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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is form Study report

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Principles of method if other than guideline:
To determine the toxicity likely to arise from the repeated exposure of the test chemical over a relatively limited period of time (28 days) in male and female SD rats.
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report):Methyl 2-naphthyl ether (MNE) (Batch no. 0001)
- Molecular formula (if other than submission substance): C11H10O
- Molecular weight (if other than submission substance): 158.19g/mol
- Substance type: Organic
- Physical state: Solid
- Impurities (identity and concentrations):Impurity : 1.34 %

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Central Animal Facility (CAF), NIPER, Sector-67, S.A.S. Nagar, Punjab, India.

- Age at study initiation: 7 to 8 weeks old

- Weight at study initiation: Male 184.38-234.56 g, Female 176.90-208.56 g

- Fasting period before study: No data available

- Housing: Four rats per sex per cage were housed in sterilized solid bottom polypropylene cages with stainless steel grill tops with bedding of clean paddy husk. The cages were suspended on stainless steel racks in a controlled environment.

- Diet (e.g. ad libitum): Standard laboratory sterile extruded pelleted rodent feed (Provimi Animal Nutrition India Pvt. Ltd, Bangalore, India), ad libitum

- Water (e.g. ad libitum): Potable tap water filtered through Reviva Reverse Osmosis System (water filter cum purifier), ad libitum.

- Acclimatization period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3°C
- Humidity (%):30-70 %
- Air changes (per hr): 25 ± 5 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours dark/12 hours light

IN-LIFE DATES:
From: 11.04.2014 (Male), 13.04.2014 (Female);
To: 17.05.2014 (Male), 19.05.2014 (Female)

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: Groundnut oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: 125, 250 and 500 mg was weighed and dissolved in 10 ml of groundnut oil by vortexing the resultant (concentration / dose).

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Groundnut oil
- Concentration in vehicle: 0, 125, 250 and 500 mg/kg body weight/day
- Amount of vehicle (if gavage): 5 ml/kg body weight
- Lot/batch no. (if required): No data available
- Purity: No data available
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Doses were analyzed by using HPLC system.
Duration of treatment / exposure:
28 days
Frequency of treatment:
Daily
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (actual dose received)
Dose / conc.:
125 mg/kg bw/day (actual dose received)
Dose / conc.:
250 mg/kg bw/day (actual dose received)
Dose / conc.:
500 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
Total animals: 56
Control: 7 males, 7 females
125 mg/kg/day: 7males, 7 females
250 mg/kg/day: 7 males, 7 females
500 mg/kg/day: 7 males, 7 females
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: No data available
- Rationale for animal assignment (if not random): Animals were randomized by body weight and sex.
- Rationale for selecting satellite groups: No data available
- Post-exposure recovery period in satellite groups: No data available
- Section schedule rationale (if not random): No data available

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily
- Cage side observations performed: Visual inspection

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily for to characterize the onset and duration of clinical signs. However, detailed physical and clinical observations were conducted before the first exposure of test item and at least once a week thereafter till the end of the study.

BODY WEIGHT: Yes
- Time schedule for examinations: On day 1, 8, 15, 22, 28 and on day 29 before sacrifice.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data available
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data available

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data available

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: Once weekly (on days 2, 9, 16 & 23)

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Fourth week of chemical treatment.
- Dose groups that were examined: All 56 animals were examined.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: On completion of 28 days of treatment and prior to necropsy.
- Anaesthetic used for blood collection: Yes
- Animals fasted: Yes, overnight fasting.
- How many animals: All 56 animals were examined.
- Parameters examined: Haemoglobin (Hb) , RBC Count, Total and differential leucocyte count, Haematocrit (Hct /PCV), Mean corpuscular volume (MCV), Mean corpuscular haemoglobin (MCH), Mean corpuscular haemoglobin concentration (MCHC) and Platelet Count.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: On completion of 28 days of treatment and prior to necropsy.
- Animals fasted: Yes, overnight fasting.
- How many animals: All 56 animals were examined.
- Parameters examined: Sodium, Potassium, Glucose, Total Cholesterol, Blood Urea, Creatinine, Total Protein Albumin, SGPT (Serum glutamic pyruvic transaminase)/ALT, SGOT (Serum glutamic oxaloacetic transaminase)/AST, Hormones analysis (testosterone and estrogen) and Total Bile acids.

