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:

The results of the two-generation reproductive and developmental toxicity study on Wistar rats revealed no adverse effects on reproductive performance of the F0 (P) generation. The animals were fed with the test chemical at dietary levels of 0, 25, 100 or 500 mg/kg body weight. Maternal toxicity, indicated by a significant increase of the liver weight, was seen at 500 mg/kg body weight and day. No differences in mating success were found in treated dams and pregnancy proceeded normally in all groups. There was no alteration in numbers of resorption sites and no statistically significant change was seen in the number of fetuses per dam. The number of pups per litter did not differ. According to the findings of this study, no adverse effects on reproduction were found. The NOAEL value for Reproductive toxicity is considered to be 500 mg/kg/bw/day for females and 100 mg/kg/bw/day for male test animals. NOAEL for Developmental toxicity in the offspring is set as 750 mg/kg/bw/day.

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
Justification for type of information:
Data is from a publication.
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
no guideline followed
Principles of method if other than guideline:
The above experiments were performed to evaluate and assess reproductive toxicity of the test chemicals in test animals.
GLP compliance:
not specified
Justification for study design:
No Data Available
Specific details on test material used for the study:
- Molecular weight (if other than submission substance): 1011.3956 g/mol
- Substance type: Organic
- Physical state: No Data Available
- Impurities (identity and concentrations): No Data Available
Species:
other: Study 2, 3 and 5: Rat, Study 4 and 6: Mice
Strain:
other: Study 2, 3 and 5: Wistar, Study 4: ICR, Study 6: Crj: CD(SD)
Details on species / strain selection:
No Data Available
Sex:
male/female
Details on test animals and environmental conditions:
Study 2:
TEST ANIMALS
- Source: Bantin and Kingman (Hull, UK)
- Age at study initiation: (P) x wks; (F1) x wks
- Weight at study initiation: (P) Males: 200 g; Females: 60 g;
- Fasting period before study:Not available
- Housing: Polypropylene cages with sterilized sawdust as bedding male rats were housed singly, females in groups of seven or eight.
- Diet (e.g. ad libitum): ad libitum standard rodent breeding diet (CRM, Labsure)
- Water (e.g. ad libitum): ad libitum
- Acclimation period:2 wk

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3°C
- Humidity (%):30-70%
- Photoperiod (hrs dark / hrs light): 12-hr light dark cycle

Study 3:
TEST ANIMALS
- Source: Charles River Japan
- Age at study initiation: 4-week-old
- Weight at study initiation: 57.7 to 70.6 g
- Fasting period before study: Not available
- Housing: individually
- Diet (e.g. ad libitum):CE-2, ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period:Not available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 25±1 deg C
- Humidity (%):55±5%
- Air changes (per hr):Not available
- Photoperiod (hrs dark / hrs light): 11:13 h light: dark cycle

Study 4:
TEST ANIMALS
- Source: Charles River Japan
- Age at study initiation: 4-week-old
- Weight at study initiation: 19.4-22.8 g
- Fasting period before study:Not available
- Housing: individually
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Not available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 25±1 deg C
- Humidity (%):55±5%
- Air changes (per hr): Not available
- Photoperiod (hrs dark / hrs light): 11:13 h light: dark cycle

Study 5:
TEST ANIMALS
- Source: Charles River Japan
- Age at study initiation: 4-week-old
- Weight at study initiation: 57.7 to 70.6 g
- Fasting period before study: Not available
- Housing: individually
- Diet (e.g. ad libitum):CE-2 ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period:Not available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 25±1 deg C
- Humidity (%):55±5%
- Air changes (per hr):Not available
- Photoperiod (hrs dark / hrs light): 11:13 h light: dark cycle

Study 6:
TEST ANIMALS
- Source: Charles River Japan
- Age at study initiation: 4-week-old
- Weight at study initiation: 19.4-22.8 g
- Fasting period before study:Not available
- Housing: individually
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Not available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 25±1 deg C
- Humidity (%):55±5%
- Air changes (per hr): Not available
- Photoperiod (hrs dark / hrs light): 11:13 h light: dark cycle
Route of administration:
other: Study 2 to 6: oral: feed
Vehicle:
other: Study 2 to 6: Test chemical contained in the diet
Details on exposure:
Study 2: DIET PREPARATION:
- Rate of preparation of diet (frequency):twice a week
- Mixing appropriate amounts with (Type of food): standard rodent breeding diet
VEHICLE: Test chemical contained in the diet

Study 3: DIET PREPARATION
- Rate of preparation of diet (frequency):not available
- Mixing appropriate amounts with (Type of food): not available
- Storage temperature of food: not available
VEHICLE
Mixed with diet.

Study 4: DIET PREPARATION
- Rate of preparation of diet (frequency):not available
- Mixing appropriate amounts with (Type of food): not available
- Storage temperature of food: not available
VEHICLE
Mixed with diet.

Study 5: DIET PREPARATION
- Rate of preparation of diet (frequency):not available
- Mixing appropriate amounts with (Type of food): not available
- Storage temperature of food: not available
VEHICLE
Mixed with diet.

Study 6:DIET PREPARATION
- Rate of preparation of diet (frequency):not available
- Mixing appropriate amounts with (Type of food): not available
- Storage temperature of food: not available
VEHICLE
Mixed with diet.
Details on mating procedure:
Study 2:- M/F ratio per cage:1/8
- Length of cohabitation:5 week after first offering the test chemical containing diet and continuing until sufficient pregnancies obtained.
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy Not available
- After … days of unsuccessful pairing replacement of first male by another male with proven fertility. Not available
- Further matings after two unsuccessful attempts: [no / yes (explain)] Not available
- After successful mating each pregnant female was caged (how): singly on sterilized wood chips
- Any other deviations from standard protocol:-Not available

Study 3 to 6: Mating of animals were not performed
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No Data Available
Duration of treatment / exposure:
Study 2: Exposure period: male: 5 weeks (F0)
4 weeks (F1) and 22 months (F1)
female: 8 weeks (F0)
Premating exposure period (males): 3 weeks
Premating exposure period (females): 3 weeks

Study 3 to 6: 2 months
Frequency of treatment:
Study 2 to 6: Daily
Details on study schedule:
Study 2: Animals were offered diets containing the test chemical ,concentration of the test chemical adjusted every 2 weeks. Male rats were allowed access to female rats at commencing 5 wk after first offering the test chemical containing diet and continuing until sufficient pregnancies were obtained and ensure number of pups of the F1, generation. Throughout pregnancy and lactation, dams continued to receive the dose of the test chemical.

