Registration Dossier

Administrative data

Key value for chemical safety assessment

Effects on fertility

Description of key information

Reproductive Toxicity Study:

In the present repeated dose toxicity study for 28 days, male and female Wistar rats in group of 5/sex were exposed to test chemical orally in the concentration of 0, 125, 250 and 500 mg/kg/day body weight. Recovery groups were dosed with 0 and 500mg/kg of test chemical. The animals were observed for clinical signs of toxicity; mortality and morbidity; feed consumption, haematological and clinical chemistry parameter analysis; urinalysis; and at the end of the study necropsy were performed in alll survived animals and change in organ weight, gross pathology and histopathology examination were performed. The results of the observed paramters revealed,No mortality and morbidity were observed among all the groups of animals throughout the study period; No apparent treatment related clinical signs were observed in any of the animals throughout the treatment and recovery period. Detailed clinical examinations like Home cage observation, handling observation and open field observation all animals were observed normal during study period at 125, 250 and 500mg/kg dose groups; No change in body weight were seen in animals of 125 and 250mg/kg doses howver 500mg/kg group females shows significant decreased in body weight compared to control animals. No change in food consumption and opthalmological examination in treated male and female rat at 125, 250 and 500mg/kg dose groups in treatment and recovery period were observed. The haematological examination revealed statistically significant increase neutrophil count in 500 mg/kg in female rats; statistically significant decrease was observed in MCV and WBC and increase observed in PLT in females and statistically significant decrease in APTT in male rat as compare to control. The clinical biochemistry paramters changes were observed. In male rat statistically significant increase were observed in Sodium (Na) and decrease in Lactate Dehydrogenase (LD) and Creatine Kinase (CK) when treated with 125 and 500 mg/kg/day body weight. When treated with 250 and 500 mg/kg/day, Statistically significant increase was observed in Alkaline phosphatase (ALP) in male rat. When treated with 500 mg/kg/day body weight, Alanine amino transferase (ALT) increased in male rat as compare to control. Urine parameters did not show any significant difference, except statistical significant increase observed in volume in (500 mg/kg body weight) in females as compared to control group the change were not attributed to the effect of test item administration. The organ weight study revealed, not statistically significant in both sexes compared to control group among all organs like  testes, prostate with seminal vesicle, epididymis, ovaries with oviduct and uterus (g) and relative weights (%). However, absolute and relative organ weight S.V. with coagulating gland and prostrate as whole was decreased in (250 mg/kg body weight) male as compared to control animals. No external and internal gross pathological changes were seen at 125, 250 and 500 mg/kg body weight. In histopatholgy study, When treated with 500 mg/kg/day body weight, hyperplasia of BALT in Lung and MNC infiltration in trachea were observed in male and female this effect was reversed in 14 days recovery period. S.V. with coagulating gland and prostrate as whole was a incidental finding as histopathologically tissues were normal. Therefore, NOAEL was considered to be 500 mg/kg/day body weight when rats are exposed to test chemical orally for 28 days. 

Link to relevant study records
Reference
Endpoint:
toxicity to reproduction
Remarks:
Repeated Dose 28 Days Toxicity Testing swith additional testing on reproductive organs
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from a study report.
Qualifier:
according to
Guideline:
other: OECD No. 407
Principles of method if other than guideline:
The above study was performed to assess and evaluate the effects of the test chemical on reproductive organs of Sprague Dawley rats.
GLP compliance:
yes
Limit test:
no
Justification for study design:
No Data Available
Specific details on test material used for the study:
- Molecular weight (if other than submission substance): 127.19 g/mole
- Substance type: Organic
- Physical state: Solid
- Impurities (identity and concentrations): 0.29 %
Species:
rat
Strain:
Wistar
Details on species / strain selection:
No Data Available
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: In-House Bred at sa-FORD, Animal Facility
- Age at study initiation: 6 - 8 weeks
- Weight at study initiation: Male : 137-191 g, Female: 137- 165 g
- Fasting period before study: No data available
- Housing: Animals were housed in Polycarbonate cages. Cage rotation was carried out weekly and cleaned at regular intervals. Animals were bedded on sterilized corn cob produced from pure corn, dried and free from dust. Floor of the experimental room and work tops were swept and mopped with disinfectant solution every day or as on requirement.
- Diet (e.g. ad libitum): A conventional laboratory pelleted diet, ad libitum.
- Water (e.g. ad libitum): Aqua guard filtered drinking water in bottles, ad libitum.
- Acclimation period: Male: 6 days, Female: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.00 to 23.10 °C
- Humidity (%): 49.90 to 69.40%.
- Air changes (per hr): Adequately filtered air 12 times per hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark artificial light.

IN-LIFE DATES: From: September 22, 2014
To: October 20, 2014
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Test chemical was weighed and dissolved in a vehicle (corn oil) to achieve desired concentration of test item. Dose formulation was freshly prepared daily. At the time of dosing, dose formulation was kept on the magnetic stirrer to maintain the homogeneity of test item.

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

VEHICLE
- Justification for use and choice of vehicle (if other than water): Corn oil was used as a vehicle based on the solubility testing.
- Concentration in vehicle: 0,125,250 and 500 mg/kg/day
- Amount of vehicle (if gavage): 1 ml/100g body weight
- Lot/batch no. (if required): MKBQ9948V and MKBG9426V
- Purity: No data available
Details on mating procedure:
Reproductive organ weight was observed.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Stability and concentration of the test chemical in dose formulation was analysed by a validated analytical method, at the start of treatment (on day 1) and on day 21
Duration of treatment / exposure:
28 days
Frequency of treatment:
Daily
Dose / conc.:
0 other: mg/kg/day
Dose / conc.:
125 other: mg/kg/day
Dose / conc.:
250 other: mg/kg/day
Dose / conc.:
500 other: mg/kg/day
No. of animals per sex per dose:
Total : 60
0 mg/kg/day: 5 male, 5 female
125 mg/kg/day: 5 male, 5 female
250 mg/kg/day: 5 male, 5 female
500 mg/kg/day: 5 male, 5 female

