Registration Dossier

Administrative data

Description of key information

Sub-chronic toxicity: NOAEL - 800 mg/kg bw/day

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 2017-05-18 to 2017-09-21
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Version / remarks:
Adopted 1998-09-21
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.26 (Sub-Chronic Oral Toxicity Test: Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Commercial sample. Lot No. 3606217009
- Expiration date of the lot/batch: 2018-01-09
- Purity: 97.76%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient conditions
- Stability under test conditions: Stable
- Solubility and stability of the test substance in the solvent/vehicle: Soluble/miscible and stable
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: None

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: None
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS srl, San Pietro al Natisone, Italy
- Females (if applicable) nulliparous and non-pregnant: Yes
- Age at study initiation: 42 - 44 days
- Weight at study initiation: Males: 181-226 g; Females: 144-177 g
- Fasting period before study: None
- Housing: Group caged
- Diet (e.g. ad libitum): Commercial laboratory rodent dient, ad-libitum
- Water (e.g. ad libitum): Municipal drinking water, ad-libitum
- Acclimation period: 15 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24 deg C
- Humidity (%): 40 - 70 %
- Air changes (per hr): 15-20
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 2017-05-25 To: 2017-09-21
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
- PREPARATION OF DOSING SOLUTIONS:

- VEHICLE
- Justification for use and choice of vehicle (if other than water): Commonly used vehicle for non-water miscible materials
- Concentration in vehicle: 0, 10, 40 and 160 mg/mL
- Amount of vehicle (if gavage): 5 mL/kg bw
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The formulation procedure was checked in the range from 1 to 250 mg/mL by chemical analysis for concentration and homogeneity to confirm that the method was suitable.
Samples of the formulations prepared on Week 1 and last week (Week 13) of the dosing phase were analysed to check the homogeneity and concentration.
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Daily
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Vehicle control
Dose / conc.:
50 mg/kg bw/day (nominal)
Dose / conc.:
200 mg/kg bw/day (nominal)
Dose / conc.:
800 mg/kg bw/day (nominal)
No. of animals per sex per dose:
10 males/10 females per dose
Additional 5 males/5 females in control and high dose groups to examine recovery from treatment
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: Based on data from shorter-term studies
- Rationale for animal assignment: Random
- Rationale for selecting satellite groups: Groups pre-assigned, animal assignment with groups random as for main study groups
- Post-exposure recovery period in satellite groups: 4 weeks
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least daily, at same time interval
- Cage side observations included. Mortality/morbidity, gross clinical signs/response to treatment

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly
- Observations included: Detailed clinical examination in an open arena for changes in gait and posture, reactivity to handling, presence of clonic or tonic movements, stereotypies or bizarre behaviour
and effects on the autonomic nervous system (e.g. lachrymation, piloerection, unusual respiratory pattern). Changes in fur, skin, eyes, mucous membranes, occurrences of secretions and excretions.

BODY WEIGHT: Yes
- Time schedule for examinations: On the day of allocation to treatment, on the day before treatment commenced, weekly thereafter and just prior to necropsy.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes - Weight of food consumed by each cage of rats recorded at weekly
intervals following allocation. Group mean daily intake per rat calculated.

WATER CONSUMPTION AND COMPOUND INTAKE: No

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Pre-dose and during Week 13 of treatment.
- Dose groups that were examined: All animals pre-dose. Control and high-dose animals during Week 13 of treatment.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: During Week 13 of treatment and Week 4 of recovery
- Anaesthetic used for blood collection: Yes (isofluorane)
- Animals fasted: Yes
- How many animals: All animals
- Parameters examined: Haematocrit, Haemoglobin, Red blood cell count, Reticulocyte count, Mean red blood cell volume, Mean corpuscular haemoglobin, Mean corpuscular haemoglobin concentration,
White blood cell count , Differential leucocyte count, Platelets, Prothrombin time.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: During Week 13 of treatment and Week 4 of recovery
- Anaesthetic used for blood collection: Yes (isofluorane)
- Animals fasted: Yes
- How many animals: All animals
- Parameters examined: Alkaline phosphatase, Alanine aminotransferase, Aspartate aminotransferase, Gamma –glutamyltransferase, Urea, Creatinine, Glucose, Triglycerides, Phosphorus,
Total bilirubin, Total cholesterol, Total protein, Albumin, Globulin, A/G Ratio, Sodium, Potassium, Calcium, Chloride

URINALYSIS: Yes
- Time schedule for collection of blood: During Week 13 of treatment and Week 4 of recovery
- Animals fasted: Yes
- How many animals: All animals
- Parameters examined: Appearance, Volume, Specific gravity, pH, Protein, Glucose, Ketones, Bilirubin, Urobilinogen, Blood, sediment obtained from centrifugation examined microscopically for
epithelial cells, leucocytes, erythrocytes, crystals, spermatozoa and precursors, other abnormal components

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Once during Week 12 of treatment and once during Week 4 of recovery
- Dose groups that were examined: All groups at end of treatment phase, Control and high-dose animals at end of recovery phase.
- Battery of functions tested: Evaluation of sensory reactivity to stimuli of different modalities (e.g. auditory, visual and proprioceptive stimuli) and an assessment of grip strength. Motor activity
assessed using an automated activity recorder.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Detailed post mortem examination including examination of the external surface and orifices. Changes noted, the requisite organs weighed and the required tissue samples preserved in fixative.

Organ weights - Adrenal glands, Brain, Coagulating glands, Epididymides, Heart, Kidneys, Liver, Ovaries, Parathyroid glands, Prostate gland, Seminal vesicles, Spleen, Testes, Thymus (where
present), Thyroid, Uterus – cervix.

Tissues fixed and preserved – Abnormalities, Adrenal glands, Aorta, Bone marrow (from sternum), Brain (cerebrum, cerebellum, medullalpons), Caecum, Coagulating glands, Colon, Duodenum,
Epididymides, Eyes, Femur with joint, Heart, Ileum, Jejunum (including Peyer’s patches), Kidneys, Larynx, Liver, Lungs (including mainstem bronchi), Lymph nodes — cervical, Lymph nodes — mesenteric,
Mammary area, Nasopharynx, Oesophagus, Optic nerves, Ovaries, Oviducts, Pancreas, Parathyroid glands, Pituitary gland, Prostate gland, Rectum, Salivary glands, Sciatic nerve, Seminal vesicles,
Skeletal muscle, Skin, Spinal column, Spinal cord (cervical, mid-thoracic, lumbar), Spleen, Stomach, Testes, Thymus (where present), Thyroid, Trachea, Urinary bladder, Uterus – cervix, Vagina


