Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Remarks:
28-days repeated dose toxicity study with extended 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 a 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:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Version / remarks:
Adopted in 3 October, 2008
GLP compliance:
yes (incl. QA statement)
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)cyclohexanol
Test material form:
liquid
Details on test material:
- Name of test material (as cited in study report): Iso bornyl cyclo hexanol (or 3-(5,5,6-trimethylbicyclo[2.2.1]hept-2-yl)cyclohexan-1-ol)
- Molecular Formula: C16H28O
- Molecular Weight: 236.396 g/mol
- Substance type:Organic
- Physical state:Liquid
-SMILES:C1(C)(C)C(C)C2C(C3CC(O)CCC3)CC1C2

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, Punjab, India.
- Age at study initiation: 7 to 8 weeks old
- Weight at study initiation: Males: 200.20-241.22 g; Female: 194.30-220.28 g
- Fasting period before study: Yes, 2 hrs fasted before dose administration.
- Housing: Animals were housed in sterilized solid bottom polypropylene cages with stainless steel grill tops with bedding of clean paddy husk in a controlled environment.
- 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, 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): 12-hrs dark/12-hrs light

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral 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
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:
Once per day.
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:
7 rats per sex per dose.
Control animals:
yes, concurrent vehicle
Details on study design:
No data available
Positive control:
No positive control was included.

Examinations

Observations and examinations performed and frequency:
Mortality, clinical signs (once per day), detailed clinical signs (once per week), body weight (treatment days 1, 8, 15, 22, 28, 29) food intake (once per week), water intake (once per week), ophthalmology (4th week of treatment), locomotor activity (4th week of treatment), hematology (end of treatment), clinical chemistry (end of treatment)

Hematologic parameters: Haemoglobin, RBC, Total and differential leucocyte count, Haematocrit, Mean corpuscular volume (MCV), Mean corpuscular haemoglobin (MCH), Mean corpuscular haemoglobin concentration (MCHC), and Platelet count

Clinical chemistry parameters: Sodium, Potassium, Glucose, Total cholesterol, Blood urea, Creatinine, Total protein, Albumin, SGPT (Serum glutamic pyruvic transaminase), ALT, SGOT (Serum glutamic oxaloacetic transaminase), AST, Hormones analysis (testosterone and estrogen) and Total bile acids.
Sacrifice and pathology:
The following organs were weighed: liver, adrenals, spleen, heart, kidney, brain, testes, epididymides, ovaries, thymus. The following organs/tissues were examined microscopically: brain, stomach, large intestine, small intestine, liver, kidney, adrenal gland, spleen, heart, thymus, lungs, testis, ovaries, uterus, lymph nodes, perihperal nerve, bone marrow, and gross lesions (if any).
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.

Results and discussion

Results of examinations

Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
There were no clinical signs attributed to the test item. 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.
Mortality:
no mortality observed
Description (incidence):
All animals survived to planned death.
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
No significant changes in body weight were observed, except for a slight increase in male body weight at 1125 mg/kg on day 8 of treatment (mean, 248 g) compared to the control group (mean 237 g). This effect was considered incidental and not toxicologically significant.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No significant effects were observed in the study.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
No significant effects were observed in the study.
Ophthalmological findings:
no effects observed
Description (incidence and severity):
No significant effects were observed in the study.
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Significant changes in haematology were as follows: significant increase in % monocytes in males treated at 125 mg/kg; significant increase in % basophils in males treated at 125 mg/kg; significant increases in white blood cell counts in females treated at 375 and 1125 mg/kg; significant decreases in red blood cell counts in females treated at 375 and 1125 mg/kg; significant decrease in haematocrit counts in females treated at 1125 mg/kg; significant increase in MCV in females treated at 1125 mg/kg; and significant increases in MCH in females treated at 375 and 1125 mg/kg. The observed changes in haematology were considered not toxicologically relevant due to lack of dose relationship and due to lack of associated histopathological findings.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Significant changes in clinical chemistry were as follows: significant increase in total cholesterol in male rats treated at 1125 mg/kg; significant increase in creatinine in male rats treated at 125 mg/kg; significant increase in glucose in male rats treated at 375 mg/kg; significant increases in total proteins in male rats treated at 375 and 1125 mg/kg; significant increase in potassium in female rats treated at 1125 mg/kg; significant increase in sodium in female rats treated at 1125 mg/kg; significant increase in SGPT in female rats treated at 375 and 1125 mg/kg; significant decrease in glucose in female rats treated at 1125 mg/kg; significant increase in total protein in female rats treated at 1125 mg/kg; significant increases in urea nitrogen in female rats treated at 375 and 1125 mg/kg; and a significant increase in bile acid in female rats treated at 375 mg/kg. No significant changes in serum testosterone or serum oestrogen were observed.
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
No significant changes in locomotor activity were observed.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
No significant changes in absolute organ weight were observed, with the exception of significant increases in liver weight in female rats treated at 375 and 1125 mg/kg. Significant changes in relative organ weight included a significant decrease in relative heart weight in male rats treated at 125 mg/kg and two significant increases in relative liver weight in female rats treated at 375 and 1125 mg/kg.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
In male rats, no gross pathological findings were observed. In female rats, gross pathological findings were limited to a slightly increased stomach at 1125 mg/kg and an enlarged uterus with uterine fluid at 1125 mg/kg.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Histopathology performed at 0 and 1125 mg/kg showed excess of lymphocytes in the lungs of 2/7 control males, 1/7 high-dosed males, 1/7 control females, and 1/7 high-dosed females and reactive spleens in 3/7 control males, 1/7 high-dosed males, 2/7 control females, and 1/7 high-dosed females. These histopathological findings were consistent with the historical control data of the test facility and were therefore not considered to be toxicologically relevant. The histopathologic examinations did not reveal any significant effects on spermatogenesis.
Histopathological findings: neoplastic:
no effects observed