Recovery:
0 mg/kg/day: 5 male, 5 female
500 mg/kg/day: 5 male, 5 female
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The dose levels were selected based on the information provided by Sponsor.
- Rationale for animal assignment (if not random): No data available
- Rationale for selecting satellite groups: No data available
- Post-exposure recovery period in satellite groups: No data available
- Section schedule rationale (if not random): No data available
Positive control:
No data available
Parental animals: Observations and examinations:
Animals of all dose groups were observed for Clinical signs/ symptoms daily during the experimental period. Mortality and morbidity were observed and recorded twice a day Sensory reactivity to stimuli, assessment of grip strength, hind limb foot splay and motor activity assessment were conducted for rats from main groups. Animals were weighed during randomization, at start of treatment and thereafter weekly, till the end of experimental period. Clinical pathology evaluation of all surviving rats from the main groups and recovery groups was performed just prior to necropsy. Hematological and clinical chemistry parameters were analyzed at the end of the treatment and recovery period. Urine samples were collected from all the survived rats of main and recovery group and urinary parameters were determined.
Oestrous cyclicity (parental animals):
No data available
Sperm parameters (parental animals):
No data available
Litter observations:
No data available
Postmortem examinations (parental animals):
Reproductive organ weights were recorded in all treated animals at 0, 125, 250 and 500mg/kg dose levels, gross pathology and histopathology were examined.
Postmortem examinations (offspring):
No data available
Statistics:
Statistical analysis of raw data was performed by using statistical software Sigma Plot 11.0. The mean and standard deviation was calculated using the software and all data was summarized in tabular form. All continuous data (body weight, feed consumption, haematology, clinical chemistry, absolute and relative organ weights) were checked for their homogeneity using Bartlett’s test. All homogenous data was analysed using ANOVA and data showing significance in their variances was subjected to Dunnett’s t-test. All heterogeneous data was analysed using F test and Student’s t-test.
Reproductive indices:
No data available
Offspring viability indices:
No data available
Clinical signs:
no effects observed
Description (incidence and severity):
No apparent treatment related clinical signs were observed in any of the animals throughout the treatment and recovery period.
Detailed clinical examinations like Home cage observation, Handling observation and Open field observation all animals were observed normal during study period at 125, 250 and 500mg/kg dose groups
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
No mortality and morbidity were observed among all the groups of animals throughout the study period
Body weight and weight changes:
no effects observed
Description (incidence and severity):
In treated group with 500 mg/kg/day body weight, significant decreased in female rats were observed as compare to control.

No change in body weight were observed at 125 and 250 mg/kg/day in treated male and female rats as compare to control.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No change was observed in food consumption of treated male and female rat at 125, 250 and 500mg/kg dose groups in treatment and recovery period as compare to control.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
no effects observed
Description (incidence and severity):
No abnormalities were observed in treated 125, 250 and 500 mg/kg male and female rat in treatment groups at and recovery period as compare to control.
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Animalswhen treated with 500 mg/kg/day body weight, in female rat statistically significant increase neutrophil were observed.
In recovery, statistically significant decrease was observed in MCV and WBC and increase observed in PLT in females and statistically significant decrease in APTT in male rat as compare to control.
Above changes were not related to the test substence and may due to the preanalytical and analytical variables.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Clinical chemistry: When treated with 125, 250 and 500 mg/kg/day body weight, in male rat statistically significant increase were observed in Sodium (Na) and decrease in Lactate Dehydrogenase (LD) and Creatine Kinase (CK) when treated with 125 and 500 mg/kg/day body weight.
When treated with 250 and 500 mg/kg/day, Statistically significant increase was observed in Alkaline phosphatase (ALP) in male rat.
When treated with 500 mg/kg/day body weight, Alanine amino transferase (ALT) increased in male rat as compare to control.
In recovery, Statistically significant increase were observed in Blood urea nitrogen (BUN), Urea, Albumin (ALB), Alkaline phosphatase (ALP), Potassium (K), Chloride (Cl) and Bile acids and statistically significant decrease in Cholesterol (Chol) in male rat and Sodium (Na) in female rat as compare to control.
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
Urine parameters did not show any significant difference among all the experimental animals of both the sexes and were comparable to animals of control group, except statistical significant increase observed in volume in (500 mg/kg body weight) in females as compared to control group the change were not attributed to the effect of test item administration.
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
Among all organs like testes, prostate with seminal vesicle, epididymis, ovaries with oviduct and uterus (g) and relative weights (%) were not statistically significant in both sexes compared to control group. Absolute and relative organ weight S.V. with coagulating gland and prostrate as whole was decreased in (250 mg/kg body weight) male as compared to control animals.
Gross pathological findings:
no effects observed
Description (incidence and severity):
Gross necropsy of all male and female animals did not show any extermal and internal abnormal changes in any animal at 125, 250 and 500mg/kg.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
When treated with 500 mg/kg/day body weight, hyperplasia of BALT in Lung and MNC infiltration in trachea were observed in male and female this effect was reversed in 14 days recovery period. Significant decrease in absolute and relative organ weight of S.V. with coagulating gland and prostrate as whole was a incidental finding as histopathologically tissues were normal.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
not specified
Dose descriptor:
NOAEL
Effect level:
500 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
ophthalmological examination
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
other: Effect on reproductive organ weight, gross pathology and histopathology.
Critical effects observed:
not specified
System:
other: Not Spcified
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
not specified
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings:
not specified
Other effects:
not specified
Behaviour (functional findings):
not specified
Developmental immunotoxicity:
not specified
No Data Availab;e
Remarks on result:
not measured/tested
Reproductive effects observed:
not specified

Absolute Organ Weight (g)

Sex:Male

Organs

 

Group 1

Group 2

Group 3

Group 4

Mean

SD

Mean

SD

Mean

SD

Mean

SD

Body weight (g)

251.80

21.09

239.20

24.78

240.60

18.37

234.60

11.48

Brain

1.910

0.062

1.935

0.053

1.966

0.051

1.900

0.057

Adrenals

0.046

0.013

0.059

0.007

0.053

0.009

0.051

0.010

S.V. With Coagulating gland and prostrate

1.1831

0.22882

1.11382

0.167243

0.8658↓

0.1214

1.1246

0.12127

Testes

3.003

0.246

2.857

0.296

2.966

0.167

2.810

0.150

Epdidymides

0.902

0.164

1.004

0.135

0.958

0.106

0.954

0.045

Heart

1.044

0.088

1.007

0.078

0.998

0.069

0.911

0.043

Liver

10.450

0.919

11.804↑

1.553

13.062↑

1.580

13.492↑

1.428

Kidneys

2.294

0.219

2.271

0.261

2.575

0.202

2.427

0.192

Spleen

1.356

0.434

1.142

0.332

1.136

0.196

0.905

0.198

Thymus

0.301

0.071

0.311

0.044

0.299

0.071

0.241

0.056

Organs

 

Group 1-R

Group 4-R

Mean

SD

Mean

SD

Body weight (g)

273.40

20.21

280.80

15.39

Brain

1.935

0.120

1.927

0.067

Adrenals

0.056

0.009

0.064

0.008

S.V. With Coagulating gland and prostrate

1.5541

0.4754

1.4925

0.260007

Testes

3.210

0.166

3.168

0.254

Epdidymides

1.145

0.063

1.112

0.153

Heart

1.148

0.080

1.175

0.125

Liver

10.861

0.878

10.788

0.658

Kidneys

2.417

0.304

2.368

0.199

Spleen

1.002

0.106

1.048

0.285

Thymus

0.296

0.106

0.349

0.090

Sex:Female

Organs

Group 1

Group 2

Group 3

Group 4

Mean

SD

Mean

SD

Mean

SD

Mean

SD

Body weight (g)

185.40

15.13

173.00

4.00

178.40

7.37

169.60

5.41

Brain

1.791

0.076

1.692

0.091

1.728

0.040

1.789

0.087

Adrenals

0.059

0.011

0.051

0.011

0.061

0.011

0.064

0.012

Ovary

0.092

0.012

0.106

0.032

0.111

0.024

0.091

0.015

Uterus

0.720

0.120

0.645

0.107

0.561

0.152

0.553

0.115

Heart

0.853

0.118

0.725

0.027

0.731

0.063

0.725

0.079

Liver

8.231

0.754

7.106↓

0.640

7.743

0.646

8.403

0.586

Kidneys

1.582

0.132

1.412

0.098

1.470

0.097

1.585

0.177

Spleen

0.719

0.279

0.502

0.122

0.485

0.120

0.516

0.225

Thymus

0.300

0.049

0.266

0.081

0.235

0.036

0.242

0.042

Organs

Group 1-R

Group 4-R

Mean

SD

Mean

SD

Body weight (g)

