Registration Dossier

Administrative data

Key value for chemical safety assessment

Toxic effect type:
concentration-driven

Effects on fertility

Description of key information

Reproductive toxicity study:

In a repeated dose toxicity study, the effect of test chemical was evaluated in male and female Sprague Dawley rats. The test chemical was administered by oral gavage in the concentration of 0, 125, 375 or 1125 mg/kg/day. The results showed that test chemical significantly change the percentage of monocytes and basophils, WBC count, MCV and MCH levels in hematology, total protein and total cholesterol, sodium, potassium, total proteins, BUN levels in clinical biochemistry. Absolute and relative liver and heart weight was changed as compared to control while no effects were observed in water consumption, opthalmoscopic examination or locomotor activity. Histopathology performed on reproductive organs after treatment with 1125 mg/kg/day did not reveal any toxic lesions as compared to control. Hence, NOAEL was considered to be 1125 mg/kg/day when male and female Sprague Dawley rats were exposed daily to test chemical by oral route for 28 days.

Link to relevant study records
Reference
Endpoint:
reproductive toxicity, other
Remarks:
REPEATED DOSE 28- DAYS ORAL TOXICITY STUDY EXTENDED WITH REPRODUCTIVE PARAMETERS
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 study report
Qualifier:
according to guideline
Guideline:
other: According to OECD Guideline 407 with additional examinations for reproductive performance.
Principles of method if other than guideline:
The effect of repeated exposure of the test chemical on the reproductive systems of Sprague Dawley (SD) Rats
GLP compliance:
yes
Limit test:
no
Justification for study design:
No Data Available
Specific details on test material used for the study:
- Name of the test material: 3-(5,5,6-trimethylbicyclo[2.2.1]hept-2-yl)cyclohexan-1-ol
- IUPAC name: 3-(5,5,6-trimethylbicyclo[2.2.1]hept-2-yl)cyclohexanol
- Molecular formula: C16H28O
- Moleclar weight: 236.396 g/mol
- Substance type: Organic
Species:
rat
Strain:
Sprague-Dawley
Details on species / strain selection:
No Data Available
Sex:
male/female
Details on test animals or test system and environmental conditions:
- Source: Central Animal Facility (CAF), NIPER, Sector-67, S.A.S. Nagar, 160 062, Punjab, India.
- Age at study initiation: (P) x wks; (F1) x wks: (P) 7 to 8 weeks old
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: No data available
- Fasting period before study: Yes, 2 hrs fasted before dose admonition.
- Housing: Animals were housed in sterilized solid bottom polypropylene cages with stainless steel grill tops with facilities for food, water bottles and bedding of clean paddy husk. The cages were suspended on stainless steel racks. Animal was identified by a unique identification (ID) number. The animals were identified by individual numerical number written on the tail, also specified on individual cage tag.
- Diet (e.g. ad libitum): Standard laboratory sterile extruded pelleted rodent feed, ad libitum.
- Water (e.g. ad libitum): Potable tap water filtered through Reviva Reverse Osmosis System (water filter cum purifier) of Eureka Forbes was provided in polypropylene bottles with stainless steel sipper tubes, ad libitum.
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3º C
- Humidity (%): 30-70 %
- Air changes (per hr): 25±5 air changes per hour
- Photoperiod (hrs dark / hrs light): 12hrs dark/ light cycle
Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
corn oil
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: The test chemical was dissolved in corn oil prior to treatment.

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
- Concentration in vehicle: 0, 125, 375 or 1125 mg/kg/day
- Amount of vehicle (if gavage): 5 ml/kg
- Lot/batch no. (if required): No data available
- Purity: No data available
Details on mating procedure:
No data available
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The test item (125, 375 and 1125 mg) was weighed on balance and dissolved in 5 ml of corn oil. An aliquot was taken from three different layers of the dose mixtures and was extracted in methanol for GCMS analysis. The concentration of the test item in each layer was calculated using the standard curve (5 mg IBC / ml methanol).
Duration of treatment / exposure:
28 days
Frequency of treatment:
Daily
Details on study schedule:
No data available
Dose / conc.:
0 mg/kg bw/day (actual dose received)
Dose / conc.:
125 mg/kg bw/day (actual dose received)
Dose / conc.:
375 mg/kg bw/day (actual dose received)
Dose / conc.:
1 125 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
Total : 56
0 mg/kg/day: 7 male, 7 female
125 mg/kg/day: 7 male, 7 female
375 mg/kg/day: 7 male, 7 female
1125 mg/kg/day: 7 male, 7 female
Control animals:
yes, concurrent vehicle
Details on study design:
No data available
Positive control:
No data available
Parental animals: Observations and examinations:
The parental generation was examined for mortality, changes in body weight, food consumption, water consumption, hematology, clinical chemistry, and ophthalmological examination and locomotor activity were examined.
Oestrous cyclicity (parental animals):
No data available
Sperm parameters (parental animals):
No data available
Litter observations:
No data available
Postmortem examinations (parental animals):
The testes, ovaries and uterus were pathologically examined after termination. Examinations also included weight of brain, testes and ovaries (incl. paired ovaries and uterus, with cervix included).
Postmortem examinations (offspring):
No data available
Statistics:
Statistical analysis was carried out by using Microsoft Excel and IBM SPSS statistics version-20.0. All analyses and comparisons were evaluated at the 5 % level, statistically significant differences (p≤0.05) indicated by appropriate notation. The focus was to examine the mean differences and their significance between control and low dose group, control and mid dose group and control and high dose group. The statistical decision was taken by preparing the univariant GLM MODEL procedure was used to check the significance between above mentioned groups. For multiple comparisons Turkey’s HSD test was applied.
Reproductive indices:
No data available
Offspring viability indices:
No data available
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Some abnormalities like nasal discharge, slight dullness, hunched posture, fore paw stained red, sneezing, red crust around the nostrils, perineum wet were common to all the groups.
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Description (incidence):
All animals survived throughout the treatment period of 28 days.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
In male animals, a significant increase has been noticed in the body weight of high-dose group as compared to control animals on day 8. However, no change was noticed in the body weight of female animals taken at different time periods.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No difference was observed among any groups of male as well as female animals.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
No difference was observed among any groups of male as well as female animals.
Ophthalmological findings:
no effects observed
Description (incidence and severity):
No abnormalities were found in the ophthalmological examination (control vs. high-dose treated group).
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
When treated with 1125 mg/kg/day, in male rat significant increased were observed in percentage of monocytes and basophils.

