Disodium 2-(2,4,5,7-tetrabromo-6-oxido-3-oxoxanthen-9-yl)benzoate

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 1000mg/kg /day for reproductive toxicity, when male and female 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:

one-generation reproductive 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

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

1.TEST ANIMALS
- Source: Laboratory Supply Co., Indianapolis, Indiana, USA
- Age at study initiation: adult rats
- Weight at study initiation: (P) Males: 200-220 g; Females: 200-220 g
- Fasting period before study: No data available
- Housing: No data available
- Use of restrainers for preventing ingestion (if dermal): Not applicable
- Diet (e.g. ad libitum): The experimental diets were freely available throughout postnatal development to the offspring of all delivering dams (up to the end of the experiments at 90-110 days of age of the offspring).
- Water (e.g. ad libitum): No data available
- Acclimation period: 5 days before assignment to treatment groups

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
2.Details on test animal
TEST ANIMALS
- Source: Charles River Breeding Laboratories
- Age at study initiation: (P) x wks; (F1) x wks: P- 18 weeks
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: P-246 to 379 g
- Fasting period before study: No data available
- Housing: Animals were housed individually and identified by ear tag.
- Diet (e.g. ad libitum): Animals were housed individually
- Water (e.g. ad libitum): water, ad lib.
- Acclimation period: 4 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.22 ± 15 °C
- Humidity (%):50 ± 15%
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): 12-hr light/dark cycle.

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

Route of administration:

other: 1.oral: feed 2.oral: gavage

Type of inhalation exposure (if applicable):

not specified

Vehicle:

not specified

Details on exposure:

1.Details on exposure
PREPARATION OF DOSING SOLUTIONS: Test chemical mixed in powdered Purina rat chow.

VEHICLE
- Justification for use and choice of vehicle (if other than water):Purina rat chow were used.
- Concentration in vehicle: 0.0, 0.25, 0.5 and 1.0 %
2.PREPARATION OF DOSING SOLUTIONS: The test chemical was mixed with vehicle at dose levels of 0, 100, 500 or 1500 mg/kg body weight and prepared daily

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

VEHICLE
- Justification for use and choice of vehicle (if other than water):No data
- Concentration in vehicle:0, 100, 500 or 1500 mg/kgbw
- Amount of vehicle (if gavage):10 mL/Kg
- Lot/batch no. (if required):No data
- Purity:No data

Details on mating procedure:

Study 1
- M/F ratio per cage:1:1
- Length of cohabitation:14 days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:The presence of sperm in the daily vaginal lavage of the females and designated as day zero of gestation.
Study 2.- M/F ratio per cage: 1: 1.
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy Copulatory plug or presence of sperm in vaginal washings was designated as day 0 of gestation

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

study 1
For male:fed diets containing the dye for 2 weeks
For female:The dye was fed to each treatment group for 2 weeks prior to breeding, 1-14 days during breeding then to the females during gestation and lactation.
Study 2
14 days (days 6-19 of gestation)

Frequency of treatment:

Daily

Details on study schedule:

On the day following birth two males and two females from each litter were designated for preweaning testing. Two additional males and two additional females per litter which were not tested
prior to weaning were designated for postweaning testing at weaning.

Usually the first 5-15 pairs of animals enrolled in the study in each group

Remarks:

Study 1
Doses / Concentrations:
0.0, 0.25, 0.5, or 1.0%( 250, 500 and 1000 mg/kg bw/d)

Study 2
0.0,100,500,1500mg/kg bw /d

No. of animals per sex per dose:

Study 1
Total: 196
0.0%: 19 male, 19 female
0.25%: 22 male, 22 female
0.5 %: 18 male, 18 female
1.0%: 21 male, 21 female
50 mg/kg (positive control): 18 male, 18 female
Study 2
Total: 100 females
0 mg/Kg bw: 25 females
100 mg/Kg bw: 25 females
500 mg/Kg bw: 25 females
1500 mg/Kg bw: 25 females

Control animals:

yes

Details on study design:

No data available

Positive control:

For experiment 1:offspring injected daily with 50 mg/kg of hydroxyurea on postnatal days 2-10 of life as a positive control
For experiment 2: no positive control were used.

