Registration Dossier

Administrative data

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

Effects on fertility

Description of key information

Reproductive toxicity study

Based on the data available from different studies, the No Observed Adverse Effect Level (NOAEL) for test material was considered to be 1000 mg/kg/day for reproductive toxicity, when rodents were treated with test material orally by gavage. 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 data of the read-across chemicals.
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
WoE report is based on 3 reproductive toxicity studies i.e. WoE-2, WoE-3 and WoE-4.
GLP compliance:
not specified
Limit test:
no
Justification for study design:
No data
Species:
rat
Strain:
other: 2. Crj: CD(SD) 3. Wistar 4. HanBrl:WIST (SPF)
Sex:
male/female
Details on test animals and environmental conditions:
2. No Data Available
3. No data available
4. No data available
Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
other: 2. 0.5% w/v Methylcellulose aqueous solution (suspended) 3. 1% carboxymethylcellulose in water 4. bi-distilled water containing 1% carboxymethylcellulose
Details on exposure:
2. VEHICLE
- Justification for use and choice of vehicle (if other than water): 0.5 % Methylcellulose solution was used because the test chemical was insoluble in water.
- Concentration in vehicle: 5 mL/kg
3. - Concentration in vehicle: 0, 100, 300 and 1000 mg/kg bw/day (0, 94, 282 or 940 mg active dye/kg bw/day)
- Amount of vehicle (if gavage): 10 ml/kg
4. PREPARATION OF DOSING SOLUTIONS: The test substance mixed in bi-destilled water containing 1% carboxymethylcellulose
VEHICLE
- Justification for use and choice of vehicle (if other than water): The test substance mixed in bi-destilled water containing 1% carboxymethylcellulose
- Concentration in vehicle :0, 100, 300 and 1000 mg/kg bw/day (0, 86, 258, and 861 mg active dye/kg bw/day)
- Amount of vehicle (if gavage): 10 ml/kg bw
Details on mating procedure:
2. .- M/F ratio per cage: 12 pairs/dose
- Length of cohabitation: Male and female were allowed to live in a male cage one to one.
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy
3. No data available
4. No data available
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
2. The prepared solution was confirmed by UV-visible absorbance method
Duration of treatment / exposure:
2. Males: 42 Days
Females: 41-45 days(from 14 days before mating to day 4 of lactation)
3. 12 days (6-17 of gestation period)
4. 14 days (from day 6 (implantation) through to day 20 post coitum )
Frequency of treatment:
Daily
Details on study schedule:
2. No Data Available
3. A prenatal developmental study was conducted on female Wistar rats.
4. No data available
Remarks:
2. 0 (Control), 0 (Recovery), 40 (Low Dose), 200 (Mid-Dose), 1000 (High dose), 1000 (Recovery) mg/kg/day
3. 0, 100, 300 and 1000 mg/kg bw/day (0, 94, 282 or 940 mg active dye/kg bw/day)
4. 0, 100, 300 and 1000 mg/kg bw/day (0, 86, 258, and 861 mg active dye/kg bw/day)
No. of animals per sex per dose:
2. Males: 12 animals/group/sex (5 animals for recovery group)
Females: 12 animals/group/sex (5 animals for recovery group)
3. Total number of animals-88
0 mg /kg bw/day -22 female rats
94 mg /kg bw/day -22 female rats
282 mg /kg bw/day -22 female rats
940 mg /kg bw/day-22 female rats
4. Total:88
0 mg/kg bw/day:22
100mg/kg bw/day:22
300mg/kg bw/day:22
1000 mg/kg bw/day:22
Control animals:
yes, concurrent vehicle
Details on study design:
2. No Data Available
3. No data
4. No data available
Positive control:
No Data Available
Parental animals: Observations and examinations:
2. CAGE SIDE OBSERVATIONS: Yes
- Time schedule: No Data Available

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: No Data Available

BODY WEIGHT: Yes
- Time schedule for examinations: No Data Available

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
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes

OTHER: No Data Available
3. Parental animal: observation and examination- Clinical sign, body weight and food intake was observed.
Histopathology- About one half of the foetuses were examined for soft tissue anomalies whereas remaining foetuses were examined for skeletal anomalies following alizarin red staining.
4. CAGE SIDE OBSERVATIONS: yes

DETAILED CLINICAL OBSERVATIONS: Yes

Time schedule: twice daily.