In female rat, significant increase were observed in WBC count, MCV and MCH levels and significant decreased in RBC count and Hct level as compare to control.

When treated with 375 mg/kg/day, in female rat significant increase were observed in WBC count and MCH level and significant decreased in RBC count as compare to control.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
When treated with 1125 mg/kg/day, in male rat significant increase were observed in total protein and total cholesterol.

In female rat, significant increasewere observed in sodium, potassium, total proteins, BUN levels. And activity of SGPT and significant decreased were observed in Glucose level as compare to control.

When treated with 375 mg/kg/day, in male rat significant increase were observed in glucose and total proteins leve.
In female rat, significant increase were observed in SGPT activity, BUN level and decreased TBA level as compare to control.

When treated with 125 mg/kg/day, in male rat significant increase were observed in creatinine level as compare to control.
Urinalysis findings:
not specified
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
No statistically significant difference was observed in the locomotor activity scores for any dose group when compared with the control animals.
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
No remarkable changes were observed in control and in 1125 mg/kg /day-treated animals.

A few microscopic findings were observed in 1125 mg/kg/day-treated animals included reactive spleen and excess of lymphocytes in lungs.

These types of findings may be considered to be within the range of normal background lesions which may be seen in rats of this strain and age of this study type and were considered incidental in nature with carbon dioxide inhalation and terminal changes at sacrifice, reflecting the usual individual variability.

However, 1125 mg/kg/day group animals did not reveal any toxic lesions on histological examinations when compared with the respective control group, hence further histological investigation was not extended to the two other lower dose (low-dose and mid-dose) treatment groups
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Reproductive function: oestrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
Description (incidence and severity):
Histopathology performed on reproductive organs after treatment with 1125 mg/kg/day did not reveal any toxic lesions as compared to control.
Dose descriptor:
NOAEL
Effect level:
1 125 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
ophthalmological examination
haematology
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
Remarks on result:
other: No toxic effects were observed
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality / viability:
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
Dose descriptor:
other: not specified
Generation:
other: not specified
Based on:
not specified
Sex:
not specified
Basis for effect level:
other: not specified
Remarks on result:
other: not specified
Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Reproductive effects observed:
not specified
Treatment related:
not specified

SUMMARY OF ABSOLUTE ORGAN WEIGHTS (g) Male and Female

Dose

 

Testes

Epididymides

Ovaries

Uterus

0 (mg/kg)

Mean

2.6556

1.0201

0.0944

 

0.5236

SD

0.6146

0.1210

0.0322

0.1078

SEM

0.2323

0.0457

 

0.0122

 

0.0407

  125 (mg/kg)

Mean

2.7550

0.9950

0.0953

0.5293

SD

0.2371

0.1474

0.0382

0.1607

SEM

0.0896

0.0557

0.0144

0.0607

375

(mg/kg)

Mean

2.8762

0.9389

 

0.0725

 

0.4847

SD

0.1901

0.1180

0.0209

0.0706

SEM

0.0718

0.0446

0.0079

0.0267

1125

(mg/kg)

Mean

2.6205

0.9031

 

0.0693

 

0.5155

SD

0.4577

0.2122

0.0290

0.1438

SEM

0.1730

0.0802

0.0110

0.0543

 

 SUMMARY OF RELATIVE ORGAN WEIGHTS Male and Female

Dose

 

Testes

Epididymides

Ovaries

Uterus

 0

(mg/kg)

Mean

0.9291

0.3560

0.0383

0.2180

SD

0.2331

0.0436

 

0.0136

 

0.0385

SEM

0.0881

0.0165

 

0.0051

 

0.0146

375

(mg/kg)

Mean

0.9504

0.3462

0.0384

0.2135

SD

0.1070

0.0756

0.0158

0.0687

SEM

0.0405

0.0286

0.00601

0.0260

1125

(mg/kg)

Mean

0.9617

     0.3139

0.0297

0.1994

SD

0.0908

0.0451

0.0083

0.0375

SEM

0.0343

0.0171

0.0031

0.0135

1000

(mg/kg)

Mean

0.9235

0.3176

0.0295

0.2213

SD

0.1438

0.0678

0.0116

0.0614

SEM

0.0544

0.0256

0.0044

0.0232

Clinical chemistry:

Dose

 

Testesterone

nmol/L

Estrogen

ng/L

0 (mg/kg)

Mean

14.11

9.01

SD

2.94

0.87

SEM

1.11

0.33

125 (mg/kg)

Mean

15.19

9.14

SD

1.07

1.14

SEM

0.40

0.43

375 (mg/kg)

Mean

12.30

10.00

SD

1.80

2.03

SEM

0.68

0.77

1125 (mg/kg)

Mean

13.64

10.39

SD

1.94

1.35

SEM

0.73

0.51

Conclusions:
NOAEL was considered to be 1125 mg/kg body weight/day for test chemical . when test chemical administered orally by gavage to male and female Sprague-Dawley rats.
Executive summary:

In a repeated dose toxicity study, the effect of test chemical was evaluated in male and female Sprague Dawley rats. The test chemical was administered by oral gavage in the concentration of 0, 125, 375 or 1125 mg/kg/day. The results showed that test chemical significantly change the percentage of monocytes and basophils, WBC count, MCV and MCH levels in hematology, total protein and total cholesterol, sodium, potassium, total proteins, BUN levels in clinical biochemistry. Absolute and relative liver and heart weight was changed as compared to control while no effects were observed in water consumption, opthalmoscopic examination or locomotor activity. Histopathology performed on reproductive organs after treatment with 1125 mg/kg/day did not reveal any toxic lesions as compared to control. Hence, NOAEL was considered to be 1125 mg/kg/day when male and female Sprague Dawley rats were exposed daily to test chemical by oral route for 28 days.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 125 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Data is Klimicsh 1 and 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

