Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Repeated dose toxicity: via oral route:

Study 1:

In a study performed according to OECD 407 (adopted in 2008), the study-derived NOAEL for the registered substance was 1000 mg/kg bw/day. This effect level was based on the absence of any toxic effect when the substance was tested up to 1000 mg/kg bw/day.

Repeated dose toxicity via inhalation route:

According to Annex IX of the REACH regulation, testing by the inhalation route is appropriate only if exposure of humans via inhalation is likely. The test chemical has a low vapour pressure value; it is reported to be 6.638046e-20 mmHg at 25°C. The particle size distribution of the test substance is in the range of 150-10 micron, which is larger than the inhalable particulate size. Thus, exposure to inhalable dust, mist and vapour of the test chemical is highly unlikely. Therefore, the repeated dose toxicity study via inhalation route is considered for waiver.

Repeated dose toxicity via dermal route:

The acute toxicity value for the test chemical (as provided in section 7.2.3) is >2000 mg/kg body weight. Given the use of the chemical, repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Thus, it is expected that the test chemical does not exhibit toxicity by dermal route after repeated exposure. In addition, there is no data available that suggests that test chemical exhibit repeated dose toxicity by the dermal route. Hence this endpoint was considered for waiver.

Key value for chemical safety assessment

Toxic effect type:
dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: Manufactured by Sustainability Support Services (Europe) AB Shaanxi Pioneer Biotech co.,Ltd
- Expiration date of the lot/batch: 05/12/2021
- Purity test date: ≥98 %
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: The test item was store in sealed containers at cool and dry place. It was protected from light, moisture, and pest infestation.
- Stability under storage conditions:
- Vehicle: Distilled water
- Solubility and stability of the test substance in the solvent/dispersant/vehicle/test medium: : Samples in triplicate for low and high dose formulation were taken for stability analysis. The stability in the dose formulation was checked at 0 and 6 hours post dose formulation preparation.
- Reactivity of the test substance with the solvent/vehicle /test medium (if applicable): NA
Species:
rat
Strain:
Wistar
Details on species / strain selection:
The rat is a standard laboratory rodent species and is also a recommended species.
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Sipra Labs Limited, Plot No.13, IDA, Balanagar, Hyderabad, India.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 11 to 12 weeks
- Weight at study initiation: Male: 131.24 g -143.54 g; Female: 134.19 g -143.22 g
- Fasting period before study: No
- Housing: Animals were housed in autoclaved polycarbonate cages with stainless steel top grill having facilities for holding pellet feed and drinking water in polycarbonate bottles. Autoclaved corn cob was used as bedding material, and it was changed at least once a week.
- Diet (e.g. ad libitum): Rodent feed VRK Nutritional solutions was available ad libitum.
- Water (e.g. ad libitum): Aquaguard purified water was available ad libitum.
- Acclimation period: Nineteen days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.1– 23.4°C
- Humidity (%): 51 – 61 %.
- Air changes (per hr): 10 Air Changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle per day
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
The test item was administered by oral gavage once daily.
Dose volume: 10 ml/kg bodyweight
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Dose formulation samples were taken immediately after diluting the concentrates in vehicle (Distilled water) on the first day of treatment and on the last day of treatment for homogeneity analysis and. On the third week, samples from all dose formulations were taken for dose concentration analysis.The analyses were performed based on a validated analytical method.
Duration of treatment / exposure:
28 days
Frequency of treatment:
Daily
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Vehicle control
Dose / conc.:
250 mg/kg bw/day (nominal)
Remarks:
Low dose
Dose / conc.:
500 mg/kg bw/day (nominal)
Remarks:
Middle dose
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
High dose
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
Recovery group
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Recovery group
No. of animals per sex per dose:
Main groups: 5 rats per sex per dose
Total of 70 rats (35 males and 35 females)
Recovery group: 5 rats per sex per dose
Control animals:
yes, concurrent vehicle
Details on study design:
In a dose-range finding study, the test chemical was administered to 5 rats /sex/dose by oral gavage at doses of 0 (vehicle control), 100, 300, and 1000 mg/kb bw/day for 14 days. All animals were observed for mortality, clinical signs of toxicity and body weight. At the end of treatment all animals were sent to necropsy for gross pathological examinations. No adverse effects of treatment were observed; hence, the doses of 250, 500 and 1000 mg/kg body weight were selected for the main study.
Observations and examinations performed and frequency:
MORTALITY: Twice daily
CAGE SIDE OBSERVATIONS: Yes
Acclimatization period: Once daily
Treatment period: Twice daily on first 3 days of treatment; once daily thereafter
Recovery period: Once daily
DETAILED CLINICAL OBSERVATIONS: Yes
Detailed clinical examinations were done prior to initiation of treatment and on day 8, 15, 22, and 28 of treatment (main study) and on day 8, 15, 22, 28, 36, and 42 (recovery groups). All animals were observed for changes in skin and fur, eyes, mucous membrane, occurrence of secretions and excretions, autonomic activity, changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypes and bizarre behaviour.
BODY WEIGHT: Yes
Treatment Period: Once weekly
Recovery Period: Once Weekly
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes (gram/animal/day)
Acclimatization and before randomization: Once
Treatment Period: Once weekly
FOOD EFFICIENCY: Not examined
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Not examined.
OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: once during acclimatization and once during week 4
- Dose groups that were examined:
Main groups: 0 and 1000 mg/kg bw/day
Recovery groups: 0 and 1000 mg/kg bw/day
HAEMATOLOGY: Yes
- Time schedule for collection of blood: at week 4 and 6
- Anaesthetic used for blood collection: Yes, under isoflurane
- Animals fasted: Yes, rats were fasted overnight.
- How many animals: all animals in the main groups (0, 250, 500 and 1000 mg/kg bw/day) and in the recovery groups (0 and 1000 mg/kg bw/days)
- Parameters checked in table: Yes. See Chapter 23.8.
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at week 4 and 6
- Animals fasted: Yes
- How many animals: all animals in the main groups (0, 250, 500 and 1000 mg/kg bw/day) and in the recovery groups (0 and 1000 mg/kg bw/days)
- Parameters checked in table [No.?] were examined. Yes, See Chapter 23.9.
URINALYSIS: Yes
- Time schedule for collection of urine: at week 4 and 6
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked in table [No.?] were examined. Yes. See Chapter 23.10.
NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: on day 28 (main groups), and on day 42 (recovery group)
- Dose groups that were examined: all animals in the main groups (0, 250, 500 and 1000 mg/kg bw/day) and in the recovery groups (0 and 1000 mg/kg bw/days)
- Battery of functions tested: sensory activity / grip strength / motor activity / other: Yes
The examinations for sensory reactivity, grip strength, and motor activity assessment were performed once on day 28 for all animals (main study) and on day 42 for all animals (recovery groups).
IMMUNOLOGY: Not examined
- Time schedule for examinations: NA
- How many animals:NA
- Dose groups that were examined:NA
- Parameters checked in table [No.?] were examined.NA
Sacrifice and pathology:
GROSS PATHOLOGY: Yes .
All animals of the main groups (0, 250, 500 and 1000 mg/kg bw /day) were sacrificed at the end of week 4. Rats from the recovery groups (0 and 1000 mg/kg bw/day) were sacrificed at the end of week 6 of experimental period by carbon dioxide asphyxiation. All animals were weighed and necropsied. Descriptions of all macroscopic abnormalities were recorded.
Samples of the following tissues and organs were collected and fixed:
Adrenal glands, Aorta, Bone marrow (Sternum), Brain (cerebrum, cerebellum, pons), Cecum, Cervix, Colon, Duodenum, Epididymides, Eye, Heart, Ileum, with Peyer's patches, Jejunum, Kidneys, Liver, Lungs (inflated with NBF at necropsy), Lymph nodes (mesenteric, axillary), Oesophagus, Ovaries, Pancreas, Rectum, Prostate + Seminal vesicle with coagulating glands, Pituitary, Sciatic nerve. Skeletal muscle, Spinal cord (cervical, mid-thoracic, lumbar), Spleen, Stomach, Testes , Thymus, Thyroid, Trachea, Urinary bladder (inflated with NBF at necropsy), Uterus, Vagina, All Gross lesions
ORGAN WEIGHTS MEASUREMENTS: Yes.
The following organs were weighed and recorded on the day of necropsy: Brain, Spleen, Thymus Epididymides, Heart, Kidneys, Testes, Ovaries, Liver, Adrenals, Uterus, Prostate + Seminal vesicle with coagulating glands
HISTOPATHOLOGY: Yes.
All organ and tissue samples collected during necropsy were processed, embedded and cut to 3-5µm sections then stained with haematoxylin and eosin and examined under light microscope. Full histopathology was performed on the preserved organs and tissues of all animals in the control and high dose groups (main study). Lesions were graded as mild, moderate, or severe based on vascular/degenerative/inflammatory changes in specific organs. Mild=less than 10% of the region affected; Moderate=less than 25% of the region were affected; Severe=more than 25% of the region affected.
Statistics:
The following statistical methods were used to analyse the body weight, feed consumption, organ weights as well as clinical pathology data:
Data was summarized in tabular form. Statistical analysis was performed using SAS/STAT® Software (Version 9.3).
All the data were analysed for homogeneity using Levene’s test
The difference between the means was analysed using Dunnet’s t- test
Significant differences between control and treated groups analysed using one-way ANOVA statistics. Values were summarized as mean ± standard deviation (SD).
Clinical signs:
no effects observed
Description (incidence and severity):
No clinical signs of toxicity were observed inany of the groups.In addition, no abnormal findings were observed in any of the animals during the detailed clinical examinations.
Mortality:
no mortality observed
Description (incidence):
All animals survived to planned death.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
No significant changes in body weight or body weight gain were observed in any of the groups.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No signficanct changes in feed consumption were observed in any of the groups.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Description (incidence and severity):
No abnormal findings were observed in any of the animals treated at at 0 or 1000 mg/kg bw/day.
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
No treatment-related adverse effects were observed at any dose level.
Significant changes were observed in the haematology parameters; however, these were within the historical control ranges of the test facility.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
No treatment-related adverse effects were observed at any dose level.
Significant changes were observed in the clinical biochemistry parameters; however, these were within the historical control ranges.
Urinalysis findings:
no effects observed
Description (incidence and severity):
No significant changes in urinalysis were observed in any of the groups.
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
No significant changes in sensory reactivity, motor activity, or grip strength were observed in any of the groups.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
No adverse effect was observed at any dose level tested.
Gross pathological findings:
no effects observed
Description (incidence and severity):
No treatment-related gross pathological effects were observed at the end of the treatment period (main study) or at the end of the recovery period (recovery groups).
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
No treatment-related histopathological effects were observed at 1000 mg/kg bw/day in the main study.
Histopathological findings: neoplastic:
no effects observed
Description (incidence and severity):
No treatment-related histopathological effects were observed at 1000 mg/kg bw/day in the main study.
Key result
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (nominal)
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
urinalysis
Key result
Critical effects observed:
no