URINALYSIS: No data available
- Time schedule for collection of urine: No data available
- Metabolism cages used for collection of urine: No data available
- Animals fasted: No data available
- Parameters examined: No data available

NEUROBEHAVIOURAL EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available
- Battery of functions tested: sensory activity / grip strength / motor activity / other: Yes, Locomotor activity was observed.

OTHER:
Organ weight were measured.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
On completion of treatment, all surviving rats were sacrificed and a complete necropsy was carried out on all animals. Tissues were collected from brain, stomach, large and small intestine, liver, kidneys, adrenal gland(s), spleen, heart, thymus, lungs, testis/ovary, uterus, lymph nodes, peripheral nerve (sciatic), bone marrow, and other gross lesions, if any.
Tissues were preserved in 10% formal saline. However testes, ovaries and uterus were first fixed in Bouin’s fixative for two hours then transferred to 10% formal saline.

HISTOPATHOLOGY: Yes
Histological examination was conducted on tissues/organs from the control and the high-dose group animals Organ examined: lung, liver, kidney, heart, spleen, testis, ovaries, adrenal glands, large and small intestine, brain, sciatic nerve, lymph nodes, bone marrow, stomach and thymus were examined.
Other examinations:
The collected organs were also weighed.
Statistics:
Statistical analysis was carried out by using Microsoft Excel and IBM SPSS statistics version -20. All analyses and comparisons were evaluated at the 5 % level, statistically significant differences (p0.05) indicated by appropriate notation. PAIRED T-Testing procedure was used to check the significance between above mentioned groups. For multiple comparisons Turkey’s HSD test was applied.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
Abnormalities such as nasal discharge, red crust around the nostrils was observedin all treatment groups. In addition, a few cases of snuffling, eye lid swelling and hunched back posture were also observed in the treated groups.
Mortality:
no mortality observed
Description (incidence):
All animals except animal No. 25 & 26 of high dose survived throughout the treatment period.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Treatment with 500 mg/kg body weight/day showed a significant reduction in body weights in male rats on the first, second and fourth week of treatment. In female rats, a significant decrease in body weight was observed on the first, second and third week of treatment.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No difference was observed among the groups.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
No difference in water consumption was observed among the groups.
Ophthalmological findings:
no effects observed
Description (incidence and severity):
No difference was observed in opthalmoscopic examination.
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
In male rats, significant increase in the platelet count was observed along with decrease in MCV and MCH levels at 125 mg/kg body weight/day.

Treatment with 250 mg/kg body weight/day showed significantly decreased values of hemoglobin and MCHC in male rats. The level of MCHC was also found to be significantly decreased when treated with 500 mg/kg body weight/day.

In female rats, treatment with 125 mg/kg body weight/day showed significant alterations in platelet count and MCV.

At 250 mg/kg body weight/day, a significant decrease was noticed in neutrophils along with altered values of MCV in female rats.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
In female animals, the level of total bile acid was decreased at 250 mg/kg body weight/day.

A significant increase was noticed in the level of estrogen in the same group. A similar trend was also noticed at 125 and 500 mg/kg body weight/day., but was statistically non-significant.

At 250 and 500 mg/kg body weight/day the levels of potassium and albumin were found to be significantly increased. The cholesterol level in the 250 mg/kg body weight/day group was noticed to be significantly elevated in the mid-dose group.

In male rats, the level of testosterone was significantly increased after treatment with 500 mg/kg body weight/day.
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
The weight of spleen was decreased in the 250 mg/kg body weight/day groups while treatment with 500 mg/kg body weight/day resulted in the decreased weight of kidney in both female and male rats. However, the absolute organ weight of male animals does not show any significant difference.

In male rats, the relative weight of liver was noticed to be increased at 125 mg/kg body weight/day. Significant increase in the relative weights of heart and testes was also noticed at 500 mg/kg body weight/day.