Reproductive toxicity study:

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

Reproductive ToxicityStudy 1:

In a repeated dose toxicity study, the effect of test chemical was evaluated in male and female Sprague Dawley rats. The test chemical was administered by oral gavage in the concentration of 0, 125, 375 or 1125 mg/kg/day. The results showed that test chemical significantly change the percentage of monocytes and basophils, WBC count, MCV and MCH levels in hematology, total protein and total cholesterol, sodium, potassium, total proteins, BUN levels in clinical biochemistry. Absolute and relative liver and heart weight was changed as compared to control while no effects were observed in water consumption, opthalmoscopic examination or locomotor activity. Histopathology performed on reproductive organs after treatment with 1125 mg/kg/day did not reveal any toxic lesions as compared to control. Hence, NOAEL was considered to be 1125 mg/kg/day when male and female Sprague Dawley rats were exposed daily to test chemical by oral route for 28 days.

Reproductive Toxicity Study 2:

In a Embryotoxicity study, Fischer 344 female rat treated with test chemical in the concentration of 0, 250, 500 and 1000 mg/kg/day orally by gavage in corn oil for 10 days (6 through 15 of gestation). Tremors and mild convulsions were observed in treated female rats at 1000 mg/kg bw. Decrease in body weight gain were observed in females rat at 1000 mg/kg bw and from day 6 to 10 of gestation at 500 mg/kg bw. No effect on body weight gain of treated female rats were observed as compared to control at 250 mg/kg bw. Similarly, Statistically significant increase in litters containing resorptions or implants at 1000 mg/kg bw. Increase in resorptions/litter although elevated was not significant. In addition, Increase in embryolethality were observed at 1000 mg/kg bw. Significant decrase in fetal weight were observed at 500 mg/kg bw and No effect on fetal body weight were observed as compared to control at 1000 mg/kg bw. Nanoidism (unusually small fetus) at 500 and 1000 mg/kg bw. This abnormality did not occur in the oral control or 250 mg/kg bw dose group in this study and has occurred in other control litters historically in this laboratory at a rate similar to that observed in the higher treatment groups. Hydroinephrosis, dilated ureter, and agenesis of the bladder which occurred in 3 fetuses from 1 litter at 1000 mg/kg bw. Since this malformation was observed in the fetuses from only one litter it probably arose spontaneously rather than as a result of treatment. Therefore, NOAEL was considered to be 500 mg/kg/day for P and 1000 mg/kg bw for F1 generation when Fischer 344 female rat treated with test chemical orally by gavage for 10 days (6 through 15 of gestation).

Reproductive Toxicity Study 3:

In aTeratogenicity study,ICR female mice were treated with test chemical in the concentration of 0, 188, 376, 569 and 752 mg/kg/day orally by gavage in Soybean oil for 4 days (6, 7, 8 and 9 of gestation). Increase in body weight gain were observed in the females mice at 188, 376 and 752 mg/kg bw as compared to control and 569 mg/kg bw treated mice.The increased maternal weight in the188 and 376 mg/kg is apparently due to increased fetal weight and the increased maternal weight gain in the 752 mg/kg group is due to slightly larger average litter size. Similarly,No significant effect on mean implants/female, mean resorptions/litter and frequency of resorptions of treated female mice were observed as compared to control. In addition, No effect on viability of pups were observed as compared to control. Increase in mean fetal weights were observed in male and female pups at 188 and 376 mg/kg bw as compared to control and 569 and 752 mg/kg bw treated mice.The increased fetal weight in the mouse is not easily explained particularly when no such increase occurred in the groups receiving 569 and 752 mg/kg. No external abnormalities, Soft Tissue and or Skeletal findings were observed in any pup as compared to control. Therefore, NOAEL was considered to be 752 mg/kg/day for P and F1 generation when ICR female mice treated with test chemical orally by gavage for 4 days (6, 7, 8 and 9 of gestation).

Reproductive Toxicity Study 4:

In a Reproductive Toxicity study,Syrian female hamsters were treated with test chemical the concentration of 4.05, 21.15, 98.2 and 405.0 mg/kg bw/day orally by gavage during gestation days 6-10. No effect on survival of treated female hamsters and fetus were observed. Similarly, No effect on Reproductive performance of treated female hamsters was observed.In addition, No dose-related number of abnormalities in soft or skeletal tissues were observed in fetus of treated hamsters were observed.Therefore, NOAEL was considered to be 405.0 mg/kg bw for P and F1 generation when Syrian female hamsters were treated with test chemical orally by gavage during gestation days 6-10.

Effects on developmental toxicity

Description of key information

Developmental toxicity study

Based on the various studies available for the test chemical were reviewed to determine the developmental toxicity, NOAEL for test chemical was considered to be 500 mg /kg bw/day .When rats were treated with test chemical orally. Thus, comparing this value with the criteria of CLP regulation test chemical is not likely to classify as reproductive and developmental toxicant.

Link to relevant study records
Reference
Endpoint:
developmental toxicity
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
Experimental data from various test chemicals
Justification for type of information:
Weight of evidence approach based on the available information from various test chemicals.
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: as below
Principles of method if other than guideline:
WoE report is based on developmental toxicity studies on rats,mice ,hamster and rabbits
GLP compliance:
not specified
Limit test:
no
Specific details on test material used for the study:
- Name of the test material: 3-(5,5,6-trimethylbicyclo[2.2.1]hept-2-yl)cyclohexan-1-ol
- IUPAC name: 3-(5,5,6-trimethylbicyclo[2.2.1]hept-2-yl)cyclohexanol
- Molecular formula: C16H28O
- Moleclar weight: 236.396 g/mol
- Substance type: Organic
Species:
other: 1&4.mouse,2&3 rats.5. hamsters 6.rabbit
Strain:
other: 1.ICR 2.Fischer 344 3.Wistar 4.albino CD-l 5.golden hamsters 6.Dutch-belted
Details on test animals or test system and environmental conditions:
study 1.
- Source: Harlan Industries.
- Age at study initiation: (P) x wks; (F1) x wks: not specified
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: not specified
- Fasting period before study: not specified
- Housing: Animals were housed in plastic cages on wood chip
Bedding. Pregnant females were moved to wire bottomed cages.
- Diet (e.g. ad libitum): Purina Mouse Chow, ad libitum
- Water (e.g. ad libitum): Water , ad libitum
- Acclimation period: not specified

