproductive organ weight was observed at 500mg/kg dose group hence,No Observed Adverse Effect Level (NOAEL) was considered to be 500mg/kg/day, Whenmale and femaleCrj:CD(SD)IGS rats were treated with test material orally.

 

Study 2

Reproductive/developmental Toxicitystudy of test materialwas performed according to OECD guideline 414 . Female Wistar rat Crl:(WI) BR (outbred, SPF-Quality) 24female/dose group were treated withtest material in dose concentration 0, 100, 300 and 1000 mg/kg bwinwater (Milli-U)by oral gavage route in dose volume 10ml/kg once daily from day 6 to day 20post-coitum.The animals were checked daily for the clinical signs. Body weights and food consumptionwas determined at periodically during pregnancy. On GD 21, all females were subjected toan examinationpost-mortemand external, thoracic and abdominal macroscopic findingswere recorded. The ovaries and uterine horns were dissected and examined for the numberofcorpora lutea, the weight of the gravid uterus, the number of implantation site scars, the number and distribution of live/dead foetuses and embryo-foetal deaths, the weight of eachlive foetus and corresponding placenta, foetal sex and externally visible foetal macroscopicabnormalities. Alternate foetuses of each litter were preserved in 96% aqueous ethanol or Bouin's fluid, and subjected to skeletal or visceral examinations respectively.

There was one death in the 1000mg/kg dose group on GD 13, but there were no relevant toxicologically clinical or pathological signs. Since the cause of death is unclear, a treatment-related effect can not be excluded. Orange to red urine was noted in all animals in the 300mg/kg and 1000mg/kg dose groups and in some of the 100mg/kg dose group. Body weight and body weight gain were comparable to the control in the 100mg/kg and 300mg/kg dose group, but there was a statistically significant decrease in the 1000mg/kg dose group. This decrease was mirrored in the food consumption in the 1000mg/kg dose group, In the 300mg/kg dose group, food consumption decreased from GD12. Macroscopic observation of enlarged spleen in one 300mg/kg dose and 5 in 1000mg/kg dose animals was observed. In addition, 2 animals in 1000mg/kg dose group had crateriform retractions in the stomach.Abdominal fat and the forestomach appeared stained yellow in some of the 1000mg/kg dose group. These were considered to be treatment related. Eight animals were not pregnant (3 control; 2 in 100mg/kg dose group, 2 in 300mg/kg dose group and 1 in 1000mg/kg dose group ) and in 300 mg/kg dose group one animal only showed implantation sites. Post-implantation losses increased at the 1000mg/kg dose resulting in decreased foetal numbers and increased foetal deaths. Embryonic resorption was not affected. No treatment related effects were seen at the other doses. The foetal sex ratio was not affected by treatment. At the 1000mg/kg dose, there was a high incidence of foetuses with major visceral abnormalities, i.e. severe umbilical hernia and associated visceral changes, including displacement of organs and absence of diaphragm. A number of the abnormal foetuses also exhibited malrotated hind limbs, atypical skeletal ossification. Some ossification parameters showed slight retardation that could be explained by the reduction in mean foetal weight recorded, whilst other parameters were slightly better ossified. The toxicological significance of the latter finding is unclear. HenceNo Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was considered to be 300 mg/kg bw/day (237 mg/kg bw/day corrected for dye content).on the bases of reproductive parameter and developmental effects. When female wistarrats were treated with test material orally.

 

Based on the data available from different studies ,test material did not showedreproductive toxicityat dose concentration 300 mg/kg body weight/day,when male and female rats were treated with test material orally,Thus, comparing this value with the criteria ofCLP regulationtest materialis not likely to classify as reproductive toxicant.

 

 

 

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

300 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Data is Klimicsh 2 and from authoritative database

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Reproductive toxicity study

Data available from different studies were reviewed to determine the reproductive toxicity of testchemical.The studies are as mentioned below:

 Study 1

The reproductive organ toxicity study of test material was performed using twenty-eight-day Repeat Dose Oral Toxicity study of test material in Crj: CD (SD)IGS male and female rats. The test material was suspended using a 0.5% methyl cellulose aqueous solutionand administeried in dose concentration 0, 250, 500, and 1000 mg / kg orally for 28 days. Each dose group and recovery group contain 7 male and 7 female. Doses were selected on preliminary study in which male and female SD rats were administered 0, 250, 500 and 1000 mg / kg for 14 days, Body weight and food intake of individual animals were measured on days 1, 7, and 14 before administration on days 1, 2, 7, 14, 21 and 28, and on the autopsy day of each day after the end of each period, All the cases were autopsied on day 28 of administration or the next day after recovery 14 days.

No abnormality was observed in both sexes in each administration group in either administration period or recovery period. Only the males in the 250 mg / kg administration group showed significant increase in body weight gain and body weight gain during 28 days of administration, but not a dose-dependent change. Only the males in the 250 mg / kg administration group showed a significant increase in food intake compared with the control group on the 28th administration but not the dose-dependent change. There was no significant change in Urinalysis each treatment group of males and females as compared with the control group in either the administration period or the recovery period. Male of 250 mg / kg or higher administration group and female of 500 mg / kg or more administration group were reddish purple jejunum ileum contents, dark greening of contents of cecum in males and females of 250 mg/ kg or more administered group , Pale reddening of jejunum ileum and intraabdominal fat tissue were observed. These changes showed an increase in the number of cases as the dose increased and it was judged that the change in color tone itself had no toxicological significance.