In female rats, at all dose levels, the relative weight of ovaries was decreased in comparison to the control group animals. At 250 mg/kg body weight/day, the relative weights of brain and heart were significantly decreased. The relative weight of uterus was also decreased in the 125 or 250 mg/kg body weight/day groups.A significant decrease in the relative weight of spleen was observed at250 or 500 mg/kg body weight/day.

The absolute organ weight of ovaries from all treated groups was found to be decreased significantly in female rats.

The weight of uterus was found to be significantly decreased in 250 or 500 mg/kg body weight/day-treated male and female rats.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Nasal discharge, red crust around nostril, perineum wet, bloated stomach, viscous oily secretion was found inside the thoracic cavity.

Small white solid mass slightly firm in consistency floating inside the urinary bladder was observed.

However, due to adaptive metabolic and physiological changes, anoxic/ hypoxic conditions during anesthesia and terminal sacrifice, these findings were considered to be of no toxicological significance.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
No remarkable changes observed in control and the 500 mg/kg body weight/day-treated animals.

A few microscopic findings observed in 500 mg/kg body weight/day-treated animals included collapsed lung with focal inflammation, focal fatty change and excess of lymphocytes in liver, inflammation in small intestine, reactive spleen and excess of mucous in colon. However, these findings were also observed in the control animals.
Histopathological findings: neoplastic:
not specified
Other effects:
not specified

Effect levels

open allclose all
Dose descriptor:
LOAEL
Effect level:
250 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
haematology
clinical biochemistry
organ weights and organ / body weight ratios
other: Effects observed
Dose descriptor:
NOAEL
Effect level:
125 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
water consumption and compound intake
ophthalmological examination
haematology
clinical biochemistry
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
other: No effect observed

Target system / organ toxicity

Critical effects observed:
not specified
System:
other: not specified
Organ:
not specified

Any other information on results incl. tables

SUMMARY OF BODY WEIGHTS(g)

Male

 

 

Group

 

Dose mg/kg

 

 

 

Day1

 

 

Day8

 

 

Day15

 

 

Day22

 

 

Day28

 

Terminal

Day

 

 

1

 

 

Vehicle

Mean

211.80

242.34

262.97

283.01

300.76

296.93

S.D.

14.03

14.91

16.29

24.57

26.41

27.19

 

S.E.M.

6.62

6.15

6.20

8.68

8.78

9.16

 

 

2

 

 

125

Mean

218.02

243.85

260.27

279.35

296.61

291.98

S.D.

9.56

11.42

11.59

14.54

14.88

14.86

 

S.E.M.

4.38

4.68

4.45

5.21

5.02

5.09

 

 

3

 

 

250

Mean

210.57

221.01

241.60

267.42

286.08

282.17

S.D.

9.82

26.98

30.11

23.54

25.05

24.52

 

S.E.M.

4.67

12.21

12.46

8.80

8.76

8.69

 

 

4

 

 

500

Mean

214.35

212.06

223.41

240.97

256.81

256.24

S.D.

10.64

10.08

11.64

13.72

6.86

9.27

 

S.E.M.

4.96

4.76

5.21

5.70

     2.67

3.62

 

SUMMARY OF BODYWEIGHTS(g)

Female

 

 

Group

 

Dose mg/kg

 

 

 

Day1

 

 

Day8

 

 

Day15

 

 

Day22

 

 

Day28

 

Terminal

Day

 

 

1

 

 

Vehicle

Mean

191.17

200.82

208.91

215.94

227.15

219.85

S.D.

7.12

12.39

16.56

15.30

17.64

19.32

 

S.E.M.

3.72

6.17

7.93

7.09

7.77

8.79

 

 

2

 

 

125

Mean

192.17

201.14

208.56

214.38

220.79

216.51

S.D.

9.31

10.16

8.58

7.62

10.76

10.09

 

S.E.M.

4.85

5.05

4.11

3.55

4.87

4.66

 

 

3

 

 

250

Mean

194.94

202.08

209.12

216.24

228.85

226.38

S.D.

9.45

8.89

8.24

7.85

7.51

8.75

 

S.E.M.

4.85

4.40

3.94

3.63

3.28

3.87

 

 

4

 

 

500

Mean

190.49

182.52

192.46

205.74

216.14

212.37

S.D.