Parental animals: Observations and examinations:

Study 1 & 2
Parental animals observation and examinations
CAGE SIDE OBSERVATIONS: No data
- Time schedule:No data
- Cage side observations checked in table [No.?] were included.No data

DETAILED CLINICAL OBSERVATIONS: No data
- Time schedule:No data

BODY WEIGHT: Yes
- Time schedule for examinations:weekly except during breeding

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

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

OTHER: No data

Oestrous cyclicity (parental animals):

No data available

Sperm parameters (parental animals):

No data available

Litter observations:

Study 1 & 2.Litters were examined and data collected on litter size, sex distribution, weight, and number of dead or malformed offspring.
The offspring were assessed on a series of tests using the Cincinnati Psychoteratogenicity Screening Test Battery, plus weight, food consumption, physical landmarks of development, and brain weight.

Postmortem examinations (parental animals):

Study 1 & 2
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.]

GROSS NECROPSY: yes
- Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]

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

Postmortem examinations (offspring):

Malformed offspring were observed.

Statistics:

1.Frequency data (e.g., mortality) were analyzed by Fisher's test
2.Differences in the fetal sex distribution and the number of litters with malformations between control and treated groups wcre compared using the Chi-square test criterion with Yates" correction for 2 x 2 contingency lables and, or Fisher's exact probability test as described by Sicgel (1956). The numbers of early and late resorptions, dead foetuses and post-implantation losses were compared between groups by the Mann Whitney U test as described by Siegel (1956) and Well (1970). The mean numbers of viable foetuses, total implantations, corpora lutca and mean t\)etal weights were compared between groups by analysis of variance (oneway classification), Bartlett's test l\~r homogeneity of variances, and the appropriate t test (for equal or unequal variances) as described by Steel & Torric (1960) using Dunnctt's multiple comparison tables (1964).

Reproductive indices:

No data available

Offspring viability indices:

Viability indice on day 1-90 were examined.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

2.Orange discoloration of the urine was observed in all the treated rats as compared to control.

Dermal irritation (if dermal study):

not specified

Mortality:

mortality observed, treatment-related

Description (incidence):

2.When treated with 1500 mg/kg bw, Six rats died during the dosing period as compared to control.
Survival was 100% in the controls and the groups receiving 100 and 500 mg/kg of dye.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

1.no effects observed
2.When treated with 1500 mg/kg bw, slight reductions in body-weight gains as compared to controls, throughout the dosing period

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

There were no significant differences found among the groups in the experiment for food consumption prior to breeding or during gestation or lactation in the parental animals.

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:

effects observed, treatment-related

Description (incidence and severity):

2.Orange discoloration of the urine was noted in all treated rats during the treatment period.

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:

no effects observed

Description (incidence and severity):

1.No significant effects of any of the measures of reproductive performance were found in either experiment
2.No effect on Viable and non-viable foetuses, early and late resorptions, total implantations and corpora lutea and sex ratio of fetuses were observed as compared to control.

Body weight: effects were not, however, reflected in any significant changes in body weights.
Food consumption: There were no significant differences found among the groups in the first Red-3 experiment for food consumption prior to breeding or during gestation or lactation in the parental animals.
Reproductive performance: No significant effects of any of the measures of reproductive performance were found in either experiment

Dose descriptor:

NOAEL

Effect level:

> 1 000 - < 1 500 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No compound related effects observed on body weight,food consumption and reproductive performance

Remarks on result:

other: No toxic effects were observed

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

no mortality observed

Description (incidence and severity):

No effect on Viable and non-viable fetuses were observed as compared to control.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No effect on Body weight of fetuses were observed as compared 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:

no effects observed

Description (incidence and severity):

1.No malformations were seen upon external examination
2.When treated with 1500 mg/kg bw, A slight increase in the number of litters with unossified sternebrae (sternebrae nos 1-6) and rudimentary 14th rib(s) was observed however, these values fell within the ranges of historical control data.

No biologically meaningful or statistically significant differences in the number of litters and number of fetuses with malformations and number of fetuses or litters with developmental variations were observed in treated rats as compared to control.

Histopathological findings:

not specified

Other effects:

not specified

Behaviour (functional findings):

no effects observed

Description (incidence and severity):

No advers effect were observed onPsychotoxicity of treated pups as compared to control.