HISTOPATHOLOGY: Yes
Tissues examined - Abnormalities, Adrenal glands, Aorta, Bone marrow (from sternum), Brain (cerebrum, cerebellum, medullalpons), Caecum, Coagulating glands, Colon, Duodenum,
Epididymides, Eyes, Femur with joint, Heart, Ileum, Jejunum (including Peyer’s patches), Kidneys, Larynx, Liver, Lungs (including mainstem bronchi), Lymph nodes — cervical, Lymph nodes — mesenteric,
Mammary area, Nasopharynx, Oesophagus, Optic nerves, Ovaries, Oviducts, Pancreas, Parathyroid glands, Pituitary gland, Prostate gland, Rectum, Salivary glands, Sciatic nerve, Seminal vesicles,
Skeletal muscle, Skin, Spinal cord (cervical, mid-thoracic, lumbar), Spleen, Stomach, Testes, Thymus (where present), Thyroid, Trachea, Urinary bladder, Uterus – cervix, Vagina
After dehydration and embedding in paraffin wax, sections were cut at 5 micrometre thickness and stained with haematoxylin and eosin. In the first instance the examination was limited to: All abnormalities in all main
phase groups and tissues listed above from all animals in the control and high dose groups at the end of the 13 weeks of treatment. Examination of the liver and spleen was extended to intermediate groups and
recovery animals. In addition, the testes and epididymides of main group animals were cut at 2-3 micrometre thickness and stained with Periodic Acid Schiff (PAS). The morphological evaluation of the seminiferous
epithelium (staging of spermatogenic cycle) was performed in all animals of the control and high dose groups at the end of the 13 weeks of treatment.
Other examinations:
The testes and epididymides of main group animals were cut at 2-3 micrometre thickness and stained with Periodic Acid Schiff (PAS). A morphological evaluation of the seminiferous epithelium (staging of
spermatogenic cycle) was performed on all animals in the control and high dose groups killed at the end of the 13 weeks of treatment. Should effects have been observed, examination would have been
extended to animals of intermediate groups and recovery groups.
Statistics:
For continuous variables the significance of the differences amongst group means was assessed by Dunnett’s test or a modified t test, depending on the homogeneity of data.
Statistical analysis of histopathological findings was carried out by means of the non parametric Kolmogorov Smirnov test.
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Ophthalmological findings:
no effects observed
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Leucocytosis, mainly represented by increased lymphocytes and monocytes, was noted in a number of animals from all treated groups.
Changes were with no dose-relation and males were more sensitive than females
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Fluctuations of some parameters (alkaline phosphatase, total bilirubin, triglycerides) were recorded in treated animals.
Due to the slight severity and/or absence of dose-relation, the severity of these findings was not considered to be suggestive of tissue/organ injury.
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Absolute and relative liver weights were increased approximately 25% in high dose (800 mg/kg bw/day males and females.
The relative weight (to terminal body weight) of the liver was also increased in Group 3 animals (200 mg/kg bw/day).
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Treatment-related findings were only seen in the liver of most males and females treated at 800 mg/kg bw/day.
Minimal, hepatocellular hypertrophy with centrilobular distribution, morphologically represented by a minimal increase in liver cell size, accompanied by an increase in liver weight,
when compared with controls, was commonly associated with microsomal enzyme induction and considered in this case as an adaptive response
Other effects:
no effects observed
Description (incidence and severity):
Seminiferous tubules were evaluated with respect to their stage in the spermatogenic cycle and to the integrity of the various cell types within the different stages.
Regular layering in the germinal epithelium was noted and the cycle of spermatogenesis in the high dose males was regarded as normal with no treatment-related effect apparent.

Normal physiology of the oestrous cycle (oestrus, metestrus, diestrus and proestrus) was noted in control and treated females. The morphological changes seen were normal when
compared to each “oestrus phase” in the ovaries, uterus/cervix and vagina.
Key result
Dose descriptor:
NOAEL
Effect level:
800 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
histopathology: non-neoplastic
Critical effects observed:
no
Lowest effective dose / conc.:
800 mg/kg bw/day (nominal)
System:
hepatobiliary
Organ:
liver

Body weight (g) - Group mean data

Males

 

 

 

 

 

 

 

Treatment Day

 

 

 

 

 

 

 

 

1

8

15

22

29

36

43

50

57

64

71

78

85

92

 

(n)

15

15

15

15

15

15

15

15

15

15

15

15

15

15

Control

Mean

206.10

257.26

297.97

330.65

354.21

375.46

391.38

407.69

417.38

426.77

434.11

444.59

452.85

443.87

 

SD

9.186

12.573

16.945

18.495

21.101

23.511

25.325

26.277

27.477

29.018

30.732

31.424

33.114

37.744

 

(n)

10

10

10

10

10

10

10

10

10

10

10

10

10

10

50 mg/kg

Mean

205.66

257.19

301.54

332.12

354.93

376.75

394.49

411.29

420.31

430.56

439.41

448.09

456.96

442.66

 

SD

8.035

14.045

21.965

25.789

26.152

28.406

29.724

30.992

32.992

33.890

35.674

37.576

39.497

40.577

 

(n)

10

10

10

10

10

10

10

10

10

10

10

10

10

10

200 mg/kg

Mean

207.75

261.09

305.55

336.02

358.37

378.18

394.96

412.41

420.61

430.64

438.59

449.21

456.70

438.66

 

SD

10.601

15.130

19.735

23.387

27.611

30.222

32.541

34.286

32.617

33.558

37.864

41.566

38.985

39.664

 

(n)

15

15

15

15

15

15

15

15

15

15

15

15

15

15

800 mg/kg

Mean

206.79

256.69

297.54

326.23

345.90

364.95

378.73

391.86

400.69

408.26

415.77

421.59

429.58

417.81

 

SD

13.059

17.097

19.175

21.858

23.728

24.501

24.476

25.001

27.528

27.927

26.854

28.853

29.291

33.514

Males

 

 

Recovery Day

 

 

 

8

15

22

29

 

(n)

5

5

5

5

Control

Mean

478.40

487.86

493.92

475.86

 

SD

41.003

46.580

55.650

54.009

 

(n)

5

5

5

5

800 mg/kg

Mean

442.90

451.70

458.62

446.44

 

SD

41.771

38.104

40.245

37.303

 

Females

 

 

 

 

 

 

 

Treatment Day

 

 

 

 

 

 

 

 

1

8

15

22

29

36

43

50

57

64

71

78

85

92

 

(n)

15

15

15

15

15

15

15

15

15

15

15

15

15

15

Control

Mean

159.50

181.24

194.11

210.43

220.31

229.25

239.47

244.41

249.87

253.57

255.67

264.56

268.06

265.77

 

SD

7.993

7.875

9.302

11.329

13.014

12.911

13.028

13.157

15.413

15.064

13.315

15.595

18.342

16.741

 

(n)

10

10

10

10

10

10

10

10

10

10

10

10

10

10

50 mg/kg

Mean

160.13

180.02

193.70

209.24

222.45

230.78

236.94

243.94

248.02

251.94

255.20

265.59

271.77

264.01

 

SD

6.158

7.218

9.808

12.724

13.319

14.007

12.756

14.412

15.970

14.168

17.436

11.912

12.745

14.810

 

(n)

10

10

10

10

10

10

10

10

10

10

10

10

10

10

200 mg/kg

Mean

158.94

177.94

187.39

201.06

214.10

221.05

229.99

236.13

239.18

242.74

245.50

261.86

265.89

258.18

 

SD

7.389

14.173

18.364

20.167

23.240

20.702

22.514

22.849

21.424

21.844

20.503

22.838

20.176

20.051

 

(n)