ANALYSIS OF DOSE FORMULATIONS

Dose concentration verifications were carried out for the test item fortified in distilled water at 25, 50 and 100 mg/ml. The recovery values in the prepared dose formulations were within the guideline specification. The test item was found to be homogeneously dispersed in distilled water, which was evinced from the recovery of test item from different layers (top, middle and bottom layers of distilled water).

Clinical signs - Treatment groups

Day of

observation

Control (G1)

Low dose (G2)

Middle dose (G3)

High dose (G4)

M

F

M

F

M

F

M

F

1

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

2

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

3

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

4

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

6

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

7

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

8

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

9

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

10

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

11

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

12

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

13

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

14

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

15

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

16

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

17

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

18

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

19

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

20

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

21

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

22

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

23

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

24

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

25

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

26

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

27

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

28

01/5

01/5

01/5

01/5

01/5

01/5

01/5

01/5

Key: M – Male; F – Female; Observation/No. of animals; 01 – Normal

Clinical signs - Recovery groups

Day of

observation

Control Recovery

(G1R)

High dose Recovery

(G4R)

M

F

M

F

1

01/5

01/5

01/5

01/5

2

01/5

01/5

01/5

01/5

3

01/5

01/5

01/5

01/5

4

01/5

01/5

01/5

01/5

5

01/5

01/5

01/5

01/5

6

01/5

01/5

01/5

01/5

7

01/5

01/5

01/5

01/5

8

01/5

01/5

01/5

01/5

9

01/5

01/5

01/5

01/5

10

01/5

01/5

01/5

01/5

11

01/5

01/5

01/5

01/5

12

01/5

01/5

01/5

01/5

13

01/5

01/5

01/5

01/5

14

01/5

01/5

01/5

01/5

15

01/5

01/5

01/5

01/5

16

01/5

01/5

01/5

01/5

17

01/5

01/5

01/5

01/5

18

01/5

01/5

01/5

01/5

19

01/5

01/5

01/5

01/5

20

01/5

01/5

01/5

01/5

21

01/5

01/5

01/5

01/5

22

01/5

01/5

01/5

01/5

23

01/5

01/5

01/5

01/5

24

01/5

01/5

01/5

01/5

25

01/5

01/5

01/5

01/5

26

01/5

01/5

01/5

01/5

27

01/5

01/5

01/5

01/5

28

01/5

01/5

01/5

01/5

Key: M – Male; F – Female; Observation/No. of animals; 01 – Normal

Clinical signs - Recovery groups (Cont.)

Day of

observation

Control Recovery

(G1R)

High dose Recovery

(G4R)

M

F

M

F

29

01/5

01/5

01/5

01/5

30

01/5

01/5

01/5

01/5

31

01/5

01/5

01/5

01/5

32

01/5

01/5

01/5

01/5

33

01/5

01/5

01/5

01/5

34

01/5

01/5

01/5

01/5

35

01/5

01/5

01/5

01/5

36

01/5

01/5

01/5

01/5

37

01/5

01/5

01/5

01/5

38

01/5

01/5

01/5

01/5

39

01/5

01/5

01/5

01/5

40

01/5

01/5

01/5

01/5

41

01/5

01/5

01/5

01/5

42

01/5

01/5

01/5

01/5

Key: M – Male; F – Female; Observation/No. of animals; 01 – Normal

Mortality - Treatment groups

Days of

observation

G1

G2

G3

G4

M

F

M

F

M

F

M

F

1

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

2

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

3

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

4

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

6

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

7

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

8

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

9

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

10

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

11

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

12

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

13

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

14

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

15

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

16

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

17

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

18

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

19

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

20

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

21

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

22

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

23

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

24

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

25

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

26

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

27

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

28

0/5

0/5

0/5

0/5

0/5

0/5

0/5

0/5

Key: M – Male; F – Female; No. of animals found dead/Total no. of animals

Mortality - recovery group

Days of

observation

G1R

G4R

M

F

M

F

29

0/5

0/5

0/5

0/5

30

0/5

0/5

0/5

0/5

31

0/5

0/5

0/5

0/5

32

0/5

0/5

0/5

0/5

33

0/5

0/5

0/5

0/5

34

0/5

0/5

0/5

0/5

35

0/5

0/5

0/5

0/5

36

0/5

0/5

0/5

0/5

37

0/5

0/5

0/5

0/5

38

0/5

0/5

0/5

0/5

39

0/5

0/5

0/5

0/5

40

0/5

0/5

0/5

0/5

41

0/5

0/5

0/5

0/5

42

0/5

0/5

0/5

0/5

Key: M – Male; F – Female; No. of animals found dead/Total no. of animals.

Body weights - Treatment groups/males

 

Group 1

Group 2

Group 3

Group 4

Day 1

Mean

139.6

140.1

140.6

140.4

SD

4.77

3.57

2.49

2.62

N

5

5

5

5

Day 8

Mean

152.4

153.4

153.9

153.0

SD

4.85

3.88

2.36

2.28

N

5

5

5

5

Day 15

Mean

166.4

166.2

167.1

167.1

SD

5.45

2.77

1.59

2.87

N

5

5

5

5

Day 22

Mean

182.3

181.3

182.4

182.0

SD

5.85

3.24

2.02

2.30

N

5

5

5

5

Day 28

Mean

194.5

194.1

195.3

195.6

SD

5.42

3.50

2.17

3.28

N

5

5

5

5

Body weights - Treatment groups/females 

 

Group 1

Group 2

Group 3

Group 4

Day 1

Mean

139.7

139.6

139.8

140.4

SD

3.16

2.85

2.71

1.97

N

5

5

5

5

Day 8

Mean

152.6

152.9

153.5

154.3

SD

3.06

2.47

2.12

2.41

N

5

5

5

5

Day 15

Mean

165.4

164.9

166.8

168.3

SD

4.45

1.26

1.94

1.85

N

5

5

5

5

Day 22

Mean

181.9

180.4

182.1

184.0

SD

4.87

1.50

1.70

1.17

N

5

5

5

5

Day 28

Mean

194.4

193.8

196.2

197.0

SD

4.76

1.88

2.92

2.58

N

5

5

5

5

Body weights -Recovery groups/males

 

Group 1R

Group 4R

Day 1

Mean

140.7

140.9

SD

2.06

1.57

N

5

5

Day 8

Mean

154.0

153.8

SD

2.86

1.18

N

5

5

Day 15

Mean

167.9

167.0

SD

2.84

1.51

N

5

5

Day 22

Mean

182.6

182.1

SD

2.87

1.95

N

5

5

Day 29

Mean

197.6

197.8

SD

3.80

2.12

N

5

5

Day 36

Mean

212.8

212.4

SD

4.05

2.48

N

5

5

Day 42

Mean

225.1

223.8

SD

4.05

1.93

N

5

5

Body weights -Recovery groups/females  

           