196.60

9.96

188.40

8.50

Brain

1.792

0.087

1.800

0.059

Adrenals

0.077

0.007

0.090↑

0.006

Ovary

0.164

0.015

0.170

0.018

Uterus

0.777

0.305

0.723

0.120

Heart

0.846

0.054

0.782

0.045

Liver

7.429

0.677

7.394

0.560

Kidneys

1.684

0.058

1.522↓

0.110

Spleen

0.776

0.126

0.441↓

0.054

Thymus

0.281

0.062

0.320

0.037

Relative Organ Weight (%)

Sex: Male

Organs

 

Group 1

Group 2

Group 3

Group 4

Mean

SD

Mean

SD

Mean

SD

Mean

SD

Brain

0.762

0.052

0.817

0.102

0.820

0.044

0.824

0.040

Adrenals

0.019

0.006

0.025

0.004

0.022

0.005

0.022

0.004

S.V. With Coagulating gland and prostrate

0.467

0.058

0.470

0.083

0.360↓

0.045

0.480

0.055

Testes

1.196

0.100

1.196

0.062

1.237

0.090

1.217

0.062

Epdidymides

0.356

0.046

0.420

0.037

0.401

0.058

0.413

0.026

Heart

0.416

0.041

0.425

0.056

0.415

0.016

0.395

0.025

Liver

4.155

0.252

4.933

0.358

5.417↑

0.275

5.850↑

0.690

Kidneys

0.915

0.105

0.949

0.046

1.071↑

0.053

1.051↑

0.087

Spleen

0.534

0.153

0.476

0.128

0.471

0.066

0.393

0.094

Thymus

0.120

0.032

0.132

0.032

0.125

0.034

0.104

0.023

Organs

Group 1-R

Group 4-R

Mean

SD

Mean

SD

Brain

0.710

0.063

0.687

0.023

Adrenals

0.021

0.004

0.023

0.003

S.V. With Coagulating gland and prostrate

0.572

0.184

0.535

0.115

Testes

1.181

0.133

0.974

0.345

Epdidymides

0.420

0.021

0.406

0.030

Heart

0.424

0.059

1.043

1.396

Liver

3.999

0.523

3.305

1.377

Kidneys

0.893

0.174

0.779

0.143

Spleen

0.367

0.036

0.297

0.082

Thymus

0.107

0.031

0.124

0.030

Sex: Female

Organs

Group 1

Group 2

Group 3

Group 4

Mean

SD

Mean

SD

Mean

SD

Mean

SD

Brain

0.970

0.077

0.978

0.037

0.970

0.036

1.056

0.072

Adrenals

0.032

0.005

0.030

0.007

0.034

0.005

0.038

0.007

Ovary

0.0494

0.0049

0.0616

0.0189

0.0626

0.016

0.0536

0.0078

Uterus

0.392

0.081

0.373

0.062

0.313

0.081

0.325

0.061

Heart

0.463

0.071

0.419

0.015

0.410

0.038

0.428

0.050

Liver

4.443

0.280

4.107

0.341

4.342

0.342

4.954

0.298

Kidneys

0.855

0.061

0.815

0.044

0.824

0.046

0.936

0.111

Spleen

0.382

0.129

0.291

0.078

0.274

0.079

0.301

0.120

Thymus

0.162

0.026

0.153

0.046

0.131

0.016

0.143

0.026

Organs

Group 1-R

Group 4-R

Mean

SD

Mean

SD

Brain

0.913

0.053

0.956

0.043

Adrenals

0.040

0.005

0.048↑

0.004

Ovary

0.0837

0.0103

0.0901

0.0100

Uterus

0.395

0.151

0.385

0.068

Heart

0.431

0.033

0.415

0.012

Liver

3.786

0.379

3.929

0.313

Kidneys

0.859

0.060

0.809

0.069

Spleen

0.395

0.066

0.234↓

0.027

Thymus

0.144

0.036

0.170

0.025

N = Number of animals in the group, SD = Standard deviation,↓= statisticalsignificant decrease at 95% level of significance (p > 0.05),↑= statisticalsignificant increase at 95% level of significance (p < 0.05).

Conclusions:
The NOAEL was considered to be 500 mg/kg/day body weight when male and female rat were exposed to test chemical for 28 days.
Executive summary:

In the present repeated dose toxicity study for 28 days, male and female Wistar rats in group of 5/sex were exposed to test chemical orally in the concentration of 0, 125, 250 and 500 mg/kg/day body weight. Recovery groups were dosed with 0 and 500mg/kg of test chemical. The animals were observed for clinical signs of toxicity; mortality and morbidity; feed consumption, haematological and clinical chemistry parameter analysis; urinalysis; and at the end of the study necropsy were performed in alll survived animals and change in organ weight, gross pathology and histopathology examination were performed. The results of the observed paramters revealed, No mortality and morbidity were observed among all the groups of animals throughout the study period; No apparent treatment related clinical signs were observed in any of the animals throughout the treatment and recovery period. Detailed clinical examinations like Home cage observation, handling observation and open field observation all animals were observed normal during study period at 125, 250 and 500mg/kg dose groups; No change in body weight were seen in animals of 125 and 250mg/kg doses howver 500mg/kg group females shows significant decreased in body weight compared to control animals. No change in food consumption and opthalmological examination in treated male and female rat at 125, 250 and 500mg/kg dose groups in treatment and recovery period were observed. The haematological examination revealed statistically significant increase neutrophil count in 500 mg/kg in female rats; statistically significant decrease was observed in MCV and WBC and increase observed in PLT in females and statistically significant decrease in APTT in male rat as compare to control. The clinical biochemistry paramters changes were observed. In male rat statistically significant increase were observed in Sodium (Na) and decrease in Lactate Dehydrogenase (LD) and Creatine Kinase (CK) when treated with 125 and 500 mg/kg/day body weight. When treated with 250 and 500 mg/kg/day, Statistically significant increase was observed in Alkaline phosphatase (ALP) in male rat. When treated with 500 mg/kg/day body weight, Alanine amino transferase (ALT) increased in male rat as compare to control. Urine parameters did not show any significant difference, except statistical significant increase observed in volume in (500 mg/kg body weight) in females as compared to control group the change were not attributed to the effect of test item administration. The organ weight study revealed, not statistically significant in both sexes compared to control group among all organs like  testes, prostate with seminal vesicle, epididymis, ovaries with oviduct and uterus (g) and relative weights (%). However, absolute and relative organ weight S.V. with coagulating gland and prostrate as whole was decreased in (250 mg/kg body weight) male as compared to control animals. No external and internal gross pathological changes were seen at 125, 250 and 500 mg/kg body weight. In histopatholgy study, When treated with 500 mg/kg/day body weight, hyperplasia of BALT in Lung and MNC infiltration in trachea were observed in male and female this effect was reversed in 14 days recovery period. S.V. with coagulating gland and prostrate as whole was a incidental finding as histopathologically tissues were normal. Therefore, NOAEL was considered to be 500 mg/kg/day body weight when rats are exposed to test chemical orally for 28 days. 