15

15

15

15

15

15

15

15

15

15

15

15

15

15

800 mg/kg

Mean

159.35

179.51

192.91

205.89

217.01

225.83

234.09

241.49

245.33

247.59

250.87

258.26

262.08

258.97

 

SD

7.415

8.743

11.766

14.402

14.231

13.939

15.163

15.089

14.991

14.613

16.792

16.396

17.949

18.740

 

Females

 

 

Recovery Day

 

 

 

8

15

22

29

 

(n)

5

5

5

5

Control

Mean

261.52

263.24

266.16

260.64

 

SD

19.520

22.964

24.209

24.966

 

(n)

5

5

5

5

800 mg/kg

Mean

265.18

270.14

269.10

252.90

 

SD

17.092

17.966

16.195

15.697

Food consumption (g) - Group mean data

Males

 

 

 

 

 

 

 

Treatment Day

 

 

 

 

 

 

 

8

15

22

29

36

43

50

57

64

71

78

85

92

 

(n)

6

6

6

6

6

6

6

6

6

6

6

6

6

Control

Mean

23.02

23.10

22.82

22.13

21.56

21.22

20.67

19.78

19.49

18.97

18.48

18.86

15.54

 

SD

0.762

1.209

1.108

0.814

0.760

0.430

0.663

0.577

0.610

0.862

1.009

0.721

2.160

 

(n)

4

4

4

4

4

4

4

4

4

4

4

4

4

50 mg/kg

Mean

22.87

23.06

22.40

23.02

21.76

21.54

20.48

20.40

20.06

19.75

19.23

19.77

15.65

 

SD

1.299

0.867

0.579

1.044

0.604

0.941

1.179

0.995

0.731

0.807

0.936

0.724

0.876

 

(n)

4

4

4

4

4

4

4

4

4

4

4

4

4

200 mg/kg

Mean

23.15

23.80

23.28

22.30

21.96

21.69

20.72

20.44

20.12

19.98

19.64

19.66

14.97

 

SD

0.896

1.056

1.323

1.126

5.570

1.103

0.798

1.113

0.871

0.926

0.837

0.436

0.163

 

(n)

6

6

6

6

6

6

6

6

6

6

6

6

6

800 mg/kg

Mean

21.37*D

22.45

22.79

22.31

21.75

21.81

20.25

20.50

19.63

19.45

17.98

19.18

16.13

 

SD

1.020

1.343

1.205

1.232

1.049

1.100

1.450

1.510

1.085

1.276

2.972

1.336

2.283

 

Males

 

 

Recovery Day

 

 

 

8

15

22

29

 

(n)

2

2

2

2

Control

Mean

34.35

23.17

26.88

22.30

 

SD

2.133

3.357

2.234

1.414

 

(n)

2

2

2

2

800 mg/kg

Mean

15.21

24.79

25.45

21.58

 

SD

8.656

1.057

0.422

1.156

 

Females

 

 

 

 

 

 

 

Treatment Day

 

 

 

 

 

 

 

8

15

22

29

36

43

50

57

64

71

78

85

92

 

(n)

6

6

6

6

6

6

6

6

6

6

6

6

6

Control

Mean

16.05

16.27

15.51

15.61

15.12

15.66

14.53

14.41

14.83

13.32

13.82

14.30

11.87

 

SD

0.578

2.389

0.368

0.662

0.536

0.715

0.391

0.864

1.300

0.509

0.694

1.070

0.422

 

(n)

4

4

4

4

4

4

4

4

4

4

4

4

4

50 mg/kg

Mean

15.47

15.11

15.64

16.17

15.82

15.44

14.14

14.13

14.60

14.02

14.24

14.50

11.97

 

SD

0.555

0.466

0.664

0.468

0.745

0.760

0.500

0.642

0.915

0.559

1.399

0.616

1.095

 

(n)

4

4

4

4

4

4

4

4

4

4

4

4

4

200 mg/kg

Mean

15.24

14.45

15.12

15.58

14.13

15.17

13.65

13.52

12.94

13.21

14.16

14.04

10.75

 

SD

1.056

1.546

1.091

1.168

0.758

2.345

0.559

0.489

1.958

0.679

0.662

0.914

0.532

 

(n)

6

6

6

6

6

6

6

6

6

6

6

6

6

800 mg/kg

Mean

15.34

15.21

15.68

15.88

15.42

15.43

15.20

14.27

14.76

13.74

13.49

14.27

12.28

 

SD

0.677

0.990

0.854

0.673

1.603

1.504

2.139

0.790

1.049

1.131

1.156

1.140

1.104

 

Females

 

 

Recovery Day

 

 

 

8

15

22

29

 

(n)

2

2

2

2

Control

Mean

16.40

16.67

16.98

14.97

 

SD

0.406

0.375

0.031

0.211

 

(n)

2

2

2

2

800 mg/kg

Mean

16.82

18.22

17.59

14.00

 

SD

1.643

1.350

0.883

0.897

Haematology - Terminal sacrifice - Group mean data

Males

 

RBC

HGB

HCT

MCV

MCH

MCHC

RET

RETR

WBC

 

 

(x10E6/μL)

(g/dL)

(%)

(fL)

(pg)

(g/dL)

(x10E9/L)

(%)

(x10E3/μL)

 

(n)

10

10

10

10

10

10

10

10

10

Control

Mean

8.257

14.54

44.52

53.91

17.61

32.70

158.62

1.921

7.217

 

SD

0.2502

0.276

1.401

1.006

0.398

0.632

13.180

0.1500

1.6226

 

(n)

10

10

10

10

10

10

10

10

10

50 mg/kg

Mean

8.201

14.68

45.13

55.03

17.91

32.58

164.10

2.000

9.510+D

 

SD

0.1962

0.365

1.538

1.396

0.325

0.561

17.706

0.1988

1.2913

 

(n)

10

10

10

10

10

10

10

10

10

200 mg/kg

Mean

8.520

14.98

46.09

54.08

17.60

32.51

164.63

1.925

9.637+D

 

SD

0.5238

0.764

2.799

0.992

0.527

0.669

28.522

0.2449

1.1377

 

(n)

10

10

10

10

10

10

10

10

10

800 mg/kg

Mean

8.334

14.73

45.09

54.09

17.67

32.66

147.61

1.768

9.169+D

 

SD

0.2876

0.604

1.806

1.003

0.337

0.288

21.134

0.2301

1.4721

 

Males

 

NEU

LYM

MON

EOS

BAS

LUC

NEUR

LYMR

MONR

EOSR

BASR

LUCR

PLT

PT

 

 

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(%)

(%)

(%)

(%)

(%)

(%)

(x10E3/μL)

(sec)

 

(n)

10

10

10

10

10

10

10

10

10

10

10

10

10

10

Control

Mean

1.078

5.735

0.204

0.091

0.056

0.053

14.80

79.54

2.87

1.29

0.76

0.74

873.1

19.98

 

SD

0.6394

1.3202

0.0554

0.0281

0.0196

0.0216

6.455

6.255

0.615

0.370

0.151

0.190

61.10

0.594

 

(n)

10

10

10

10

10

10

10

10

10

10

10

10

10

10

50 mg/kg

Mean

1.133

7.821+D

0.281*D

0.105

0.093+D

0.077*C

11.65

82.42

3.00

1.12

0.98*D

0.81

863.8

19.66

 