Group 1R

Group 4R

Day 1

Mean

140.7

140.6

SD

1.49

1.69

N

5

5

Day 8

Mean

153.4

154.4

SD

2.03

2.27

N

5

5

Day 15

Mean

166.9

168.1

SD

1.88

3.35

N

5

5

Day 22

Mean

181.8

183.3

SD

1.58

3.19

N

5

5

Day 29

Mean

197.0

197.8

SD

1.78

3.74

N

5

5

Day 36

Mean

212.5

212.6

SD

1.89

4.17

N

5

5

Day 42

Mean

224.3

223.8

SD

1.90

4.01

N

5

5

Body weight gain (%) - Treatment groups/male

           

Group 1

Group 2

Group 3

Group 4

Day 1

Mean

0.0

0.0

0.0

0.0

SD

0.00

0.00

0.00

0.00

N

5

5

5

5

Day 8

Mean

9.2

9.4

9.5

8.9

SD

0.45

0.68

0.40

0.58

N

5

5

5

5

Day 15

Mean

19.2

18.6

18.9

19.0

SD

0.83

1.49

1.11

0.67

N

5

5

5

5

Day 22

Mean

30.6

29.4

29.8

29.6

SD

0.72

1.78

1.09

0.94

N

5

5

5

5

Day 28

Mean

39.4

38.5

39.0

39.3

SD

1.32

2.37

1.32

0.43

N

5

5

5

5

Body weight gain (%) - Treatment groups/females            

                       

Group 1

Group 2

Group 3

Group 4

Day 1

Mean

0.0

0.0

0.0

0.0

SD

0.00

0.00

0.00

0.00

N

5

5

5

5

Day 8

Mean

9.3

9.6

9.8

9.9

SD

0.48

0.72

0.78

0.57

N

5

5

5

5

Day 15

Mean

18.5

18.2

19.3

19.9

SD

0.79

2.07

1.01

0.85

N

5

5

5

5

Day 22

Mean

30.2

29.3

30.2

31.0

SD

0.64

2.02

1.73

1.21

N

5

5

5

5

Day 28

Mean

39.2

38.9

40.3

40.3

SD

0.39

1.68

1.52

1.83

N

5

5

5

5

Body weight gain (%) - Recovery groups/males              

                       

Group 1R

Group 4R

Day 1

Mean

0.0

0.0

SD

0.00

0.00

N

5

5

Day 8

Mean

9.5

9.1

SD

0.79

0.68

N

5

5

Day 15

Mean

19.4

18.6

SD

0.65

1.21

N

5

5

Day 22

Mean

29.8

29.3

SD

0.72

1.37

N

5

5

Day 29

Mean

40.5

40.4

SD

0.79

1.57

N

5

5

Day 36

Mean

51.3

50.8

SD

0.99

1.82

N

5

5

Day 42

Mean

60.0

58.8

SD

0.96

1.68

N

5

5

Body weight gain (%) - Recovery groups/females  

                       

Group 1R

Group 4R

Day 1

Mean

0.0

0.0

SD

0.00

0.00

N

5

5

Day 8

Mean

9.0

9.9

SD

0.63

0.43

N

5

5

Day 15

Mean

18.6

19.5

SD

1.12

1.42

N

5

5

Day 22

Mean

29.2

30.4

SD

1.31

1.26

N

5

5

Day 29

Mean

40.0

40.7

SD

1.47

1.40

N

5

5

Day 36

Mean

51.0

51.2

SD

1.56

1.65

N

5

5

Day 42

Mean

59.4

59.2

SD

1.32

1.65

N

5

5

Feed consumption (G/animal/day) – Treatment group/males

 

Group 1

Group 2

Group 3

Group 4

Week 1

Mean

15.98

15.93

15.88

16.01

SD

2.84

2.88

2.91

2.91

Week 2

Mean

15.97

15.99

15.93

16.03

SD

2.96

2.86

2.99

2.92

Week 3

Mean

15.92

15.99

16.03

15.99

SD

2.86

2.99

2.94

2.93

Week 4

Mean

18.60

18.64

18.51

18.64

SD

3.41

3.44

3.52

3.38

 

Feed consumption (G/animal/day) – Recovery group/males

 

Group 1R

Group 4R

Week 1

Mean

15.96

16.02

SD

2.83

2.92

Week 2

Mean

15.98

15.95

SD

2.86

2.88

Week 3

Mean

15.96

15.86

SD

2.83

2.96

Week 4

Mean

15.73

15.90

SD

2.71

2.95

Week 5

Mean

15.95

15.96

SD

2.91

2.83

Week 6

Mean

18.48

18.57

SD

3.56

3.45

 

Feed consumption (G/animal/day) – Treatment group/females

 

Group 1

Group 2

Group 3

Group 4

Week 1

Mean

15.98

15.93

15.96

15.93

SD

2.74

2.78

2.89

2.85

Week 2

Mean

15.95

15.94

15.89

15.97

SD

2.84

2.86

2.83

2.82

Week 3

Mean

15.91

15.91

15.90

15.93

SD

2.82

2.82

2.82

2.80

Week 4

Mean

18.65

18.59

18.60

18.57

SD

3.35

3.25

3.28

3.22

 

Feed consumption (G/animal/day) – Recovery group/females

 

 

Group 1R

Group 4R

Week 1

Mean

15.93

15.92

SD

2.85

2.84

Week 2

Mean

15.91

15.96

SD

2.83

2.83

Week 3

Mean

15.90

15.94

SD

2.81

2.85

Week 4

Mean

15.91

15.94

SD

2.82

2.86

Week 5

Mean

15.85

15.85

SD

2.83

2.87

Week 6

Mean

18.65

18.64

SD

3.40

3.45

 

OPTHALMOSCOPY

Group:Control (G1)

S. No.

Parameter

Pre Dose

Week 4

1

Cornea

1 (10)

1 (10)

2

Iris

1 (10)

1 (10)

3

Conjunctivae

1 (10)

1 (10)

4

Chemosis

1 (10)

1 (10)

Key: 1 (10) – Observation (No. of animals); 1 – Normal.

 

 

Group:Low dose (G2)

S. No.

Parameter

Pre Dose

1

Cornea

1 (10)

2

Iris

1 (10)

3

Conjunctivae

1 (10)

4

Chemosis

1 (10)

Key: 1 (10) – Observation (No. of animals); 1 – Normal.

 

 

Group:Middle dose (G3)

S. No.

Parameter

Pre Dose

1

Cornea

1 (10)

2

Iris

1 (10)

3

Conjunctivae

1 (10)

4

Chemosis

1 (10)

Key: 1 (10) – Observation (No. of animals); 1 – Normal.

 

Group:High dose (G4)

S. No.

Parameter

Pre Dose

Week 4

1

Cornea

1 (10)

1 (10)

2

Iris

1 (10)

1 (10)

3

Conjunctivae

1 (10)

1 (10)

4

Chemosis

1 (10)

1 (10)

Key: 1 (10) – Observation (No. of animals); 1 – Normal.

 

Group Vehicle Control Recovery(G1R)

S. No.

Parameter

Pre Dose

1

Cornea

1 (10)

2

Iris

1 (10)

3

Conjunctivae

1 (10)

4

Chemosis

1 (10)

Key: 1 (10) – Observation (No. of animals); 1 – Normal.

 

GroupHigh Dose Recovery(G4R)

S. No.

Parameter

Pre Dose

1

Cornea

1 (10)

2

Iris

1 (10)

3

Conjunctivae

1 (10)

4

Chemosis

1 (10)

Key: 1 (10) – Observation (No. of animals); 1 – Normal.