Study 3: Feeding Experiment: Male rats were fed 0, 0.06% or 0.25% of the test substance in diet for 2 months, followed by various examinations.

Study 4: Feeding Experiment: Male mice were fed 0, 0.06%, ,0.125 or 0.25% of the test substance in diet for 2 months, followed by various examinations.

Study 5: Feeding Experiment: Male rats were fed 0.06% and 0.25% of the test chemical in diet for 2 months,followed by various examinations.

Study 6: Feeding Experiment: Male mice were fed 0.06%, 0.125 or 0.25% of the test substance in diet for 2 months, followed by various examinations.

Remarks:
Study 2: Doses / Concentrations:
F0-0,25,100 or 500 mg/kg bw day F1: 0, 25, 100 or 250 mg/kg bw/day
Basis:
nominal in diet
Study 3:
Doses / Concentrations:
0, 0.06% (60 mg/kg/day) or 0.25% (250 mg/kg/day)
Basis:
nominal in diet
Study 4:
Doses / Concentrations:
0, 0.06%, ,0.125% or 0.25%
Basis:
nominal in diet
Study 5:
Doses / Concentrations:
0, 0.06% (60 mg/kg/day) or 0.25% (250 mg/kg/day)
Basis:
Study 6:
Doses / Concentrations:
0, 0.06%, 0.125% or 0.25%
Basis:
nominal in diet
No. of animals per sex per dose:
Study 2:
Control: 7 males and 50 females
25 mg/kg bw/day:7 males and 50 females
100 or 500 mg/kg bw/day:7 males and 50 females

Study 3:
Control-8 male rats
0.06%-8 male rats
0.25%-8 male rats

Study 4:
Control-8 male mice
0.06%-8 male mice
0.125%-8 male mice
0.25%-8 male mice

Study 5:
Control: 8 males
0.06%: 8 males
0.25%: 8 males

Study 6:
Control: 8 males
0.06%: 8 males
0.125%:8 males
0.25%:8 males
Control animals:
other: Please see 'remarks'
Details on study design:
Study 2: Dose ranging study was done for dose selection
Study 3 to 6: No further details available.
Positive control:
No Data Available
Parental animals: Observations and examinations:
Study 2: CAGE SIDE OBSERVATIONS:
Time:Daily
BODY WEIGHT: Yes
- Time schedule for examinations:Weekly
ORGAN WEIGHT: Yes
Histopathological examinations:Yes
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
liver-biochemistry.: Yes

Study 3 to 6:
Parental animal: observation and examination
Body weight gain: Yes
Time Schedule: Occasionally
Clinical signs of toxicity:Yes
Time Schedule: Daily
Clinical Biochemistry:Yes, Serum testosterone
Oestrous cyclicity (parental animals):
Study 2 to 6: No Data Available
Sperm parameters (parental animals):
Study 2: No Data Available
Study 3 to 6: Testicular sperm content and daily sperm production
Litter observations:
Study 2: Examinations included:
number of foetuses/dams, number of pups per litter, number of pups found dead.
Study 3 to 6: No Data Available
Postmortem examinations (parental animals):
Study 2: GROSS NECROPSY: Liver, kidney, thyroid, lung, adrenal and body fat (with emphasis on the liver).
Study 3 to 6: Organ weights: Preputial glands, testes, epididymides, prostate glands, seminal vesicles with coagulation glands, kidneys, spleen(0.25% group) and liver.
Postmortem examinations (offspring):
Study 2: GROSS NECROPSY: Liver, kidney, thyroid, lung, adrenal and body fat (with emphasis on the liver)
Study 3 to 6: No Data Available
Statistics:
Study 2: Statistical analysis of data was performed using Student's t-test.
Study 3 to 6: ariations of body and organ weights-as per standard deviation. Bartlett’s test, analysis of variance (ANOVA), Kruskal–Wallis test, and Dunnett’s parametric or non-parametric test were used as tests for significance of differences. Fisher’s exact test- histopathologic data.The limit for significance was set at P<0.05.
Reproductive indices:
Study 2: Pregnancy proceeded normally in all groups of animals.There was no alteration in numbers of resorption sites. No statistically significant change was seen in the number of foetuses/dams.
Study 3 to 6: No Data Available
Offspring viability indices:
Study 2: The number of pups per litter did not differ between dose groups. There was a trend to an increase in the number of pups found dead or dying soon after birth with increase in dose but the actual number of deaths in affected litters was not influenced by treatment with the test chemical.
Study 3 to 6: No Data Available
Clinical signs:
no effects observed
Description (incidence and severity):
Study 2: No effects observed
Study 3 to 6: No effects observed
Dermal irritation (if dermal study):
not specified
Mortality:
not specified
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
Study 2: Dams given diets containing the test chemical at 750 or 1000 mg/kg body weight gained less weight than controls in the last week of pregnancy and did not increase food consumption to the same extent during lactation, resulting in a fall in body weight. Dams treated with the test chemicals at nominal doses of 500 mg/kg body weight/day upwards showed a reduction in body weight gain during lactation.
Study 3 to 6: Body weight and food consumption: No significant differences in body weights at a level of 0.06% and 0.25%.
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
Study 2: Dams given diets containing the test chemical at 750 or 1000 mg/kg body weight id not increase food consumption to the same extent during lactation, resulting in a fall in body weight.
Study 3 to 6: Body weight and food consumption: No significant differences in body weights at a level of 0.06% and 0.25%. Mean intakes were almost equal to 0.06% and 0.25%.
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:
no effects observed
Description (incidence and severity):
Study 3 and 5: Serum testosterone concentration was not significantly changed.
Study 4: Serum testosterone was lower than the control in 0.25% dose group however increase at 0.125% dose but not significant difference was observed.
Study 6: 0.25% dose group: Serum testosterone was lower than the control but not significant difference was observed.
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):
Study 3: Histopathology:
0.06% (60 mg/kg bw) dose group-Histopathological observation of testes did not indicate severe injuries .
0.25% (250 mg/kg bw) - Exfoliation and sloughing of seminiferous tubules were abundantly observed.
DSP and DSP/g testis were a little reduced
Study 4: Histopathology:
0.25% dose group: severe exfoliation of seminiferous tubules was found ,Sloughing was also found in 50%
mice. Dilatated lumen was observed in 75% mice.
Dose dependent decrease was observed DSP (Daily Sperm Count) and DSP/g testis.
Study 5: Histopathology:
0.06% dose group: Histopathological observation of testes did not indicate severe injuries .
0.25%: Exfoliation and sloughing of seminiferous tubules were abundantly observed. Fatty changes in liver and liver enlargement were found in all rats fed BM. DSP and DSP/g testis were a little reduced.
Study 6: Histopathology:
0.25% dose group: severe exfoliation of seminiferous tubules was found ,Sloughing was also found in 50%
mice. Dilatated lumen was observed in 75% mice.
Dose dependent decrease was observed DSP and DSP/g testis.
Histopathological findings: neoplastic:
not specified
Other effects:
no effects observed
Description (incidence and severity):
Study 3: Other findings-
Uterotrophic activity: no consistent evidence for uterotrophic activity of TM at doses up to 1,000 mg/kg.
ER alpha competitive binding- TM competitively inhibited E2–ERa binding. IC50 of TM was found to be 1.8x10-5 M.