Developmental immunotoxicity:

not specified

Body weight and Food consumption: No significant reductions were found in offspring food consumption or body weight in either experiment from birth through day 90 of postnatal life.

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

> 1 000 - <= 1 500 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effects observed on external examination of offspring,food consumption,body weight.

Remarks on result:

other: No developmental toxic effects were observed

Critical effects observed:

not specified

System:

other: not specified

Organ:

not specified

Reproductive effects observed:

not specified

Treatment related:

not specified

Conclusions:

NOAEL was considered to be in range of 1000-1500 mg/kg/day for F0 and F1 generation when Sprague-dawley male and female rat were exposed to test chemical .

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 and developmental effects of test chemical have been investigated in a 1-generation study in rats. The test chemical was administered at dietary dose levels of 0, 0.25, 0.5and 1.0 % (250, 500 and 1000 mg /kg bw/d) togroups of male and female Sprague-Dawley rats for two weeks prior to breeding, 1-14 days during breeding, then to females during gestation and lactation. The experimental diets were freely available to the offspring throughout postnatal development up to the age of 90-110 days. Body weights were measured weekly except during breeding. On the dayfollowing birth, all litters were examined and data collected on litter size, sex distribution,weight, and number of dead or malformed offspring. A variety of behavioural tests were determined in the offspring. Further, brainweights of day 90 were determined in the offspring. The experiment was replicated after2 years using the same exposure regimen.

The test chemical produced no effects on paternal or offspring weight or food consumption. No significant effects of any of the measures ofreproductive performance (proportion of females producing litters to those bred, thenumber of small litters, gestation length, litter size, sex ratio) were observed. No malformations were seen upon external examination. Preweaning offspring mortality was significantly increased at the 1.0 % and 0.5 % dose levels in the first experiment, but not inthe second. No adverse effects on offspring growth or adult regional brain weight were observed.No adverse effects were found in the present experiments on parental weight, food consumption, or reproductive performance.Therefore,NOAEL was considered to be 1.0 % (1000 mg/kg/day) for F0 and F1 generation when Sprague-dawleymale andfemale rat were exposed to test chemical .

Study 2

In a Teratogenic Potential Test, CD Sprague-Dawley female rats treated with test material in the concentration of 0, 100, 500 or 1500 mg/kg bw orally by gavage in water for 14 days (days 6-19 of gestation). Six rats died during the dosing period at 1500 mg/kg bw as compared to control. Survival was 100% in the controls and the groups receiving 100 and 500 mg/kg of dye. Slight reductions in body-weight gains at 1500 mg/kg bw and Orange discoloration of the urine was noted in all treated rats during the treatment period. No effect on reproductive parameters such as Viable and non-viable fetuses, early and late resorptions, total implantations and corpora lutea and sex ratio of fetuses were observed in treated female rats as compared to control. Green discoloration of the amniotic fluid was observed in 1, 10 and 16 rats at 100, 500 and 1500 mg/kg/day groups, respectively, and the small intestines were green in colour in many rats at 500 mg/Kg group. In addition, No effect on Body weight of fetuses were observed as compared to control. A slight increase in the number of litters with unossified sternebrae (sternebrae nos 1-6) and rudimentary 14th rib(s) was observed at 1500 mg/kg bw however, these values fell within the ranges of historical control data. No biologically meaningful or statistically significant differences in the number of litters and number of fetuses with malformations and number of fetuses or litters with developmental variations were observed in treated rats as compared to control. Therefore, NOAEL was considered to be 1500 mg/kg/day for P and F1 generation when CD Sprague-Dawley female rats treated with test material orally by gavage for 14 days. 

Based on the data available from different studies, test material did not showedreproductive toxicityat dose concentration 1000mg/kg /day. When male and female rats were treated with test material orally,thus, comparing this value with the criteria ofCLP regulationtest materialis not likelyto classify as reproductive toxicant.

 

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

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 and developmental effects of test chemical have been investigated in a 1-generation study in rats. The test chemical was administered at dietary dose levels of 0, 0.25, 0.5and 1.0 % (250, 500 and 1000 mg /kg bw/d) togroups of male and female Sprague-Dawley rats for two weeks prior to breeding, 1-14 days during breeding, then to females during gestation and lactation. The experimental diets were freely available to the offspring throughout postnatal development up to the age of 90-110 days. Body weights were measured weekly except during breeding. On the dayfollowing birth, all litters were examined and data collected on litter size, sex distribution,weight, and number of dead or malformed offspring. A variety of behavioural tests were determined in the offspring. Further, brainweights of day 90 were determined in the offspring. The experiment was replicated after2 years using the same exposure regimen.