DETAILED CLINICAL EXAMINATIONS

SUMMARY OF DETAILED CLINICAL EXAMINATIONS

GroupCONTROL(G1)

S.NO

Parameter

Pre Dose

Day 8

Day 15

Day 22

Day 28

1

Body Posture

1(10)

1(10)

1(10)

1(10)

1(10)

2

Gait

1(10)

1(10)

1(10)

1(10)

1(10)

3

Skin

1(10)

1(10)

1(10)

1(10)

1(10)

4

Fur

1(10)

1(10)

1(10)

1(10)

1(10)

5

Eyes

1(10)

1(10)

1(10)

1(10)

1(10)

6

Mucous membrane

1(10)

1(10)

1(10)

1(10)

1(10)

7

Respiratory pattern

1(10)

1(10)

1(10)

1(10)

1(10)

8

Salivation

1(10)

1(10)

1(10)

1(10)

1(10)

9

Response to handling

1(10)

1(10)

1(10)

1(10)

1(10)

10

Convulsions

1(10)

1(10)

1(10)

1(10)

1(10)

11

Stereotypic behaviour

1(10)

1(10)

1(10)

1(10)

1(10)

12

Bizzare Behaviour

1(10)

1(10)

1(10)

1(10)

1(10)

              Key: 1(10) – Observation(No of animals), 1-Normal

SUMMARY OF DETAILED CLINICAL EXAMINATIONS

GroupLow Dose(G2)

S.NO

Parameter

Pre Dose

Day 8

Day 15

Day 22

Day 28

1

Body Posture

1(10)

1(10)

1(10)

1(10)

1(10)

2

Gait

1(10)

1(10)

1(10)

1(10)

1(10)

3

Skin

1(10)

1(10)

1(10)

1(10)

1(10)

4

Fur

1(10)

1(10)

1(10)

1(10)

1(10)

5

Eyes

1(10)

1(10)

1(10)

1(10)

1(10)

6

Mucous membrane

1(10)

1(10)

1(10)

1(10)

1(10)

7

Respiratory pattern

1(10)

1(10)

1(10)

1(10)

1(10)

8

Salivation

1(10)

1(10)

1(10)

1(10)

1(10)

9

Response to handling

1(10)

1(10)

1(10)

1(10)

1(10)

10

Convulsions

1(10)

1(10)

1(10)

1(10)

1(10)

11

Stereotypic behaviour

1(10)

1(10)

1(10)

1(10)

1(10)

12

Bizzare Behaviour

1(10)

1(10)

1(10)

1(10)

1(10)

Key: 1(10) – Observation(No of animals), 1-Normal

SUMMARY OF DETAILED CLINICAL EXAMINATIONS

GroupIntermediate Dose(G3)

S.NO

Parameter

Pre Dose

Day 8

Day 15

Day 22

Day 28

1

Body Posture

1(10)

1(10)

1(10)

1(10)

1(10)

2

Gait

1(10)

1(10)

1(10)

1(10)

1(10)

3

Skin

1(10)

1(10)

1(10)

1(10)

1(10)

4

Fur

1(10)

1(10)

1(10)

1(10)

1(10)

5

Eyes

1(10)

1(10)

1(10)

1(10)

1(10)

6

Mucous membrane

1(10)

1(10)

1(10)

1(10)

1(10)

7

Respiratory pattern

1(10)

1(10)

1(10)

1(10)

1(10)

8

Salivation

1(10)

1(10)

1(10)

1(10)

1(10)

9

Response to handling

1(10)

1(10)

1(10)

1(10)

1(10)

10

Convulsions

1(10)

1(10)

1(10)

1(10)

1(10)

11

Stereotypic behaviour

1(10)

1(10)

1(10)

1(10)

1(10)

12

Bizzare Behaviour

1(10)

1(10)

1(10)

1(10)

1(10)

Key: 1(10) – Observation(No of animals), 1-Normal

SUMMARY OF DETAILED CLINICAL EXAMINATIONS

GroupHigh Dose(G4)

S.NO

Parameter

Pre Dose

Day 8

Day 15

Day 22

Day 28

1

Body Posture

1(10)

1(10)

1(10)

1(10)

1(10)

2

Gait

1(10)

1(10)

1(10)

1(10)

1(10)

3

Skin

1(10)

1(10)

1(10)

1(10)

1(10)

4

Fur

1(10)

1(10)

1(10)

1(10)

1(10)

5

Eyes

1(10)

1(10)

1(10)

1(10)

1(10)

6

Mucous membrane

1(10)

1(10)

1(10)

1(10)

1(10)

7

Respiratory pattern

1(10)

1(10)

1(10)

1(10)

1(10)

8

Salivation

1(10)

1(10)

1(10)

1(10)

1(10)

9

Response to handling

1(10)

1(10)

1(10)

1(10)

1(10)

10

Convulsions

1(10)

1(10)

1(10)

1(10)

1(10)

11

Stereotypic behaviour

1(10)

1(10)

1(10)

1(10)

1(10)

12

Bizzare Behaviour

1(10)

1(10)

1(10)

1(10)

1(10)

Key: 1(10) – Observation(No of animals), 1-Normal

SUMMARY OF DETAILED CLINICAL EXAMINATIONS

Group Vehicle Control Recovery(G1R)

S.NO

Parameter

Pre Dose

Day 8

Day 15

Day 22

Day 28

Day 36

Day 42

1

Body Posture

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

2

Gait

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

3

Skin

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

4

Fur

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

5

Eyes

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

6

Mucous membrane

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

7

Respiratory pattern

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

8

Salivation

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

9

Response to handling

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

10

Convulsions

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

11

Stereotypic behaviour

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

12

Bizzare Behaviour

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

Key: 1(10) – Observation(No of animals), 1-Normal

SUMMARY OF DETAILED CLINICAL EXAMINATIONS

GroupHigh Dose Recovery(G4R)

S.NO

Parameter

Pre Dose

Day 8

Day 15

Day 22

Day 28

Day 36

Day 42

1

Body Posture

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

2

Gait

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

3

Skin

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

4

Fur

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

5

Eyes

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

6

Mucous membrane

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

7

Respiratory pattern

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

8

Salivation

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

9

Response to handling

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

10

Convulsions

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

11

Stereotypic behaviour

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

12

Bizzare Behaviour

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

1(10)

Key: 1(10) – Observation(No of animals), 1-Normal

SUMMARY OFSENSORY REACTIVITY ASSESSMENT

Group:Control (G1)

S. No.

Activity

Day 28

1

Pupil reflex

1 (10)

2

Pinna reflex

1 (10)

3

Proprioceptive reflex

1 (10)

4

Approach reflex

1 (10)

5

Nociceptive reflex

1 (10)

6

Righting reflex

1 (10)

Key: 1 (10) – Observation (No. of animals); 1 – Normal.

 

 

Group:Low dose (G2)

S. No.

Activity

Day 28

1

Pupil reflex

1 (10)

2

Pinna reflex

1 (10)

3

Proprioceptive reflex

1 (10)

4

Approach reflex

1 (10)

5

Nociceptive reflex

1 (10)

6

Righting reflex

1 (10)

Key: 1 (10) – Observation (No. of animals); 1 – Normal.

 

SUMMARY OF SENSORY REACTIVITY ASSESSMENT(Contd.,)

Group:Middle dose (G3)

S. No.

Activity

Day 28

1

Pupil reflex

1 (10)

2

Pinna reflex

1 (10)

3

Proprioceptive reflex

1 (10)

4

Approach reflex

1 (10)

5

Nociceptive reflex

1 (10)

6

Righting reflex

1 (10)

Key: 1 (10) – Observation (No. of animals); 1 – Normal.

 

 

Group:High dose (G4)

S. No.

Activity

Day 28

1

Pupil reflex

1 (10)

2

Pinna reflex

1 (10)

3

Proprioceptive reflex

1 (10)

4

Approach reflex

1 (10)

5

Nociceptive reflex

1 (10)

6

Righting reflex

1 (10)

Key: 1 (10) – Observation (No. of animals); 1 – Normal.

 

SUMMARY OF GRIP STRENGTH MEASUREMENT

              MALES

TREATMENT & RECOVERY

Grip Strength

Fore limb

Hind limb

Group 1

Mean

206.00

209.33

SD

30.311

34.595

Group 2

Mean

213.33

219.33

SD

33.914

34.831

Group 3

Mean

204.00

202.67

SD

31.922

14.606

Group 4

Mean

213.33

184.67

SD

14.142

32.371

Group 1R

Mean

211.33

193.93

SD

18.649

39.555

Group 4R

Mean

218.67

198.67

SD

20.494

25.990

           

 

SUMMARY OF GRIP STRENGTH MEASUREMENT(Contd.,)

FEMALES

TREATMENT & RECOVERY

Grip Strength

Fore limb

Hind limb

Group 1

Mean

182.67

153.73

SD

47.690

75.098

Group 2

Mean

232.00

216.67

SD

27.348

16.499

Group 3

Mean

193.33

216.67

SD

44.785

27.489

Group 4

Mean

222.67

241.20

SD

43.297

14.923

Group 1R

Mean

191.27

192.67

SD

34.055

33.283

Group 4R

Mean

174.67

207.33

SD

29.871

22.659

 

SUMMARY OF MOTOR ACTIVITY ASSESSMENT

MALES

TREATMENT & RECOVERY

IR Activity Meter

Slow

Movements

(cm/seconds)

Fast

movements

(cm/seconds)

Slow Stereotypic movements

(seconds)

Fast stereotypic movements (seconds)