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
500 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
The data is K1 level as the data has been obtained from Study report .
Effect on fertility: via inhalation route
Endpoint conclusion:
no study available
Effect on fertility: via dermal route
Endpoint conclusion:
no study available
Additional information

Toxicity To Reproduction:

The summaries for assessment of reproductive toxicity of the test chemical are as follows:

Reproductive Toxicity Study 1:

In the present repeated dose toxicity study for 28 days, male and female Wistar rats in group of 5/sex were exposed to test chemical orally in the concentration of 0, 125, 250 and 500 mg/kg/day body weight. Recovery groups were dosed with 0 and 500mg/kg of test chemical. The animals were observed for clinical signs of toxicity; mortality and morbidity; feed consumption, haematological and clinical chemistry parameter analysis; urinalysis; and at the end of the study necropsy were performed in alll survived animals and change in organ weight, gross pathology and histopathology examination were performed. The results of the observed paramters revealed,No mortality and morbidity were observed among all the groups of animals throughout the study period; No apparent treatment related clinical signs were observed in any of the animals throughout the treatment and recovery period. Detailed clinical examinations like Home cage observation, handling observation and open field observation all animals were observed normal during study period at 125, 250 and 500mg/kg dose groups; No change in body weight were seen in animals of 125 and 250mg/kg doses howver 500mg/kg group females shows significant decreased in body weight compared to control animals. No change in food consumption and opthalmological examination in treated male and female rat at 125, 250 and 500mg/kg dose groups in treatment and recovery period were observed. The haematological examination revealed statistically significant increase neutrophil count in 500 mg/kg in female rats; statistically significant decrease was observed in MCV and WBC and increase observed in PLT in females and statistically significant decrease in APTT in male rat as compare to control. The clinical biochemistry paramters changes were observed. In male rat statistically significant increase were observed in Sodium (Na) and decrease in Lactate Dehydrogenase (LD) and Creatine Kinase (CK) when treated with 125 and 500 mg/kg/day body weight. When treated with 250 and 500 mg/kg/day, Statistically significant increase was observed in Alkaline phosphatase (ALP) in male rat. When treated with 500 mg/kg/day body weight, Alanine amino transferase (ALT) increased in male rat as compare to control. Urine parameters did not show any significant difference, except statistical significant increase observed in volume in (500 mg/kg body weight) in females as compared to control group the change were not attributed to the effect of test item administration. The organ weight study revealed, not statistically significant in both sexes compared to control group among all organs like  testes, prostate with seminal vesicle, epididymis, ovaries with oviduct and uterus (g) and relative weights (%). However, absolute and relative organ weight S.V. with coagulating gland and prostrate as whole was decreased in (250 mg/kg body weight) male as compared to control animals. No external and internal gross pathological changes were seen at 125, 250 and 500 mg/kg body weight. In histopatholgy study, When treated with 500 mg/kg/day body weight, hyperplasia of BALT in Lung and MNC infiltration in trachea were observed in male and female this effect was reversed in 14 days recovery period. S.V. with coagulating gland and prostrate as whole was a incidental finding as histopathologically tissues were normal.Therefore, NOAEL was considered to be 500 mg/kg/day body weight when rats are exposed to test chemical orally for 28 days. 

Reproductive Toxicity Study 2:

In the present study, 10 weeks repeated dose toxicity to ddY male mice was evaluated by using test chemical orally in the concentration of 317.975 mg/kg bw in DMSO for 10 weeks. The results revealed no significant effect on body weight, testicular weights and Neurotoxicity of treated mice were observed as compared to control. Therefore, NOAEL was considered to be 317.975 mg/kg bw when ddY male mice were treated with test chemical orally for 10 weeks.

Reproductive Toxicity Study 3:

In the present study, the effect of test chemical onrepeated administration and reproductive developmental toxicity study performed by oral administration in the test animals.The test chemical was administered to rats at doses 0, 70, 155, 350 mg/kg bw/day for approximately 8 weeks.The animals observed for clinical signs of toxicity; mortality; feed consumption; change in body weight; behaviour such as locomotor activity and grip strength; gross necropsy examination histopathological examination and reproductive performance were examined in the present study.The results of study revealed, no mortality during the period of study and one 350mg/kg female, on days 4 and 5 of dosing, showed post-dosing signs of subdued behaviour, twitching, semi-closed eyes, piloerection, was cold and showed staggered movement.The body weight results revealedmajority of males in 350mg/kg group lost weight during the first week of dosing. Although mean body weights increased every week after that, the high dose males were approximately 9% lighter than control males by the end of the study. Group mean body weight gain was lower in high 350mg/kg dose females than controls during gestation. 155 and 350mg/kg dose females had lower mean body weight gain than controls during lactation. Feed consumption was observed to be reduced in 350mg/kg group animals as compared to control animals. Forelimb grip strength in males during Week 7 but there were no other dose-related observations on functional effects including locomotor activity at dose levels of 70, 155 and 350mg/kg. Organ weight change were seen in kidney, liver and adrenal weight, significantly increase were observed in high dose 350mg/kg group animals as compared to control animals.   No gross necropsy findings were seen that attributed to be treatment-related at any dose group at 70, 155 and 350mg/kg.The reproductive findings of the study showed no indication of any qualitative abnormality in the various cell types present within the different stages of the spermatogenic cycle and No treatment-related effects on numbers of males and females mating or on male or female fertility indices. The F1 generation results from parent rats revealed reduction in the pups viability in the 350mg/kg dose group. The litter data shown numbers of implantations, litter sizes and Day 4 weights were all lower than controls in the high dose 350mg/kg. Total litter loss occurred for two females in the 350mg/kg and one from each of the 70 and 155mg/kg dose groups.Total embryo foetal loss occurred for one female from 350mg/kg group. Malformed/ shortened limbs occurred in one pup in the 155mg/kg and one pup in the 350mg/kg dose groups, however as this effect occurred in only one pup in one litter in these groups, the toxicological significance is not clear. From the observations and result of the study, The NOAEL in parent rats was considered to be 350 mg/kg/day and LOAEL for the F1 generation was considered to be 350mg/kg when test chemical adminisrtered in male and female rats for 8 weeks.