The test chemical produced no effects on paternal or offspring weight or food consumption. No significant effects of any of the measures ofreproductive performance (proportion of females producing litters to those bred, thenumber of small litters, gestation length, litter size, sex ratio) were observed. No malformations were seen upon external examination. Preweaning offspring mortality was significantly increased at the 1.0 % and 0.5 % dose levels in the first experiment, but not inthe second. No adverse effects on offspring growth or adult regional brain weight were observed.No adverse effects were found in the present experiments on parental weight, food consumption, or reproductive performance.Therefore,NOAEL was considered to be 1.0 % (1000 mg/kg/day) for F0 and F1 generation when Sprague-dawleymale andfemale rat were exposed to test chemical .

Study 2

In a Teratogenic Potential Test, CD Sprague-Dawley female rats treated with test material in the concentration of 0, 100, 500 or 1500 mg/kg bw orally by gavage in water for 14 days (days 6-19 of gestation). Six rats died during the dosing period at 1500 mg/kg bw as compared to control. Survival was 100% in the controls and the groups receiving 100 and 500 mg/kg of dye. Slight reductions in body-weight gains at 1500 mg/kg bw and Orange discoloration of the urine was noted in all treated rats during the treatment period. No effect on reproductive parameters such as Viable and non-viable fetuses, early and late resorptions, total implantations and corpora lutea and sex ratio of fetuses were observed in treated female rats as compared to control. Green discoloration of the amniotic fluid was observed in 1, 10 and 16 rats at 100, 500 and 1500 mg/kg/day groups, respectively, and the small intestines were green in colour in many rats at 500 mg/Kg group. In addition, No effect on Body weight of fetuses were observed as compared to control. A slight increase in the number of litters with unossified sternebrae (sternebrae nos 1-6) and rudimentary 14th rib(s) was observed at 1500 mg/kg bw however, these values fell within the ranges of historical control data. No biologically meaningful or statistically significant differences in the number of litters and number of fetuses with malformations and number of fetuses or litters with developmental variations were observed in treated rats as compared to control. Therefore, NOAEL was considered to be 1500 mg/kg/day for P and F1 generation when CD Sprague-Dawley female rats treated with test material orally by gavage for 14 days. 

Based on the data available from different studies, test material did not showed reproductive toxicity at dose concentration 1000mg/kg /day. When male and female 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.

 

Effects on developmental toxicity

Description of key information

Developmental toxicity study

Based on the various studies available for the test chemical were reviewed to determine the developmental toxicity, NOAELfor test chemical was considered to be 1000 mg /kg bw/day .When rats were treated with test chemical orally. Thus, comparing this value with the criteria of CLP regulation test chemical is not likely to classify as reproductive and developmental toxicant.

Link to relevant study records

Reference

Endpoint:

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 developmental toxicity studies on rats

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

1.TEST ANIMALS
- Source:Laboratory Supply Co., Indianapolis, Indiana, USA
- Age at study initiation:90-110 days of age of the offspring
- Weight at study initiation:Male (200-220 g) and female (200-220 g)
- Fasting period before study:No data available
- Housing:No data available
- Diet (e.g. ad libitum):ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period:5 days
2.Details on test animal
TEST ANIMALS
- Source: Charles River Breeding Laboratories
- Age at study initiation: (P) x wks; (F1) x wks: P- 18 weeks
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: P-246 to 379 g
- Fasting period before study: No data available
- Housing: Animals were housed individually and identified by ear tag.
- Diet (e.g. ad libitum): Animals were housed individually
- Water (e.g. ad libitum): water, ad lib.
- Acclimation period: 4 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.22 ± 15 °C
- Humidity (%):50 ± 15%
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): 12-hr light/dark cycle.

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

Route of administration:

other: 1.oral: feed 2.oral: gavage

Type of inhalation exposure (if applicable):

not specified

Vehicle:

water

Details on exposure:

1.Details on exposure
PREPARATION OF DOSING SOLUTIONS: Test chemical mixed in powdered Purina rat chow.