SD

0.4925

0.9709

0.0536

0.0378

0.0216

0.0149

3.686

3.924

0.741

0.343

0.132

0.173

102.29

0.440

 

(n)

10

10

10

10

10

10

10

10

10

10

10

10

10

10

200 mg/kg

Mean

1.216

7.816+D

0.332+D

0.083

0.097+D

0.087*C

12.72

81.05

3.43

0.89*D

1.01*D

0.89

836.0

19.55

 

SD

0.2718

1.0413

0.0846

0.0200

0.0275

0.0340

2.945

3.153

0.682

0.26

0.238

0.311

46.44

0.544

 

(n)

10

10

10

10

10

10

10

10

10

10

10

10

10

10

800 mg/kg

Mean

0.959

7.637+D

0.316+D

0.063

0.083

0.113+C

10.47

83.32

3.45

0.66+D

0.88

1.22*C

865.5

20.41

 

SD

0.2335

1.2057

0.0793

0.0283

0.0323

0.0531

2.110

2.478

0.615

0.222

0.193

0.437

57.09

0.808

 

 

Females

 

RBC

HGB

HCT

MCV

MCH

MCHC

RET

RETR

WBC

 

 

(x10E6/μL)

(g/dL)

(%)

(fL)

(pg)

(g/dL)

(x10E9/L)

(%)

(x10E3/μL)

 

(n)

10

10

10

10

10

10

10

10

10

Control

Mean

7.426

13.83

42.63

57.42

18.63

32.42

174.78

2.353

4.966

 

SD

0.2726

0.523

1.758

1.225

0.350

0.361

32.840

0.4335

1.0897

 

(n)

10

10

10

10

10

10

10

10

10

50 mg/kg

Mean

7.523

13.91

42.53

56.56

18.50

32.73

145.89

1.943

5.946

 

SD

0.2931

0.479

1.748

0.906

0.383

0.356

20.732

0.2886

1.4523

 

(n)

10

10

10

10

10

10

10

10

10

200 mg/kg

Mean

7.349

13.58

41.80

56.92

18.50

32.51

148.03

2.015

6.169

 

SD

0.3193

0.365

1.473

1.737

0.577

0.345

28.280

0.3853

1.0246

 

(n)

10

10

10

10

10

10

10

10

10

800 mg/kg

Mean

7.500

13.83

42.54

56.74

18.43

32.53

158.59

2.123

5.924

 

SD

0.3016

0.521

1.833

1.326

0.350

0.347

29.226

0.4272

0.7587

 

Females

 

NEU

LYM

MON

EOS

BAS

LUC

NEUR

LYMR

MONR

EOSR

BASR

LUCR

PLT

PT

 

 

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(%)

(%)

(%)

(%)

(%)

(%)

(x10E3/μL)

(sec)

 

(n)

10

10

10

10

10

10

10

10

10

10

10

10

10

10

Control

Mean

0.448

4.305

0.089

0.079

0.024

0.021

9.18

86.48

1.81

1.62

0.45

0.44

985.9

23.44

 

SD

0.1440

1.0019

0.0360

0.0208

0.0117

0.0099

2.296

2.409

0.656

0.478

0.158

0.143

89.05

0.908

 

(n)

10

10

10

10

10

10

10

10

10

10

10

10

10

10

50 mg/kg

Mean

0.461

5.168

0.145*D

0.112

0.033

0.028

7.94

86.75

2.54

1.80

0.50

0.47

965.3

22.73

 

SD

0.1225

1.3316

0.0438

0.1203

0.0170

0.0140

1.731

2.848

0.844

1.702

0.183

0.134

117.68

1.106

 

(n)

10

10

10

10

10

10

10

10

10

10

10

10

10

10

200 mg/kg

Mean

0.440

5.464

0.127

0.072

0.029

0.036

7.10

88.58

2.12

1.18

0.45

0.58

935.7

22.98

 

SD

0.1438

0.9283

0.0469

0.0270

0.0088

0.0126

1.573

2.134

0.761

0.405

0.071

0.193

188.66

1.583

 

(n)

10

10

10

10

10

10

10

10

10

10

10

10

10

10

800 mg/kg

Mean

0.510

5.121

0.146*D

0.071

0.032

0.043+D

8.67

86.41

2.46

1.24

0.51

0.73+D

965.4

22.70

 

SD

0.2047

0.7039

0.0386

0.0264

0.0140

0.0195

3.666

3.266

0.638

0.357

0.179

0.271

106.27

1.347

Haematology - Recovery sacrifice - Group mean data

Males

 

RBC

HGB

HCT

MCV

MCH

MCHC

RET

RETR

WBC

 

 

(x10E6/μL)

(g/dL)

(%)

(fL)

(pg)

(g/dL)

(x10E9/L)

(%)

(x10E3/μL)

 

(n)

5

5

5

5

5

5

5

5

5

Control

Mean

9.348

15.82

51.04

54.60

16.94

31.02

171.30

1.828

7.022

 

SD

0.1883

0.148

1.210

1.334

0.251

0.531

27.001

0.2621

1.2996

 

(n)

5

5

5

5

5

5

5

5

5

800 mg/kg

Mean

8.882

15.52

49.34

55.58

17.48*D

31.42

197.68

2.238

6.442

 

SD

0.4775

0.716

2.082

1.904

0.444

0.522

46.671

0.5832

0.5558

 

Males

 

NEU

LYM

MON

EOS

BAS

LUC

NEUR

LYMR

MONR

EOSR

BASR

LUCR

PLT

 

 

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(%)

(%)

(%)

(%)

(%)

(%)

(x10E3/μL)

 

(n)

5

5

5

5

5

5

5

5

5

5

5

5

5

Control

Mean

0.746

5.738

0.274

0.144

0.080

0.046

10.78

81.28

3.94

2.22

1.08

0.64

942.6

 

SD

0.1623

1.2240

0.0472

0.0650

0.0224

0.0134

2.286

3.761

0.760

1.472

0.110

0.089

100.50

 

(n)

5

5

5

5

5

5

5

5

5

5

5

5

5

800 mg/kg

Mean

0.808

5.172

0.238

0.124

0.054*D

0.046

12.50

80.28

3.66

1.98

0.88*D

0.74

932.4

 

SD

0.1344

0.4123

0.0978

0.0635

0.0089

0.0089

1.505

1.424

1.246

1.123

0.084

0.114

56.64

 

Females

 

RBC

HGB

HCT

MCV

MCH

MCHC

RET

RETR

WBC

 

 

(x10E6/μL)

(g/dL)

(%)

(fL)

(pg)

(g/dL)

(x10E9/L)

(%)

(x10E3/μL)

 

(n)

4

4

4

4

4

4

4

4

4

Control

Mean

8.438

15.15

46.18

54.73

17.98

32.80

148.98

1.765

4.293

 

SD

0.2534

0.480

1.576

0.670

0.250

0.082

16.391

0.1686

0.4586

 

(n)

5

5

5

5

5

5

5

5

5

800 mg/kg

Mean

8.420

15.20

46.20

54.86

18.04

32.86

136.50

1.622

4.380

 