Group 1

Mean

74.25

331.40

22.67

205.07

SD

18.872

132.768

5.157

62.752

Group 2

Mean

69.60

267.33

21.05

184.80

SD

33.065

152.481

8.190

71.821

Group 3

Mean

69.00

238.27

24.93

165.20

SD

21.710

117.110

5.410

73.774

Group 4

Mean

70.13

289.20

19.73

174.73

SD

33.574

143.031

7.188

87.180

Group 1R

Mean

66.40

258.60

24.40

169.80

SD

23.860

117.622

5.376

55.035

Group 4R

Mean

56.13

178.20

24.20

133.73

SD

24.819

92.009

7.453

59.448

 

SUMMARY OF MOTOR ACTIVITY ASSESSMENT(Contd.,)

              FEMALES

TREATMENT & RECOVERY

IR Activity Meter

Small movements (cm/seconds)

Fast movements (cm/seconds)

Small Stereotypic movements

(seconds)

Fast stereotypic movements (seconds)

Group 1

Mean

70.27

205.47

26.27

162.53

SD

18.432

109.393

6.493

68.649

Group 2

Mean

65.80

187.07

26.20

156.40

SD

19.921

79.657

5.324

43.531

Group 3

Mean

47.07

100.40

25.67

97.27

SD

14.152

45.687

5.234

23.325

Group 4

Mean

63.06

251.54

24.20

171.40

SD

22.179

126.300

2.140

57.951

Group 1R

Mean

72.67

265.67

25.93

181.67

SD

19.323

84.563

3.971

42.345

Group 4R

Mean

57.20

173.20

23.80

138.80

SD

12.238

71.867

3.632

33.738

HEMATOLOGY SUMMARY – MALES

Week -04

 

RBC

Hb

PCV

MCV

MCH

MCHC

Reticulocyte

x106Cells/µL

g/dL

%

fL

pg

g/dL

%

Group 1

Mean

8.87

15.92

49.80

56.14

17.96

31.96

1.84

SD

0.245

0.438

1.510

0.550

0.336

0.422

0.036

Group 2

Mean

8.80

15.90

49.28

56.00

18.08

32.28

1.79

SD

0.223

0.424

1.008

0.758

0.327

0.327

0.036

Group 3

Mean

8.54

14.94

47.06@

55.10

17.50

31.76

1.86

SD

0.151

0.365

1.011

0.616

0.265

0.251

0.045

Group 4

Mean

8.73

15.20

48.14

55.20

17.52

31.74

1.86

SD

0.517

0.560

2.142

1.525

0.712

0.541

0.070

@Significant at p ≤ 0.05 level with group 1

HEMATOLOGY SUMMARY – MALES

Week -04

 

PLT

WBC

DC (%)

PT

APTT

X103cells/µl

X103cells/ µl

Neut

Lymph

Mono

Eos

Baso

Sec

Sec

Group 1

Mean

947.20

19.92

13.00

83.40

2.60

1.00

0.00

16.40

44.00

SD

114.220

5.308

2.550

2.191

0.548

0.000

0.000

0.548

1.581

Group 2

Mean

884.20

17.16

13.20

83.00

2.80

1.00

0.00

17.00

44.80

SD

126.476

3.421

2.588

2.739

0.837

0.000

0.000

1.000

1.483

Group 3

Mean

960.40

17.46

15.20

81.20

2.40

1.20

0.00

16.20

44.00

SD

108.367

6.159

2.280

2.168

0.548

0.447

0.000

1.095

2.000

Group 4

Mean

766.20#

18.40

15.00

81.20

2.80

1.00

0.00

16.40

43.00

SD

72.651

6.499

2.550

2.588

0.447

0.000

0.000

1.140

2.121

#Significant at p ≤ 0.05 level with group 1

 

HEMATOLOGY SUMMARY – MALES

Week -06

 

RBC

Hb

PCV

MCV

MCH

MCHC

Reticulocyte

x106Cells/µL

g/dL

%

fL

pg

g/dL

%

Group 1R

Mean

9.32

15.78

49.42

53.04

16.95

31.94

1.79

SD

0.610

0.726

2.685

1.081

0.417

0.513

0.038

Group 4R

Mean

9.13

15.72

49.54

54.36

17.26

31.74

1.87

SD

0.589

0.349

1.638

2.813

0.956

0.378

0.068

HEMATOLOGY SUMMARY – MALES

Week -06

 

PLT

WBC

DC (%)

PT

APTT

x103cells/µL

x103cells/ µL

Neut

Lymph

Mono

Eos

Baso

Sec

Sec

Group 1R

Mean

982.60

18.00

13.80

81.80

3.00

1.40

0.00

15.80

42.40

SD

26.293

2.055

2.588

3.033

0.000

0.548

0.000

0.837

2.074

Group 4R

Mean

812.00#

14.62

16.20

79.20

2.80

1.60

0.20

16.20

43.40

SD

63.589

5.286

3.962

4.817

0.447

0.548

0.447

1.304

2.408

#Significant at p ≤ 0.05 level with group 1

HEMATOLOGY SUMMARY – FEMALES

Week -04

 

RBC

Hb

PCV

MCV

MCH

MCHC

Reticulocyte

x106Cells/µL

g/dL

%

fL

pg

g/dL

%

Group 1

Mean

8.37

15.76

47.10

56.38

18.94

33.48

1.82

SD

0.512

0.439

1.996

2.519

0.802

0.614

0.029

Group 2

Mean

8.31

15.82

47.08

56.66

19.04

33.60

1.79

SD

0.335

0.460

1.835

1.016

0.451

0.561

0.059

Group 3

Mean

7.56@

14.88

43.48@

57.58

19.70

34.26

1.85

SD

0.397

0.342

1.572

1.171

0.704

0.581

0.076

Group 4

Mean

8.00

15.50

45.38

56.84

19.42

34.16

1.78

SD

0.582

0.620

2.213

1.504

0.698

0.451

0.087

@Significant at p ≤ 0.05 level with group 1,

HEMATOLOGY SUMMARY – FEMALES

Week -04

 

PLT

WBC

DC (%)

PT

APTT

X103cells/µl

X103cells/ µl

Neut

Lymph

Mono

Eos

Baso

Sec

Sec

Group 1

Mean

990.80

23.08

12.60

83.20

2.80

1.40

0.00

16.40

43.60

SD

85.920

2.772

2.074

2.168

0.447

0.548

0.000

1.140

1.817

Group 2

Mean

965.00

15.74

14.00

81.40

3.00

1.60

0.00

16.00

43.00

SD

109.968

4.039

2.550

2.510

0.000

0.548

0.000

0.707

2.236

Group 3

Mean

948.20

20.88

11.40

84.40

3.00

1.20

0.00

15.80

42.40

SD

117.647

7.134

4.450

6.189

1.732

0.447

0.000

0.837

1.673

Group 4

Mean

866.20

17.62

19.60

76.60

2.60

1.20

0.00

16.80

43.40

SD

96.650

3.259

18.501

18.188

0.548

0.447

0.000

1.095

1.140

 

HEMATOLOGY SUMMARY – FEMALES

Week -06

 

RBC

Hb

PCV

MCV

MCH

MCHC

Reticulocytes

x106Cells/µL

g/dL

%

fL

pg

g/dL

%

Group 1R

Mean

8.52

15.04

45.68

53.64

17.66

32.92

1.81

SD

0.223

0.351

1.052

1.754

0.709

0.482

0.054

Group 4R

Mean

8.58

15.28

46.72

54.46

17.80

32.72

1.83

SD

0.381

0.614

2.087

0.483

0.265

0.526

0.013

 

 

PLT

WBC

DC (%)

PT

APTT

x103cells/µL

x103cells/ µL

Neut

Lymph

Mono

Eos

Baso

Sec

Sec

Group 1R

Mean

803.60

21.78

14.40

81.20

3.20

1.20

0.00

16.20

42.40

SD

105.296

1.942

2.510

3.271

0.447

0.447

0.000

0.837

2.510

Group 4R

Mean

1012.80#

13.32#

22.20

73.80

2.80

1.20

0.00

15.60

41.60

SD

113.601

4.966

11.054

10.710

0.447

0.447

0.000

0.548

2.408

#Significant at p≤0.05 level with grou

CLINICAL BIOCHEMISTRY SUMMARY – MALES

WEEK -04

 

GLU

Urea

CREA

BLI

ALT

AST

ALP

TPO

mmol /L

mmol /L

µmol /L

µmol /L

U/L

U/L

U/L

g/L

Group 1

Mean

102.40

42.00

0.70

0.08

57.80

128.80

352.80

7.24

SD

10.78

6.60

0.00

0.01

7.40

6.06

41.15

0.17

Group 2

Mean

100.60

39.00

0.68

0.10!

54.60

133.60

357.00

7.22

SD

12.10

4.30

0.04

0.01

12.12

8.65

76.78

0.16

Group 3

Mean

102.80

45.60

0.66

0.08

56.40

128.40

442.80

7.18

SD

6.98

7.20

0.05

0.01

8.35

12.70

42.53

0.23

Group 4

Mean

95.60

41.20

0.66

0.10#

60.40

150.80#

411.60

7.50

SD

12.78

5.89

0.05

0.01

6.35

11.65

88.60

0.28

#Significant at p0.05 level with group 1.