VEHICLE
- Justification for use and choice of vehicle (if other than water):Purina rat chow were used.
- Concentration in vehicle: 0.0, 0.25, 0.5 and 1.0 %
2.PREPARATION OF DOSING SOLUTIONS:
The test material dissolved in water.

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

1.- Impregnation procedure: [artificial insemination / purchased timed pregnant / cohoused]:cohoused
- If cohoused:
- M/F ratio per cage:1:1
- Length of cohabitation:Till the confirmation of breeding
- Further matings after two unsuccessful attempts: [no / yes (explain)]:No data
- Verification of same strain and source of both sexes: [yes / no (explain)]:No data
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:The presence of sperm in the daily vaginal lavage of the females and designated as day zero of gestation.
- Any other deviations from standard protocol:No data
2.- Impregnation procedure: [artificial insemination / purchased timed pregnant / cohoused]
- If cohoused:
- M/F ratio per cage: 1:1
- Length of cohabitation:
- After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.
- Further matings after two unsuccessful attempts: [no / yes (explain)]
- Verification of same strain and source of both sexes: [yes / no (explain)]
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:Copulatory plug or presence of sperm in vaginal washings day 0 of pregnancy
- Any other deviations from standard protocol:

Duration of treatment / exposure:

1.For male : fed diets containing the dye for 2 weeks.
For female: The dye was fed to each treatment group for 2 weeks prior to breeding, 1-14 days during breeding , then to the females during gestation and lactation.
2.14 days

Frequency of treatment:

Daily

Duration of test:

1.From parent to offspring generation
2. 20 days

Remarks:

Study 1.
0.0, 0.25, 0.5, or 1.0%( 250, 500 and 1000 mg /kg bw/day )
Study 2.
0, 100, 500 or 1500 mg/kg body weight/day

No. of animals per sex per dose:

Study 1
Total: 196
0.0%: 19 male, 19 female
0.25%: 22 male, 22 female
0.5 %: 18 male, 18 female
1.0%: 21 male, 21 female
50 mg/kg (positive control): 18 male, 18 female
Study 2.
Total: 100
0 mg/kgbw/day: 25 female
100 mg/kgbw/day: 25 female
500 mg/kgbw/day: 25 female
1500 mg/kgbw/day: 25 female

Control animals:

yes

Details on study design:

No data available

Maternal examinations:

Survival, clinical sign and body weight were examined

Ovaries and uterine content:

he ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other:

Fetal examinations:

- External examinations: Yes: all per litter
- Soft tissue examinations: Yes
- Skeletal examinations: Yes
- Head examinations: No data

Statistics:

1.Analysis of variance for unequal group sizes, with split-plot designs where repeated measures were made, were performed on the majority of the data. Duncan a posteriori multiple range comparison tests for unequal group sizes (Kramer 1956) were used on those measures on which significant F-ratios occurred. Frequency data (e.g., mortality) were analyzed by Fisher's test for uncorrelated proportions (Guilford 1965). On preweaning tests, data on individual subjects were averaged for the entire litter or within sex and the litter was, therefore, the unit of analysis. On postweaning tests, only one male and one female was used from each litter on any given test,therefore, litter was again the unit of analysis, but no litter averaging was required. Because of the large number of tests done, and therefore, the number of statistical tests required, effects were only accepted as significant if the overall test (i.e., usually an F-test) occurred at a level ofp < 0.01 or beyond. Follow-up a posteriori individual group comparisons, however, were less stringent (p < 0.05) under the assumption that the F-test provided an adequate protection level when used at p < 0.01.

Indices:

No data available

Historical control data:

No data available

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Orange discoloration of the urine was observed in all the treated rats as compared to control.

Dermal irritation (if dermal study):

not specified

Mortality:

mortality observed, treatment-related

Description (incidence):

1.Survival was 100% in the controls and the groups receiving 100 and 500 mg/kg of dye.When treated with 1500 mg/kg bw/day, Six rats died during the dosing period as compared to control.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

When treated with 1500 mg/kg bw/day, decrease in body weight gain was observed in treated rats as compared to control.