Study 4: Uterotrophic activity:
In immature female mice no consistent evidence for uterotrophic activity of TM at doses up to 1,000 mg/kg.
In OVX mice no significant effect were seen at doses of 60 and 300 mg/kg/day or dietary administration given at levels of 0.06% and 0.25% for 2 months.
Estrogenic activities: Dietary administration of 0.25% TM for 2 months significantly decreased in parametrial adipose tissue weight. TM decreased body radiation temperature.
ER alpha competitive binding- TM competitively inhibited E2–ERa binding. IC50 of TM was found to be 1.8x10-5 M.

Study 5: Uterotrophic activity: no consistent evidence for uterotrophic activity of BM at doses up to 1,000 mg/kg.
ER alpha competitive binding- The test chemical competitively inhibited E2–ERa binding. IC50 of the test chemical was found to be 7.3x10-6 M.

Study 6: Uterotrophic activity:
In immature female mice no consistent evidence for uterotrophic activity of BM at doses up to 1,000 mg/kg.
In OVX mice no significant effect were seen at doses of 60 and 300 mg/kg/day or dietary administration given at levels of 0.06% and 0.25% for 2 months.
Estrogenic activities: Dietary administration of 0.25% BM for 2 months significantly decreased in parametrial adipose tissue weight. BM decreased body radiation temperature.
ER alpha competitive binding- The test chemical competitively inhibited E2–ERa binding. IC50 of the test chemical was found to be 7.3x10-6 M.
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Study 2: Dams receiving 500 mg of the test chemical/kg body weight and day showed a significant increase in liver weight. There was no alteration in numbers of resorption sites. No statistically significant change was seen in the number of foetuses/dam.
There were no differences in food consumption or body weight gain between female control rats and female rats fed with the test chemical-containing diet. Reduced body weight gain observed, in male rats treated with 500 mg/kg body weight with the test chemical.

Study 5:Uterotrophic activity: no consistent evidence for uterotrophic activity of BM at doses up to 1,000 mg/kg.
ER alpha competitive binding- The test chemical competitively inhibited E2–ERa binding. IC50 of the test chemical was found to be 7.3x10-6 M.
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effect on body weight and food consumption,no clinical signs of toxicity.
Critical effects observed:
not specified
Clinical signs:
effects observed, treatment-related
Dermal irritation (if dermal study):
not specified
Mortality / viability:
no mortality observed
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:
no effects observed
Histopathological findings:
no effects observed
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
No significant effects on foetus development except pups belonging to 500 mg/kg bw/day failed to gain weight at the same rate as control pups.
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
500 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No differences in mating success or pregnancy were found. No alteration in numbers of resorption sites and no statistically significant change was seen in the number of fetuses per dam. The number of pups per litter did not differ
Critical effects observed:
not specified
Reproductive effects observed:
not specified
Treatment related:
not specified
Conclusions:
The NOAEL for toxicity to reproduction was assessed to be 500 mg/kg body weight and day; the NOAEL for parental toxicity was assessed to be 100 mg/kg body weight and day.
Executive summary:

Reproductive Toxicity Study:

Data from the reproductive toxicity studies is as follows:

Reproductive Toxicity Study 2:

The results of this two-generation reproductive and developmental toxicity study on Wistar rats revealed no adverse effects on reproductive performance of the F0 (P) generation. The animals were fed with the test chemical at dietary levels of 0, 25, 100 or 500 mg/kg body weight. Maternal toxicity, indicated by a significant increase of the liver weight, was seen at 500 mg/kg body weight and day. No differences in mating success were found in treated dams and pregnancy proceeded normally in all groups. There was no alteration in numbers of resorption sites and no statistically significant change was seen in the number of fetuses per dam. The number of pups per litter did not differ.According to the findings of this study, no adverse effects on reproduction were found. The NOAEL for toxicity to reproduction was assessed to be 500 mg/kg body weight and day; the NOAEL for parental toxicity was assessed to be 100 mg/kg body weight and day.