8.82

9.74

12.73

11.46

11.61

12.10

 

S.E.M.

4.63

5.34

6.61

5.57

5.37

5.70

SUMMARY OF HEMATOLOGY DATA Male

Group/Dose

 

WBC

3

(x10/

mm3)

RBC

6

(x10/

mm3)

PLT (x10/mm3)

Hct

(%)

Hgb

(g/dl)

MCV(fl)

MCH (pg)

MCHC(g/dl)

Lym

%

Mon

%

Nut

%

Eos

%

 

Bas%

 

 

1/0

 

Mean

12.06

69.49

4.34

23.27

1.17

0.99

10.15

52.77

53.54

16.40

31.07

16.67

600.2

9

±S.D.

2.24

9.56

1.11

9.04

1.07

0.33

0.63

1.40

2.92

0.51

0.97

1.00

76.78

± S.E.M.

18.59

13.76

25.46

38.85

91.51

33.35

6.17

2.65

5.45

3.11

3.12

5.99

12.79

 

 

 

2/125mg/kg

 

Mean

12.09

69.73

4.57

22.04

2.19

0.71

10.16

50.24*

51.00

15.49*

30.83

15.73

1118.

00*

 

±S.D.

2.18

5.30

0.71

6.81

2.70

0.17

0.53

1.93

2.36

0.71

0.83

0.80

383.1

9

±

S.E.M.

0.8

2.0

0.3

2.6

1.0

0.1

0.2

0.7

0.9

0.3

0.3

0.3

144.8

 

 

 

3/250mg/kg

 

Mean

14.97

63.19

4.66

28.04

2.36

0.93

9.93

52.76

52.41

15.81

29.99

15.70

614.8

6

 

±S.D.

2.34

11.26

0.75

12.16

2.83

0.20

0.24

2.50

3.03

0.80

0.18

0.96

173.0

5

±

S.E.M.

0.9

4.3

0.3

4.6

1.1

0.1

0.1

0.9

1.1

0.3

0.1

0.4

65.4

 

 

 

4/ 500mg/kg

 

Mean

9.10

53.17

3.28

16.91

2.29

0.60

7.32

38.41

39.25

11.61

21.62

11.87

476.4

9

 

±S.D.

5.76

31.42

2.26

6.52

1.66

0.35

4.89

25.12

25.52

7.57

14.70

7.70

295.4

8

± S.E.M.

2.6

14.1

1.0

2.9

0.7

0.2

2.2

11.2

11.4

3.4

6.6

3.4

132.1

 

 

SUMMARY OF HEMATOLOGY DATA Female

emaleGroup/Dose

 

WBC (x103/mm3)

RBC (x106/mm3)

PLT (x103/mm3)

 

Hct

(%)

 

Hgb

(g/dl)

 

MCV(fl)

 

MCH (pg)

 

MCHC (g/dl)

 

Lym

%

 

Mon

%

 

Nut

%

 

Eos

%

 

Bas%

 

 

Vehicle

Mean

11.66

62.20

4.41

30.31

1.39

0.87

10.18

49.90

50.74

15.44

30.97

15.71

994.57

±S.D.

2.19

8.62

1.09

8.69

1.61

0.17

0.55

2.34

2.56

0.68

0.31

0.84

520.25

± S..E..M.

 

0.8

 

3.3

 

0.4

 

3.3

 

0.6

 

0.1

 

0.2

 

0.9

 

1.0

 

0.3

 

0.1

 

0.3

 

196.6

 

 

2      /

125mg/kg

 

Mean

 

12.28

 

67.97

 

4.34

 

25.10

 

0.94

 

0.90

 

10.13

 

51.66*

 

52.26

 

15.86

 

30.71

 

16.06

518.71

±S.D.

4.51

6.98

0.78

6.78

0.47

0.18

0.47

1.36

1.39

0.73

0.71

0.45

114.08

±S.E.M.

 

1.7

 

2.6

 

0.3

 

2.6

 

0.2

 

0.1

 

0.2

 

0.5

 

0.5

 

0.3

 

0.3

 

0.2

 

43.1

 

 

3    / 250mg/kg

Mean

11.94

69.24

4.13

21.83*

2.93

1.07

10.18

51.76*

52.76

15.71

30.37

15.99

585.00

±S.D.