Food consumption and compound intake (if feeding study):

no effects observed

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):

no effects observed

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

When treated with 1500 mg/kg bw/day, green discoloration of the amniotic fluid and green colored small intestines were observed in treated rats as compared to control. When treated with 100 and 500 mg/kg bw/day, green discoloration of the amniotic fluid was observed in treated rats as compared to control.

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

not specified

Early or late resorptions:

not specified

Dead fetuses:

no effects observed

Changes in pregnancy duration:

no effects observed

Changes in number of pregnant:

no effects observed

Other effects:

not specified

Dose descriptor:

NOAEL

Effect level:

> 1 000 - <= 1 500 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Remarks on result:

other: No reproductive toxic effects were observed

Abnormalities:

not specified

Localisation:

not specified

Fetal body weight changes:

no effects observed

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

no effects observed

Changes in litter size and weights:

no effects observed

Changes in postnatal survival:

no effects observed

External malformations:

no effects observed

Skeletal malformations:

no effects observed

Description (incidence and severity):

2.When treated wtih 1500 mg/kg bw/day, slight increase in the number of litters with unossified sternebrae (sternebrae nos 1-6) and rudimentary 14th rib(s) was observed in feotus of treated rats as compared to control, but the observed effect fell within the ranges of historical control data.

Visceral malformations:

not specified

Other effects:

no effects observed

Description (incidence and severity):

1.No advers effect were observed onPsychotoxicity of treated pups as compared to control.
2.There were no biologically meaningful or statistically significant differences in number of fetuses with malformations, number of foetuses or litters with developmental variations in any of the treated groups.

Dose descriptor:

NOAEL

Effect level:

> 1 000 - <= 1 500 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

fetal/pup body weight changes

changes in postnatal survival

external malformations

Remarks on result:

other: No developmental toxic effects were observed

Abnormalities:

not specified

Developmental effects observed:

not specified

Treatment related:

not specified

Conclusions:

NOAEL was considered to be in range of 1000-1500 mg/kg/day for F0 and F1 generation when Sprague-dawley male and female rat were exposed to test chemical .

Executive summary:

Data available from different studies for test chemicals were reviewed to determine the developmental toxicity of test chemical. The studies are as mentioned below:

Study 1

The reproductive and developmental effects of test chemical have been investigated in a 1-generation study in rats. The test chemical was administered at dietary dose levels of 0, 0.25, 0.5and 1.0 % (250, 500 and 1000 mg /kg bw/d) togroups of male and female Sprague-Dawley rats for two weeks prior to breeding, 1-14 days during breeding, then to females during gestation and lactation. The experimental diets were freely available to the offspring throughout postnatal development up to the age of 90-110 days. Body weights were measured weekly except during breeding. On the dayfollowing birth, all litters were examined and data collected on litter size, sex distribution,weight, and number of dead or malformed offspring. A variety of behavioural tests were determined in the offspring. Further, brainweights of day 90 were determined in the offspring. The experiment was replicated after2 years using the same exposure regimen.

The test chemical produced no effects on paternal or offspring weight or food consumption. No significant effects of any of the measures ofreproductive performance (proportion of females producing litters to those bred, thenumber of small litters, gestation length, litter size, sex ratio) were observed. No malformations were seen upon external examination. Preweaning offspring mortality was significantly increased at the 1.0 % and 0.5 % dose levels in the first experiment, but not inthe second. No adverse effects on offspring growth or adult regional brain weight were observed.No adverse effects were found in the present experiments on parental weight, food consumption, or reproductive performance.Therefore,NOAEL was considered to be 1.0 % (1000 mg/kg/day) for F0 and F1 generation when Sprague-dawleymale andfemale rat were exposed to test chemical .

