Registration Dossier

Administrative data

Description of key information

Repeated dose toxicity: Oral

The No Observed Adverse Effect Level (NOAEL) was considered to be 1000 mg/kg/day, when rodents were treated with the given test chemical during repeated dose toxicity study.

 

Repeated dose toxicity: Inhalation

A short-term toxicity study does not need to be conducted because exposure of humans via inhalation in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment. Also, the given test chemical has very low vapor pressure 6.79E-42 mm Hg, so the potential for the generation of inhalable vapors is very low. The normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be highly unlikely and therefore this end point for repeated inhalation toxicity was considered for waiver.

 

Repeated dose toxicity: Dermal

A short-term toxicity study does not need to be conducted because exposure of humans via dermal in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment. Also, the acute dermal toxicity value for 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 test chemical shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no data available that suggests that test chemical shall exhibit repeated dose toxicity by the dermal route. Hence this end point 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:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Remarks:
Experimental data from various test chemicals
Justification for type of information:
Weight of evidence approach based on the available information from various test chemicals.
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Qualifier:
according to guideline
Guideline:
other: As mentioned below
Principles of method if other than guideline:
WoE report is based on 2 repeated dose toxicity studies i.e. WoE-2 and WoE-3.
GLP compliance:
not specified
Limit test:
no
Species:
rat
Strain:
other: 2. Sprague-Dawley 3. Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
2. Rats were caged individually.
3. TEST ANIMALS
- Source: Specified-pathogen-free breeding colony (Olac (1976) Ltd, Bicester, Oxon)
- Age at study initiation: No data available
- Weight at study initiation: 88 to 91 g male, 79 to 83 g female
- Fasting period before study: No data available
- Housing: Animals were housed in stainless steel and polypropylene grid-floored cages at all stages of the study apart from females in the latter stages of pregnancy and lactation where solid-floored cages were used.
- Diet (e.g. ad libitum): Spratt's Laboratory Animal Diet No. 2 (Spratt's Patent, Barking, Essex), ad libitum
- Water (e.g. ad libitum): Tap-water, ad libitum
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22°C
- Humidity (%): 50-70%
- Air changes (per hr): 15-20 air changes/hr with no recirculation.
- Photoperiod (hrs dark / hrs light): No data available

IN-LIFE DATES: From: To: No data available
Route of administration:
oral: feed
Details on route of administration:
2. The test chemical was mixed into a commercial laboratory chow diet for each feeding study.
3. not specified
Vehicle:
other: 2. commercial laboratory chow diet 3. not specified
Details on oral exposure:
2. The test chemical was mixed into a commercial laboratory chow diet for each feeding study.
3. PREPARATION OF DOSING SOLUTIONS: Animals were fed diets containing test chemical at concentrations calculated to provide daily intakes of 0 (control), 50, 250 or 1250 mg/kg body weight.

DIET PREPARATION
- Rate of preparation of diet (frequency): Diets were adjusted at intervals to maintain a constant daily dosage.
- Mixing appropriate amounts with (Type of food): Spratt's Laboratory Animal Diet No. 2
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Spratt's Laboratory Animal Diet No. 2
- Concentration in vehicle: 0, 50, 250 or 1250 mg/kg body weight.
- Amount of vehicle (if gavage): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
2. 2yrs
3. 111 week for male and 112 week for female
Frequency of treatment:
Daily
Remarks:
2. 0, 500, 1000 mg/kg bw
3. Doses / Concentrations: 0, 50, 250 and 1250 mg/kg body weight/day :Nominal dose level (male and female)
Calculated intake of test chemical: male:47,242,1260 mg/kg/day
female:49,246,1260 mg/kg/day
No. of animals per sex per dose:
2. control - 200(100/ sex)
500 mg/kgbw/day - 100 (50/sex)
1000 mg/kgbw/day - 100 (50/sex)
3. Total: 624
0 mg/kg bw /day: 114 male, 114 female
50 mg/kg bw /day: 66 male, 66 female
250 mg/kg bw /day: 66 male, 66 female
1250 mg/kg bw /day: 66 male, 66 female
Control animals:
yes, concurrent vehicle
Details on study design:
2. Two-year tests were run in which 400 rats were distributed into various treatment groups. The test chemical was mixed into a commercial laboratory chow diet for each feeding study.Rats were individually caged. Each animal received in the Pathology Laboratory was given a careful gross examination, and all lesions were recorded. Of those not received, most were discarded because of advanced postmortem autolysis. So far as can be determined, none of the discarded animals had tumors. Lesions which could not clearly be identified as neoplastic or non-neoplastic, and tumors, the type of which was uncertain or uncommon, were also sectioned and examined microscopically. Tissues sectioned include the following: liver, lung, heart, spleen, pancreas, stomach, small in testine, colon, kidney, adrenal, thyroid, testis (or ovary and uterus), leg muscles, leg bones with included marrow, and any tumor or other unusual condition.Sections of the mammary glands were also observed for tumours.
3. No data available
Observations and examinations performed and frequency:
2. The animals were observed for survival, growth, or weights at autopsy of heart, liver, kidney, spleen, or testes.
3. CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily
- Cage side observations checked in table [No.?] were included.: Survival were examined.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly for the first 17 week and then at 2-week intervals for the remainder of the study.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