BODY WEIGHT: Yes
Time schedule for examinations: daily
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes on 3-day intervals
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):
2. Yes, Oestrus cycle was examined.
3. No data
4. No data available
Sperm parameters (parental animals):
2. Parameters examined in [P] male parental generations: testis weight, epididymis weight
3. No data
4. No data available
Litter observations:
2. Yes, The effect of the test chemical was observed on the litter parameters.
3. Foetuses were sexed and weighed.
4. No data available
Postmortem examinations (parental animals):
2. SACRIFICE
- Male animals: All surviving animals.
- Maternal animals: All surviving animals

GROSS NECROPSY
- Yes, gross necropsy was performed on all the organs in abdomen, viscera and reproductive organs.

HISTOPATHOLOGY / ORGAN WEIGHTS: Yes, Organs weights and Histopathology was performed.
3. Embryonic resorptions and implantation sites was observed.
4. SACRIFICE: On day 21 post-coitum, the dams were sacrificed
Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]: No data
Maternal animals: yes (subjected to macroscopic examination.)
GROSS NECROPSY:
HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.: macroscopic examination was performed
Postmortem examinations (offspring):
2. No Data Available
3. Foetuses were observed externely.
4. 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: The foetuses were removed by Caesarean section, sexed, weighed, examined for gross external abnormalities, killed, and allocated to either visceral or skeletal (about one half of the foetuses for each examination).
Statistics:
2. No Data Available
3. No data
4. No data available
Reproductive indices:
2. Gestational Index, estrous cycle examination, Copulation Index, Copulation interval, fertility index, gestation period, no of corpora lutea, number of implantations, implantation index, delivery index, nursing behaviour.
3. No data
4. No data available
Offspring viability indices:
2. Pup Viability Index
3. No data
4. No data available
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
2. effects observed, treatment-related - Test item coloured fecal matter in both male and female animals was observed in both the dose groups.
3. effects observed, non-treatment-related - Discoloured faeces were observed at 940 mg/kg bw/day.
4. no effects observed - Clinical signs included darker faeces (all treated groups, from day 7-21 post coitum), and discolouration of skin, eyes and bedding (generally only in the 1000mg/kg bw dose group)
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
2. no mortality observed - No Mortality was observed in all animals in either of the dose groups.
3. no mortality observed
4. no mortality observed - No mortalities occurred.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
2. no effects observed - No abnormal effects were observed in body weight changes in males or females due to the test chemical administration.
3. not specified
4. effects observed, treatment-related - The mean body weight gain was slightly decreased and the corrected body weight gain (corrected for gravid uterus weight) was marginally lower in 1000mg/kg bw /day dose females compared to control females
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
2. no effects observed - No abnormal effects were observed in feed consumption patterns in males or females due to the test chemical administration.
3. not specified
4. no effects observed - Food consumption was similar in treated and control groups.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Description (incidence and severity):
2. no effects observed - No abnormal effects were observed in hematological parameters in males or females due to the test chemical administration.
3. not specified
4. not specified
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
2. effects observed, non-treatment-related - Significant decrease in Albumin content, Total Protein, and significant increase in α2-Globulin (%) in males of 1000 mg/kg bw was observed. No effects in females were observed. However, these effects were considered to be toxicolgically insignificant since, therse effects were observed to be reversed in the recovery group.
3. not specified
4. not specified
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
2. effects observed, treatment-related - Abnormal coloration in the urine was observed in males and females at 200 mg/kg bw and 1000 mg/kg bw dose groups. This coloration was attributed to the test chemical or a metabolite of the test chemical.
3. not specified
4. not specified
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
2. no effects observed - No effect of the test chemical was observed on the grip strength, locomotor activity and other functional observation battery parameters in either sex at all the dose groups.
3. not specified
4. not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
2. no effects observed - No histopathological abnormalities were observed in any animals of either sex and at all the dose levels, including the high dose recovery groups.
3. not specified
4. not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
2. no effects observed - No effects on the estrous cyclicity due to the test chemical was observed at any dose groups.
3. not specified
4. not specified
Reproductive function: sperm measures:
no effects observed
Description (incidence and severity):
2. no effects observed
3. not specified
4. not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