3.21

7.96

0.54

7.77

2.71

0.24

0.45

1.55

3.36

0.75

1.44

1.17

213.78

±S.E.M.

 

1.2

 

3.0

 

0.2

 

2.9

 

1.0

 

0.1

 

0.2

 

0.6

 

1.3

 

0.3

 

0.5

 

0.4

 

80.8

 

 

4       /

500mg/kg

Mean

11.37

66.26

4.36

26.57

1.06

1.03

10.00

51.77

51.79

15.79

30.47

15.79

614.29

±S.D.

2.10

9.91

0.99

9.88

1.34

0.19

0.73

0.85

3.90

0.57

0.92

1.41

139.13

±S.E

.M.

 

0.8

 

3.7

 

0.4

 

3.7

 

0.5

 

0.1

 

0.3

 

0.3

 

1.5

 

0.2

 

0.3

 

0.5

 

52.6

 

SUMMARY OF CLINICAL BIOCHEMISTRY DATA Male Rats

Group/Dose

 

Sod.(mmol/L)

Pot.(mmol/L)

ALB(g%)

CHOL(mg%)

CRT(mg%)

SGOT(IU/L)

SGPT (IU/L)

GLU(mg%)

TP(g%)

BUN(mg%)

TBA

Test

ng/L

nmol/L

 

 

1   /  0

Mean

131.5

7.0

3.9

83.9

0.9

33.1

32.9

117.0

6.5

12.5

490.1

4.363

±S.D.

8.6

0.5

0.3

18.1

0.2

4.0

5.1

7.1

0.4

3.0

44.82

1.02

±S.E.M.

3.2

0.2

0.1

6.8

0.1

1.5

1.9

2.7

0.1

1.1

16.94

0.387

 

 

 

2  /125 mg/kg

Mean

130.0

7.6

3.7

87.6

0.9

33.9

32.5

119.1

6.5

14.4

485.4

5.421

±S.D.

3.3

0.7

0.3

10.7

0.2

8.2

7.6

4.9

0.5

3.0

36.17

1.03

±S.E.M.

1.2

0.3

0.1

4.0

0.1

3.1

2.9

1.9

0.2

1.1

13.67

0.392

 

 

 

 

 

3 /250 mg/kg

Mean

130.1

7.1

3.7

87.7

0.9

65.4

30.0

121.0

6.7

12.1

465.4

4.716

±S.D.

9.8

1.0

0.2

17.4

0.1

90.0

7.0

9.5

0.5

2.8

35.10

1.28

±S.E.M.

3.7

0.4

0.1

6.6

0.1

34.0

2.6

3.6

0.2

1.1

13.26

0.487

 

 

 

4 / 500 mg/kg

 

Mean

 

124.5

 

11.0

 

3.7

 

96.2

 

0.9

 

31.1

 

30.5

 

121.6

 

6.4

 

13.7

485.2

*

 

6.289*

±S.D.

11.1

8.4

0.3

12.1

0.2

7.5

6.4

13.8

0.5

4.0

23.33

0.398

 

±S.E.M.

 

5.0

 

3.8

 

0.1

 

5.4

 

0.1

 

3.3

 

2.8

 

6.2

 

0.2

 

1.8

 

10.43

 

0.178

 

SUMMARY OF CLINICAL BIOCHEMISTRY DATA - Female

 

Group

/Dose

 

Sod.

Pot.

ALB

CHOL

CRT

SGOT

SGPT

GLU

TP

BUN

TBA

Est

(mmol/L)

(mmol/L)

(g%)

(mg%)

(mg%)

(IU/L)

(IU/L)

(mg%)

(g%)

(mg%)

ng/L

ng/L

 

1        /

0 mg/kg

Mean

139.2

7.2

3.3

93.0

1.1

32.3

39.9

110.4

7.0

12.9

485

9.156

±S.D.

9.3

0.6

0.3

14.5

0.2

15.3

21.7

14.2

0.5

4.3

30.60

1.47

±S.E.M.