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-24 °C
- Humidity (%):not specified
- Air changes (per hr):not specified
- Photoperiod (hrs dark / hrs light): 12 hour light cycle.
study 2.
- Source: Charles River Breeding Laboratories
- Age at study initiation: (P) x wks; (F1) x wks: not specified
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: not specified
- Fasting period before study: not specified
- Housing: not specified
- Diet (e.g. ad libitum): Ralston Purina Co. feed, ad libitum
- Water (e.g. ad libitum): Water , ad libitum
- Acclimation period: not specified

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-24 °C
- Humidity (%): not specified
- Air changes (per hr): not specified
- Photoperiod (hrs dark / hrs light): 12 hour light cycle.
Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
other: 1.soya oil 2.corn oil
Details on exposure:
study 1.PREPARATION OF DOSING SOLUTIONS:The test chemical diluted with soybean oil at doses of 0, 188, 376, 564 and 752 mg/kg/day
DIET PREPARATION
- Rate of preparation of diet (frequency):not specified
- Mixing appropriate amounts with (Type of food): not specified
- Storage temperature of food:not specified

VEHICLE
- Justification for use and choice of vehicle (if other than water): soybean oil
- Concentration in vehicle: 0, 188, 376, 564 and 752 mg/kg/day
- Amount of vehicle (if gavage): not specified
- Lot/batch no. (if required): not specified
- Purity:not specified
study 2.PREPARATION OF DOSING SOLUTIONS: The test chemical was diluted with corn oil and controls received an equivalent volume of corn oil (2.0 ml/kg).

DIET PREPARATION
- Rate of preparation of diet (frequency):
- Mixing appropriate amounts with (Type of food):
- Storage temperature of food:

VEHICLE
- Justification for use and choice of vehicle (if other than water): corn oil
- Concentration in vehicle: 0, 250, 500 and 1000 mg/kg/day
- Amount of vehicle (if gavage): 2.0 ml/kg
- Lot/batch no. (if required):
- Purity:
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No Data Available
Details on mating procedure:
study 1.
- M/F ratio per cage: Four to five females were kept with each male.
- Length of cohabitation: not specified
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy The day that a vaginal plug was found was designated as day zero of pregnancy.
study 2.- Length of cohabitation: Overnight
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy The day sperm in vaginal smear was found was designated as day zero of pregnancy.
Duration of treatment / exposure:
study 1.&4
4 days (6, 7, 8 and 9 of gestation)
study 2&3.10 days (6 through 15 of gestation)
study 5.
5 days ( on Day 6 and continuing daily through Day 10 of gestation)
study 6.
12days ( on Day 6 and continuing daily through Day 18of gestation)
Frequency of treatment:
Daily
Duration of test:
1.17 days
2.20 days
5.15 days
6.29 days
Remarks:
1.
0.0,188.0,376.0,564.0,752.0 mg/kg bw /day
2.0.0,250,500.0,1000.0 mg/kg bw/day
3.0,2.18,10.15,47.05,218.0mg/kg bw/day
4.0,1.85,8.59,39.9,185.0 mg/kg bw/day
5.0,4.05,21.15,98.2,405.0mg/kg bw/day
6.0,4.25,19.75,91.7,425.0 mg/kg bw/day
No. of animals per sex per dose:
study 1.
Total: 46
0 mg/kg/day: 8 female
188 mg/kg/day: 9 female
376 mg/kg/day: 9 female
564 mg/kg/day: 9 female
752 mg/kg/day: 11 female
study 2.Total: 86
0 mg/kg/day: 23 female
250 mg/kg/day: 21 female
500 mg/kg/day: 21 female
1000 mg/kg/day: 21 female
study 3.Total:150
0 mg/kg bw/day:25 female
2.18mg/kg bw/day:25female
10.15mg/kg bw/day:25female
47.05mg/kg bw/day:25 female
218.0mg/kg bw/day:25 female
Positive control :
Aspirin 250mg/kg:25 female
study 4.Total:151
0 mg/kg bw/day:25 female
1.85mg/kg bw/day:28 female
8.59mg/kg bw/day:25 female
39.9mg/kg bw/day:23 female
185.0mg/kg bw/day:25female
Positive control :
Aspirin 150mg/kg: 25female
study 5.Total:149
0 mg/kg bw/day:25 female
4.05mg/kg bw/day:24female
21.15mg/kg bw/day:25female
98.2mg/kg bw/day:25 female
405.0mg/kg bw/day:25female
Positive control :
Aspirin 250mg/kg:25 female
study 6.Total:103
0 mg/kg bw/day:16female
4.25mg/kg bw/day:17female
19.75mg/kg bw/day:19 female
91.7mg/kg bw/day:15 female
425.0mg/kg bw/day:19 female
Positive control :
2.5 mg/kg 6-amino nicotinamide dosed on Day 9. To 17 females




Control animals:
yes, concurrent vehicle
Details on study design:
not specified
Maternal examinations:
Body weight were examined.
Ovaries and uterine content:
Resorptions and Implants were examined.
Fetal examinations:
Viable fetuses, Number and position of each fetus, Number of litters and Body weight and Gross pathology, visceral and skeletal abnormalities were examined.
Statistics:
Using the methods from Olson and Back (1978) and Wilson and Warkany (1965), the number and position of each fetus was recorded, weighed and examined for abnormalities.
The Students T test was used to test for statistical significance
between mean values and the Fisher exact test was used to test for significant differences in frequency of resorption and skeletal and soft tissue abnormalities using the litter as the experimental unit.
Indices:
Number and position of each fetus, Number of litters with resorptions and weigh were observed.
Historical control data:
not specified
Clinical signs:
not specified
Dermal irritation (if dermal study):
not specified
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
study 1.When treated with 188, 376 and 752 mg/kg bw, increase in body weight gain were observed in the females mice as compared to control and 569 mg/kg bw teated mice.