2. no effects observed - No effects on the reproductive performance due to the test chemical was observed at any dose groups.
3. no effects observed - In the group who received 100 mg /kg bw/day one female had only embryonic resorptions and in the highest dose group two females were not pregnant, one female had only empty implantation sites and a further one only embryonic resorptions at Caesarean section. These findings were considered to be incidental as a dose relation was missing.
4. no effects observed - One low-dose female was not pregnant. The incidence of pre-implantation loss was slightly higher (statistically significant) in the high-dose group; this difference was considered as being incidental by the study authors because pre-implantation loss mainly occurred prior to onset of treatment. Post-implantation loss and number of foetuses per dam was not affected.
2. Based on all the data of observations and results, the test chemical was not considered as toxic to reproductive parameters.
3. No data
4. not specified
Dose descriptor:
NOAEL
Remarks:
2
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
haematology
clinical biochemistry
urinalysis
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
reproductive function (oestrous cycle)
reproductive function (sperm measures)
reproductive performance
Remarks on result:
other: No effects observed
Dose descriptor:
NOAEL
Remarks:
3
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
female
Basis for effect level:
clinical signs
mortality
reproductive performance
Remarks on result:
other: overall no effects on reproductive performance was observed
Remarks:
Maternal toxicity was not observed at all dose level.
Dose descriptor:
NOAEL
Remarks:
4
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
female
Basis for effect level:
reproductive performance
Remarks on result:
other: overall no effects on reproductive parameters
Dose descriptor:
NOAEL
Remarks:
4
Effect level:
300 mg/kg bw/day
Based on:
test mat.
Sex:
female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
gross pathology
Remarks on result:
other: maternal toxicity
Critical effects observed:
not specified
Clinical signs:
no effects observed
Description (incidence and severity):
2. no effects observed
3. no effects observed - No significannt change were observed in treated group compare to control group.
4. not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
Description (incidence and severity):
2. no mortality observed
3. not specified
4. not specified
Body weight and weight changes:
no effects observed
Description (incidence and severity):
2. no effects observed
3. no effects observed - No significannt change were observed in treated group compare to control group.
4. no effects observed - No effects on foetal weight were observed.
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:
no effects observed
Description (incidence and severity):
2. no effects observed
3. not specified
4. not specified
Anogenital distance (AGD):
not specified
Nipple retention in male pups:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Description (incidence and severity):
2. no effects observed
3. no effects observed - No significant change was observed on external soft tissue and skeletal anomalies based in treated group compare to control group .
4. no effects observed - Incidences of external, visceral and skeletal findings were similar for control and treated groups.
Histopathological findings:
no effects observed
Description (incidence and severity):
2. no effects observed
3. no effects observed
4. not specified
Other effects:
no effects observed
Description (incidence and severity):
2. no effects observed - No effects were observed on number of pups delivered, number of live pups, live birth index, viability index and sex ratio on day 0 and day 4 at any dose groups.
3. not specified
4. not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
2. Based on the observation and results of the test chemical, no effects were observed on the developmental parameters of the pups at any dose levels.
3. not specified
4. not specified
Dose descriptor:
NOAEL
Remarks:
2
Generation:
F1
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
sexual maturation
clinical signs
mortality
body weight and weight gain
gross pathology
histopathology: non-neoplastic
other: number of pups delivered, number of live pups, live birth index, viability index and sex ratio on day 0 and day 4
Remarks on result:
other: No effects observed
Dose descriptor:
NOAEL
Remarks:
3
Generation:
F1
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
body weight and weight gain
gross pathology
Remarks on result:
other: overall no developmental toxic effects observed
Dose descriptor:
NOAEL
Remarks:
4
Generation:
F1
Effect level:
1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
body weight and weight gain
gross pathology
Remarks on result:
other: overall no effects on developmental parameters were observed
Critical effects observed:
not specified
Reproductive effects observed:
not specified
Conclusions:
Based on the data available from different studies, the No Observed Adverse Effect Level (NOAEL) for test material was considered to be 1000 mg/kg/day for reproductive toxicity, when rodents were treated with test material orally by gavage. Thus, comparing this value with the criteria of CLP regulation test material is not likely to classify as reproductive toxicant.
Executive summary:

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

 