Effect levels

Key result
Dose descriptor:
NOAEL
Effect level:
1 125 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
behaviour (functional findings)
body weight and weight gain
clinical biochemistry
clinical signs
food consumption and compound intake
gross pathology
haematology
histopathology: neoplastic
histopathology: non-neoplastic
mortality
ophthalmological examination
organ weights and organ / body weight ratios
water consumption and compound intake

Target system / organ toxicity

Key result
Critical effects observed:
no

Any other information on results incl. tables

SUMMARY OF DAY-WISE MORTALTY DATA

MALE

Groups

1

2

3

4

Days

 

 

 

 

 

1

0/7

0/7

0/7

0/7

2

0/7

0/7

0/7

0/7

3

0/7

0/7

0/7

0/7

4

0/7

0/7

0/7

0/7

5

0/7

0/7

0/7

0/7

6

0/7

0/7

0/7

0/7

7

0/7

0/7

0/7

0/7

8

0/7

0/7

0/7

0/7

9

0/7

0/7

0/7

0/7

10

0/7

0/7

0/7

0/7

11

0/7

0/7

0/7

0/7

12

0/7

0/7

0/7

0/7

13

0/7

0/7

0/7

0/7

14

0/7

0/7

0/7

0/7

15

0/7

0/7

0/7

0/7

16

0/7

0/7

0/7

0/7

17

0/7

0/7

0/7

0/7

18

0/7

0/7

0/7

0/7

19

0/7

0/7

0/7

0/7

20

0/7

0/7

0/7

0/7

21

0/7

0/7

0/7

0/7

22

0/7

0/7

0/7

0/7

23

0/7

0/7

0/7

0/7

24

0/7

0/7

0/7

0/7

25

0/7

0/7

0/7

0/7

26

0/7

0/7

0/7

0/7

27

0/7

0/7

0/7

0/7

28

0/7

0/7

0/7

0/7

FEMALE

Groups

5

6

7

8

Days

 

 

 

 

 

1

0/7

0/7

0/7

0/7

2

0/7

0/7

0/7

0/7

3

0/7

0/7

0/7

0/7

4

0/7

0/7

0/7

0/7

5

0/7

0/7

0/7

0/7

6

0/7

0/7

0/7

0/7

7

0/7

0/7

0/7

0/7

8

0/7

0/7

0/7

0/7

9

0/7

0/7

0/7

0/7

10

0/7

0/7

0/7

0/7

11

0/7

0/7

0/7

0/7

12

0/7

0/7

0/7

0/7

13

0/7

0/7

0/7

0/7

14

0/7

0/7

0/7

0/7

15

0/7

0/7

0/7

0/7

16

0/7

0/7

0/7

0/7

17

0/7

0/7

0/7

0/7

18

0/7

0/7

0/7

0/7

19

0/7

0/7

0/7

0/7

20

0/7

0/7

0/7

0/7

21

0/7

0/7

0/7

0/7

22

0/7

0/7

0/7

0/7

23

0/7

0/7

0/7

0/7

24

0/7

0/7

0/7

0/7

25

0/7

0/7

0/7

0/7

26

0/7

0/7

0/7

0/7

27

0/7

0/7

0/7

0/7

28

0/7

0/7

0/7

0/7

  