SD

0.1719

0.485

1.263

0.416

0.336

0.428

28.175

0.3432

1.2468

 

Females

 

NEU

LYM

MON

EOS

BAS

LUC

NEUR

LYMR

MONR

EOSR

BASR

LUCR

PLT

 

 

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(x10E3/μL)

(%)

(%)

(%)

(%)

(%)

(%)

(x10E3/μL)

 

(n)

4

4

4

4

4

4

4

4

4

4

4

4

4

Control

Mean

0.493

3.488

0.148

0.105

0.033

0.028

11.38

81.38

3.43

2.48

0.70

0.63

1060.3

 

SD

0.1320

0.3457

0.0403

0.0191

0.0096

0.0050

2.398

2.484

0.814

0.492

0.216

0.126

46.29

 

(n)

5

5

5

5

5

5

5

5

5

5

5

5

5

800 mg/kg

Mean

0.422

3.654

0.146

0.082

0.042

0.036

9.54

83.64

3.32

1.86

0.90

0.74

997.6

 

SD

0.1452

1.0038

0.0573

0.0286

0.0239

0.0182

1.274

1.942

0.476

0.483

0.255

0.251

116.77

Clinical chemistry - Terminal sacrifice - Group mean data

Males

 

ALP

ALT

AST

GGT

TBIL

CHOL

TRIG

GLU

UREA

 

 

(U/L)

(U/L)

(U/L)

(U/L)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

 

(n)

10

10

10

0@

10

10

10

10

10

Control

Mean

185.54

38.76

77.68

-

0.057

72.59

29.49

120.51

28.01

 

SD

21.942

15.476

15.759

-

0.0170

3.566

9.029

18.760

5.368

 

(n)

10

10

10

0@

10

10

10

10

10

50 mg/kg

Mean

192.63

37.07

75.02

-

0.053

76.57

49.09+D

156.00*D

29.74

 

SD

37.139

9.523

22.362

-

0.0164

10.797

16.901

36.559

6.259

 

(n)

10

10

10

1@

10

10

10

10

10

200 mg/kg

Mean

214.29

37.25

67.29

0.90

0.041

81.56

55.40+D

125.83

26.07

 

SD

43.789

3.955

6.330

-

0.0137

12.757

9.497

18.217

4.781

 

(n)

10

10

10

4@

10

10

10

10

10

800 mg/kg

Mean

358.20+C

47.34

64.72

0.45

0.022+D

94.77#C

59.07+D

150.54

26.73

 

SD

139.244

10.907

7.801

0.412

0.0123

10.482

15.711

35.365

4.719

 

Males

 

CREA

Cl

Ca

Na

K

IP

PROT

ALB

GLO

AG

 

 

(mg/dL)

(mmol/L)

(mmol/L)

(mmol/L)

(mmol/L)

(mg/dL)

(g/dL)

(g/dL)

(g/dL)

 

 

(n)

10

10

10

10

10

10

10

10

10

10

Control

Mean

0.391

101.05

2.651

144.54

4.010

6.095

6.45

3.72

2.73

1.36

 

SD

0.0357

1.099

0.0924

0.769

0.2593

0.6497

0.201

0.140

0.082

0.052

 

(n)

10

10

10

10

10

10

10

10

10

10

50 mg/kg

Mean

0.412

100.50

2.650

144.41

4.097

6.137

6.31

3.66

2.65

1.38

 

SD

0.0605

1.101

0.0478

0.610

0.2863

0.4611

0.238

0.158

0.118

0.063

 

(n)

10

10

2.733

10

10

10

10

10

10

10

200 mg/kg

Mean

0.401

100.81

0.0887

144.41

4.049

6.244

6.32

3.70

2.62

1.42

 

SD

0.0626

1.009

 

0.610

0.2465

0.4127

0.215

0.156

0.103

0.079

 

(n)

10

10

10

10

10

10

10

10

10

10

800 mg/kg

Mean

0.394

102.46*D

2.709

144.78

3.904

6.732*D

5.73+D

3.43+D

2.30+D

1.50+D

 

SD

0.0749

0.9950

0.0755

0.691

0.2738

0.4101

0.323

0.183

0.156

0.047

 

 

Females

 

ALP

ALT

AST

GGT

TBIL

CHOL

TRIG

GLU

UREA

 

 

(U/L)

(U/L)

(U/L)

(U/L)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

 

(n)

10

10

10

8@

10

10

10

10

10

Control

Mean

136.06

25.92

58.13

0.78

0.056

78.36

37.50

131.20

31.54

 

SD

25.430

3.469

6.285

0.453

0.0158

13.220

6.846

15.874

4.913

 

(n)

10

10

10

8@

10

10

10

10

10

50 mg/kg

Mean

140.90

26.81

66.69*C

0.79

0.041

72.34

36.55

118.08

35.19

 

SD

42.704

3.542

7.029

0.795

0.0120

12.370

6.196

16.967

6.686

 

(n)

10

10

10

8@

10

10

10

10

10

200 mg/kg

Mean

140.08

23.06

50.26+C

0.53

0.050

64.30*D

41.05

157.42

29.42

 

SD

39.369

4.656

4.093

0.486

0.0271

9.302

10.554

39.192

2.929

 

(n)

9

9

9

8@

9

9

9

9

9

800 mg/kg

Mean

179.60

34.08

65.94

0.70

0.034

76.93

45.07

169.66*C

29.60

 

SD

39.862

11.452

25.231

0.457

0.0188

5.754

7.744

31.189

4.677

 

Females

 

CREA

Cl

Ca

Na

K

IP

PROT

ALB

GLO

AG

 

 

(mg/dL)

(mmol/L)

(mmol/L)

(mmol/L)

(mmol/L)

(mg/dL)

(g/dL)

(g/dL)

(g/dL)

 

 

(n)

10

10

10

10

10

10

10

10

10

10

Control

Mean

0.377

100.90

2.604

141.85

3.300

5.465

5.95

3.59

2.36

1.52

 

SD

0.0254

1.757

0.1230

1.628

0.1719

0.6427

0.443

0.288

0.165

0.063

 

(n)

10

10

10

10

10

10

10

10

10

10

50 mg/kg

Mean

0.414

102.27

2.718*D

143.20

3.635*D

6.314+D

5.96

3.60

2.36

1.51

 

SD

0.0718

1.061

0.0787

1.566

0.2331

0.4998

0.357

0.221

0.151

0.057

 

(n)

10

10

10

10

10

10

10

10

10

10

200 mg/kg

Mean

0.393

101.72

2.627

141.90

3.781+D

6.285+D

5.48*D

3.33*D

2.15*D

1.55

 

SD

0.0330

1.062

0.0595

0.751

0.2824

0.6196

0.358

0.221

0.172

0.108

 

(n)

9

9

9

9

9

9

9

9

9

9

800 mg/kg

Mean

0.394

101.060

2.616

142.40

3.740+D

5.999

5.49*D

3.36

2.13*D

1.58

 

SD

0.0482

1.788

0.0930

1.663

0.2844

0.4411

3.48

0.188

0.173

0.067

Clinical chemistry - Recovery sacrifice - Group mean data

Males

 