Reproductive Toxicity Study 3:

In a two month feeding study with male Slc:Wistar rats, the test chemical was administered in the test animals to verify if the test chemical induced toxic effects in the testes.The test chemical was administered through the diet. A total of 8 rats per groups were used including the control group. The test chemical was administered in two concentrations:0.06% test substance (approx.60 mg/kg/day) and 0.25% test substance (approx. 250 mg/kg/day). Animals feed with 0.06% test substance (approx.60 mg/kg/day) showed no toxic effects. Hence 0.06% is considered to be the NOAEL value.Animals feed with 0.25% test substance (approx. 250 mg/kg/day)  showed a decrease in absolute weight of epididymides, seminal vesicles, prostate glands, and preputial glands Moreover, the daily sperm production (DSP) showed a decrease in treated animals. Hence 0.25% is considered to be the LOAEL value; whereas the serum testosterone levels were not significantly changed in treated animals compared to control. Estrogenic activity of the animals after induction of the test chemical was noted in an in vitro ERa-binding assay (Estrogen receptor alpha). Thus, it might be assumed that, the test chemical is weakly toxic to male reproductive organs in rats. From, all the above observations, it was concluded that,the NOAEL and LOAEL value of the test chemical  was 0.06% and 0.25%, respectively ( approx. 60 mg/kg/day and approx. 250 mg/kg/day).

Reproductive Toxicity Study 4:

In a two month feeding study with male ICR mice, the test chemical was administered in the test animals to verify if the test chemical induced toxic effects in the testes.The test chemical was administered through the diet. A total of 8 mice per group were used including the control group. The test chemical was administered in three concentrations:0.06% test substance (approx.100 mg/kg/day), 0.125% (approx. 208 mg/kg bw) and 0.25% test substance (approx. 417 mg/kg/day). Animals fed with 0.06% test substance (approx.100 mg/kg/day) showed no toxic effects. Hence, 0.06% is considered to be the NOAEL value. In animals fed with 0.25% test substance (approx. 417 mg/kg/day) absolute weights of epididymides, seminal vesicles, prostate glands, and preputial glands were significantly decreased. Liver weight was significantly increased (liver enlargement).Moreover, the daily sperm production (DSP) showed a decrease in treated animals. Hence 0.25% is considered to be the LOAEL value.Serum testosterone was lower than the control in 0.25% dose group however increase at 0.125% dose but not significant difference was observed.Dietary administration of 0.25% test chemical for 2 months significantly decreased in parametrial adipose tissue weight. Test chemical decreased body radiation temperature.Thus, it might be assumed that, the test chemical is weakly toxic to male reproductive organs in mice. From, all the above observations, it was concluded that,the NOAEL and LOAEL value of the test chemical was 0.06% and 0.25%, respectively ( approx. 100 mg/kg/day and approx. 417 mg/kg/day).

Reproductive Toxicity Study 5:

In a two month feeding study with male Slc:Wistar rats, the test chemical induced toxic effects in the testes. Animals feed with 0.06% test chemical (approx. 60 mg/kg/day) showed no toxic effects. Hence 0.06% is considered to be the NOAEL value.Animals feed with 0.25% test chemical (approx. 250 mg/kg/day) showed a decrease in absolute weight of epididymides, seminal vesicles, prostate glands, and preputial glands. Moreover, the daily sperm production (DSP) was significant decreased in treated animals. Hence 0.25% is considered to be the LOAEL value; whereas the serum testosterone levels were not significant changed in treated animals compared to control. Estrogenic activity of the test substance was noted in an in vitro ERa-binding assay(Estrogen receptor alpha), which might be the reason for the test chemical being weakly toxic to male reproductive organs in rats.

Reproductive Toxicity Study 6:

In a two month feeding study with male Crj:CD-1 mice, the test chemical induced toxic effects in the testes. The animals were fed with the test chemical through diet. 8 animals per group were used in this study, including the control group.The animals were administered with the test chemical in the concentrations:0.06% (Approx.100 mg/kg bw, 0.125% (Approx. 208 mg/kg bw) and 0.25%(Approx. 417 mg/kg bw). Animals fed with 0.06% test chemical (approx.100 mg/kg/day) showed no toxic effects. Therefore this is considered to be the NOAEL value.In the 0.125% dose group a significant decrease in body weight gain was observed.Animals fed with 0.25% test substance (approx. 417 mg/kg/day) showed a decrease in absolute weights of epididymides, seminal vesicles, prostate glands, and preputial glands. Moreover, histopathological changes were seen related to reproductive organ. Hence 0.25% is considered to be the LOAEL value for male reproductive organ toxicity. Estrogenic activity of the test substance was noted in an in vitro ERa-binding assay, which might be the reason of the test chemical being weakly toxic to male reproductive organs in mice. Thus, from all the above observations,it led to the conclusion that the NOAEL and LOAEL value was 0.06% and 0.25%, respectively (approx.100 mg/kg/day and approx. 417 mg/kg/day).

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
100 mg/kg bw/day
Study duration:
chronic
Species:
rat
Quality of whole database:
Data is from a Klimisch 2 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 from the reproductive toxicity studies is as follows:

Reproductive Toxicity Study 2:

The results of this two-generation reproductive and developmental toxicity study on Wistar rats revealed no adverse effects on reproductive performance of the F0 (P) generation. The animals were fed with the test chemical at dietary levels of 0, 25, 100 or 500 mg/kg body weight. Maternal toxicity, indicated by a significant increase of the liver weight, was seen at 500 mg/kg body weight and day. No differences in mating success were found in treated dams and pregnancy proceeded normally in all groups. There was no alteration in numbers of resorption sites and no statistically significant change was seen in the number of fetuses per dam. The number of pups per litter did not differ.According to the findings of this study, no adverse effects on reproduction were found. The NOAEL for toxicity to reproduction was assessed to be 500 mg/kg body weight and day; the NOAEL for parental toxicity was assessed to be 100 mg/kg body weight and day.