In a combined repeated and reproductive toxicity study according to OECD 422 guideline, the test chemical was administered to Crl:CD (SD) male and female rats, aged 10 weeks old at the initiation of dosing. The rats were dosed at concentrations of 0, 40, 200, 1000 mg/kg/day with two reversal groups of 0 and 1000 mg/kg bw/day as control and high dose group recovery group, respectively. The vehicle used in the study was 0.5% w/v Methylcellulose aqueous suspended solution. Male animals were dosed for 42 Days, while females were dosed for 41-45 days. After the administration of the test chemical, test item colored fecal matter in both male and female animals was observed in both the dose groups. However, no mortality was observed in all animals in either of the dose groups. No abnormal effects were observed in body weight changes and feed consumption in males or females due to the test chemical administration. In hematological parameters, no abnormal effects were observed in hematological parameters in males or females due to the test chemical administration. In blood chemistry parameters, significant decrease in Albumin content, Total Protein, and significant increase in α2-Globulin (%) in males of 1000 mg/kg bw was observed. No effects in females were observed. However, these effects were considered to be toxicolgically insignificant since, these effects were observed to be reversed in the recovery group. Also, abnormal coloration in the urine was observed in males and females at 200 mg/kg bw and 1000 mg/kg bw dose groups. This coloration was attributed to the test chemical or a metabolite of the test chemical. No effect of the test chemical was observed on the grip strength, locomotor activity and other functional observation battery parameters in either sex at all the dose groups. No adverse effects were observed on any abdominal, visceral or reproductive organs / 100 grams of body weight due to the test chemical in either of the sex at any dose groups. In gross necropsy, colored aqueous content in the alimentary tract was observed in males, and in males and females both 40 mg/kg bw dose group and 200 mg/kg bw dose group, respectively. Also, mucosal discoloration of alimentary tract in males of 200 mg/kg bw was also observed. Colored aqueous content in the alimentary tract and mucosal discoloration of alimentary tract in both males and females were observed in 1000 mg/kg bw dose group. However, these effects were reversed in the recovery group doses and thus were not considered to be of toxicological significance. No histopathological abnormalities were observed in any animals of either sex and at all the dose levels, including the high dose recovery groups. No effects on the estrous cyclicity due to the test chemical were observed at any dose groups. No effects on the reproductive performance due to the test chemical were observed at any dose groups. No effects were observed on number of pups delivered, number of live pups, live birth index, viability index and sex ratio on day 0 and day 4 at any dose groups. Also, no test chemical changes were noted in the number of pups delivered, number of live pups, live birth index, viability index or sex ratio on day 0 and 4, clinical signs, body weights, external examinations, body weights or necropsy. Thus, based on all the observations and results it was concluded that the NOAEL for the test chemical was considered to be 1000 mg/kg bw.

 

In another Prenatal Developmental Toxicity Study, the given test chemical was exposed at the concentration of 0, 100, 300 and 1000 mg/kg bw/day (0, 94, 282 or 940 mg active dye/kg bw/day) through 6-17of gestation period by oral (gavage) to 22 females Wistar rats. The test substance (in 1% carboxymethylcellulose in water) was given daily at dose volumes of 10 ml/kg bw by oral gavage. The doses were selected on the basis of the results of a preliminary study in rats. Maternal evaluations and measurements included daily clinical signs and body weight/food intake recorded at designated intervals. The females were killed on gestation day 21, subjected to macroscopic examination, and foetuses were removed by Caesarean section. Common litter parameters were recorded and foetuses were sexed, weighed and submitted to external examination. About one half of the foetuses were also examined for soft tissue anomalies whereas remaining foetuses were examined for skeletal anomalies following alizarin red staining. There were 20 to 22 pregnant females per group. In the group who received 100 mg /kg bw/day one female had only embryonic resorptions and in the highest dose group two females were not pregnant, one female had only empty implantation sites and a further one only embryonic resorptions at Caesarean section. These findings were considered to be incidental as a dose relation was missing. No deaths were reported, and clinical signs were limited to discoloured faeces at 1000 mg/kg bw/day. No effects on litter parameters or foetal weight were observed. Foetal and litter incidences of external, soft tissue and skeletal anomalies were similar for control and treated groups. Under the condition of the study, NOAEL was considered to be 1000 mg /kg bw/day for reproductive toxicity in P0 female Wistar rats, when they were exposed with test material at the concentration of 0, 100, 300 and 1000 mg/kg bw/day through 6-17 of gestation period by oral (gavage).