3.5

0.2

0.1

5.5

0.1

5.8

8.2

5.4

0.2

1.6

11.56

0.558

 

 

2     /

125 mg/kg

Mean

136.6

7.8

3.4

98.0

1.1

28.8

39.6

100.0

7.3

12.5

475.1

9.869

±S.D.

4.3

0.9

0.1

17.1

0.4

7.4

9.0

8.9

0.5

2.1

30.59

1.27

±S.E.M.

1.6

0.3

0.1

6.5

0.1

2.8

3.4

3.4

0.2

0.8

11.56

0.482

 

 

 

 

3     /

250 mg/kg

Mean

141.6

8.1

3.8

119.9

1.1

27.9

40.0

100.2

7.0

12.5

425.5*

10.478*

±S.D.

7.3

0.9

0.4

24.7

0.2

6.0

8.1

9.0

0.6

2.8

52.38

1.33

±S.E.M.

2.8

0.4

0.2

9.3

0.1

2.3

3.0

3.4

0.2

1.1

19.79

0.505

 

 

4     /

500 mg/kg

Mean

137.0

8.1

3.7

113.4

1.1

32.3

43.5

100.1

7.0

14.1

448.5

9.842

±S.D.

5.9

0.8

0.2

23.6

0.2

8.4

9.8

5.8

0.6

2.3

31.16

1.93

 

±S.E.M.

 

2.2

 

0.3

 

0.1

 

8.9

 

0.1

 

3.2

 

3.7

 

2.2

 

0.2

 

0.9

 

13.93

 

0.73

 

SUMMARY OF ABSOLUTE ORGAN WEIGHTS(g) - Male Rats

Group/ Dose

 

 

Brain

 

Thymus

 

Liver

 

Kidneys

 

Adrenals

 

Ovaries

 

Spleen

 

Heart

 

Epididymides

 

 

1

Vehicle

Mean

1.79

0.30

10.72

1.98

0.03

2.79

1.04

0.55

0.88

±S.D.

0.28

0.06

1.48

0.30

0.01

0.27

0.15

0.11

0.09

±S.E.M.

0.11

0.02

0.56

0.11

0.01

0.10

0.05

0.04

0.03

 

 

2

(125) mg/kg

 

Mean

1.89

0.46

11.54

2.09

0.07

2.75

1.05

0.54

0.89

 

±S.D.

0.12

0.16

0.70

0.11

0.08

0.22

0.06

0.13

0.08

±S.E.M.

0.05

0.06

0.26

0.04

0.03

0.08

0.02

0.05

0.03

 

3

(250)mg/kg

 

Mean

1.87

0.32

11.25

2.04

0.04

2.81

1.10

0.63

0.91

 

±S.D.

0.07

0.06

1.96

0.23

0.01

0.14

0.12

0.19

0.10

±S.E.M.

0.02

0.02

0.74

0.09

0.00

0.05

0.05

0.07

0.04

 

 

4

 (500)

mg/kg

Mean

1.79

0.28

10.61

1.93

0.04

2.87

1.03

0.54

0.88

±S.D.

0.07

0.05

0.51

0.17

0.01

0.13

0.06

0.11

0.05

±S.E.M.

0.03

0.02

0.19

0.06

0.00

0.05

0.02

0.04

0.02

 SUMMARY OF ABSOLUTE ORGAN WEIGHTS(g)

 Femalerats

Group/ Dose

 

 

Brain

 

Thymus

 

Liver

 

Kidneys

 

Adrenals

 

Ovaries

 

Spleen

Heart

 

Uterus

1/0

   (mg/kg)

Mean

1.79639

0.69686

8.13916

1.66367

0.0451

0.14289

0.63041

0.79684

0.46711

±S.D.

0.13458

0.547

0.47047

0.18627

0.01172

0.02302

0.11848

0.11455

0.07792

±S.E.M.

0.05087

0.20675

0.17782

0.0704

0.00443

0.0087

0.04478

0.04329

0.02945

 

2/(120)

mg/kg

Mean

1.78147

0.28884

8.15586

1.55246

0.04734

0.08319*

0.63869

0.77013

0.39303

±S.D.

0.11628

0.1304

1.06437

0.16655

0.00937

0.01815

0.24329

0.06459

0.08246

±S.E.M.