Reproductive Toxicity Study 3:

In a two month feeding study with male Slc:Wistar rats, the test chemical was administered in the test animals to verify if the test chemical induced toxic effects in the testes.The test chemical was administered through the diet. A total of 8 rats per groups were used including the control group. The test chemical was administered in two concentrations:0.06% test substance (approx.60 mg/kg/day) and 0.25% test substance (approx. 250 mg/kg/day). Animals feed with 0.06% test substance (approx.60 mg/kg/day) showed no toxic effects. Hence 0.06% is considered to be the NOAEL value.Animals feed with 0.25% test substance (approx. 250 mg/kg/day)  showed a decrease in absolute weight of epididymides, seminal vesicles, prostate glands, and preputial glands Moreover, the daily sperm production (DSP) showed a decrease in treated animals. Hence 0.25% is considered to be the LOAEL value; whereas the serum testosterone levels were not significantly changed in treated animals compared to control. Estrogenic activity of the animals after induction of the test chemical was noted in an in vitro ERa-binding assay (Estrogen receptor alpha). Thus, it might be assumed that, the test chemical is weakly toxic to male reproductive organs in rats. From, all the above observations, it was concluded that,the NOAEL and LOAEL value of the test chemical  was 0.06% and 0.25%, respectively ( approx. 60 mg/kg/day and approx. 250 mg/kg/day).

Reproductive Toxicity Study 4:

In a two month feeding study with male ICR mice, the test chemical was administered in the test animals to verify if the test chemical induced toxic effects in the testes.The test chemical was administered through the diet. A total of 8 mice per group were used including the control group. The test chemical was administered in three concentrations:0.06% test substance (approx.100 mg/kg/day), 0.125% (approx. 208 mg/kg bw) and 0.25% test substance (approx. 417 mg/kg/day). Animals fed with 0.06% test substance (approx.100 mg/kg/day) showed no toxic effects. Hence, 0.06% is considered to be the NOAEL value. In animals fed with 0.25% test substance (approx. 417 mg/kg/day) absolute weights of epididymides, seminal vesicles, prostate glands, and preputial glands were significantly decreased. Liver weight was significantly increased (liver enlargement).Moreover, the daily sperm production (DSP) showed a decrease in treated animals. Hence 0.25% is considered to be the LOAEL value.Serum testosterone was lower than the control in 0.25% dose group however increase at 0.125% dose but not significant difference was observed.Dietary administration of 0.25% test chemical for 2 months significantly decreased in parametrial adipose tissue weight. Test chemical decreased body radiation temperature.Thus, it might be assumed that, the test chemical is weakly toxic to male reproductive organs in mice. From, all the above observations, it was concluded that,the LOAEL value of the test chemical was 0.06% and 0.25%, respectively ( approx. 100 mg/kg/day and approx. 417 mg/kg/day).

Reproductive Toxicity Study 5:

In a two month feeding study with male Slc:Wistar rats, the test chemical induced toxic effects in the testes. Animals feed with 0.06% test chemical (approx. 60 mg/kg/day) showed no toxic effects. Hence 0.06% is considered to be the NOAEL value.Animals feed with 0.25% test chemical (approx. 250 mg/kg/day) showed a decrease in absolute weight of epididymides, seminal vesicles, prostate glands, and preputial glands. Moreover, the daily sperm production (DSP) was significant decreased in treated animals. Hence 0.25% is considered to be the LOAEL value; whereas the serum testosterone levels were not significant changed in treated animals compared to control. Estrogenic activity of the test substance was noted in an in vitro ERa-binding assay(Estrogen receptor alpha), which might be the reason for the test chemical being weakly toxic to male reproductive organs in rats.

Reproductive Toxicity Study 6:

In a two month feeding study with male Crj:CD-1 mice, the test chemical induced toxic effects in the testes. The animals were fed with the test chemical through diet. 8 animals per group were used in this study, including the control group.The animals were administered with the test chemical in the concentrations:0.06% (Approx.100 mg/kg bw, 0.125% (Approx. 208 mg/kg bw) and 0.25%(Approx. 417 mg/kg bw). Animals fed with 0.06% test chemical (approx.100 mg/kg/day) showed no toxic effects. Therefore this is considered to be the NOAEL value.In the 0.125% dose group a significant decrease in body weight gain was observed.Animals fed with 0.25% test substance (approx. 417 mg/kg/day) showed a decrease in absolute weights of epididymides, seminal vesicles, prostate glands, and preputial glands. Moreover, histopathological changes were seen related to reproductive organ. Hence 0.25% is considered to be the LOAEL value for male reproductive organ toxicity. Estrogenic activity of the test substance was noted in an in vitro ERa-binding assay, which might be the reason of the test chemical being weakly toxic to male reproductive organs in mice. Thus, from all the above observations,it led to the conclusion that the NOAEL and LOAEL value was 0.06% and 0.25%, respectively (approx.100 mg/kg/day and approx. 417 mg/kg/day).

Effects on developmental toxicity

Description of key information

Developmental Toxicity Study:

In an experimental study, groups of mice of strain Evans No.1 were administered with the test chemical in oral dosage regime, with a daily dose of 0 and 750 mg/kg bw during the period before mating, continuing through mating and gestation up to the time when the animals were killed, and during pregnancy (1-18 days) only. The vehicle used for dosing the test chemical was Archias oil. The maternal animals were observed forMean body weight,fertility index and mating index.In uterine observations, the animals were observed for the number of corpora lutea, numbers and positions of implantation sites (including resorption sites). After performing all the observations, Nosignificant maternal toxicity and no significant embryotoxic effects were observed, as judged by examination of the skeletal or gross abnormalities of fully-developed fetuses. Thus, from all the observations, it was concluded that the NOAEL for embryotoxicity and teratogenicity was therefore considered to be 750 mg/kg bw.