Effects on developmental toxicity

Description of key information

Developmental Toxicity Study:

In the present study, developmental Toxicity of test chemical was evaluated by adinistration of test chemical by Gavage to Swiss CCD-I (trade name)) Mice on Gestational Days 6 through 17. The dose levels of 0, 30, 60, 120, 180mg/kg were selected and administered in pregnant female mice at day 6 to 17 of gestation period. Dams were observed for clinical signs, change in body weight, feed and water consumption and organ weight change specially liver and gravid uterus. The fetuses were observed for external, visceral and skeletal abnormalities. The results of the study revealed clinical toxicity life, Cyst on right ovary,Left ovary enclosed in blood· filled cyst  were observed at 30mg/kg group; Cyst on left ovary was observed in 60mg/kg group; Uterus enlarged, Rough coat,Vaginal bleeding,Cyst on right ovary were observed in 120mg/kg group females perioral wetness, Vaginal discharge (brownish red), Lethargic,Convulsing, Cyst on right ovary, Curling hind limbs under bodywere obseved in treated females but this effects were non significant compared to control female mice but no change in body weight change, feed consumption and water consumption were observed maternal animals at 30,60,120 and 180mg/kg groups. The fetuses observed for external , visceral and skeletal abnormalities, the results were Cleft Palate, Ectocardia, Gastroschisis and Short Tail were obseved in 2 fetuses. Skeletal malformations were observed such as asymmetrical Fusion of Sternal Plate, Cleft Sternum, Perforated Sternum, Fused Rib Cartilage, Incomplete Ossification, Cartilage Present, Extra Ossification Site(s), Hematoma: Head, Face, Jaw, Neck, Abdomen, Thorax, Hindlimb. the visceral abnormalities also obseved in treated animals including, Situs Inversus Viscerum and left hydronephrosis. From the observations and results of the study, The NOAEL value from the maternal observations was considered to be 180mg/kg/day howeveer the NOAEL value from the fetuses observation considered to be 120mg/kg.

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is based on weight of evidence approach prepared from various publication.
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 414 (Prenatal Developmental Toxicity Study)
Principles of method if other than guideline:
The above experiments were performed to evaluate and assess the developmental toxicity of the test chemical in the test animals.
GLP compliance:
not specified
Specific details on test material used for the study:
- Molecular weight (if other than submission substance): 127.1857g/mol
- Substance type: Organic
- Physical state: No Data Available
- Impurities (identity and concentrations): No Data Available
Species:
other: Study 2: Mouse; Study 3: rat; study 4: rat
Strain:
other: Study 2: Crl:CD-le(ICR) BR VAF/Plus; Study 2: Crl:WI(Glx/BRL/Han)BR; Study 4: Sprague-Dawley
Details on test animals and environmental conditions:
Study 2: TEST ANIMALS
- Source: Charles River laboratories, Inc. (Raleigh, NC).
- Age at study initiation: Females (8-10 weeks); males (10-12 weeks
- Weight at study initiation: Weight Range for Females: 20-35 g on gestation day 0 (day of vaginal plug detection
- Fasting period before study: No data
- Housing: Solid bottom polycarbonate cages with stainless steel wire lids (Laboratory Products, Rochelle Park, NJ).
- Use of restrainers for preventing ingestion (if dermal):no data
- Diet (e.g. ad libitum): Purina ·Certified Rodent Chowe (15002) (Ralston Purina Co., St. Louis, Missouri) available ad libitum to males and females (pelleted chow)
- Water (e.g. ad libitum): deionized/filtered water provided ad libitum
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 69-75 F
- Humidity (%): 30-70% relative humidity
- Air changes (per hr): 12-14 air changes per hour
- Photoperiod (hrs dark / hrs light):12/12hrs of light and dark

Study 3: - At study initiation the rats were 13 to 15 weeks old.

Study 4: TEST ANIMALS
- Source: the Charles River Laboratories. Inc.
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 201 g to 263 g
- Housing: Males were housed singly in solid bottom polycarbonate cages (19" x 10 1/2" x 8") and females were group housed (maximum, 3 per cage) in solid bottom polycarbonate cages (19" x 10 1/2" x 8") with stainless steel wire lids (Laboratory Products. Rochelle Park. NJ).
- Diet (e.g. ad libitum):Purin Certified Rodent Chow, ad libitum
- Water (e.g. ad libitum): deionized/filtered water, ad libitum
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): ranged between 67.7 and 76.8 deg F
- Humidity (%): maintained at an average of 49.0%
- Air changes (per hr): Air in each animal room was exchanged at least 12 to 14 times per hour.
- Photoperiod (hrs dark / hrs light): 12-hrs light/12-hrs dark
Route of administration:
other: Study 2, 3 and 4: Oral; gavage
Vehicle:
other: study 2 and 4: Distliled water; study 3: not specified
Details on exposure:
Study 2: PREPARATION OF DOSING SOLUTIONS: Test chemical will be
dissolved in distilled water to a concentration determined by the following
formula:
concentration (mg/ml) = dose level (mg/kg)/ dose volume (10.0 ml/kg)
Test chemical will be weighed into a volumetric flask and distilled water will be added to slightly below the mark. test chemical will be dispersed into a visually uniform solution and the flask will then be filled to the mark and agitated a second time. The solution will be sonicated, if necessary, to dissolve the chemical. 'Each concentration of test chemical will be formulated independently once during each' study replicate in a quantity sufficient for use across the entire dosing period for each replicate.
- Storage condition of formulation: Dose formulations will be stored at refrigeratea temperatures in 60 ml amber vials

Study 3: Dose volume was 5mL/kg.

Study 4: VEHICLE
- Justification for use and choice of vehicle (if other than water): Distilled water
- Concentration in vehicle: 0, 2.5. 7.5 or 15 mg/kg/day
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Study 2: Prior to administration and after administration, each formulation were analyzed to ensure a concentration between 90 and 110% of the target concentration.

Study 3: Achieved concentrations were 90 to 100% of expected when measured on Week 1 and Week 8

Study 4: Prior to initiation of dosing, an aliquot of the control and each ACRL formulation was analyzed for the test chemical. Formulations that assayed at 90-110% of the nominal concentration were considered acceptable for use. All other formulations were remixed and reanalyzed prior to administration. Additional samples of each formulation were analyzed after use at the end of the dosing period to confirm stability. Duplicate aliquots of the predosing and postdosing samples were refrigerated and stored as archival samples.
Details on mating procedure:
Study 2: Monogamous breeding pairs were cohabited over night in the home cage of the male. On the following morning, the female was examined for a copulation plug. The day of plug detection was designated as gd 0. Plug-negative females was returned to the males cage and checked for plugs on successive mornings until insemina tion occurs or the treatment groups are filled, whichever comes first.

Study 3: No data
- Impregnation procedure: cohoused
- M/F ratio per cage: 1/1 of same treatment group
- Length of cohabitation: 15 days
- Proof of pregnancy: Mating was confirmed by the presence of a vaginal plug in situ or by sperm in a vaginal washing. The day on which mating was confirmed was designated Day 0 of gestation

Study 4: - M/F ratio per cage: 1:1
- Length of cohabitation: overnight
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy : Female were examined by vaginal smear for the presence of sperm.
- After successful mating each pregnant female was caged (how): Sperm-positive females were individually housed until scheduled sacrifice on gestation day 20.
Duration of treatment / exposure:
Study 2: 11 days (from gestation period day 6 to 17)

Study 3: Males - 2 weeks before pairing, throughout pairing and until the day before necropsy (at least 4 weeks of treatment)
Females- 2 weeks before pairing, throughout pairing and until Day 4 postpartum, inclusive