The increased maternal weight in the 188 and 376 mg/kg is apparently due to increased fetal weight and the increased maternal weight gain in the 752 mg/kg group is due to slightly larger average litter size.
study 2.When treated with 1000 mg/kg bw, decrase in body weight gain were observed in females rat as compared to control.

When treated with 500 mg/kg bw, decrase in body weight gain were observed in females rat as compared to control from day 6 to 10 of gestation. When treated with 250 mg/kg bw, No effect on body weight gain of treated female rats were observed as compared to control.
study 3.No effects on body weight was observed
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
not specified
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Number of abortions:
not specified
Pre- and post-implantation loss:
effects observed, treatment-related
Description (incidence and severity):
study 2.Statistically significant increase implants were observed at 1000 mg/kg bw.
Total litter losses by resorption:
no effects observed
Description (incidence and severity):
study 1.No significant effect on mean implants/female were observed as compared to control.
study 2.Statistically significant increase in litters containing resorptions were observed at 1000 mg/kg bw. Increase in resorptions/litter although elevated was not significant.
Early or late resorptions:
no effects observed
Description (incidence and severity):
study No effect on frequency of resorptions of treated female mice were observed as compared to control.
Dead fetuses:
effects observed, treatment-related
Description (incidence and severity):
study 2.Increase in embryolethality were observed at 1000 mg/kg bw
Changes in pregnancy duration:
not specified
Changes in number of pregnant:
not specified
Other effects:
not specified
Dose descriptor:
NOAEL
Effect level:
> 500 - <= 752 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
body weight and weight gain
early or late resorptions
maternal abnormalities
total litter losses by resorption
other: No effect observed
Remarks on result:
other: No toxic effects were observed
Abnormalities:
not specified
Fetal body weight changes:
no effects observed
Description (incidence and severity):
study 1.When treated with 188 and 376 mg/kg bw, increase in mean fetal weights were observed as compared to control and 569 and 752 mg/kg bw treated mice. The increased fetal weight in the mouse is not easily explained particularly when no such increase occurred in the groups receiving 569 and 752 mg/kg.
study 2.When treated with 500 mg/kg bw, significant decrase in fetal weight were observed as compared to control.
When treated with 1000 mg/kg bw, No effect on fetal body weight were observed as compred to control.

Migrated Data from removed field(s)
Field "Fetal/pup body weight changes" (Path: ENDPOINT_STUDY_RECORD.DevelopmentalToxicityTeratogenicity.ResultsAndDiscussion.ResultsFetuses.FetalPupBodyWeightChanges): not specified
Reduction in number of live offspring:
no effects observed
Description (incidence and severity):
study No effect on viability of pups were observed as compared to control.
Changes in sex ratio:
not specified
Changes in litter size and weights:
not specified
Changes in postnatal survival:
not specified
External malformations:
no effects observed
Description (incidence and severity):
study 1.No external abnormalities were observed in any as compared to control.
study 2.Nanoidism (unusually small fetus) at 500 and 1000 mg/kg bw.
This abnormality did not occur in the oral control or 250 mg/kg bw dose group in this study and has occurred in other control litters historically in this laboratory at a rate similar to that observed in the higher treatment groups.
Skeletal malformations:
no effects observed
Description (incidence and severity):
study 1.No Skeletal findings were observed in any as compared to control.
study 2.Hydroinephrosis, dilated ureter, and agenesis of the bladder which occurred in 3 fetuses from 1 litter at 1000 mg/kg bw.
Since this malformation was observed in the fetuses from only one litter it probably arose spontaneously rather than as a result of treatment.
Visceral malformations:
no effects observed
Description (incidence and severity):
study No Soft Tissue were observed in any as compared to control.
Other effects:
not specified
Dose descriptor:
NOAEL
Effect level:
> 752 - <= 1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
reduction in number of live offspring
fetal/pup body weight changes
external malformations
skeletal malformations
visceral malformations
other: No effect observed
Remarks on result:
other: No effects on developmental parameters were observed
Abnormalities:
not specified
Localisation:
other: not specified
Description (incidence and severity):
not specified
Developmental effects observed:
not specified
Treatment related:
not specified
Conclusions:
NOAEL was considered to be 752 mg/kg/day for P and F1 generation when ICR female mice treated with test chemical orally by gavage for 4 days (6, 7, 8 and 9 of gestation).
Executive summary:

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

study1.

In aTeratogenicity study, ICR female mice were treated with test chemical in the concentration of 0, 188, 376, 569 and 752 mg/kg/day orally by gavage in Soybean oil for 4 days (6, 7, 8 and 9 of gestation). Increase in body weight gain were observed in the females mice at 188, 376 and 752 mg/kg bw as compared to control and 569 mg/kg bw treated mice.The increased maternal weight in the188 and 376 mg/kg is apparently due to increased fetal weight and the increased maternal weight gain in the 752 mg/kg group is due to slightly larger average litter size. Similarly, No significant effect on mean implants/female, mean resorptions/litter and frequency of resorptions of treated female mice were observed as compared to control. In addition, No developmental effect such as viability of pups were observed as compared to control. Increase in mean fetal weights were observed in male and female pups at 188 and 376 mg/kg bw as compared to control and 569 and 752 mg/kg bw treated mice.The increased fetal weight in the mouse is not easily explained particularly when no such increase occurred in the groups receiving 569 and 752 mg/kg. No external abnormalities, Soft Tissue and or Skeletal findings were observed in any pup as compared to control. Therefore, NOAEL was considered to be 752 mg/kg/day for P and F1 generation when ICR female mice treated with test chemical orally by gavage for 4 days (6, 7, 8 and 9 of gestation).

study2.