Significant higher values of absolute and relative weights of liver in male of 250 mg / kg or more in comparison with control group, significant lower value of absolute and relative weight of thymus in males and females of 500 mg / kg administered group Significantly high values of relative weight of liver, high value trend of absolute weight, significant low value of ovarian relative weight, low value trend of absolute weight were observed in females at low trend, 1000 mg / kg administration group, recovery At the end of the period, a significant high value of the absolute weight of the liver, a high value tendency of the relative weight, a low tendency of the absolute weight of the spleen, and a significant low value of the relative weight were observed in the male of 1000 mg / kg administration group. Besides, a significant high value of the absolute weight of the adrenal glands was seen in the males of the 250 mg / kg administration group, but it was not a dose-dependent change, and at the end of the recovery period female of 1000 mg / kg administration group received pituitary Significant lower values of absolute and relative weights were observed, but not changed at the end of the administration. Only mild changes such as kidney, liver, pituitary gland and the like were observed in one males and two females in the 1000 mg / kg administration group. Even at the end of the recovery period, only mild changes were observed in the lungs, liver, kidneys, etc. of 3 males in the 1000 mg / kg administration group. As no effects on reproductive organ weight was observed at 500mg/kg dose group hence,No Observed Adverse Effect Level (NOAEL) was considered to be 500mg/kg/day, Whenmale and femaleCrj:CD(SD)IGSrats were treated with test materialorally.

 

Study 2

Reproductive/developmental Toxicitystudy of test materialwas performed according to OECD guideline 414 . Female Wistar rat Crl:(WI) BR (outbred, SPF-Quality) 24female/dose group were treated withtest material in dose concentration 0, 100, 300 and 1000 mg/kg bwinwater (Milli-U)by oral gavage route in dose volume 10ml/kg once daily from day 6 to day 20post-coitum.The animals were checked daily for the clinical signs. Body weights and food consumptionwas determined at periodically during pregnancy. On GD 21, all females were subjected toan examinationpost-mortemand external, thoracic and abdominal macroscopic findingswere recorded. The ovaries and uterine horns were dissected and examined for the numberofcorpora lutea, the weight of the gravid uterus, the number of implantation site scars, thenumber and distribution of live/dead foetuses and embryo-foetal deaths, the weight of eachlive foetus and corresponding placenta, foetal sex and externally visible foetal macroscopicabnormalities. Alternate foetuses of each litter were preserved in 96% aqueous ethanol orBouin's fluid, and subjected to skeletal or visceral examinations respectively.

There was one death in the 1000mg/kg dose group on GD 13, but there were no relevant toxicologically clinical or pathological signs. Since the cause of death is unclear, a treatment-related effect can not be excluded. Orange to red urine was noted in all animals in the 300mg/kg and 1000mg/kg dose groups and in some of the 100mg/kg dose group. Body weight and body weight gain were comparable to the control in the 100mg/kg and 300mg/kg dose group, but there was a statistically significant decrease in the 1000mg/kg dose group. This decrease was mirrored in the food consumption in the 1000mg/kg dose group, In the 300mg/kg dose group, food consumption decreased from GD12. Macroscopic observation of enlarged spleen in one 300mg/kg dose and 5 in 1000mg/kg dose animals was observed. In addition, 2 animals in 1000mg/kg dose group had crateriform retractions in the stomach.Abdominal fat and the forestomach appeared stained yellow in some of the 1000mg/kg dose group. These were considered to be treatment related. Eight animals were not pregnant (3 control; 2 in 100mg/kg dose group, 2 in 300mg/kg dose group and 1 in 1000mg/kg dose group ) and in 300 mg/kg dose group one animal only showed implantation sites. Post-implantation losses increased at the 1000mg/kg dose resulting in decreased foetal numbers and increased foetal deaths. Embryonic resorption was not affected. No treatment related effects were seen at the other doses. The foetal sex ratio was not affected by treatment. At the 1000mg/kg dose, there was a high incidence of foetuses with major visceral abnormalities, i.e. severe umbilical hernia and associated visceral changes, including displacement of organs and absence of diaphragm. A number of the abnormal foetuses also exhibited malrotated hind limbs, atypical skeletal ossification. Some ossification parameters showed slight retardation that could be explained by the reduction in mean foetal weight recorded, whilst other parameters were slightly better ossified. The toxicological significance of the latter finding is unclear. HenceNo Observed Adverse Effect Level (NOAEL) for reproductive and developmental toxicity was considered to be 300 mg/kg bw/day (237 mg/kg bw/day corrected for dye content).on the bases of reproductive parameter and developmental effects. When female wistarrats were treated with test material orally.

 

Based on the data available from different studies ,test material did not showedreproductive toxicityat dose concentration 300 mg/kg body weight/day,when male and female rats were treated with test material orally,Thus, comparing this value with the criteria ofCLP regulationtest materialis not likely to classify as reproductive toxicant.

 

 

 

Effects on developmental toxicity

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Justification for classification or non-classification

Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.