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
Justification for type of information:
Data is from a handbook or a collection of data.
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
no guideline available
Principles of method if other than guideline:
The above experiments were performed to assess and evaluate the effects of the test chemical on the developmental parameters of the test animals.
GLP compliance:
not specified
Specific details on test material used for the study:
- Molecular weight (if other than submission substance): 1011.3956 g/mol
- Substance type: Organic
- Physical state: No Data Available
- Impurities (identity and concentrations): No Data Available
Species:
other: Study 2: Mouse; Study 3: Rat
Strain:
other: Study 2: Evans No. 1; Study 3: Tuck albino
Details on test animals and environmental conditions:
Study 2: TEST ANIMALS
- Source: Not available
- Age at study initiation:
- Weight at study initiation: Not available
- Fasting period before study: Not available
- Housing: Not available
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Not available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not available
- Humidity (%):Not available
- Air changes (per hr): Not available
- Photoperiod (hrs dark / hrs light): Not available

Study 3: TEST ANIMALS
- Source: Not available
- Age at study initiation:
- Weight at study initiation: Not available
- Fasting period before study: Not available
- Housing: Not available
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: Not available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Not available
- Humidity (%):Not available
- Air changes (per hr): Not available
- Photoperiod (hrs dark / hrs light): Not available

IN-LIFE DATES: From: To:

Study 4: TEST ANIMALS
- Source: Bantin and Kingman (Hull, UK)
- Age at study initiation: Not available
- Weight at study initiation: (P) Males: 200 g; Females: 60 g
- Fasting period before study: Not available
- Housing: polypropylene cages with sterilized sawdust as bedding male rats were housed singly, females in groups of seven or eight.
- Diet (e.g. ad libitum): ad libitum standard rodent breeding diet (CRM, Labsure)
- Water (e.g. ad libitum): ad libitum
- Acclimation period:2 wk

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3°C
- Humidity (%):30-70%
- Photoperiod (hrs dark / hrs light): 12-hr light dark cycle
Route of administration:
other: Study 2 and 3: Oral: gavage; Study 4: Oral: feed
Vehicle:
other: Study 2 and 3: Arachis Oil; Study 4: The test chemical contained in the diet
Details on exposure:
Study 2:PREPARATION OF DOSING SOLUTIONS: Not available
DIET PREPARATION
- Rate of preparation of diet (frequency): Not available
- Mixing appropriate amounts with (Type of food): Not available
- Storage temperature of food: Not available

VEHICLE
- Justification for use and choice of vehicle (if other than water): arachis oil
- Concentration in vehicle: Not available
- Amount of vehicle (if gavage): Not available
- Lot/batch no. (if required): Not available
- Purity: Not available

Study 3: PREPARATION OF DOSING SOLUTIONS: Not available
DIET PREPARATION
- Rate of preparation of diet (frequency): Not available
- Mixing appropriate amounts with (Type of food): Not available
- Storage temperature of food: Not available

VEHICLE
- Justification for use and choice of vehicle (if other than water): arachis oil
- Concentration in vehicle: Not available
- Amount of vehicle (if gavage): Not available
- Lot/batch no. (if required): Not available
- Purity: Not available

Study 4: DIET PREPARATION
- Rate of preparation of diet (frequency):twice a week
- Mixing appropriate amounts with (Type of food): standard rodent breeding diet
VEHICLE: Test chemical contained in the diet
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No Data Available
Details on mating procedure:
Study 2 and 3: No details of mating available
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy: Presence of vaginal plug.

Study 4:- M/F ratio per cage:1/8
- Length of cohabitation: 5 week after first offering the test chemical containing diet and continuing until sufficient pregnancies obtained.
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy: No Data Available
- After … days of unsuccessful pairing replacement of first male by another male with proven fertility: No Data Available
- Further matings after two unsuccessful attempts: [no / yes (explain)] No Data Available
- After successful mating each pregnant female was caged (how): singly on sterilized wood chips
- Any other deviations from standard protocol: No data Available
Duration of treatment / exposure:
Study 2:
Dose regime 1: Daily administration for 32-46 days prior to mating and up to day 18 of pregnancy.
Dose regime 2: Daily administrationon days 1-18 of pregnancy

Study 3: 10 weeks before mating up to 20th day of gestation

Study 4: Exposure period: male: 5 weeks (F0)
4 weeks (F1) and 22 months (F1)
female: 8 weeks (F0)
Premating exposure period (males): 3 weeks
Premating exposure period (females): 3 weeks
Frequency of treatment:
Study 2: Daily for both the dose regimes
Study 3: Daily administration for 70 days before pairing, continuing through pregnancy
Study 4: daily
Duration of test:
Study 2: Dose regime 1: 50-64 days
Dose regime 2: 18-21 days
Study 3: 85 days
Remarks:
Study 2:
Doses / Concentrations:
0, 750 mg/kg body weight
Basis:
no data

Study 3: Doses / Concentrations:
0, 750 mg/kg body weight
Basis:
no data

Study 4: Doses / Concentrations:
F0: 0, 25, 100 or 500 mg/kg bw day F1: 0, 25, 100 or 250 mg/kg bw/day
Basis:
nominal in diet
No. of animals per sex per dose:
Study 2:
Dose regime 1:10 females,
Dose regime 2:10 females(Including arachis oil control and untreated control.)
Study 3:
47 females(Including animals treated with the test chemical, arachis oil control, untreated control and positive control (treated with massive oral doses of vitamin A)
Study 4:
Each treatment group.
Control: 7 virgin males and 50 virgin females
25 mg/kg bw/day: 7 virgin males and 50 virgin females
100 or 250 mg/kg bw/day: 7 virgin males and 50 virgin females
Control animals:
other: Please see 'Remarks'
Details on study design:
Study 2: The test chemical were administered as a solution in arachis oil by oral intubation. Daily administartion was given for :
1.Daily 32-46 days prior to mating and during pregnancy.
2.Daily only during pregnancy.