In a Embryotoxicity study, Fischer 344 female rat treated with Fuel test chemical in the concentration of 0, 250, 500 and 1000 mg/kg/day orally by gavage in corn oil for 10 days (6 through 15 of gestation). Tremors and mild convulsions were observed in treated female rats at 1000 mg/kg bw. Decrease in body weight gain were observed in females rat at 1000 mg/kg bw and from day 6 to 10 of gestation at 500 mg/kg bw. No effect on body weight gain of treated female rats were observed as compared to control at 250 mg/kg bw. Similarly, statistically significant increase in litters containing resorptions or implants at 1000 mg/kg bw. Increase in resorptions/litter although elevated was not significant. Increase in embryolethality were observed at 1000 mg/kg bw. In addition, no developmental effect were observed in fetuses, some effect observed which are not treatment related such as Significant decrease in fetal weight were observed at 500 mg/kg bw and No effect on fetal body weight were observed as compared to control at 1000 mg/kg bw. Nanoidism (unusually small fetus) at 500 and 1000 mg/kg bw. This abnormality did not occur in the oral control or 250 mg/kg bw dose group in this study and has occurred in other control litters historically in this laboratory at a rate similar to that observed in the higher JP-10 treatment groups. Hydroinephrosis, dilated ureter, and agenesis of the bladder which occurred in 3 fetuses from 1 litter at 1000 mg/kg bw. Since this malformation was observed in the fetuses from only one litter it probably arose spontaneously rather than as a result of JP-10 treatment. Therefore, NOAEL was considered to be 500 mg/kg/day for P and 1000 mg/kg bw for F1 generation when Fischer 344 female rat treated with test chemical orally by gavage for 10 days (6 through 15 of gestation)

study3.

The development toxicity study oftest material was performed onVirgin adult femalealbino rats (Wistar derived stock).The test material in dose concentration0,2.18,10.15,47.05,218.0 mg/kg bw/day was administered via oralintubationon Day 6 and continuing daily through Day 15 of gestation. Sham treated group administered only vechicle and positive control Aspirin in dose concentration 250mg/kg also administered . Body weights were recorded on Days 0,6, 11, 15, and 20 of gestation. All animals were observed daily for appearance and behavior withparticularattention to food consumption and weight, in order to rule out any abnormalities which may have occurred as a result of anorexic effects in the pregnant female animal. On Day 20 all dams were subjected to Caesarean section under surgical anesthesia, and the nuniliers of implantation sites, resorption sites, and live and dead fetuses were recorded. The body weights of the live pups were also recorded.Theurogenital tract of each dam was examined in detail for anatomical normality. All fetuses were examined grossly for the presence of external congenital abnormalities.One-third of the' fetuses of each litter underwent detailed visceral examinations employing 10X magnification. The remaining two-thirds were cleared in potassium hydroxide (KOB), stained with alizarin red S dye and examined for skeletal defects.No effects onNo effects on total pregnant rats , Number of implant sites, Number of fetuses alive, Number of resorptions were observed . no clearly discernible effect on nidation or on maternal or fetal survival. The number of abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the number occurring spontaneously in the sham treated controls. HenceThe No Observed Adverse Effect Level (NOAEL) for developmental toxicity was considered to be 218.0mg/kg When female ratswere treated with test chemical orally.

study4.

The development toxicity study oftest material was performedVirgin adult female albino CD-l outbred mice.The test material in dose concentration0,1.85,8.59,39.9,185.0 mg/kg bw/day mg/kg bw/day was administered via oralintubationon Day 6 and continuing daily through Day 15 of gestation. Sham treated group administered only vechicle and positive control Aspirin in dose concentration 150mg/kg also administered . Body weights were recorded on Days 0,6, 11, 15, and 17 of gestation. All animals were observed daily for appearance and behavior withparticularattention to food consumption and weight, in order to rule out any abnormalities which may have occurred as a result of anorexic effects in the pregnant female animal. On Day 17 all dams were subjected to Caesarean section under surgical anesthesia, and the nuniliers of implantation sites, resorption sites, and live and dead fetuses were recorded. The body weights of the live pups were also recorded.The urogenital tract of each dam was examined in detail for anatomical normality. All fetuses were examined grossly for the presence of external congenital abnormalities.One-third of the' fetuses of each litter underwent detailed visceral examinations employing lOX magnification. The remaining two-thirds were cleared in potassium hydroxide (KOB), stained with alizarin red S dye and examined for skeletal defects.No effects onNo effects on total pregnant mice, Number of implant sites, Number of fetuses alive, Number of resorptions were observed . no clearly dis-cernible effect on nidation or on maternal or fetal survival. The number of abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the number occurring spontaneously in the sham treated controls. HenceThe No Observed Adverse Effect Level (NOAEL) for developmental toxicity was considered to be 185.0 mg/kg When female micewere treated with test chemical orally.

study5.

The development toxicity study oftest material was performedgolden hamsters from an outbred strain.The test material in dose concentration0,4.05,21.15,98.2,405.0mg/kg bw/day was administered via oralintubationon Day 6 and continuing daily through Day 10 of gestation. Sham treated group administered only vechicle and positive control Aspirin in dose concentration 250mg/kg also administered . Body weights were recorded on Days 0,8,10and 14 of gestation. All animals were observed daily for appearance and behavior withparticularattention to food consumption and weight, in order to rule out any abnormalities which may have occurred as a result of anorexic effects in the pregnant female animal. On Day 14 all animals were subjected to Caesarean section under surgical anesthesia, and the nuniliers of implantation sites, resorption sites, and live and dead fetuses were recorded. The body weights of the live pups were also recorded.Theurogenital tract of each dam was examined in detail for anatomical normality. All fetuses were examined grossly for the presence of external congenital defects and one-third of each litter underwent detailed visceralexaminationunder 10x magnification. The remaining two-thirds of the pups were cleared in potassium hydroxide, stained with alizarin red dye, and examined for the presence of sketal abnormalities.No effects onNo effects on total pregnant animals , Number of implant sites, Number of fetuses alive, Number of resorptions were observed . no clearly dis-cernible effect on nidation or on maternal or fetal survival. The number of abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the number occurring spontaneously in the sham treated controls. HenceThe No Observed Adverse Effect Level (NOAEL) for developmental toxicity was considered to be 405 mg/kg When female hamster were treated with test chemical orally.

study6.