CLINICAL BIOCHEMISTRY SUMMARY – MALES

WEEK -04

 

ALB

GLB

CHOL

TRIGL

Na

K

Cl

GGT

A/G

g/L

g/L

mmol /L

mmol /L

mmol /L

mmol /L

mmol /L

U/L

_

Group 1

Mean

2.33

4.92

60.60

69.40

139.22

4.85

98.20

2.20

0.48

SD

0.03

0.19

5.55

14.86

0.79

0.20

1.12

0.64

0.03

Group 2

Mean

2.41

4.82

52.00

71.20

138.80

4.89

99.64

1.74

0.50

SD

0.10

0.22

14.35

20.73

0.43

0.29

1.17

1.30

0.03

Group 3

Mean

2.31

4.88

57.40

62.60

139.42

32.14

80.96

21.64

0.47

SD

0.06

0.20

7.57

10.06

1.32

60.86

42.44

44.04

0.02

Group 4

Mean

2.27

5.24

57.60

80.40

139.70

4.74

99.78

2.84

0.43#

SD

0.11

0.31

9.21

22.68

0.75

0.16

0.73

2.34

0.04

#Significant at p0.05 level with group 1.

CLINICAL BIOCHEMISTRY SUMMARY – MALES - RECOVERY

WEEK -06

 

GLU

Urea

CREA

BLI

ALT

AST

ALP

TPO

mmol /L

mmol /L

µmol /L

µmol /L

U/L

U/L

U/L

g/L

Group 1R

Mean

115.20

42.20

0.62

0.09

55.20

113.20

404.20

7.46

SD

16.16

5.50

0.08

0.02

4.71

7.36

122.76

0.25

Group 4R

Mean

116.80

38.80

0.66

0.08

46.80#

119.00

214.80#

7.26

SD

19.14

5.97

0.05

0.01

5.45

4.30

48.53

0.30

 

 

ALB

GLB

CHOL

TRIGL

Na

K

Cl

GGT

A/G

g/L

g/L

mmol /L

mmol /L

mmol /L

mmol /L

mmol /L

U/L

_

Group 1R

Mean

2.23

5.24

49.40

63.60

140.64

4.98

100.84

2.12

0.42

SD

0.15

0.21

9.63

15.84

0.82

0.08

1.47

1.31

0.03

Group 4R

Mean

2.21

5.04

47.20

51.80

140.72

4.98

101.32

1.02

0.47

SD

0.17

0.23

12.91

24.26

1.02

0.40

1.56

0.58

0.07

#Significant at p ≤ 0.05 level with group 1

CLINICAL BIOCHEMISTRY SUMMARY – FEMALES

WEEK -04

 

GLU

Urea

CREA

BLI

ALT

AST

ALP

TPO

mmol /L

mmol /L

µmol /L

µmol /L

U/L

U/L

U/L

g/L

Group 1

Mean

90.00

47.60

0.74

0.11

56.60

134.40

272.00

7.34

SD

4.69

8.73

0.05

0.03

10.45

11.26

93.73

0.15

Group 2

Mean

89.00

47.60

0.74

0.10

55.80

143.80

261.20

7.70

SD

10.56

4.83

0.05

0.01

8.96

27.00

71.51

0.51

Group 3

Mean

101.00

49.40

0.70

0.10

53.80

144.20

292.60

7.50

SD

9.54

9.66

0.00

0.03

6.34

15.30

85.39

0.22

Group 4

Mean

89.20

50.00

0.76

0.10

57.60

139.00

327.60

7.28

SD

17.22

4.74

0.13

0.02

3.78

15.10

38.42

0.24

 

CLINICAL BIOCHEMISTRY SUMMARY – FEMALES

WEEK -04

 

ALB

GLB

CHOL

TRIGL

Na

K

Cl

GGT

A/G

g/L

g/L

mmol /L

mmol /L

mmol /L

mmol /L

mmol /L

U/L

_

Group 1

Mean

2.44

4.90

68.20

80.40

139.32

4.54

99.78

2.44

0.49

SD

0.09

0.16

7.43

27.21

0.63

0.35

1.57

2.01

0.03

Group 2

Mean

2.45

5.26

81.40

64.20

140.28

4.49

101.34

1.72

0.46

SD

0.14

0.40

12.50

21.25

1.70

0.30

2.53

1.60

0.02

Group 3

Mean

2.36

5.16

86.20

61.40

140.00

4.83

101.80

2.68

0.45

SD

0.09

0.26

12.05

11.87

1.09

0.09

1.14

1.31

0.03

Group 4

Mean

2.27

5.02

74.80

68.60

139.76

4.50

101.98

2.36

0.46

SD

0.31

0.56

11.65

15.63

1.11

0.34

1.55

1.24

0.10

 

CLINICAL BIOCHEMISTRY SUMMARY – FEMALES - RECOVERY

WEEK -06

 

GLU

Urea

CREA

BLI

ALT

AST

ALP

TPO

mmol /L

mmol /L

µmol /L

µmol /L

U/L

U/L

U/L

g/L

Group 1R

Mean

111.80

35.60

0.64

0.12

50.60

123.40

308.60

7.20

SD

8.35

5.68

0.05

0.06

7.33

8.35

84.15

0.25

Group 4R

Mean

136.20#

43.60

0.76#

0.09

56.60

130.40

176.40#

7.48

SD

19.77

7.50

0.05

0.02

5.22

12.28

44.00

0.54

 

 

ALB

GLB

CHOL

TRIGL

Na

K

Cl

GGT

A/G

g/L

g/L

mmol /L

mmol /L

mmol /L

mmol /L

mmol /L

U/L

_

Group 1R

Mean

2.18

5.02

57.80

64.80

139.06

4.53

100.90

1.16

0.43

SD

0.20

0.22

9.88

12.07

0.83

0.17

0.49

0.90

0.05

Group 4R

Mean

2.46

5.00

61.20

41.40

140.74#

4.60

102.44#

1.18

0.49

SD

0.20

0.37

14.32

30.22

0.81

0.21

1.18

1.00

0.03

  #Significant at p ≤0.05 level with group 1R

URINANALYSIS – SUMMARY MALES

TREATMENT (Week 4)

 

 

pH

SPECIFIC GRAVITY

VOLUME

(pH)

(SG)

mL

Group 1

Mean

7.40

1.010

12.40

SD

1.02

0.01

1.67

Group 2

Mean

6.80

1.017

12.40

SD

0.45

0.01

1.67

Group 3

Mean

7.40

1.007

10.40

SD

0.55

0.01

1.67

Group 4

Mean

6.90

1.020

11.00

SD

0.55

0.01

1.41

 

RECOVERY (Week 6)

 

 

pH

SPECIFIC GRAVITY

VOLUME

(pH)

(SG)

mL

Group 1R

Mean

7.20

1.011

12.40

SD

0.76

0.01

1.67

Group 4R

Mean

7.00

1.013

12.40

SD

0.71

0.01

1.67

 

 

 

URINANALYSIS – SUMMARY FEMALES

TREATMENT (Week 4)

           

 

pH

SPECIFIC GRAVITY

VOLUME

(pH)

(SG)

mL

Group 1

Mean

6.90

1.015

9.60

SD

0.55

0.01

1.67

Group 2

Mean

6.90

1.017

10.00

SD

0.55

0.01

1.67

Group 3

Mean

6.90

1.015

10.80

SD

0.55

0.01

2.28

Group 4

Mean

7.00

1.016

9.60

SD

0.71

0.01

1.14

 

RECOVERY (Week 6)

           

 

pH

SPECIFIC GRAVITY

VOLUME

(pH)

(SG)

mL

Group 1R

Mean

7.50

1.009

10.00

SD

0.00

0.00

1.67

Group 4R

Mean

6.90

1.011

11.40

SD

0.55

0.01

1.95

ORGAN WEIGHTS (GRAM) – SUMMARY MALES

 

Group 1

Group 2

Group 3

Group 4

Group 1R

Group 4R

Body Weight (G)

Mean

188.0

187.8

190.0

190.4

225.5

226.6

SD

1.98

1.75

2.94

3.41

2.64

2.26

N

5

5

5

5

5

5

Brain

 

Mean

1.5208

1.7276

1.6722

1.7332

1.7308

1.7912

SD

0.22996

0.18949

0.20302

0.16879

0.0703

0.1121

N

5

5

5

5

5

5

Heart

Mean

1.3622

1.1748

1.1898

1.1592

1.5672

1.4318

SD

0.55108

0.13315

0.08900

0.19831

0.6597

0.2005

N

5

5

5

5

5

5

Liver

Mean

10.7768

11.9598

12.3826

11.6450

12.2516

12.1372

SD

0.96180

1.42941

1.38330

2.27308

1.9339

1.6098

N

5

5

5

5

5

5

Spleen

Mean

0.8832

0.8862

0.9058

1.0700

0.9642

0.8568

SD

0.15267

0.18838

0.23423

0.22823

0.2554

0.1664

N

5

5

5

5

5

5

 

Kidneys

 

Mean

1.9130

2.3662

2.2266

2.1862

2.3560

2.4162

SD

0.09315

0.31861

0.29819

0.35972

0.2038

0.2492

N

5

5

5

5

5

5

ORGAN WEIGHTS (GRAM) – SUMMARY MALES (CONTD.)