Study 3: The test chemical were administered as a solution in arachis oil by oral intubation.Daily administration was given for 10 weeks continuing through pregnancy. Pregnancy was terminated on day 20. Rats were anaesthetized and examinations were performed.

Study 4: Dose ranging study was done for dose selection.
Maternal examinations:
Study 2 and 3: Mean body weight,fertility index and mating index.
Study 4: BODY WEIGHT: Yes
Histopathological examinations:Yes
Clinical biochemistry:Yes
Ovaries and uterine content:
Study 2 and 3: The number of corpora lutea, numbers and positions of implantation sites (including resorption sites).
Study 4: Uterus of the dam was examined for resorption site.
Fetal examinations:
Study 2 and 3: Examined for gross abnormalities, skeletal abnormalities and sexed (by measuring the anogenital distance), weighed.
Study 4: External examinations: Yes
Histopathological examinations :Yes
Clinical biochemistry:Yes
Statistics:
Study 4: Student's t-test, analysis of variance and regression analysis. For pathological findings the trend for analysis.
Indices:
Study 2 and 3: fertility index and mating index.
Historical control data:
No Data Available
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
not specified
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Study 4: There were no differences in food consumption or body weight gain between female control rats and female rats fed with the test chemical contained diet. Reduced body weight gain was observed in male rats treated with 500 mg/kg body weight and day.
Rats treated with the test chemical showed no significant differences in weight gain or food consumption during pregnancy and lactation, compared with untreated control rats.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Study 4: There were no differences in food consumption or body weight gain between female control rats and female rats fed with the test chemical contained diet. Reduced body weight gain was observed in male rats treated with 500 mg/kg body weight and day.
Rats treated with the test chemical showed no significant differences in weight gain or food consumption during pregnancy and lactation, compared with untreated control rats.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Dams receiving 500 mg of the test chemical/kg body weight and day showed a significant increase in liver weight.
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
No Data Available
Number of abortions:
no effects observed
Description (incidence and severity):
Study 2: No fetuses were aborted in both the dose regimes at any dose levels in the study.
Pre- and post-implantation loss:
no effects observed
Total litter losses by resorption:
no effects observed
Early or late resorptions:
effects observed, non-treatment-related
Description (incidence and severity):
Study 2: Some sporadic cases were observed where early or late resorptions were observed.
Dead fetuses:
no effects observed
Changes in pregnancy duration:
not specified
Changes in number of pregnant:
not specified
Other effects:
not specified
Details on maternal toxic effects:
Study 2: Maternal toxic effects:no effects
Details on maternal toxic effects:
No significant change in body weight.
Dose regime 1:Mating index=88% and Fertility index =29%.
Dose regime 2: Mating index=Not available and Fertility index=25 %.

Study 3: Maternal toxic effects:no effects

Details on maternal toxic effects:999
No significant change in body weight.Mating index=89% and Fertility index =57%.

Study 4: Maternal toxic effects:yes

Details on maternal toxic effects:
There was no alteration in numbers of resorption sites. No statistically significant change was seen in the number of foetuses/dam.
Dose descriptor:
LOAEL
Effect level:
750 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
total litter losses by resorption
early or late resorptions
Abnormalities:
not specified
Fetal body weight changes:
no effects observed
Description (incidence and severity):
Study 4: No significant effects on foetus development except that pups belonging to 500 mg/kg bw/day failed to gain weight at the same rate as control pups.
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
Description (incidence and severity):
Study 4: The total litter weight was significantly decreased for dams treated with the high dose of the test chemical.
Changes in postnatal survival:
no effects observed
External malformations:
effects observed, treatment-related
Description (incidence and severity):
Microphthalmia was observed in the treated groups.
Skeletal malformations:
not specified
Visceral malformations:
not specified
Other effects:
not specified
Details on embryotoxic / teratogenic effects:
Study 2: Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No Embryonic or teratogenic effects observed in foetuse or offspring.

Study 3: Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No Embryotic or teratogenic effects observed in foetuse or offspring.




Dose descriptor:
NOAEL
Effect level:
750 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Teratoginicity:
Abnormalities:
not specified
Developmental effects observed:
not specified
Treatment related:
not specified
Conclusions:
The test chemical administered to female mice at a dose of 750 mg/kg bw daily for 70 days premating, during pregnancy and only during pregnancy showed no adverse .effects either on parent or fetuses.
Executive summary:

Developmental Toxicity Study:

Data from the developmental toxicity studies is as follows:

Developmental Toxicity Study 2:

In an experimental study, groups of mice of strain Evans No.1 were administered with the test chemical in oral dosage regime, with a daily dose of 0 and 750 mg/kg bw during the period before mating, continuing through mating and gestation up to the time when the animals were killed, and during pregnancy (1-18 days) only. The vehicle used for dosing the test chemical was Archias oil. The maternal animals were observed forMean body weight,fertility index and mating index.In uterine observations, the animals were observed for the number of corpora lutea, numbers and positions of implantation sites (including resorption sites).After performing all the observations, Nosignificant maternal toxicity and no significant embryotoxic effects were observed, as judged by examination of the skeletal or gross abnormalities of fully-developed fetuses. Thus, from all the observations, it was concluded that the NOAEL for embryotoxicity and teratogenicity was therefore considered to be 750 mg/kg bw.