Study 4: Gestation day 6 through 20
Frequency of treatment:
Study 2, 3 and 4: daily
Duration of test:
Study 2: 17 days (examination of fetuses were performed on day 17)
Study 3: Approximately 8 weeks
Study 4: 16 days
Remarks:
Study 2: 0, 30, 60, 120, and 180 mg/kg/day
Remarks:
Study 3: Doses / Concentrations:
0, 70, 155, 350 mg/kg bw/day
Basis:
no data
Remarks:
Study 4: Doses / Concentrations:
0, 2.5. 7.5 and 15 mg/kg/day
Basis:
nominal in water
No. of animals per sex per dose:
Study 2: 20 pregnant mice in each group
Study 3: 10 animals/sex/dose
Study 4: Control: 30 females
2.5 mg/kg/day: 29 females
5 mg/kg/day: 30 females
15 mg/kg/day: 29 females
Control animals:
other: Study 2 and 4: yes, concurrent vehicle; study 3: yes, 5% v/v etahnol in corn oil
Details on study design:
Study 2: Administration by gavage on the mornings of gd 6 through 17, using an 18-gauge 1.5 inch stainless steel dosing tube, dose volume was 10ml/kg. On gestational day 17, approximately one to one and a half days before expected parturition. pregnant dams were anesthetized by carbon dioxide gas and sacrificed by cervical dislocation. The uterus and right ovary were removed and fetus examination were performed

Study 3: No data available

Study 4: The oral route of administration corresponds to one of the potential routes of exposure to ACRL in the human population. Gavage was selected as the method of oral administration to allow accurate control of the dose administered. The doses selected for the dose range finding study were 0, 2.5, 5.0, 15, 30 and 60 mg/kg/day. The upper end of the proposed dosage range was expected to equal or exceed the rat LD10 based on rat mortality data from the literature. The lower end of the range included doses predicted to have no toxic effects on maternal or developmental parameters.
Maternal examinations:
Study 2: Feed comsumption, water consumption, Body weight, liver, and uterus of each female were recorded on the mornings of gd 0, 3, 6-17 and 1mmediately following sacrifice on gd 17.

Study 3: one 350mg/kg female, on days 4 and 5 of dosing, showed post-dosing signs of subdued behaviour, twitching, semi-closed eyes, piloerection, was cold and showed staggered movement.

Study 4: Body weight for pregnant female was recorded on the mornings of gestation day 0 and 6-20, and immediately following sacrifice on gestation day 20. Spermpositive females were observed for clinical signs at and 4 hours after dosing from gestation day 6 through gestation day 19. Before dosing on the morning of gestation day 20, clinical signs were recorded. Animals that died between gestation day 0 and scheduled sacrifice on gestation day 20 were necropsied in order to determine the cause of death.
Uteri which presented no visible implantation sites were stained with ammonium sulfide (10%) in order to visualize any implantation sites which may have undergone very early resorption. Total number of corpora lutea, liver weight, gravid uterine weight and uterine contents were examined.
Ovaries and uterine content:
Study 2: The uterus and right ovary were removed and uterine contents examined to determine the number of implantation sites, resorptions, dead fetuses, and live fetuses.

Study 4: Uteri which presented no visible implantation sites were stained with ammonium sulfide (10%) in order to visualize any implantation sites which may have undergone very early resorption. Total number of corpora lutea, liver weight, gravid uterine weight and uterine contents were examined.
Fetal examinations:
Study 2: fetal examination were performed including head, external and visceral examination.

Study 4: Live fetuses were dissected from the uterus and immediately placed on a moist paper towel over a tray of ice to induce terminal cold anesthesia. Following terminal anesthesia and gross examination, the fetuses were sexed, weighed and dissected for visceral examination.
Statistics:
Study 2: Arcsine, Square root transformation, Bartletts test, ANOVA and chi- square test were used for statistical analysis

Study 4: Parametric statistical procedures were applied to selected measures from the teratology study
Indices:
No Data Available
Historical control data:
No Data Available
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Study 2: Mouth bleeding before dosing, Cyst on right ovary,Left ovary enclosed in blood· filled cyst were observed at 30mg/kg group; Cyst on left ovary was observed in 60mg/kg group; Uterus enlarged, Rough coat,Vaginal bleeding,Cyst on right ovary were observed in 120mg/kg group females perioral wetness, Vaginal discharge (brownish red), Lethargic,Convulsing, Cyst on right ovary, Curling hind limbs under bodywere obseved in treated females but this effects were non significant compared to control female mice.
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
Study 2: No mortality were observed at 30, 60, 120 and 180mg/kg dose groups
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Study 2: No significant change in body weight were observed at 30, 60, 120 and 180mg/kg dose groups compared to control
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Study 2: No significant change in feed consumption were observed at 30, 60, 120 and 180mg/kg dose groups compared to control
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
Study 2: No significant change in water consumption were observed at 30, 60, 120 and 180mg/kg dose groups compared to control
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
effects observed, non-treatment-related
Description (incidence and severity):
Study 3: Functional observation battery: Forelimb grip strength in males during Week 7 showed a dose-related decrease. There were no other dose-related observations on functional effects.
Locomotor activity: No effects
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Study 2: Significant change in liiver and gravid uterus were observed in 180mg/kg group animals

Study 4: Organ weight effects observed in kidneys, adrenal glands and liver. Adjusted mean kidney weight was increased in high dose 350mg/kg males Adjusted mean adrenal weight was increased in high dose females( P<0.05). Adjusted mean liver weight was increased in all treated male groups at 70, 155 and 350mg/kg groups
Gross pathological findings:
no effects observed
Description (incidence and severity):
Study 3: Necropsy: No necropsy findings that were deemed to be treatment-related.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Study 3: Histopathology:There were treatment-related findings in the liver, kidney and lungs. Centrilobular hypertrophy was recorded in the majority of the high 350mg/kg dose male and female livers. In the kidney there was an increase in the level of hyaline droplets in males at 350mg/kg dose and In the lungs there was a minor increase in the level of foamy histiocytes in males and females from 350mg/kg dose group.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Number of abortions:
not specified
Pre- and post-implantation loss:
not specified
Total litter losses by resorption:
not specified
Early or late resorptions:
not specified
Dead fetuses:
not specified
Changes in pregnancy duration:
not specified
Changes in number of pregnant:
not specified
Other effects:
no effects observed
Description (incidence and severity):
Study 3: Reproductive performance: No treatment-related effects on numbers of males and females mating or on male or female fertility indices

Details on maternal toxic effects:
Study 3: Testicular staging: Qualitative testis staging did not indicate any abnormalities in the integrity of the various cell types present within the different stages of the spermatogenic cycle.