ALP

ALT

AST

GGT

TBIL

CHOL

TRIG

GLU

UREA

 

 

(U/L)

(U/L)

(U/L)

(U/L)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

 

(n)

5

5

5

1@

4@

5

5

5

5

Control

Mean

168.08

42.68

56.28

0.90

0.045

112.74

131.74

133.42

32.10

 

SD

19.615

2.739

8.854

-

0.0311

24.430

98.531

38.102

4.027

 

(n)

5

5

5

5

5

5

5

5

5

800 mg/kg

Mean

161.76

37.82

55.00

1.16

0.046

95.42

102.46

171.88

33.48

 

SD

30.229

5.136

14.137

0.594

0.0241

11.540

32.405

59.304

2.587

 

Males

 

CREA

Cl

Ca

Na

K

IP

PROT

ALB

GLO

AG

 

 

(mg/dL)

(mmol/L)

(mmol/L)

(mmol/L)

(mmol/L)

(mg/dL)

(g/dL)

(g/dL)

(g/dL)

 

 

(n)

5

5

5

5

5

5

5

5

5

5

Control

Mean

0.328

99.16

2.780

142.70

4.058

4.838

6.80

3.94

2.86

1.36

 

SD

0.0460

2.392

0.0469

1.907

0.5964

0.6305

0.141

0.152

0.114

0.089

 

(n)

5

5

5

5

5

5

5

5

5

5

800 mg/kg

Mean

0.304

99.80

2.774

141.96

4.112

5.930*D

6.52

3.78

2.74

1.38

 

SD

0.0288

2.113

0.0856

1.616

0.2872

0.7296

0.432

0.239

0.207

0.045

  

Females

 

ALP

ALT

AST

GGT

TBIL

CHOL

TRIG

GLU

UREA

 

 

(U/L)

(U/L)

(U/L)

(U/L)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

 

(n)

5

5

5

5

5

5

5

5

5

Control

Mean

112.24

43.92

65.06

0.68

0.058

101.24

64.16

97.00

42.42

 

SD

16.131

4.853

7.996

0.421

0.0084

26.033

9.259

13.258

4.209

 

(n)

5

5

5

3@

5

5

5

5

5

800 mg/kg

Mean

106.56

30.54+D

61.24

1.33

0.066

116.52

58.78

99.00

29.50+D

 

SD

12.195

4.229

5.693

0.896

0.0261

24.905

12.857

4.995

3.236

 

Females

 

CREA

Cl

Ca

Na

K

IP

PROT

ALB

GLO

AG

 

 

(mg/dL)

(mmol/L)

(mmol/L)

(mmol/L)

(mmol/L)

(mg/dL)

(g/dL)

(g/dL)

(g/dL)

 

 

(n)

5

5

5

5

5

5

5

5

5

5

Control

Mean

0.360

100.68

2.704

141.36

3.370

4.608

6.78

4.04

2.74

1.46

 

SD

0.0316

1.469

0.0391

1.193

0.3338

0.4456

0.311

0.152

0.167

0.055

 

(n)

5

5

5

5

5

5

5

5

5

5

800 mg/kg

Mean

0.330

101.86

2.716

141.36

3.564

5.290

6.48

3.88

2.60

1.50

 

SD

0.0480

1.798

0.0654

1.471

0.3378

0.8643

0.164

0.148

0.071

0.100

Absolute organ weight - Terminal sacrifice - Group mean data

Males

 

Body weight

Adrenals

Brain

Epididymides

Heart

Kidneys

Liver

Spleen

Testes

Thymus

Thyroid

 

 

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

 

(n)

10

10

10

10

10

10

10

10

10

10

10

Control

Mean

427.89

0.0590

1.862

1.3769

1.442

2.721

11.433

0.7690

3.9619

0.3236

0.0253

 

SD

32.077

0.01232

0.0552

0.10262

0.0952

0.2481

1.2470

0.10846

0.20651

0.04725

0.00957

 

(n)

10

10

10

10

10

10

10

10

10

10

10

50 mg/kg

Mean

440.59

0.0608

1.891

1.4550

1.499

2.783

12.235

0.8295

3.9352

0.3813

0.0210

 

SD

40.093

0.01069

0.0860

0.09291

0.1611

0.3009

1.3782

0.08300

0.21285

0.09731

0.00455

 

(n)

10

10

10

10

10

10

10

10

10

10

10

200 mg/kg

Mean

436.84

0.0554

1.900

1.3520

1.431

2.685

12.571

0.7816

3.6699

0.3447

0.0236

 

SD

40.037

0.00698

0.0910

0.20774

0.1081

0.2733

1.4254

0.07942

0.89508

0.09736

0.00591

 

(n)

10

10

10

10

10

10

10

10

10

10

10

800 mg/kg

Mean

404.34

0.0570

1.834

1.3379

1.346

2.568

13.490+D

0.7262

4.0157

0.3054

0.0195

 

SD

21.486

0.00920

0.0837

0.06782

0.1192

0.4162

0.6409

0.08242

0.28532

0.03642

0.00584

 

 

Females

 

Body weight

Adrenals

Brain

Heart

Kidneys

Liver

Ovaries

Spleen

Thymus

Thyroid

Uterus

 

 

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

 

(n)

10

10

10

10

10

10

10

10

10

10

10

Control

Mean

253.73

0.0585

1.762

0.959

1.557

6.897

0.1323

0.6248

0.2409

0.0264

1.0999

 

SD

11.839

0.00829

0.0770

0.0558

0.1198

0.5130

0.02362

0.10721

0.05145

0.00272

0.53173

 

(n)

10

10

10

10

10

10

10

10

10

10

10

50 mg/kg

Mean

249.08

0.0598

1.763

0.932

1.577

7.098

0.1319

0.6015

0.2374

0.0244

0.5784*D

 

SD

13.140

0.00779

0.1084

0.0623

0.0886

0.6474

0.02185

0.07545

0.04729

0.00363

0.21805

 

(n)

10

10

10

10

10

10

10

10

10

10

10

200 mg/kg

Mean

242.72

0.0559

1.741

0.917

1.504

7.298

0.1310

0.6123

0.2723

0.0261

0.8027

 

SD

19.348

0.00808

0.1010

0.0964

0.1732

0.6378

0.01500

0.05412

0.04361

0.00375

0.39748

 

(n)

10

10

10

10

10

10

10

10

10

10

10

800 mg/kg

Mean

240.31

0.0605

1.746

0.908

1.504

8.235+D

0.1133

0.5864

0.2223

0.0280

0.6016*C

 

SD

19.257

0.01318

0.0434

0.0814

0.1030

0.9561

0.01256

0.06275

0.03579

0.00386

0.2180

Absolute organ weight - Recovery sacrifice - Group mean data

Males

 

Body weight

Adrenals

Brain

Epididymides

Heart

Kidneys

Liver

Spleen

Testes

Thymus

Thyroid

 

 

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

 

(n)

5

5

5

5

5

5

5

5

5

5

5

Control

Mean

472.18

0.0552

1.826

1.3678

1.507

3.204

13.349

0.7900

3.9638

0.3148

0.0270

 

SD

57.045

0.00554

0.1223

0.09981

0.1462

0.3125

2.1647

0.09222

0.15317

0.06683

0.00245

 