0.04395

0.04929

0.40229

0.06295

0.00354

0.00686

0.09196

0.02441

0.03117

 

2/

 

250 mg/kg

Mean

1.6069

0.3289

8.3645

1.566

0.0395

0.0935*

0.4718

0.7185

0.4667*

 

±S.D.

 

0.27569

 

0.12711

 

0.78925

 

0.15522

 

0.01286

 

0.02423

 

0.11176

 

0.06134

 

0.07708

±S.E.M.

0.1042

0.04804

0.29831

0.05867

0.00486

0.00916

0.04224

0.02319

0.02913

 

4  /

(500) mg/kg

Mean

1.75207

0.42151

7.9445

1.525*

0.04407

0.07481

0.49059*

0.70961

0.34757*

±S.D.

0.12692

0.20921

0.63133

0.13336

0.00665

0.01518

0.09178

0.02657

0.10007

±S.E.M.

0.04797

0.07907

0.23862

0.05041

0.00251

0.00574

0.03469

0.01004

0.03782

SUMMARY OF RELATIVE ORGAN WEIGHTS Male Rats

 

Group/Dose

 

 

Brain

 

Thymus

 

Liver

 

Kidneys

 

Adrenals

 

Testes

 

Splee n

 

Heart

 

epididymide s

 

1/

 

0

Mean

0.6051

0.1009

3.6170

0.6680

0.0110

0.9447

0.1846

0.2970

0.3521

±S.D.

 

0.1132

 

0.0236

 

0.4211

 

0.0880

 

0.0052

 

0.1114

 

0.0407

 

0.0210

 

0.0527

±S.E.M.

 

0.0428

 

0.0089

 

0.1592

 

0.0333

 

0.0020

 

0.0421

 

0.0154

 

0.0079

 

0.0199

 

 

 

 

2/

125 mg/kg

Mean

 

0.6466

 

0.1549

 

3.9569*

 

0.7236

 

0.0243

 

0.9377

 

0.1856

 

0.3060

 

0.3599

±S.D.

 

0.0330

 

0.0515

 

0.2711

 

0.0369

 

0.0274

 

0.0434

 

0.0428

 

0.0262

 

0.0299

±S.E.M.

 

0.0125

 

0.0195

 

0.1025

 

0.0139

 

0.0104

 

0.0164

 

0.0162

 

0.0099

 

0.0113

 

 

 

3  /

250 mg/kg

Mean

 

0.6674

 

0.1140

 

3.9886

 

0.7207

 

0.0143

 

1.0007

 

0.2209

 

0.3220

 

0.3883

±S.D.

 

0.0627

 

0.0252

 

0.6023

 

0.0431

 

0.0021

 

0.0816

 

0.0608

 

0.0367

 

0.0346

±S.E.M.

 

0.0237

 

0.0095

 

0.2277

 

0.0163

 

0.0008

 

0.0309

 

0.0230

 

0.0139

 

0.0131

 

 

 

4   /

500 mg/kg

Mean

 

0.5317

 

0.0868

 

3.1877

 

0.5684

 

0.0110

 

0.8401

 

0.1709

 

0.2618*

 

0.3071*

±S.D.

 

0.3033

 

0.0461

 

1.7161

 

0.3359

 

0.0062

 

0.4873

 

0.0898

 

0.1468

 

0.1775

±S.E.M.

 

0.1357

 

0.0206

 

0.7675

 

0.1502

 

0.0028

 

0.2179

 

0.0402

 

0.0656

 

0.0794

SUMMARY OF RELATIVE ORGAN WEIGHTS- Females

Group/Dose

 

 

Brain

Thymu s

 

Liver

 

Kidneys

Adrenal s

 

Ovaries

 

uterus

 

Spleen

 

Heart

 

 

1/0

Mean

 

0.8181

 

0.3176

 

3.7199

 

0.7573

 

0.0200

 

0.0654

 

0.2107

 

0.2849

 

0.3614

±S.D.

0.0380

0.2566

0.3170

0.0667

0.0045

0.0141

0.0202

0.0354

0.0334

±S.E.M.