FOOD EFFICIENCY: No data available
- 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: Twice-weekly

OPHTHALMOSCOPIC EXAMINATION: No data available
- Time schedule for examinations: No data available
- Dose groups that were examined: No data available

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At 3 and 6 months from control and 1250 mg/kg bw/day, from all groups at 12 and 18 months and at the study termination
- Anaesthetic used for blood collection: No data available
- Animals fasted: No data available
- How many animals: From 20 male and 20 female at 3 and 6 months from control and 1250 mg/kg bw/day and from 20 male and 20 female of all treated rats at 12 and 18 months.
- Parameters checked in table [No.?] were examined: Haemoglobin concentration, packed-cell volume, methaemoglobin concentration, total erythrocyte and leucocyte counts were examined.

Reticulocyte and differential leucocyte
counts were examined in 0 and 1250 mg/kg bw/day dose group

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At termination of study
- Animals fasted: No data available
- How many animals: All treated animals were examined.
- Parameters checked in table [No.?] were examined: Content of urea, glucose, albumin and total protein and for the activities of glutamic-oxalacetic transaminase, glutamic-pyruvic transaminase, lactic dehydrogenase and alkaline phosphatase were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: at 3, 6 and 9 months from 0 and 1250 mg/kg bw/day dose group and at 12, 18 and 24 months from all treated groups.
- Metabolism cages used for collection of urine: No data available
- Animals fasted: No data available
- Parameters checked in table [No.?] were examined: Appearance, microscopic constituents, semiquantitative tests for the content of protein, glucose, ketones, bile salts and blood, refractive index, volume of urine and number of cells were examined.

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: No data available

OTHER:
Organ weight: Brain, Heart, Liver, Spleen, Kidneys, Stomach, Full and Empty Caecum, Adrenals and Gonads were weighted.
Sacrifice and pathology:
2. GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
3. GROSS PATHOLOGY: Yes
Gross abnormalities were examined at termination of study. Organs were preserved in 10% buffered formalin. Paraffin-wax sections of all tissues, except nasal bones and spinal cord, from all treatment groups were stained with haematoxylin and eosin and examined by light microscopy. Sections of the nasal bones and spinal cord were prepared in the same manner from just the control animals and those on the highest dietary level.