 

Both the above studies were supported with another Prenatal Developmental Toxicity Study performed by using the given test chemical on 22 mated females per dose group HanBrl:WIST (SPF) rats at the concentration of 0, 100, 300 and 1000 mg/kg bw/day (0, 86, 258, and 861 mg active dye/kg bw/day). The test substance (in bi-distilled water containing 1% carboxymethylcellulose) was given daily at dose volumes of 10 ml/kg bw by oral gavage. Mortality, morbidity, signs of abortion, and clinical signs and/or symptoms was checked at least twice daily. Food consumption was recorded on 3-day intervals and body weights were recorded daily. The females were sacrificed on gestation day 21, subjected to macroscopic examination. The foetuses were removed by Caesarean section, sexed, weighed, examined for gross external abnormalities, killed, and allocated to either visceral or skeletal (about one half of the foetuses for each examination). No mortalities occurred. Clinical signs included darker faeces (all treated groups, from day 7-21 post coitum), and discolouration of skin, eyes and bedding (generally only in the highdose group). The mean body weight gain was slightly decreased and the corrected body weight gain (corrected for gravid uterus weight) was marginally lower in high-dose females compared to control females. Food consumption was similar in treated and control groups. One low-dose female was not pregnant. The incidence of pre-implantation loss was slightly higher (statistically significant) in the high-dose group; this difference was considered as being incidental by the study authors because pre-implantation loss mainly occurred prior to onset of treatment. Post-implantation loss and number of foetuses per dam was not affected. At necropsy discolouration of intestinal contents, kidneys, fatty tissues, intrauterine fluids and foetuses was noted in all high-dose dams. No effects on litter parameters or foetal weight were observed. Incidences of external, visceral and skeletal findings were similar for control and treated groups. Under the condition of the study, the No Observed Adverse Effect Level (NOAEL) for maternal toxicity is considered to be 300 mg/kg bw/day (258 mg active dye/kg bw/day) and the NOAEL for reproductive and developmental toxicity is considered to be 1000 mg/kg bw/day (861 mg active dye/kg bw/day), when female HanBrl:WIST (SPF) rats were treated with test material orally.

 

Based on the data available from different studies, NOAEL for test material was considered to be 1000 mg/kg/day for reproductive toxicity, when rodents 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.

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 handbook or collection of data.
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 test chemical. The studies are as mentioned below:

 

In a combined repeated and reproductive toxicity study according to OECD 422 guideline, the test chemical was administered to Crl:CD (SD) male and female rats, aged 10 weeks old at the initiation of dosing. The rats were dosed at concentrations of 0, 40, 200, 1000 mg/kg/day with two reversal groups of 0 and 1000 mg/kg bw/day as control and high dose group recovery group, respectively. The vehicle used in the study was 0.5% w/v Methylcellulose aqueous suspended solution. Male animals were dosed for 42 Days, while females were dosed for 41-45 days. After the administration of the test chemical, test item colored fecal matter in both male and female animals was observed in both the dose groups. However, no mortality was observed in all animals in either of the dose groups. No abnormal effects were observed in body weight changes and feed consumption in males or females due to the test chemical administration. In hematological parameters, no abnormal effects were observed in hematological parameters in males or females due to the test chemical administration. In blood chemistry parameters, significant decrease in Albumin content, Total Protein, and significant increase in α2-Globulin (%) in males of 1000 mg/kg bw was observed. No effects in females were observed. However, these effects were considered to be toxicolgically insignificant since, these effects were observed to be reversed in the recovery group. Also, abnormal coloration in the urine was observed in males and females at 200 mg/kg bw and 1000 mg/kg bw dose groups. This coloration was attributed to the test chemical or a metabolite of the test chemical. No effect of the test chemical was observed on the grip strength, locomotor activity and other functional observation battery parameters in either sex at all the dose groups. No adverse effects were observed on any abdominal, visceral or reproductive organs / 100 grams of body weight due to the test chemical in either of the sex at any dose groups. In gross necropsy, colored aqueous content in the alimentary tract was observed in males, and in males and females both 40 mg/kg bw dose group and 200 mg/kg bw dose group, respectively. Also, mucosal discoloration of alimentary tract in males of 200 mg/kg bw was also observed. Colored aqueous content in the alimentary tract and mucosal discoloration of alimentary tract in both males and females were observed in 1000 mg/kg bw dose group. However, these effects were reversed in the recovery group doses and thus were not considered to be of toxicological significance. No histopathological abnormalities were observed in any animals of either sex and at all the dose levels, including the high dose recovery groups. No effects on the estrous cyclicity due to the test chemical were observed at any dose groups. No effects on the reproductive performance due to the test chemical were observed at any dose groups. No effects were observed on number of pups delivered, number of live pups, live birth index, viability index and sex ratio on day 0 and day 4 at any dose groups. Also, no test chemical changes were noted in the number of pups delivered, number of live pups, live birth index, viability index or sex ratio on day 0 and 4, clinical signs, body weights, external examinations, body weights or necropsy. Thus, based on all the observations and results it was concluded that the NOAEL for the test chemical was considered to be 1000 mg/kg bw.