 

Group 1

Group 2

Group 3

Group 4

Group 1R

Group 4R

Adrenals

Mean

0.0786

0.0736

0.0730

0.0846

0.0900

0.0762

SD

0.01876

0.02028

0.01166

0.02758

0.0294

0.0199

N

5

5

5

5

5

5

Testes

Mean

2.6902

2.5696

2.5204

2.7464

2.4740

2.5856

SD

0.43755

0.28320

0.25850

0.18720

0.1792

0.1397

N

5

5

5

5

5

5

Epididymes

Mean

1.2890

1.1456

1.2376

1.3560

1.5204

1.2406

SD

0.18165

0.09788

0.21020

0.17377

0.3331

0.1656

N

5

5

5

5

5

5

Thymus

Mean

1.0384

0.8776

1.2882

1.0558

1.2958

1.6642

SD

0.32879

0.21025

0.25352

0.19113

0.3700

0.5540

N

5

5

5

5

5

5

Pro + Sem. Vesical with Coagulating glands

Mean

0.5738

0.4596

0.4090

0.6230

0.4750

0.4074

SD

0.24899

0.14134

0.11354

0.63532

0.1850

0.1806

N

5

5

5

5

5

5

ORGAN WEIGHTS (GRAM) – SUMMARY FEMALES

 

Group 1

Group 2

Group 3

Group 4

Group 1R

Group 4R

Body Weight (G)

Mean

183.6

183.6

185.9

184.7

222.7

227.2

SD

3.08

3.56

3.72

3.85

2.59

2.61

N

5

5

5

5

5

5

Brain

 

Mean

1.5868

1.6318

1.7080

1.6560

1.5964

1.6218

SD

0.11644

0.18147

0.16832

0.11120

0.1598

0.1828

N

5

5

5

5

5

5

Heart

Mean

0.7894

0.8420

0.8842

1.0262#

0.8322

0.9012

SD

0.07433

0.07927

0.08066

0.19578

0.0669

0.1068

N

5

5

5

5

5

5

Liver

Mean

7.5688

6.8344

8.5234

8.9648

7.3988

7.1562

SD

1.39591

0.66916

1.24858

0.70799

1.0276

0.6172

N

5

5

5

5

5

5

Spleen

Mean

0.8416

0.6292

0.7840

0.8542

0.6492

0.6170

SD

0.23081

0.07286

0.20496

0.18374

0.1478

0.1407

N

5

5

5

5

5

5

 

Kidneys

 

Mean

1.4710

1.4272

1.5918

1.7734#

1.5310

1.5238

 

SD

0.15456

0.10609

0.20817

0.15517

0.1065

0.0870

 

N

5

5

5

5

5

5

 

#Significant at p  0.05 level with group 1.

ORGAN WEIGHTS (GRAM) – SUMMARY FEMALES (CONTD.)

 

Group 1

Group 2

Group 3

Group 4

Group 1R

Group 4R

Adrenals

Mean

0.0888

0.0738

0.1058

0.1138

0.0840

0.0864

SD

0.01925

0.01375

0.01791

0.01671

0.0354

0.0100

N

5

5

5

5

5

5

Ovaries

Mean

0.4436

0.4368

0.4282

0.3584

0.2512

0.4246

SD

0.24545

0.14870

0.06111

0.08873

0.0887

0.1754

N

5

5

5

5

5

5

Uterus

Mean

0.1386

0.2108

0.2110

0.1978

0.2238

0.1784

SD

0.03879

0.19715

0.11964

0.04727

0.1753

0.0963

N

5

5

5

5

5

5

Thymus

Mean

0.5072

0.3848

0.4190

0.4840

0.3182

0.3098

SD

0.23162

0.09232

0.08428

0.05829

0.1121

0.0822

N

5

5

5

5

5

5

 

ORGAN WEIGHT RATIO – SUMMARY

ORGAN WEIGHTS RATIO (%) – SUMMARY MALES

 

Group 1

Group 2

Group 3

Group 4

Group 1R

Group 4R

Body Weight (G)

Mean

188.0

187.8

190.0

190.4

225.5

226.6

SD

1.98

1.75

2.94

3.41

2.64

2.26

N

5

5

5

5

5

5

Brain

 

Mean

0.8097

0.9197

0.8811

0.9105

0.7677

0.7905

SD

0.12965

0.09784

0.11473

0.08662

0.0325

0.0519

N

5

5

5

5

5

5

Heart

Mean

0.7234

0.6257

0.6267

0.6093

0.6970

0.6318

SD

0.28820

0.07217

0.05489

0.10704

0.3007

0.0888

N

5

5

5

5

5

5

Liver

Mean

5.7312

6.3700

6.5133

6.1234

5.4337

5.3526

SD

0.50647

0.76796

0.67993

1.22570

0.8610

0.6795

N

5

5

5

5

5

5

Spleen

Mean

0.4699

0.4724

0.4760

0.5639

0.4273

0.3778

SD

0.08238

0.10296

0.12012

0.12963

0.1112

0.0713

N

5

5

5

5

5

5

ORGAN WEIGHTS RATIO (%) – SUMMARY MALES (CONTD.)

 

Group 1

Group 2

Group 3

Group 4

Group 1R

Group 4R

Kidneys

 

Mean

1.0176

1.2608

1.1713

1.1494

1.0450

1.0658

SD

0.05456

0.17507

0.15281

0.19641

0.0917

0.1047

N

5

5

5

5

5

5

Adrenals

Mean

0.0417

0.0393

0.0384

0.0445

0.0400

0.0336

SD

0.00971

0.01104

0.00616

0.01438

0.0133

0.0087

N

5

5

5

5

5

5

Testes

 

Mean

1.4320

1.3682

1.3275

1.4429

1.0976

1.1409

SD

0.24391

0.14935

0.14716

0.09710

0.0841

0.0605

N

5

5

5

5

5

5

Epididymes

Mean

0.6850

0.6099

0.6505

0.7128

0.6751

0.5476

SD

0.09126

0.04943

0.10455

0.09461

0.1519

0.0745

N

5

5

5

5

5

5

Thymus

Mean

0.5532

0.4668

0.6775

0.5538

0.5738

0.7345

SD

0.17793

0.10902

0.13025

0.09314

0.1594

0.2452

N

5

5

5

5

5

5

Pro + Sem. Vesical with Coagulating glands

Mean

0.3049

0.2449

0.2147

0.3284

0.2107

0.1796

SD

0.13065

0.07613

0.05707

0.33245

0.0826

0.0790

N

5

5

5

5

5

5

ORGAN WEIGHTS RATIO (%) – SUMMARY FEMALES

 

Group 1

Group 2

Group 3

Group 4

Group 1R

Group 4R

Body Weight (G)

Mean

183.6

183.6

185.9

184.7

222.7

227.2

SD

3.08

3.56

3.72

3.85

2.59

2.61

N

5

5

5

5

5

5

Brain

 

Mean

0.8645

0.8900

0.9186

0.8970

0.7163

0.7142

SD

0.06122

0.10638

0.08771

0.06611

0.0652

0.0832

N

5

5

5

5

5

5

Heart

Mean

0.4300

0.4584

0.4756

0.5562

0.3737

0.3971

SD

0.03948

0.03897

0.04171

0.11063

0.0302

0.0508

N

5

5

5

5

5

5

Liver

Mean

4.1145

3.7229

4.5865

4.8520

3.3202

3.1529

SD

0.69366

0.35077

0.68639

0.35283

0.4416

0.3069

N

5

5

5

5

5

5

Spleen

Mean

0.4570

0.3428

0.4212

0.4612

0.2914

0.2720

SD

0.11788

0.03867

0.10886

0.09231

0.0654

0.0649

N

5

5

5

5

5

5

 

Kidneys

 

Mean

0.8006

0.7783

0.8574

0.9602

0.6875

0.6711

SD

0.07398

0.06938

0.12291

0.08426

0.0484

0.0439

N

5

5

5

5

5

5

 

ORGAN WEIGHTS RATIO (%) – SUMMARY FEMALES (CONTD.)