Study 2

In a Teratogenic toxicity study, CD Sprague-Dawley female rats were treated with test material in the concentration of 0, 100, 500 and 1500 mg/kg body weight/day by oral gavage. Six rats died during the dosing period and at 1500 mg/kg bw/day and Orange discoloration of the urine was observed in all the treated rats as compared to control. Similarly, green discoloration of the amniotic fluid and green colored small intestines were observed at 1500 mg/kg bw/day and green discoloration of the amniotic fluid was observed at 100 and 500 mg/kg bw/day treated rats as compared to control. No effect on Post-implantation loss/dam, Total implantations/dam, Corpora lutea/dam and There were no biologically meaningful or statistically significant differences in the number of litters and Foetal sex of treated rats were observed as compared to control. In addition, No effect on viability of foetuse, number of fetuses with malformations, number of foetuses or litters with developmental variations was observed in any of the treated groups. Slight increase in the number of litters with unossified sternebrae (sternebrae nos 1-6) and rudimentary 14^thrib(s) was observed in feotus of 1500 mg/kg bw/day treated rats as compared to control, but the observed effect fell within the ranges of historical control data. Therefore, NOAEL was considered to be 1500 mg/kg body weight /day for F0 and F1 generation when CD Sprague-Dawley female rats were treated with test material orally.

Thus, based on the data available fortest chemical,No Observed Adverse Effect Level (NOAEL) was considered to be 1000mg /kg bw .When rats were treated with test chemical orally. Thus, comparing this value with the criteria of CLP regulationtest chemicalis not likely to classify as reproductive and developmental toxicant.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Data is Klimicsh 2 and from authoritative database

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 study

Data available from different studies for test chemicals were reviewed to determine the developmental toxicity of test chemical. The studies are as mentioned below:

Study 1

The reproductive and developmental effects of test chemical have been investigated in a 1-generation study in rats. The test chemical was administered at dietary dose levels of 0, 0.25, 0.5and 1.0 % (250, 500 and 1000 mg /kg bw/d) togroups of male and female Sprague-Dawley rats for two weeks prior to breeding, 1-14 days during breeding, then to females during gestation and lactation. The experimental diets were freely available to the offspring throughout postnatal development up to the age of 90-110 days. Body weights were measured weekly except during breeding. On the dayfollowing birth, all litters were examined and data collected on litter size, sex distribution,weight, and number of dead or malformed offspring. A variety of behavioural tests were determined in the offspring. Further, brainweights of day 90 were determined in the offspring. The experiment was replicated after2 years using the same exposure regimen.

The test chemical produced no effects on paternal or offspring weight or food consumption. No significant effects of any of the measures ofreproductive performance (proportion of females producing litters to those bred, thenumber of small litters, gestation length, litter size, sex ratio) were observed. No malformations were seen upon external examination. Preweaning offspring mortality was significantly increased at the 1.0 % and 0.5 % dose levels in the first experiment, but not inthe second. No adverse effects on offspring growth or adult regional brain weight were observed.No adverse effects were found in the present experiments on parental weight, food consumption, or reproductive performance.Therefore,NOAEL was considered to be 1.0 % (1000 mg/kg/day) for F0 and F1 generation when Sprague-dawleymale andfemale rat were exposed to test chemical .

Study 2

In a Teratogenic toxicity study, CD Sprague-Dawley female rats were treated with test material in the concentration of 0, 100, 500 and 1500 mg/kg body weight/day by oral gavage. Six rats died during the dosing period and at 1500 mg/kg bw/day and Orange discoloration of the urine was observed in all the treated rats as compared to control. Similarly, green discoloration of the amniotic fluid and green colored small intestines were observed at 1500 mg/kg bw/day and green discoloration of the amniotic fluid was observed at 100 and 500 mg/kg bw/day treated rats as compared to control. No effect on Post-implantation loss/dam, Total implantations/dam, Corpora lutea/dam and There were no biologically meaningful or statistically significant differences in the number of litters and Foetal sex of treated rats were observed as compared to control. In addition, No effect on viability of foetuse, number of fetuses with malformations, number of foetuses or litters with developmental variations was observed in any of the treated groups. Slight increase in the number of litters with unossified sternebrae (sternebrae nos 1-6) and rudimentary 14^thrib(s) was observed in feotus of 1500 mg/kg bw/day treated rats as compared to control, but the observed effect fell within the ranges of historical control data. Therefore, NOAEL was considered to be 1500 mg/kg body weight /day for F0 and F1 generation when CD Sprague-Dawley female rats were treated with test material orally.

Thus, based on the data available fortest chemical,No Observed Adverse Effect Level (NOAEL) was considered to be 1000mg /kg bw .When rats were treated with test chemical orally. Thus, comparing this value with the criteria of CLP regulationtest chemicalis not likely to classify as reproductive and developmental toxicant.

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.