The development toxicity study oftest material was performedDutch-belted female rabbits. The test material in dose concentration0,4.25,19.75,91.7,425.0 mg/kg bw/day was administered via oralintubationon Day 6 and continuing daily through Day 18of gestation. Sham treated group administered only vechicle and positive control2.5mg of 6-amino nicotinamide dosed on Day 9.On Day 0each doe was given an injection of 0.4 ml of human chorionic gonadotropin(400 IU) via the marginal ear vein. Three hours later, eachdoe was inseminated artificially with 0.3 ml of diluted semen froma proven donor buck using approximately 20 x 106motile sperm according to the procedure described by Vogin et al.Body weights were recorded on Days 0,6,12,18, and 29 of gestation.All animals were observed daily for appearance and behavior, withparticular attention to food consumption and body weight in order torule out any abnormalities which may have occurred as a result ofanorexic effects in the pregnant female animaL

On Day 29 all does were subjected to Caesarean section under surgical anesthesia, and the numbers of corpora lutea, implantationsites ,resorption sites and live and dead fetuses were recorded. Body weights of the live pups were also recorded. The urogenital tract of each animal was examined in detail for normality. In addition all fetuses underwent a detailed gross examination for the presence of external congenital abnormalities. The live fetuses of each Ltitter were ,then placed in an incubator for 24 hours for the evaluation of neonatal survival. All surviving pups were sacrificed, and all pups examined for visceral abnormalities (by dissection). All fetuses were thenclearedin potassium hydroxide (KOH), stained with alizarin red'S dye and examined for skeletal defects No effects onNo effects on total pregnant animals , Number of implant sites, Number of fetuses alive, Number of resorptions were observed . no clearly discernible effect on nidation or on maternal or fetal survival. The number of abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the number occurring spontaneously in the sham treated controls. HenceThe No Observed Adverse Effect Level (NOAEL) for developmental toxicity was considered to be 425.0mg/kg When female rabbits were treated with test chemical orally.

 

Based on the various studies available for the test chemical were reviewed to determine the developmental toxicity, NOAEL for test chemical was considered to be 500 mg /kg bw/day .When rats were treated with test chemical orally. Thus, comparing this value with the criteria of CLP regulation test chemical is not likely to classify as reproductive and developmental toxicant.

 

 

Effect on developmental toxicity: 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:
Data is Klimicsh 2 and from authoritative database

Effect on developmental toxicity: via inhalation route
Endpoint conclusion:
no study available
Effect on developmental toxicity: via dermal route
Endpoint conclusion:
no study available
Additional information

Developmental toxicity study:

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

Developmental Toxicity Study 1:

In aTeratogenicity study, ICR female mice were treated with test chemical in the concentration of 0, 188, 376, 569 and 752 mg/kg/day orally by gavage in Soybean oil for 4 days (6, 7, 8 and 9 of gestation). Increase in body weight gain were observed in the females mice at 188, 376 and 752 mg/kg bw as compared to control and 569 mg/kg bw treated mice.The increased maternal weight in the188 and 376 mg/kg is apparently due to increased fetal weight and the increased maternal weight gain in the 752 mg/kg group is due to slightly larger average litter size. Similarly, No significant effect on mean implants/female, mean resorptions/litter and frequency of resorptions of treated female mice were observed as compared to control. In addition, No developmental effect such as viability of pups were observed as compared to control. Increase in mean fetal weights were observed in male and female pups at 188 and 376 mg/kg bw as compared to control and 569 and 752 mg/kg bw treated mice.The increased fetal weight in the mouse is not easily explained particularly when no such increase occurred in the groups receiving 569 and 752 mg/kg. No external abnormalities, Soft Tissue and or Skeletal findings were observed in any pup as compared to control. Therefore, NOAEL was considered to be 752 mg/kg/day for P and F1 generation when ICR female mice treated with test chemical orally by gavage for 4 days (6, 7, 8 and 9 of gestation).

Developmental Toxicity Study 2:

In a Embryotoxicity study, Fischer 344 female rat treated with Fuel test chemical in the concentration of 0, 250, 500 and 1000 mg/kg/day orally by gavage in corn oil for 10 days (6 through 15 of gestation). Tremors and mild convulsions were observed in treated female rats at 1000 mg/kg bw. Decrease in body weight gain were observed in females rat at 1000 mg/kg bw and from day 6 to 10 of gestation at 500 mg/kg bw. No effect on body weight gain of treated female rats were observed as compared to control at 250 mg/kg bw. Similarly, statistically significant increase in litters containing resorptions or implants at 1000 mg/kg bw. Increase in resorptions/litter although elevated was not significant. Increase in embryolethality were observed at 1000 mg/kg bw. In addition, no developmental effect were observed in fetuses, some effect observed which are not treatment related such as Significant decrease in fetal weight were observed at 500 mg/kg bw and No effect on fetal body weight were observed as compared to control at 1000 mg/kg bw. Nanoidism (unusually small fetus) at 500 and 1000 mg/kg bw. This abnormality did not occur in the oral control or 250 mg/kg bw dose group in this study and has occurred in other control litters historically in this laboratory at a rate similar to that observed in the higher JP-10 treatment groups. Hydroinephrosis, dilated ureter, and agenesis of the bladder which occurred in 3 fetuses from 1 litter at 1000 mg/kg bw. Since this malformation was observed in the fetuses from only one litter it probably arose spontaneously rather than as a result of JP-10 treatment. Therefore, NOAEL was considered to be 500 mg/kg/day for P and 1000 mg/kg bw for F1 generation when Fischer 344 female rat treated with test chemical orally by gavage for 10 days (6 through 15 of gestation).