(n)

5

5

5

5

5

5

5

5

5

5

5

800 mg/kg

Mean

442.12

0.0530

1.930

1.4830

1.474

3.172

12.198

0.8484

4.0182

0.3410

0.0244

 

SD

38.069

0.00742

0.1424

0.11812

0.0521

0.2520

2.1146

0.10889

0.34339

0.07041

0.00598

  

Females

 

Body weight

Adrenals

Brain

Heart

Kidneys

Liver

Ovaries

Spleen

Thymus

Thyroid

Uterus

 

 

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

(g)

 

(n)

5

5

5

5

5

5

5

5

5

5

5

Control

Mean

256.34

0.0584

1.735

0.979

1.651

6.607

0.1254

0.5970

0.2410

0.0210

0.5406

 

SD

24.342

0.00971

0.1379

0.1623

0.2396

1.4251

0.01383

0.06660

0.04632

0.00200

0.13906

 

(n)

5

5

5

5

5

5

5

5

5

5

5

800 mg/kg

Mean

248.80

0.0588

1.814

0.949

1.529

6.406

0.1364

0.6016

0.2134

0.0232

0.6300

 

SD

15.680

0.01028

0.0431

0.1295

0.1772

0.7354

0.02962

0.11855

0.02692

0.00396

0.13957

Relative (to body weight) organ weight - Terminal sacrifice - Group mean data

Males

 

Body weight

Adrenals

Brain

Epididymides

Heart

Kidneys

Liver

Spleen

Testes

Thymus

Thyroid

 

 

(g)

 

 

 

 

 

 

 

 

 

 

 

(n)

10

10

10

10

10

10

10

10

10

10

10

Control

Mean

427.89

0.0138

0.4377

0.3226

0.3382

0.6360

2.6677

0.1796

0.9299

0.0754

0.0059

 

SD

32.077

0.00247

0.03881

0.02272

0.02820

0.03698

0.14363

0.02022

0.07382

0.00754

0.00220

 

(n)

10

10

10

10

10

10

10

10

10

10

10

50 mg/kg

Mean

440.59

0.0138

0.4322

0.3324

0.3410

0.6327

2.7753

0.1889

0.8978

0.0863

0.0048

 

SD

40.093

0.00215

0.04174

0.03362

0.03073

0.05399

0.13949

0.01817

0.06920

0.02107

0.00126

 

(n)

10

10

10

10

10

10

10

10

10

10

10

200 mg/kg

Mean

436.84

0.0128

0.4369

0.3120

0.3285

0.6148

2.8736+D

0.1796

0.8480

0.0782

0.0055

 

SD

40.037

0.00196

0.02737

0.05419

0.01543

0.03024

0.10664

0.01801

0.21473

0.01786

0.00152

 

(n)

10

10

10

10

10

10

10

10

10

10

10

800 mg/kg

Mean

404.34

0.0141

0.4542

0.3320

0.3327

0.6351

3.3421+D

0.1796

0.9929

0.0756

0.0048

 

SD

21.486

0.00215

0.02057

0.02776

0.02388

0.09630

0.18709

0.01741

0.04181

0.00851

0.00139

  

Females

 

Body weight

Adrenals

Brain

Heart

Kidneys

Liver

Ovaries

Spleen

Thymus

Thyroid

Uterus

 

 

(g)

 

 

 

 

 

 

 

 

 

 

 

(n)

10

10

10

10

10

10

10

10

10

10

10

Control

Mean

253.73

0.0231

0.6956

0.3783

0.6137

2.7177

0.0520

0.2461

0.0944

0.0104

0.4311

 

SD

11.839

0.00343

0.04290

0.02443

0.04321

0.14280

0.00810

0.03969

0.01673

0.00100

0.20526

 

(n)

10

10

10

10

10

10

10

10

10

10

10

50 mg/kg

Mean

249.08

0.0241

0.7091

0.3752

0.6335

2.8534

0.0529

0.2412

0.0950

0.0098

0.2305*C

 

SD

13.140

0.00362

0.04777

0.03439

0.02515

0.26763

0.00768

0.02514

0.01594

0.00125

0.07765

 

(n)

10

10

10

10

10

10

10

10

10

10

10

200 mg/kg

Mean

242.72

0.0232

0.7208

0.3781

0.6185

3.0089+D

0.0541

0.2532

0.1118

0.0108

0.3402

 

SD

19.348

0.00373

0.06257

0.02967

0.03461

0.14404

0.00636

0.02482

0.01210

0.00175

0.18950

 

(n)

10

10

10

10

10

10

10

10

10

10

10

800 mg/kg

Mean

240.31

0.0252

0.7306

0.3785

0.6283

3.4195+D

0.0474

0.2444

0.0934

0.0117

0.2515*C

 

SD

19.257

0.00517

0.06155

0.02593

0.05162

0.16480

0.00612

0.02270

0.02063

0.00206

0.09083

Relative (to body weight) organ weight - Recovery sacrifice - Group mean data

Males

 

Body weight

Adrenals

Brain

Epididymides

Heart

Kidneys

Liver

Spleen

Testes

Thymus

Thyroid

 

 

(g)

 

 

 

 

 

 

 

 

 

 

 

(n)

5

5

5

5

5

5

5

5

5

5

5

Control

Mean

472.18

0.0118

0.3889

0.2916

0.3213

0.6833

2.8227

0.1676

0.8465

0.0662

0.0058

 

SD

57.045

0.00153

0.02800

0.02619

0.03577

0.07627

0.24615

0.01076

0.07623

0.00716

0.00110

 

(n)

5

5

5

5

5

5

5

5

5

5

5

800 mg/kg

Mean

442.12

0.0121

0.4400

0.3375

0.3358

0.7205

2.7434

0.1919*D

0.9090

0.0774

0.0055

 

SD

38.069

0.00264

0.05862

0.03936

0.03789

0.06950

0.30758

0.01829

0.01457

0.01623

0.00095

  

Females

 

Body weight

Adrenals

Brain

Heart

Kidneys

Liver

Ovaries

Spleen

Thymus

Thyroid

Uterus

 

 

(g)

 

 

 

 

 

 

 

 

 

 

 

(n)

5

5

5

5

5

5

5

5

5

5

5

Control

Mean

256.34

0.0227

0.6783

0.3808

0.6420

2.5582

0.0490

0.2331

0.0935

0.0082

0.2121

 

SD

24.342

0.00274

0.03887

0.03945

0.03783

0.33112

0.00415

0.01823

0.01146

0.00077

0.05635

 

(n)

5

5

5

5

5

5

5

5

5

5

5

800 mg/kg

Mean

248.80

0.0235

0.7309

0.3802

0.6151

2.5726

0.0544

0.2404

0.0863

0.0094

0.2558

 

SD

15.680

0.00318

0.03464

0.03481

0.06943

0.23437

0.00890

0.03293

0.01510

0.00168

0.06799

Macroscopic findings - Group incidence

 

Males

  

Terminal sacrifice

 

Recovery sacrifice

 

 Treatment:

Control

50 mg/kg

200 mg/kg

800 mg/kg

 

Control

800 mg/kg

 