 

In another Prenatal Developmental Toxicity Study, the given test chemical was exposed at the concentration of 0, 100, 300 and 1000 mg/kg bw/day (0, 94, 282 or 940 mg active dye/kg bw/day) through 6-17of gestation period by oral (gavage) to 22 females Wistar rats. The test substance (in 1% carboxymethylcellulose in water) was given daily at dose volumes of 10 ml/kg bw by oral gavage. The doses were selected on the basis of the results of a preliminary study in rats. Maternal evaluations and measurements included daily clinical signs and body weight/food intake recorded at designated intervals. The females were killed on gestation day 21, subjected to macroscopic examination, and foetuses were removed by Caesarean section. Common litter parameters were recorded and foetuses were sexed, weighed and submitted to external examination. About one half of the foetuses were also examined for soft tissue anomalies whereas remaining foetuses were examined for skeletal anomalies following alizarin red staining. There were 20 to 22 pregnant females per group. In the group who received 100 mg /kg bw/day one female had only embryonic resorptions and in the highest dose group two females were not pregnant, one female had only empty implantation sites and a further one only embryonic resorptions at Caesarean section. These findings were considered to be incidental as a dose relation was missing. No deaths were reported, and clinical signs were limited to discoloured faeces at 1000 mg/kg bw/day. No effects on litter parameters or foetal weight were observed. Foetal and litter incidences of external, soft tissue and skeletal anomalies were similar for control and treated groups. Under the condition of the study, NOAEL was considered to be 1000 mg /kg bw/day for reproductive toxicity in P0 female Wistar rats, when they were exposed with test material at the concentration of 0, 100, 300 and 1000 mg/kg bw/day through 6-17 of gestation period by oral (gavage).

 

Both the above studies were supported with another Prenatal Developmental Toxicity Study performed by using the given test chemical on 22 mated females per dose group HanBrl:WIST (SPF) rats at the concentration of 0, 100, 300 and 1000 mg/kg bw/day (0, 86, 258, and 861 mg active dye/kg bw/day). The test substance (in bi-distilled water containing 1% carboxymethylcellulose) was given daily at dose volumes of 10 ml/kg bw by oral gavage. Mortality, morbidity, signs of abortion, and clinical signs and/or symptoms was checked at least twice daily. Food consumption was recorded on 3-day intervals and body weights were recorded daily. The females were sacrificed on gestation day 21, subjected to macroscopic examination. The foetuses were removed by Caesarean section, sexed, weighed, examined for gross external abnormalities, killed, and allocated to either visceral or skeletal (about one half of the foetuses for each examination). No mortalities occurred. Clinical signs included darker faeces (all treated groups, from day 7-21 post coitum), and discolouration of skin, eyes and bedding (generally only in the highdose group). The mean body weight gain was slightly decreased and the corrected body weight gain (corrected for gravid uterus weight) was marginally lower in high-dose females compared to control females. Food consumption was similar in treated and control groups. One low-dose female was not pregnant. The incidence of pre-implantation loss was slightly higher (statistically significant) in the high-dose group; this difference was considered as being incidental by the study authors because pre-implantation loss mainly occurred prior to onset of treatment. Post-implantation loss and number of foetuses per dam was not affected. At necropsy discolouration of intestinal contents, kidneys, fatty tissues, intrauterine fluids and foetuses was noted in all high-dose dams. No effects on litter parameters or foetal weight were observed. Incidences of external, visceral and skeletal findings were similar for control and treated groups. Under the condition of the study, the No Observed Adverse Effect Level (NOAEL) for maternal toxicity is considered to be 300 mg/kg bw/day (258 mg active dye/kg bw/day) and the NOAEL for reproductive and developmental toxicity is considered to be 1000 mg/kg bw/day (861 mg active dye/kg bw/day), when female HanBrl:WIST (SPF) rats were treated with test material orally.

 

Based on the data available from different studies, NOAEL for test material was considered to be 1000 mg/kg/day for reproductive toxicity, when rodents 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

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

Comparing the NOAEL value and effects observed on rodents, with the criteria of CLP regulation the given test chemical is not likely to classify as reproductive toxicant.