Study 4: Maternal toxic effects:no effects
Details on maternal toxic effects:
Maternal toxicity during test chemical exposure was expressed primarily by decreased weight gain at the high dose. Maternal weight gain during treatment and gestation as well as weight gain corrected for gravid uterine weight exhibited decreasing trends. Maternal weight gain during treatment was reduced below control values in the high dose group, and corrected weight gain was reduced in the 7.5 and 15 mg/kg dose groups. Gravid uterine and liver weights were not measurably affected by test chemical treatment. In addition, other clinical signs were observed infrequently and exhibited no clear-cut relationship to test chemical dose.
Dose descriptor:
NOAEL
Effect level:
180 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
behaviour (functional findings)
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Abnormalities:
not specified
Fetal body weight changes:
effects observed, treatment-related
Description (incidence and severity):
Study 3: For the 350mg/kg dose group, Day 4 weights were lower than controls
Reduction in number of live offspring:
effects observed, treatment-related
Description (incidence and severity):
Study 3: Total embryo foetal loss occurred for one high 350mg/kg dose female.
Changes in sex ratio:
not specified
Changes in litter size and weights:
effects observed, treatment-related
Description (incidence and severity):
Study 3: litter sizes was lower in 350mg/kg group than controls. .
Total litter loss occurred for two females in the high 350mg/kg dose group and one from each of the 70 and 155mg/kg and dose groups
Changes in postnatal survival:
effects observed, treatment-related
Description (incidence and severity):
Study 3: a reduction in the pups viability in the high dose 350mg/kg group was observed when compared to control animals
External malformations:
effects observed, non-treatment-related
Description (incidence and severity):
Study 2: Cleft Palate, Ectocardia, Gastroschisis and Short Tail were obseved in 2 fetuses

Study 3: Malformed/ shortened limbs occurred in one pup in the 155mg/kg and one pup in the 350mg/kg groups, however as this effect occurred in only one pup in one litter in these groups, the toxicological significance is not clear.
Skeletal malformations:
effects observed, treatment-related
Description (incidence and severity):
Study 2: Asymmetrical Fusion of Sternal Plate, Cleft Sternum, Perforated Sternum, Fused Rib Cartilage, Incomplete Ossification, Cartilage Present, Extra Ossification Site(s), Hematoma: Head, Face, Jaw, Neck, Abdomen, Thorax, Hindlimb were observed.
Visceral malformations:
effects observed, treatment-related
Description (incidence and severity):
Study 2: Situs Inversus Viscerum and left hydronephrosis noted in treated animals
Other effects:
effects observed, treatment-related
Description (incidence and severity):
Study 3: For the high 350mg/kg dose, numbers of implantations was lower than control animals
Details on embryotoxic / teratogenic effects:
Study 3: Embryotoxic / teratogenic effects:yes
Details on embryotoxic / teratogenic effects:
At the 350mg/kg dose tested, the test material reduces pup viability after delivery

Study 4: Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
ACRL treatment during gestation did not alter measured endpoints of embryo/fetal viability, growth or development. The percentages of resorptions. dead fetuses or malformed fetuses per litter, as well as fetal body weights, were statistically similar among the dose groups. ACRL treatment had no effect on the incidence of skeletal. visceral or external malformations. There was a dose-related trend toward an increased incidence of variations, but there were no statistically significant pairwise comparisons.
Dose descriptor:
NOAEL
Effect level:
120 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: The results is based on the effects on teratoloical parameters.
Remarks on result:
other: Not Specified
Abnormalities:
not specified
Developmental effects observed:
not specified
Treatment related:
not specified
Conclusions:
Based on all the observations and results, it was observed that, the NOAEL for the test chemical for reproductive parameters of parental animals was found to be 180 mg/kg bw and NOAEL for the developmental parameters fetuses was found to be 120 mg/kg bw.
Executive summary:

Developmental Toxicity Study:

The data for developmental toxicity as summaries is given below:

Developmental Toxicity Study 2:

In the present study, developmental Toxicity of test chemical was evaluated by adinistration of test chemical by Gavage to Swiss CCD-I (trade name)) Mice on Gestational Days 6 through 17. The dose levels of 0, 30, 60, 120, 180mg/kg were selected and administered in pregnant female mice at day 6 to 17 of gestation period. Dams were observed for clinical signs, change in body weight, feed and water consumption and organ weight change specially liver and gravid uterus. The fetuses were observed for external, visceral and skeletal abnormalities.

The results of the study revealed clinical toxicity life, Cyst on right ovary,Left ovary enclosed in blood· filled cyst  were observed at 30mg/kg group; Cyst on left ovary was observed in 60mg/kg group; Uterus enlarged, Rough coat,Vaginal bleeding,Cyst on right ovary were observed in 120mg/kg group females perioral wetness, Vaginal discharge (brownish red), Lethargic,Convulsing, Cyst on right ovary, Curling hind limbs under bodywere obseved in treated females but this effects were non significant compared to control female mice but no change in body weight change, feed consumption and water consumption were observed maternal animals at 30,60,120 and 180mg/kg groups.

The fetuses observed for external , visceral and skeletal abnormalities, the results were Cleft Palate, Ectocardia, Gastroschisis and Short Tail were obseved in 2 fetuses. Skeletal malformations were observed such as asymmetrical Fusion of Sternal Plate, Cleft Sternum, Perforated Sternum, Fused Rib Cartilage, Incomplete Ossification, Cartilage Present, Extra Ossification Site(s), Hematoma: Head, Face, Jaw, Neck, Abdomen, Thorax, Hindlimb. the visceral abnormalities also obseved in treated animals including, Situs Inversus Viscerum and left hydronephrosis. From the observations and results of the study, The NOAEL value from the maternal observations was considered to be 180mg/kg/day howeveer the NOAEL value from the fetuses observation considered to be 120mg/kg.

Developmental Toxicity Study 3:

In the present study, the effect of test chemical on repeated administration of the test chemical and reproductive developmental toxicity study performed by oral administration. The test substance was administered to rats at doses 0, 70, 155, 350 mg/kg bw/day for approximately 8 weeks.The animals observed for clinical signs of toxicity; mortality; feed consumption; change in body weight; behaviour such as locomotor activity and grip strength; gross necropsy examination histopathological examination and reproductive performance were examined in the present study. The results of study revealed, no mortality during the period of study and one 350mg/kg female, on days 4 and 5 of dosing, showed post-dosing signs of subdued behaviour, twitching, semi-closed eyes, piloerection, was cold and showed staggered movement.The body weight results revealedmajority of males in 350mg/kg group lost weight during the first week of dosing. Although mean body weights increased every week after that, the high dose males were approximately 9% lighter than control males by the end of the study. Group mean body weight gain was lower in high 350mg/kg dose females than controls during gestation. 155 and 350mg/kg dose females had lower mean body weight gain than controls during lactation. Feed consumption was observed to be reduced in 350mg/kg group animals as compared to control animals. Forelimb grip strength in males during Week 7 but there were no other dose-related observations on functional effects including locomotor activity at dose levels of 70, 155 and 350mg/kg. Organ weight change were seen in kidney, liver and adrenal weight, significantly increase were observed in high dose 350mg/kg group animals as compared to control animals. No gross necropsy findings were seen that attributed to be treatment-related at any dose group at 70, 155 and 350mg/kg.The reproductive findings of the study showed no indication of any qualitative abnormality in the various cell types present within the different stages of the spermatogenic cycle and No treatment-related effects on numbers of males and females mating or on male or female fertility indices. The F1 generation results from parent rats revealed reduction in the pups viability in the 350mg/kg dose group. The litter data shown numbers of implantations, litter sizes and Day 4 weights were all lower than controls in the high dose 350mg/kg. Total litter loss occurred for two females in the 350mg/kg and one from each of the 70 and 155mg/kg dose groups.Total embryo foetal loss occurred for one female from 350mg/kg group. Malformed/ shortened limbs occurred in one pup in the 155mg/kg and one pup in the 350mg/kg dose group, however as this effect occurred in only one pup in one litter in these groups, the toxicological significance is not clear. From the observations and result of the study, The NOAEL and LOAEL for rats was considered to be 155 mg/kg/day  and 350mg/kg/day respectively in rats.