 Number of animals:

10

10

10

10

 

5

5

Finding 

 Number examined:

10

10

10

10

 

5

5

Epididymides

Abnormal size

0

0

1

0

 

 

 

Eyes

Staining

1

0

0

0

 

 

 

Kidneys

Cyst(s)

1

0

0

0

 

 

 

Spleen

Abnormal shape

0

1

1

0

 

 

 

Stomach

Abnormal area(s)

1

0

1

0

 

 

 

Tail

Abnormal shape

1

0

0

0

 

 

 

Testes

Abnormal size

0

0

2

0

 

 

 

Thymus

Abnormal area(s)

0

1

0

1

 

 

 

 

Abnormal colour

0

0

1

0

 

 

 

 

 

 

 

 

 

 

 

 

No abnormalities

  

8

8

7

9

 

5

5

 

 

Females

 

Terminal sacrifice

 

Recovery sacrifice

 

 Treatment:

Control

50 mg/kg

200 mg/kg

800 mg/kg

 

Control

800 mg/kg

 

 Number of animals:

10

10

10

10

 

5

5

Finding 

 Number examined:

10

10

10

10

 

5

5

Adrenals

Abnormal size

0

0

0

1

 

 

 

Cervical nodes

Abnormal colour

1

0

0

0

 

 

 

Ovaries

Abnormal size

4

4

4

0

 

0

3

Stomach

Abnormal area(s)

0

1

0

1

 

 

 

Thymus

Abnormal area(s)

0

0

1

0

 

 

 

 

Abnormal size

3

2

0

1

 

1

1

Uterus

Abnormal contents

6

1

3

1

 

 

 

 

Abnormal size

6

1

3

1

 

 

 

 

 

 

 

 

 

 

 

 

No abnormalities

 

1

3

3

7

 

4

2

 

Microscopic findings - Group incidence

 

Males

  

Terminal sacrifice

 

Recovery sacrifice

 

 Treatment:

Control

50 mg/kg

200 mg/kg

800 mg/kg

 

Control

800 mg/kg

 

 Number of animals:

10

10

10

10

 

5

5

Finding 

 Number examined:

10

10

10

10

 

5

5

Adrenals

Cortical vacuolation

0

0

0

2

 

 

 

 

Hypertrophy

1

0

0

0

 

 

 

Epididymides

Hypospermia

0

0

1

0

 

 

 

Heart

Inflammatory cell foci

2

0

0

1

 

 

 

Kidneys

Cyst(s)

1

0

0

0

 

 

 

 

Nephropothy

8

0

0

0

 

 

 

Liver

Hepatocytic hypertophy

0

0

0

8

 

 

 

 

Hepatocytic vacuolation

7

5

2

0

 

1

0

 

Inflammatory cell foci

10

10

10

10

 

3

5

Lungs

Inflammatory cell foci

0

0

0

1

 

 

 

 

Mineralisation

0

0

0

2

 

 

 

Prostate gland

Inflammatory cell foci

4

0

0

1

 

 

 

Testes

Germ cell depletion

1

0

2

0

 

 

 

Tail 

Luxation 

1

0

0

0

 

 

 

 

Females

  

Terminal sacrifice

 

Recovery sacrifice

 

 Treatment:

Control

50 mg/kg

200 mg/kg

800 mg/kg

 

Control

800 mg/kg

 

 Number of animals:

10

10

10

10

 

5

5

Finding 

 Number examined:

10

10

10

10

 

5

5

Cervical nodes

Congestion

1

0

0

0

 

 

 

Heart

Inflammatory cell foci

0

0

0

1

 

 

 

Kidneys

Nephropathy

4

0

0

1

 

 

 

Liver

Hepatocytic hypertrophy

0

0

0

8

 

 

 

 

Inflammatory cell foci

6

6

7

10

 

4

5

Lungs

Mineralisation

1

0

0

0

 

 

 

Oesophagus

Chronic inflammation

1

0

0

1

 

 

 

Stomach

Mucosal erosion

0

1

0

0

 

 

 

Thyroid gland

Follicular cyst(s)

1

0

0

0

 

 

 

Uterus

Luminal dilatation

5

1

3

1

 

 

 

Vagina

Diestrus

0

0

0

3

 

 

 

 

Estrus

6

0

0

3

 

 

 

 

Metestrus

2

0

0

2

 

 

 

 

Proestrus 

2

0

0

2

 

 

 

 

Key to tables

SD = Standard deviation

*D or * = Dunnett LSD Test Significant at the 0.05 level

+D or + = Dunnett LSD Test Significant at the 0.01 level

*C = Cochran and Cox Test Significant at the 0.05 level

+C = Cochran and Cox Test Significant at the 0.01 level

#C = Cochran and Cox Test Significant at the 0.001 level

($) = Data inhomogeneous by Bartlett’s test

@ = Number examined reduced due to excluded data

e = Excluded

- = Deceased animal or missing data or unsuitable sample

Conclusions:
There were no significant changes in clinical signs, body weight or food consumption. Minor changes in some blood chemistry parameters, together with organ weights suggest an effect in the liver.
Microscopic pathology indicated changes to the liver which could be attributed to administration of the substance.
The NO(A)EL in this study was considered to be 800 mg/kg bw/day in males & females.
Executive summary:

Sub-chronic toxicity has been investigated in a study conducted according to OECD/EU test guidelines.

There were no significant changes in clinical signs, body weight or food consumption. Minor changes in some blood chemistry parameters, together with organ weights suggest an effect in the liver. Microscopic pathology indicated changes, probably adaptive, to the liver which could be attributed to administration of the substance. Examination of sex organs revealed no changes indicative of a potential effect of the substance on reproductive capacity. In partuicular, seminiferous tubules were evaluated with respect to their stage in the spermatogenic cycle and to the integrity of the various cell types within the different stages. Regular layering in the germinal epithelium was noted and the cycle of spermatogenesis in the high dose males was regarded as normal with no treatment-related effect apparent. Normal physiology of the oestrous cycle (oestrus, metestrus, diestrus and proestrus) was noted in control and treated females. The morphological changes seen were normal when compared to each “oestrus phase” in the ovaries, uterus/cervix and vagina.

The NO(A)EL in this study was considered to be 800 mg/kg bw/day in males and females.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
800 mg/kg bw/day
Study duration:
subchronic
Species:
rat

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

A repeat-dose toxicity subchronic study has been conducted according to OECD TG 408 methods and utilising dose levels of 0, 50, 200 and 800 mg/kg bw/day.

Only mild effects were observed in the animals (essentially males) dosed at 800 mg/kg/day and this dose level was regarded as the No Observed Adverse Effect Level (NOAEL). No effects were observed at the lower doses investigated.

Justification for classification or non-classification

No significant toxicity was observed at a concentration of up to 800 mg/kg/day in a repeated-dose subchronic study of at least 90 days duration.

 

According to EU Directive 67/548/EEC and EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008, classification and labelling is not needed for repeated dose toxicity, as the effects seen in the repeated dose toxicity test do not indicate significant functional change or organ dysfunction occurring at levels below indicated cut-off values. Those effects that were observed are regarded as being of minimal toxicological significance insufficient to warrant classification.