SUMMARY OF CLINICAL FINDINGS OF ANIMALS

MALE

GROUPS

DOSE (mg/kg)

Animals ID

Total occurance/total no. of animals

Remarks

1

0

2

1/7

Abnormal

2

125

11

1/7

Abnormal

3

375

17,18,21

3/7

Abnormal

4

1125

22,24,25

3/7

Abnormal

 

Female

GROUPS

DOSE (mg/kg)

Animals ID

Total occurance/total no. of animals

Remarks

5

0

30,31

2/7

Abnormal

6

125

-

7/7

Abnormal

7

375

47

1/7

Abnormal

8

1125

51,52,55,56

4/7

Abnormal

 

SUMMARY OF body-weights (g)

Male

Groups

Dose

(mg/kg)

 

Day 1

Day 8

Day 15

Day 22

Day 28

Terminal day·

 

1

 

0

Mean

218.32

237.41

262.26

277.80

289.58

287.90

SD

13.88

19.91

23.69

24.50

28.75

28.69

SEM

5.25

7.53

8.95

9.09

10.86

10.84

 

2

 

125

Mean

222.48

243.24

266.67

285.77

293.54

291.38

SD

7.73

11.32

18.07

22.09

23.33

23.23

SEM

2.92

4.28

6.83

8.35

8.82

8.78

 

3

 

375

Mean

230.02

255.17

280.56

296.58

303.07

300.22

SD

7.50

12.42

17.61

16.75

19.13

18.54

SEM

2.83

4.69

6.66

6.33

7.23

7.01

 

4

 

1125

Mean

230.90

248.17*

263.75

277.82

285.36

283.46

SD

7.83

20.67

25.55

25.44

24.82

25.15

SEM

2.96

7.81

9.66

9.62

9.38.

9.50

·:Significant at p≤0.05 in comparison to control group

*:Terminal body weight reflect the body weights of animals after overnight fasting on the day of necropsy

Female

Groups

Dose

(mg/kg)

 

Day 1

Day 8

Day 15

Day 22

Day 28

Terminal day·

 

5

 

0

Mean

204.43

217.45

230.94

244.10

250.12

247.75

SD

8.79

11.32

12.19

11.53

11.44

11.60

SEM

3.32

4.28

4.61

4.36

4.32

4.39

 

6

 

125

Mean

202.53

210.25

230.15

244.96

250.58

250.58

SD

6.53

5.48

7.68

7.22

8.05

7.72

SEM

2.47

2.07

2.90

 

2.73

3.04

2.92

 

7

 

375

Mean

207.56

216.32

229.87

241.52

245.76

244.45

SD

7.42

10.03

8.61

8.94

10.81

10.71

SEM

2.81

3.79

3.25

3.38

4.08

4.05

 

8

 

1125

Mean

204.30

206.34

219.92

232.85

234.80

233.05

SD

6.75

6.45

10.36

6.51

7.95

7.58

SEM

2.55

2.44

3.92

2.46

3.00

2.86

Terminal body weight reflect the body weights of animals after overnight fasting on the day of necropsy

SUMMARY OF FOOD CONSUMPTION (g)

Male

Groups

 

Dose (mg/kg)

 

Average Food Intake / Animal / Day

Week 1

Week 2

Week 3

Week 4

1

0

13.29

10.29

12.43

12.71

2

125

11.86

9.14

13.00

10.57

3

375

9.00

11.57

12.43

9.43

4

1125

9.14

11.57

14.14

8.57

 

Female

Groups

 

Dose (mg/kg)

 

Average Food Intake / Animal / Day

Week 1

Week 2

Week 3

Week 4

5

0

7.86

9.86

10.57

8.57

6

125

6.71

6.86

9.86

4.14

7

375

8.14

8.14

10.00

5.71

8

1125

6.86

10.29

10.00

5.14

 

SUMMARY OF WATER CONSUMPTION (ml)

Male

Groups

 

Dose (mg/kg)

 

Average water consumption / Animal / Day

Week 1

Week 2

Week 3

Week 4

1

0

27.14

21.43

30.00

28.57

2

125

27.14

25.71

24.29

37.14

3

375

30.00

31.43

31.43

32.86

4

1125

32.86

28.57

27.14

35.71

Female

Groups

 

Dose (mg/kg)

 

Average water consumption / Animal / Day

Week 1

Week 2

Week 3

Week 4

5

0

26.43

21.43

25.71

28.57

6

125

21.43

17.14

24.29

27.14

7

375

25.71

20.00

22.86

24.29

8

1125

28.57

27.14

25.71

38.57

 SUMMARY OF LOCOMOTOR ACTIVITY SCORES

Male

Groups

Dose (mg/kg)