0.0144

0.0970

0.1198

0.0252

0.0017

0.0053

0.0076

0.0134

0.0126

 

 

2 / 125 mg/kg

Mean

 

0.8227

 

0.1324

 

3.7541

 

113.7454

 

0.0214

 

*0.0380

 

0.1824

 

0.2926

 

0.3557

±S.D.

0.0439

0.0598

0.3280

299.0822

0.0053

0.0090

0.0439

0.1052

0.0323

±S.E.M.

0.0166

0.0226

0.1240

113.0424

0.0020

0.0034

0.0166

0.0398

0.0122

 

3  /          250 mg/kg

Mean

 

0.7137

 

0.1443

 

3.6900

 

0.6917

 

0.0171

 

0.0406*

 

0.2054

 

0.2077*

 

0.3169*

±S.D.

0.1425

0.0553

0.2348

0.0629

0.0053

0.0089

0.0298

0.0487

0.0212

±S.E.M.

0.0539

0.0209

0.0887

0.0238

0.0020

0.0034

0.0113

0.0184

0.0080

 

4    /  500mg/kg

Mean

0.8253

0.1970

3.7417

0.7173

0.0203

0.0349*

0.1636

0.2304*

0.3341

±S.D.

0.0546

0.0920

0.2484

0.0423

0.0028

0.0080

0.0463

0.0405

0.0130

±S.E.M.

0.0206

0.0348

0.0939

0.0160

0.0011

0.0030

0.0175

0.0153

0.0049

Applicant's summary and conclusion

Conclusions:
NOAEL was considered to be 125 mg/kg bw and LOAEL was considered to be 250 mg/kg body weight/day when Sprague Dawley male and female rats were exposed daily to the test chemical by oral route for 28 days.
Executive summary:

In a 28 days repeated dose toxicity study, the effect of the test chemical was evaluated in male and female Sprague Dawley rats. The test chemical was administered by oral gavage in the concentration of 0, 125, 250 or 500 mg/kg body weight/day. 2 animals died at 500 mg/kg bw as compared to control.Abnormalities such as nasal discharge, red crust around the nostrils was observed in all treatment groups. In addition, a few cases of snuffling, eye lid swelling and hunched back posture were also observed in the treated groups. A significant decrease in body weights in male rats on the first, second and fourth week of treatment and in female rats, a significant decrease in body weight was observed on the first, second and third week of treatment were observed as compared to control. No effects on food and water consumption were observed as compared to control. In addition, In hematology, the test chemical resulted in significantly increased platelet count along with decreased MCV, MCH and MCHC levels. Treatment with 250 mg/kg body weight/day resulted in significantly decreased levels of hemoglobin and MCHC in male rats. At 250 mg/kg body weight/day, a significant decrease was noticed in neutrophils along with altered values of MCV in female rats. In clinical chemistry, significantly increased the level of testosterone in the 500 mg/kg body weight/day group in males, as well as it significantly increased the level of estrogen in the 250 mg/kg body weight/day group in females. Significant increased levels in cholesterol, potassium and albumin was also observed. Similarly, Changes in relative and absolute organ weight of spleen, kidney, liver, brain, heart, ovaries and uterus were observed when treated with 125, 250 or 500 mg/kg body weight/day. Nasal discharge, red crust around nostril, perineum wet, bloated stomach, viscous oily secretion was found inside the thoracic cavity. Small white solid mass slightly firm in consistency floating inside the urinary bladder was observed. However, due to adaptive metabolic and physiological changes, anoxic/ hypoxic conditions during anesthesia and terminal sacrifice, these findings were considered to be of no toxicological significance. No remarkable changes observed in control and the 500 mg/kg body weight/day-treated animals. A few microscopic findings observed in 500 mg/kg body weight/day-treated animals included collapsed lung with focal inflammation, focal fatty change and excess of lymphocytes in liver, inflammation in small intestine, reactive spleen and excess of mucous in colon. Since the histopathological changes were not observed in 125 mg/kg bw and these findings were also observed in the control animals. Hence, NOAEL was considered to be 125 mg/kg bw and LOAEL was considered to be 250 mg/kg body weight/day when Sprague Dawley male and female rats were exposed daily to the test chemical by oral route for 28 days.