Developmental Toxicity Study 3:

In an experimental study, groups of rats of strains Tuck albino rats were administered with the test chemical in an oral dosage regime, with a daily dose of 0 or 750 mg/kg bw during the period before mating, continuing through mating and gestation up to the time when the animals were sacrificed. The vehicle used for dosing was Arachis oil. The maternal animals were observed for mean body weight, fertility index and mating index. In uterine examinations, the animals were observed for The number of corpora lutea, numbers and positions of implantation sites (including resorption sites). In fetal examinations, the animals were observed for gross abnormalities, skeletal abnormalities and soft tissue abnormalities, sexed (by measuring the anogenital distance) and were weighed. After performing all the observation, it was seen that, no significant embryotoxic effect was observed, as judged by examination of the skeletal and soft tissues of fully-developed fetuses. The abnormalities encountered were considered to be spontaneous in origin.NOAEL for embryotoxicity and teratogenicity was therefore considered to be 750 mg/kg bw.

Developmental Toxicity Study 4:

A two-generation experimental study was conducted for assessment and evaluation of the changes in reproductive and the developmental parameters. In this study, the Wistar rats were administered with the test chemical via the oral feed route. The test chemical was mixed in the diet of the test animals. The test chemical was administered in the concentration of0, 25, 100 or 500 mg/kg bw to parental animals while for the F1 generation, the test chemical was administered in the concentrations of 0, 25, 100 or 250 mg/kg bw. The results of this study on Wistar rats revealed no adverse effects on reproductive performance of the F0 generation fed with the test chemical, at dietary levels of 25, 100 or 500 mg/kg body weight. Dams receiving 500 mg/kg body weight and day showed a significant increase in liver weight. The NOAEL for maternal toxicity was assumed to be 100 mg/kg body weight and day. No adverse effects on fetal development were observed, but there was a significant reduction in body weight gain of pups nursed by dams on the highest dose group (500 mg/kg body weight). In addition, the development was retarded in pups of the high dose group.The NOAEL for pups was assessed to be 100 mg/kg bw/day, and the NOAEL for maternal toxicity was assumed to be 100 mg/kg bw/day. There were no effects on fertility and no increase in foetal abnormalities at any dose of the test chemical. Thus, from all theabove observations, it was concluded that the NOAEL for maternal toxicity and developmental toxicity was found to be 100 mg/kg bw/day, while the LOAEL for maternal toxicity and developmental toxicity was found to be 500 and 250 mg/kg bw/day respectively.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
750 mg/kg bw/day
Study duration:
chronic
Species:
rat
Quality of whole database:
Data is from a Klimisch 2 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 from the developmental toxicity studies is as follows:

Developmental Toxicity Study 2:

In an experimental study, groups of mice of strain Evans No.1 were administered with the test chemical in oral dosage regime, with a daily dose of 0 and 750 mg/kg bw during the period before mating, continuing through mating and gestation up to the time when the animals were killed, and during pregnancy (1-18 days) only. The vehicle used for dosing the test chemical was Archias oil. The maternal animals were observed forMean body weight,fertility index and mating index.In uterine observations, the animals were observed for the number of corpora lutea, numbers and positions of implantation sites (including resorption sites).After performing all the observations, Nosignificant maternal toxicity and no significant embryotoxic effects were observed, as judged by examination of the skeletal or gross abnormalities of fully-developed fetuses. Thus, from all the observations, it was concluded that the NOAEL for embryotoxicity and teratogenicity was therefore considered to be 750 mg/kg bw.

Developmental Toxicity Study 3:

In an experimental study, groups of rats of strains Tuck albino rats were administered with the test chemical in an oral dosage regime, with a daily dose of 0 or 750 mg/kg bw during the period before mating, continuing through mating and gestation up to the time when the animals were sacrificed. The vehicle used for dosing was Arachis oil. The maternal animals were observed for mean body weight, fertility index and mating index. In uterine examinations, the animals were observed for The number of corpora lutea, numbers and positions of implantation sites (including resorption sites). In fetal examinations, the animals were observed for gross abnormalities, skeletal abnormalities and soft tissue abnormalities, sexed (by measuring the anogenital distance) and were weighed. After performing all the observation, it was seen that, no significant embryotoxic effect was observed, as judged by examination of the skeletal and soft tissues of fully-developed fetuses. The abnormalities encountered were considered to be spontaneous in origin.NOAEL for embryotoxicity and teratogenicity was therefore considered to be 750 mg/kg bw.

Developmental Toxicity Study 4:

A two-generation experimental study was conducted for assessment and evaluation of the changes in reproductive and the developmental parameters. In this study, the Wistar rats were administered with the test chemical via the oral feed route. The test chemical was mixed in the diet of the test animals. The test chemical was administered in the concentration of0, 25, 100 or 500 mg/kg bw to parental animals while for the F1 generation, the test chemical was administered in the concentrations of 0, 25, 100 or 250 mg/kg bw. The results of this study on Wistar rats revealed no adverse effects on reproductive performance of the F0 generation fed with the test chemical, at dietary levels of 25, 100 or 500 mg/kg body weight. Dams receiving 500 mg/kg body weight and day showed a significant increase in liver weight. The NOAEL for maternal toxicity was assumed to be 100 mg/kg body weight and day. No adverse effects on fetal development were observed, but there was a significant reduction in body weight gain of pups nursed by dams on the highest dose group (500 mg/kg body weight). In addition, the development was retarded in pups of the high dose group.The NOAEL for pups was assessed to be 100 mg/kg bw/day, and the NOAEL for maternal toxicity was assumed to be 100 mg/kg bw/day. There were no effects on fertility and no increase in foetal abnormalities at any dose of the test chemical. Thus, from all theabove observations, it was concluded that the NOAEL for maternal toxicity and developmental toxicity was found to be 100 mg/kg bw/day, while the LOAEL for maternal toxicity and developmental toxicity was found to be 500 and 250 mg/kg bw/day respectively.

Justification for classification or non-classification

The test chemical does not show any adverse effects on the reproductive and developmental parameters and thus, as per CLP classification, it may not be classified as a Reproductive and Developmental toxicant.