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Toxicological information

Toxicity to reproduction

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Administrative data

Endpoint:
reproductive toxicity, other
Remarks:
REPEATED DOSE 28- DAYS ORAL TOXICITY STUDY EXTENDED WITH REPRODUCTIVE PARAMETERS
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from study report

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report date:
2015

Materials and methods

Test guideline
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

Test material

Constituent 1
Chemical structure
Reference substance name:
3-(5,5,6-trimethylbicyclo[2.2.1]hept-2-yl)cyclohexan-1-ol
EC Number:
222-294-1
EC Name:
3-(5,5,6-trimethylbicyclo[2.2.1]hept-2-yl)cyclohexan-1-ol
Cas Number:
3407-42-9
Molecular formula:
C16H28O
IUPAC Name:
3-(5,5,6-trimethylbicyclo[2.2.1]hept-2-yl)cyclohexano
Test material form:
liquid
Details on test material:
- 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

Test animals

Species:
rat
Strain:
Sprague-Dawley
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

Administration / exposure

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
Doses / concentrationsopen allclose all
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

Examinations

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

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

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 / performance (P0)

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.

Effect levels (P0)

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

Target system / organ toxicity (P0)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Results: F1 generation

General toxicity (F1)

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

Developmental neurotoxicity (F1)

Behaviour (functional findings):
not specified

Developmental immunotoxicity (F1)

Developmental immunotoxicity:
not specified

Effect levels (F1)

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

Target system / organ toxicity (F1)

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified
Treatment related:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Overall reproductive toxicity

Reproductive effects observed:
not specified
Treatment related:
not specified
Relation to other toxic effects:
not specified
Dose response relationship:
not specified
Relevant for humans:
not specified

Any other information on results incl. tables

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

Applicant's summary and conclusion

Conclusions:
NOAEL was considered to be 1125 mg/kg body weight/day for test chemical . when 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.

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