 

Activity during 4thweek of treatment

 

1

 

0

Mean

347.71

SD

26.18

SEM

9.90

 

2

 

125

Mean

331.71

SD

20.45

SEM

7.73

 

3

 

375

Mean

365.57

SD

49.75

SEM

18.80

 

4

 

1125

Mean

376.29

SD

18.32

SEM

6.92

 

 Female

Groups

 

Dose (mg/kg)

 

 

Activity during 4thweek of treatment

 

5

 

0

Mean

387.00

SD

53.51

SEM

20.23

 

6

 

125

Mean

396.71

SD

39.46

SEM

14.92

 

7

 

375

Mean

14.92

SD

47.11

SEM

17.81

 

8

 

1125

Mean

376.57

SD

54.16

SEM

20.47

SUMMARY OF HEMATOLOGY DATA

MALE

            Groups / Dose

 

 

WBC (x103/ mm3)

 

RBC (x106/ mm3)

 

PLT (x103/ mm3)

 

Hct (%)

 

Hgb (g/dl)

 

MCV (fl)

 

MCH (pg)

 

MCHC (g/dl)

 

Lym (%)

 

Mon (%)

 

Nut (%)

 

Eos (%)

 

Bas (%)

 

 

1/0

(mg/kg)

 

Mean

9.34

7.44

549.14

36.70

14.66

49.40

19.74

39.99

63.84

4.53

28.84

1.19

0.87

SD

2.44

0.46

202.40

2.10

0.71

1.58

0.95

1.90

9.15

0.30

9.73

1.84

0.21

SEM

0.92

0.17

76.50

0.79

0.27

0.60

0.36

0.72

3.46

0.11

3.68

0.70

0.08

 

2/125

(mg/kg)

 

Mean

10.78

7.29

508.71

36.61

14.53

50.24

19.93

39.66

62.54

5.06*

29.49

1.10

1.10*

SD

0.89

0.39

77.48

2.16

0.87

0.61

0.42

0.49

9.06

0.68

9.34

2.25

0.15

SEM

0.33

0.15

29.28

0.82

0.33

0.23

0.16

0.18

3.43

0.26

3.53

0.85

0.06

 

3/375

(mg/kg)

 

Mean

10.36

7.57

653.29

37.53

14.79

49.67

19.59

39.44

73.17

4.24

20.61

0.21

1.11

SD

1.29

0.54

227.12

2.06

0.80

1.76

0.67

0.77

3.03

0.41

3.00

0.15

0.26

SEM

0.49

0.20

85.85

0.78

0.30

0.67

0.25

0.29

1.14

0.16

1.13

0.06

0.10

 

4/1125 (mg/kg)

 

Mean

9.10

7.35

715.71

36.30

14.04

49.40

19.17

38.77

54.36

4.66

39.06

0.37

0.84

SD

3.65

0.36

166.93

1.41

0.60

1.02

0.73

0.77

20.45

0.82

20.16

0.38

0.36

SEM

1.38

0.14

63.09

0.53

0.23

0.39

0.28

0.29

7.73

0.31

7.62

0.14

0.14

* Significant at p ≤ 0.05 in comparison to control group

FEMALE

            Groups / Dose

 

 

WBC (x103/ mm3)

 

RBC (x106/ mm3)

 

PLT (x103/ mm3)

 

Hct (%)

 

Hgb (g/dl)

 

MCV (fl)

 

MCH (pg)

 

MCHC (g/dl)

 

Lym (%)

 

Mon (%)

 

Nut (%)

 

Eos (%)

 

Bas (%)

 

 

5/0

(mg/kg)

Mean

6.32

7.94

523.29

39.21

14.26

49.43

17.97

36.31

67.73

3.56

26.90

0.21

1.06

SD

1.96

0.24

135.99

0.96

0.43

1.23

0.77

1.37

3.38

0.77

3.27

0.20

0.25

SEM

0.74

0.09

51.40

0.36

0.16

0.46

0.29

0.52

1.28

0.29

1.24

0.07

0.09

 

6 / 125 (mg/kg)

Mean

9.13

7.73

424.29

38.90

14.19

50.40

18.33

36.39

69.43

3.40

25.30

0.23

1.06

SD

3.84

0.28

51.81

1.14

0.45

0.65

0.48

0.59

7.04

0.42

6.47

0.19

0.14

SEM

1.45

0.11

19.58

0.43

0.17

0.24

0.18

0.22

2.66

0.16

2.45

0.07

0.05

 

7/ 375 (mg/kg)