 

Group 1

Group 2

Group 3

Group 4

Group 1R

Group 4R

Adrenals

Mean

0.0484

0.0403

0.0568

0.0616

0.0376

0.0381

SD

0.01034

0.00790

0.00892

0.00924

0.0155

0.0047

N

5

5

5

5

5

5

ovaries

Mean

0.2401

0.2373

0.2302

0.1945

0.1130

0.1864

SD

0.12868

0.07796

0.03161

0.04990

0.0406

0.0761

N

5

5

5

5

5

5

Uterus

 

Mean

0.0753

0.1152

0.1131

0.1071

0.1002

0.0788

SD

0.02016

0.10839

0.06295

0.02551

0.0781

0.0435

N

5

5

5

5

5

5

 

Thymus

 

Mean

0.2766

0.2096

0.2254

0.2619

0.1426

0.1365

SD

0.12572

0.04993

0.04549

0.03004

0.0494

0.0367

N

5

5

5

5

5

5

SUMMARY OF GROSS PATHOLOGY

           Male

Gross observation

Group

control

(G1)

Low dose

(G2)

Intermediate dose(G3)

High dose

(G4)

External

NAD

5/5

5/5

5/5

5/5

Internal

NAD

5/5

5/5

5/5

5/5

             Female

Gross observation

Group

control

(G1)

Low dose

(G2)

Intermediate dose(G3)

High dose

(G4)

External

NAD

5/5

5/5

5/5

5/5

Internal

NAD

5/5

5/5

5/5

5/5

      Key: M – Male; F – Female; No. of animals showing observation/Total no. of animals.

SUMMARY OF HISTOPATHOLOGY

ORGAN

OBSERVATIONS

Control

(G1)

High dose (G4)

M

F

M

F

Liver

Sinusoidal Haemorrhage

1/5

2/5

3/5

0/5

Foci of centrilobular/ Periportalnecrosis//Inflammation

1/5

2/5

2/5

1/5

No Abnormality Detected (NAD)

3/5

1/5

2/5

4/5

Lungs

Alveolar Haemorrhage

1/5

0/5

1/5

0/5

Alveolar wall Thickening /Peribronchiolar Lymphoid tissue hyperplastia/ Alveolar inflammation

2/5

2/5

2/5

1/5

No Abnormality Detected (NAD)

2/5

3/5

2/5

4/5

Spleen

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Heart

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Aorta

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Kidneys

Tubular /interstitial inflammation

1/5

3/5

1/5

3/5

No Abnormality Detected (NAD)

4/5

2/5

4/5

2/5

Adrenals

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Brain

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Pituitary

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Thyroid andthyroid

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Trachea

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Oesophagus

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Jejunum

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Duodenum

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

         Key: M – Male; F – Female; No. of animals showing observation/Total no. of animals.

           SUMMARY OF HISTOPATHOLOGY (Contd.,)

ORGAN

OBSERVATIONS

Control

(G1)

High dose (G4)

M

F

M

F

Ileum

Submucosal lymphoid tissue hyperplastia

1/5

1/5

0/5

0/5

No Abnormality Detected (NAD)

4/5

4/5

5/5

5/5

Colon

Sub mucosal lymphoid tissue hyperplasia

2/5

0/5

1/5

0/5

No Abnormality Detected (NAD)

3/5

5/5

4/5

5/5

Rectum

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Caecum

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Lymph nodes

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Pancreas

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Thymus

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Urinary bladder

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Testes

No Abnormality Detected (NAD)

5/5

-

5/5

-

Epididymis

No Abnormality Detected (NAD)

5/5

-

5/5

-

Prost+Seminal Vesicles+coagula

Ting glands

No Abnormality Detected (NAD)

5/5

-

5/5

-

Ovary

No Abnormality Detected (NAD)

-

5/5

-

5/5

Uterus

No Abnormality Detected (NAD)

-

5/5

-

5/5

Cervix+Vagina

No Abnormality Detected (NAD)

-

5/5

-

5/5

Stomach

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Skeletal muscle

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Sciatic nerve

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Eye

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Spinal cord

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

Sternum

No Abnormality Detected (NAD)

5/5

5/5

5/5

5/5

         Key: M – Male; F – Female; No. of animals showing observation/Total no. of animals.

 

                   

Conclusions:
The toxicity of the registered substance following repeated oral exposure was tested in a 28-day study (OECD 407) using male and female Wistar rats. No treatment related adverse effect was observed. The study-derived NOAEL was 1000 mg/kg bw/day.
Executive summary:

The study was performed according to OECD 407 and GLP. The test substance was given by oral gavage to 5 rats per sex per dose level at 0 (distilled water), 250, 500 and 1000 mg/kg bw/day. In addition, recovery groups treated with 0 (distilled water) or 1000 mg/kg bw/day were included to study the reversibility of treatment-related effects (if any). Male and female Wistar rats of the treatment groups were dosed for 28 days, rats in the recovery groups were allowed to recover for 14 days after the final dose. The observation and examinations performed in the study were mortality, clinical signs, detailed clinical signs, body weight, food consumption, sensory reactivity, assessment of grip strength, motor activity assessment, ophthalmoscopy, clinical pathology (hematology and clinical chemistry), urinalysis, gross pathology at termination, organ weight, and histopathology on the preserved organs and tissues. Results: All animals survived to planned death, and there were no clinical signs of toxicity. No test item-related changes were noted in body weight or body weight gain and feed consumption of both sexes from any of the dose levels during the experimental period. No significant changes in sensory reactivity, grip strength, or motor activity were observed in any of the animals during treatment or recovery. No abnormalities were observed during the ophthalmological examinations. No treatment-related adverse effect was observed on haematological and clinical biochemistry parameters of both sexes from any dose groups. No significant changes in urinalysis were observed in any of the groups. No changes were noted in organ weights of both sexes from any dose levels. No treatment-related histopathological effects were observed at 1000 mg/kg bw/day in the main study. Conclusion: The study-derived NOAEL was 1000 mg/kg bw/day.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Klimisch 1

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

Repeated dose toxicity via oral route:

The study was performed according to OECD 407 and GLP. The test substance was given by oral gavage to 5 rats per sex per dose level at 0 (distilled water), 250, 500 and 1000 mg/kg bw/day. In addition, recovery groups treated with 0 (distilled water) and 1000 mg/kg bw/day were included to study the reversibility of treatment-related effects (if any). Male and female Wistar rats of the treatment groups were dosed for 28 days, rats in the recovery groups were allowed to recover for 14 days after the final dose. The observation and examinations performed in the study were mortality, clinical signs, detailed clinical signs, body weight, food consumption, sensory reactivity, assessment of grip strength, motor activity assessment, ophthalmoscopy, clinical pathology (hematology and clinical chemistry), urinalysis, gross pathology at termination, organ weight, and histopathology on the preserved organs and tissues. Results: All animals survived to planned death, and there were no clinical signs of toxicity. No test item-related changes were noted in body weight or body weight gain and feed consumption of both sexes from any of the dose levels during the experimental period. No significant changes in sensory reactivity, grip strength, or motor activity were observed in any of the animals during treatment or recovery. No abnormalities were observed during the ophthalmological examinations. No treatment-related adverse effect was observed on haematological and clinical biochemistry parameters of both sexes from any dose groups. No significant changes in urinalysis were observed in any of the groups. No changes were noted in organ weights of both sexes from any dose levels. No treatment-related histopathological effects were observed at 1000 mg/kg bw/day in the main study. Conclusion: The study-derived NOAEL was 1000 mg/kg bw/day.

Repeated dose toxicity via inhalation route:

According to Annex IX of the REACH regulation, testing by the inhalation route is appropriate only if exposure of humans via inhalation is likely. The test chemical has a low vapour pressure value; it is reported to be 6.638046e-20 mmHg at 25°C. The particle size distribution of the test substance is in the range of 150-10 micron, which is larger than the inhalable particulate size. Thus, exposure to inhalable dust, mist and vapour of the test chemical is highly unlikely. Therefore, the repeated dose toxicity study via inhalation route is considered for waiver.

Repeated dose toxicity via dermal route:

The acute toxicity value for the test chemical (as provided in section 7.2.3) is >2000 mg/kg body weight. Given the use of the chemical, repeated exposure by the dermal route is unlikely since the use of gloves is common practice in industries. Thus, it is expected that the test chemical does not exhibit toxicity by dermal route after repeated exposure. In addition, there is no data available that suggests that test chemical exhibit repeated dose toxicity by the dermal route. Hence this endpoint was considered for waiver.

Justification for classification or non-classification

The registered substance was given by oral gavage to 5 rats per sex per dose level at 0 (vehicle), 250, 500 and 1000 mg/kg bw/day. The study was performed according to OECD 407 and GLP. No adverse effect was observed. The study-derived NOAEL was 1000 mg/kg bw/day. Considering the results of this OECD 407 study, the registered substance is regarded to be classified as Not Classified for repeated dose toxicity according to Regulation EC 1272/2008.

Categories Display