Developmental Toxicity Study 3:

The development toxicity study oftest material was performed onVirgin adult femalealbino rats (Wistar derived stock).The test material in dose concentration0,2.18,10.15,47.05,218.0 mg/kg bw/day was administered via oralintubationon Day 6 and continuing daily through Day 15 of gestation. Sham treated group administered only vechicle and positive control Aspirin in dose concentration 250mg/kg also administered . Body weights were recorded on Days 0,6, 11, 15, and 20 of gestation. All animals were observed daily for appearance and behavior withparticularattention to food consumption and weight, in order to rule out any abnormalities which may have occurred as a result of anorexic effects in the pregnant female animal. On Day 20 all dams were subjected to Caesarean section under surgical anesthesia, and the nuniliers of implantation sites, resorption sites, and live and dead fetuses were recorded. The body weights of the live pups were also recorded.Theurogenital tract of each dam was examined in detail for anatomical normality. All fetuses were examined grossly for the presence of external congenital abnormalities.One-third of the' fetuses of each litter underwent detailed visceral examinations employing 10X magnification. The remaining two-thirds were cleared in potassium hydroxide (KOB), stained with alizarin red S dye and examined for skeletal defects.No effects onNo effects on total pregnant rats , Number of implant sites, Number of fetuses alive, Number of resorptions were observed . no clearly discernible effect on nidation or on maternal or fetal survival. The number of abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the number occurring spontaneously in the sham treated controls. HenceThe No Observed Adverse Effect Level (NOAEL) for developmental toxicity was considered to be 218.0mg/kg When female ratswere treated with test chemical orally.

Developmental Toxicity Study 4:

The development toxicity study oftest material was performedVirgin adult female albino CD-l outbred mice.The test material in dose concentration0,1.85,8.59,39.9,185.0 mg/kg bw/day mg/kg bw/day was administered via oralintubationon Day 6 and continuing daily through Day 15 of gestation. Sham treated group administered only vechicle and positive control Aspirin in dose concentration 150mg/kg also administered . Body weights were recorded on Days 0,6, 11, 15, and 17 of gestation. All animals were observed daily for appearance and behavior withparticularattention to food consumption and weight, in order to rule out any abnormalities which may have occurred as a result of anorexic effects in the pregnant female animal. On Day 17 all dams were subjected to Caesarean section under surgical anesthesia, and the nuniliers of implantation sites, resorption sites, and live and dead fetuses were recorded. The body weights of the live pups were also recorded.The urogenital tract of each dam was examined in detail for anatomical normality. All fetuses were examined grossly for the presence of external congenital abnormalities.One-third of the' fetuses of each litter underwent detailed visceral examinations employing lOX magnification. The remaining two-thirds were cleared in potassium hydroxide (KOB), stained with alizarin red S dye and examined for skeletal defects.No effects onNo effects on total pregnant mice, Number of implant sites, Number of fetuses alive, Number of resorptions were observed . no clearly dis-cernible effect on nidation or on maternal or fetal survival. The number of abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the number occurring spontaneously in the sham treated controls. HenceThe No Observed Adverse Effect Level (NOAEL) for developmental toxicity was considered to be 185.0 mg/kg When female micewere treated with test chemical orally.

Developmental Toxicity Study 5:

The development toxicity study oftest material was performedgolden hamsters from an outbred strain.The test material in dose concentration0,4.05,21.15,98.2,405.0mg/kg bw/day was administered via oralintubationon Day 6 and continuing daily through Day 10 of gestation. Sham treated group administered only vechicle and positive control Aspirin in dose concentration 250mg/kg also administered . Body weights were recorded on Days 0,8,10and 14 of gestation. All animals were observed daily for appearance and behavior withparticularattention to food consumption and weight, in order to rule out any abnormalities which may have occurred as a result of anorexic effects in the pregnant female animal. On Day 14 all animals were subjected to Caesarean section under surgical anesthesia, and the nuniliers of implantation sites, resorption sites, and live and dead fetuses were recorded. The body weights of the live pups were also recorded.Theurogenital tract of each dam was examined in detail for anatomical normality. All fetuses were examined grossly for the presence of external congenital defects and one-third of each litter underwent detailed visceralexaminationunder 10x magnification. The remaining two-thirds of the pups were cleared in potassium hydroxide, stained with alizarin red dye, and examined for the presence of sketal abnormalities.No effects onNo effects on total pregnant animals , Number of implant sites, Number of fetuses alive, Number of resorptions were observed . no clearly dis-cernible effect on nidation or on maternal or fetal survival. The number of abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the number occurring spontaneously in the sham treated controls. HenceThe No Observed Adverse Effect Level (NOAEL) for developmental toxicity was considered to be 405 mg/kg When female hamster were treated with test chemical orally.

Developmental Toxicity Study 6:

The development toxicity study oftest material was performedDutch-belted female rabbits. The test material in dose concentration0,4.25,19.75,91.7,425.0 mg/kg bw/day was administered via oralintubationon Day 6 and continuing daily through Day 18of gestation. Sham treated group administered only vechicle and positive control2.5mg of 6-amino nicotinamide dosed on Day 9.On Day 0each doe was given an injection of 0.4 ml of human chorionic gonadotropin(400 IU) via the marginal ear vein. Three hours later, each doe was inseminated artificially with 0.3 ml of diluted semen froma proven donor buck using approximately 20 x 106motile sperm according to the procedure described by Vogin et al.Body weights were recorded on Days 0,6,12,18, and 29 of gestation.All animals were observed daily for appearance and behavior, withparticular attention to food consumption and body weight in order torule out any abnormalities which may have occurred as a result ofanorexic effects in the pregnant female animal. On Day 29 all does were subjected to Caesarean section under surgical anesthesia, and the numbers of corpora lutea, implantationsites ,resorption sites and live and dead fetuses were recorded. Body weights of the live pups were also recorded. The urogenital tract of each animal was examined in detail for normality. In addition all fetuses underwent a detailed gross examination for the presence of external congenital abnormalities. The live fetuses of each Ltitter were ,then placed in an incubator for 24 hours for the evaluation of neonatal survival. All surviving pups were sacrificed, and all pups examined for visceral abnormalities (by dissection). All fetuses were thenclearedin potassium hydroxide (KOH), stained with alizarin red'S dye and examined for skeletal defects No effects on No effects on total pregnant animals , Number of implant sites, Number of fetuses alive, Number of resorptions were observed . no clearly discernible effect on nidation or on maternal or fetal survival. The number of abnormalities seen in either soft or skeletal tissues of the test groups did not differ from the number occurring spontaneously in the sham treated controls. HenceThe No Observed Adverse Effect Level (NOAEL) for developmental toxicity was considered to be 425.0mg/kg When female rabbits were treated with test chemical orally.  

Justification for classification or non-classification

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

Additional information