Mean

10.99**

7.60*

427.57

38.27

14.43

50.36

18.94*

37.66

70.74

4.03

21.96

1.40

1.24

SD

1.58

0.25

111.75

1.77

0.67

2.27

0.88

0.86

8.05

0.99

8.03

1.96

0.21

SEM

0.60

0.09

42.24

0.67

0.25

0.86

0.33

0.33

3.04

0.37

3.03

0.74

0.08

 

8 / 1125 (mg/kg)

Mean

10.57*

7.19***

394.29

36.93*

13.97

51.40**

19.43**

37.83

68.56

4.14

25.20

0.29

1.24

SD

2.17

0.25

99.97

1.38

0.24

1.01

0.61

1.19

4.11

0.43

3.74

0.19

0.23

SEM

0.82

0.09

37.78

0.52

0.09

0.38

0.23

0.45

1.55

0.16

1.41

0.07

0.09

* Significant at p ≤ 0.05 in comparison to control group

**Significant at p ≤ 0.01 in comparison to control group

*** Significant at p ≤ 0.001 in comparison to control group

Applicant's summary and conclusion

Conclusions:
NOAEL was considered to be 1125 mg/kg bw/day in both male and female SD rats.
Executive summary:

The test chemical was given to 7 rats per sex per dose level at 0 (vehicle; corn oil), 125, 375 and 1125 mg/kg/day. The study was performed according to GLP and according to OECD 407 with some additional reproductive-related endpoints included.Results:All animals survived to planned death and there were no clinical signs attributed to the test chemical. No significant changes in body weight were observed, except for a slight increase in male body weight at 1125 mg/kg on day 8 of treatment (mean, 248 g) compared to the control group (mean 237 g). This effect was considered incidental and not toxicologically significant. No significant changes in food intake or water intake were observed. No significant changes in locomotor activity were observed. No abnormalities were found during the ophthalmic examinations. Significant changes in haematology were as follows: significant increase in % monocytes in males treated at 125 mg/kg; significant increase in % basophils in males treated at 125 mg/kg; significant increases in white blood cell counts in females treated at 375 and 1125 mg/kg; significant decreases in red blood cell counts in females treated at 375 and 1125 mg/kg; significant decrease in haematocrit counts in females treated at 1125 mg/kg; significant increase in MCV in females treated at 1125 mg/kg; and significant increases in MCH in females treated at 375 and 1125 mg/kg. The observed changes in haematology were considered not toxicologically relevant due to lack of dose relationship and due to lack of associated histopathological findings. Significant changes in clinical chemistry were as follows: significant increase in total cholesterol in male rats treated at 1125 mg/kg; significant increase in creatinine in male rats treated at 125 mg/kg; significant increase in glucose in male rats treated at 375 mg/kg; significant increases in total proteins in male rats treated at 375 and 1125 mg/kg; significant increase in potassium in female rats treated at 1125 mg/kg; significant increase in sodium in female rats treated at 1125 mg/kg; significant increase in SGPT in female rats treated at 375 and 1125 mg/kg; significant decrease in glucose in female rats treated at 1125 mg/kg; significant increase in total protein in female rats treated at 1125 mg/kg; significant increases in urea nitrogen in female rats treated at 375 and 1125 mg/kg; and a significant increase in bile acid in female rats treated at 375 mg/kg. No significant changes in serum testosterone or serum oestrogen were observed. The observed changes in clinical chemistry were considered not toxicologically relevant due to lack of dose relationship and due to lack of associated histopathological findings. No significant changes in absolute organ weight were observed, with the exception of significant increases in liver weight in female rats treated at 375 and 1125 mg/kg. Significant changes in relative organ weight included a significant decrease in relative heart weight in male rats treated at 125 mg/kg and two significant increases in relative liver weight in female rats treated at 375 and 1125 mg/kg. In male rats, no gross pathological findings were observed. In female rats, gross pathological findings were limited to a slightly increased stomach at 1125 mg/kg and an enlarged uterus with uterine fluid at 1125 mg/kg. Histopathology performed at 0 and 1125 mg/kg showed excess of lymphocytes in the lungs of 2/7 control males, 1/7 high-dosed males, 1/7 control females, and 1/7 high-dosed females and reactive spleens in 3/7 control males, 1/7 high-dosed males, 2/7 control females, and 1/7 high-dosed females. These histopathological findings were consistent with the historical control data of the test facility and were therefore not considered to be toxicologically relevant. The histopathologic examinations did not reveal any significant effects on spermatogenesis.Conclusion:NOAEL was considered at 1125 mg/kg bw/day.