Developmental Toxicity Study 4:

The test chemical, a cumulative neurotoxicant was evaluated in a 1 generation study for developmental toxicity in pregnant Sprague-Dawley (Crl:CD BR VAF/Plus) rats. Acrylamide was administered once daily by gavage on gestation day 6 through gestation day 20 in the concentration of 0, 2.5, 7.5 and 15 mg/kg. Maternal weight gain during treatment and gestation, as well as weight gain corrected for gravid uterine weight, exhibited decreasing trends. Maternal weight gain during treatment was reduced below control values in the high dose group, and corrected weight gain was reduced in the 7.5 and 15 mg/kg dose groups. The test chemical treatment during gestation did not alter measured endpoints of embryo/fetal viability, growth or development. The percentages of resorptions, dead fetuses or malformed fetuses per litter, as well as fetal body weights, were statistically similar among the dose groups. The test chemical treatment had no effect on the incidence of skeletal, visceral or external malformations. There was a dose-related trend toward an increased incidence of variations, but there were no statistically significant pairwise comparisons. through20.The No observed effect level (NOAEL) for the test chemical in rats was observed to be15 mg/kg/day. The No observed effect level (NOAEL) for acrylamide in rats was observed to be 2.5 mg/kg/day for maternal toxicity.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
120 mg/kg bw/day
Study duration:
subchronic
Species:
mouse
Quality of whole database:
The 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:

The data for developmental toxicity as summaries is given below:

Developmental Toxicity Study 2:

In the present study, developmental Toxicity of test chemical was evaluated by adinistration of test chemical by Gavage to Swiss CCD-I (trade name)) Mice on Gestational Days 6 through 17. The dose levels of 0, 30, 60, 120, 180mg/kg were selected and administered in pregnant female mice at day 6 to 17 of gestation period. Dams were observed for clinical signs, change in body weight, feed and water consumption and organ weight change specially liver and gravid uterus. The fetuses were observed for external, visceral and skeletal abnormalities. The results of the study revealed clinical toxicity life, Cyst on right ovary,Left ovary enclosed in blood· filled cyst  were observed at 30mg/kg group; Cyst on left ovary was observed in 60mg/kg group; Uterus enlarged, Rough coat,Vaginal bleeding,Cyst on right ovary were observed in 120mg/kg group females perioral wetness, Vaginal discharge (brownish red), Lethargic,Convulsing, Cyst on right ovary, Curling hind limbs under bodywere obseved in treated females but this effects were non significant compared to control female mice but no change in body weight change, feed consumption and water consumption were observed maternal animals at 30,60,120 and 180mg/kg groups. The fetuses observed for external , visceral and skeletal abnormalities, the results were Cleft Palate, Ectocardia, Gastroschisis and Short Tail were obseved in 2 fetuses. Skeletal malformations were observed such as asymmetrical Fusion of Sternal Plate, Cleft Sternum, Perforated Sternum, Fused Rib Cartilage, Incomplete Ossification, Cartilage Present, Extra Ossification Site(s), Hematoma: Head, Face, Jaw, Neck, Abdomen, Thorax, Hindlimb. the visceral abnormalities also obseved in treated animals including, Situs Inversus Viscerum and left hydronephrosis. From the observations and results of the study, The NOAEL value from the maternal observations was considered to be 180mg/kg/day howeveer the NOAEL value from the fetuses observation considered to be 120mg/kg.

Developmental Toxicity Study 3:

In the present study, the effect of test chemical onrepeated administration of the test chemical and reproductive developmental toxicity study performed by oral administration.The test substance was administered to rats at doses 0, 70, 155, 350 mg/kg bw/day for approximately 8 weeks.The animals observed for clinical signs of toxicity; mortality; feed consumption; change in body weight; behaviour such as locomotor activity and grip strength; gross necropsy examination histopathological examination and reproductive performance were examined in the present study.The results of study revealed, no mortality during the period of study and one 350mg/kg female, on days 4 and 5 of dosing, showed post-dosing signs of subdued behaviour, twitching, semi-closed eyes, piloerection, was cold and showed staggered movement.The body weight results revealedmajority of males in 350mg/kg group lost weight during the first week of dosing. Although mean body weights increased every week after that, the high dose males were approximately 9% lighter than control males by the end of the study. Group mean body weight gain was lower in high 350mg/kg dose females than controls during gestation. 155 and 350mg/kg dose females had lower mean body weight gain than controls during lactation. Feed consumption was observed to be reduced in 350mg/kg group animals as compared to control animals. Forelimb grip strength in males during Week 7 but there were no other dose-related observations on functional effects including locomotor activity at dose levels of 70, 155 and 350mg/kg. Organ weight change were seen in kidney, liver and adrenal weight, significantly increase were observed in high dose 350mg/kg group animals as compared to control animals. No gross necropsy findings were seen that attributed to be treatment-related at any dose group at 70, 155 and 350mg/kg.The reproductive findings of the study showed no indication of any qualitative abnormality in the various cell types present within the different stages of the spermatogenic cycle and No treatment-related effects on numbers of males and females mating or on male or female fertility indices. The F1 generation results from parent rats revealed reduction in the pups viability in the 350mg/kg dose group. The litter data shown numbers of implantations, litter sizes and Day 4 weights were all lower than controls in the high dose 350mg/kg. Total litter loss occurred for two females in the 350mg/kg and one from each of the 70 and 155mg/kg dose groups.Total embryo foetal loss occurred for one female from 350mg/kg group. Malformed/ shortened limbs occurred in one pup in the 155mg/kg and one pup in the 350mg/kg dose group, however as this effect occurred in only one pup in one litter in these groups, the toxicological significance is not clear. From the observations and result of the study, The NOAEL and LOAEL for rats was considered to be 155 mg/kg/day  and 350mg/kg/day respectively in rats.

Developmental Toxicity Study 4:

The test chemical, a cumulative neurotoxicant was evaluated in a 1 generation study for developmental toxicity in pregnant Sprague-Dawley (Crl:CD BR VAF/Plus) rats. The test chemical was administered once daily by gavage on gestation day 6 through gestation day 20 in the concentration of 0, 2.5, 7.5 and 15 mg/kg. Maternal weight gain during treatment and gestation, as well as weight gain corrected for gravid uterine weight, exhibited decreasing trends. Maternal weight gain during treatment was reduced below control values in the high dose group, and corrected weight gain was reduced in the 7.5 and 15 mg/kg dose groups. The test chemical treatment during gestation did not alter measured endpoints of embryo/fetal viability, growth or development. The percentages of resorptions, dead fetuses or malformed fetuses per litter, as well as fetal body weights, were statistically similar among the dose groups. The test chemical treatment had no effect on the incidence of skeletal, visceral or external malformations. There was a dose-related trend toward an increased incidence of variations, but there were no statistically significant pairwise comparisons. through20.The No observed effect level (NOAEL) for the test chemical in rats was observed to be15 mg/kg/day. The No observed effect level (NOAEL) for the test chemical in rats was observed to be 2.5 mg/kg/day for maternal toxicity.

Justification for classification or non-classification

Based on the available data for the assessment of reproductive toxicity and developmental toxicity, the test chemical may not be classified as the reproductive and developmental toxicant as per the CLP regulation.