HISTOPATHOLOGY: Yes
Organ examined: Adrenal glands, brain, caecum (with and without its contents), gonads, heart, kidneys, liver, spleen, stomach and thyroid were weighed. Samples of these tissues together with the aorta, urinary bladder, colon, epididymis, eye, Harderian gland, lungs, lymph nodes, mammary gland, skeletal muscle, nasal bones, nerve, oesophagus, pancreas, prostate, salivary glands, seminal vesicles, skin, small intestine, spinal cord, thymus, tongue, trachea, uterus, vagina and vena cava were examined.
Other examinations:
Viability of pups per litter at day 18 was examined.
Statistics:
2. No data available
3. Statistical analysis were performed by using log-rank analysis for mortality data; Student's t test for body weight, food consumption of F0, water consumption of F0,haematology, urine data, serum analysis, organ weight and relative organ weight data; chi-square test for fertility data; analysis of variance for food consumption of FI, water consumption and pup observation data; test for positive trend using 4 x 2 contingency tables for histology data, tumour incidence and total number of tumour-bearing animals. These latter data were analysed as recommended by Peto et al. (1980) for a scheduled kill. Neither the time at which findings occurred nor their contribution to the cause of death have been taken into account. Where a positive trend was identified, the findings in each treated group were compared with those of the control group.
Clinical signs:
no effects observed
Description (incidence and severity):
2. no effects observed - The ingestion of the test chemical had no effect on survival, growth, or weights at autopsy of heart, liver, kidney, spleen, or testes in the test animals
3. effects observed, treatment-related - When treated wtih 1250 mg/kg bw/day, related pink coloration of the fur and faeces moist faeces were observed in treated rats as compared to control. Dose-related pink coloration of the fur and faeces were observed in treated rats due to contamination with test substance.
Mortality:
no mortality observed
Description (incidence):
2. no mortality observed - The ingestion of the test chemical had no effect on survival, growth, or weights at autopsy of heart, liver, kidney, spleen, or testes in the test animals
3. no mortality observed - No significant effect on survival of treated rats were observed as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
2. no effects observed - The ingestion of the test chemical had no effect on survival, growth, or weights at autopsy of heart, liver, kidney, spleen, or testes in the test animals
3. effects observed, treatment-related - When treated with 1250 mg/kg bw/day, significant decrease in body weight was observed in male rats from week 4 to 73 and in female from 51 and 73 as compared to control. Slight effect on body weight was due to a reduction in the absorption or utilization of the nutrient.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
2. not specified
3. effects observed, treatment-related - When treated with 1250 mg/kg bw/day, increase in food consumption was observed in treated rats were observed as compared to control. Due to a reduction in the absorption or utilization of the nutrient.
Food efficiency:
no effects observed
Description (incidence and severity):
2. not specified
3. no effects observed
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
2. not specified
3. no effects observed - Water consumption: When treated with 1250 mg/kg bw/day, increase in water consumption was observed in treated male rats as compared to control. Increased water loss was observed due to production of softer, moister faeces and a compensatory increase in water intake.
Compound intake: Average compound intake of male rats were 47, 242 and 1250 mg/kg bw/day and for female rats 49, 246 and 1250 mg/kg bw/day.
Ophthalmological findings:
not specified
Haematological findings:
no effects observed
Description (incidence and severity):
2. not specified
3. no effects observed - When treated with 1250 mg/kg bw/day, decreased in packed cell volume was observed at month 6 and 12 months in male and at month 18 in female rats as compared to control. Statistically significant slightly increased haemoglobin concentrations were observed in treated female rats at termination of study as compared to control. Observed effect were not dose related or consistent between the sexes.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
2. not specified
3. effects observed, treatment-related - When treated with 1250 mg/kg bw/day, decrease in glutamic-oxalacetic transaminase activities was observed in male and female rats but not significant in female rats.
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
2. not specified
3. effects observed, treatment-related - When treated with 1250 mg/kg bw/day, Increase level of protein in urine at 12 months in female rats was observed as compared to control. Semi-quantitative analysis of urine for bilirubin and ketones was hindered at months 18 and 24 due to the contamination of the urine with amaranth which interfered with the colour reaction.
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
2. no effects observed - The ingestion of the test chemical had no effect on survival, growth, or weights at autopsy of heart, liver, kidney, spleen, or testes in the test animals
3. effects observed, non-treatment-related - When treated with 1250 mg/kg bw/day, statistically significant increase in absolute and relative full and empty caecum weight were observed in treated male and female rats as compared to control. When treated with 242 mg/kg bw/day, increase in absolute and relative full caecum weight was observed in treated male rats as compared to control. The observed effect was not statistically significant as compared to control.
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
2. effects observed, non-treatment-related - No effect due to ingestion of the test chemical on either tumor incidence or tumor type was noted in Sprague Dawley strain on this two-year experiment.
3. not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
2. effects observed, non-treatment-related - No effect due to ingestion of the test chemical on either tumor incidence or tumor type was noted in Sprague Dawley strain on this two-year experiment.
3. effects observed, treatment-related - Transitional-cell hyperplasia of the bladder, inflammatory cell infiltrate of seminal vesicles and testicular interstitial-cell hyperplasia were observed in 47 mg/kg bw/day male and 1250 mg/kg bw/day treated male and female rats. Statistically significant increase in renal calcification and renal pelvic epithelial hyperplasia, lung oedema and haemorrhage, of lymph-node haemorrhage and degenerative changes in brain and nerve, egenerative and inflammatory changes in heart, inflammatory changes of the thymus, aortic calcification and atrial thrombi were observed in 1250 mg/kg bw/day treated female rats.
Histopathological findings: neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
2. effects observed, non-treatment-related - No effect due to ingestion of the test chemical on either tumor incidence or tumor type was noted in Sprague Dawley strain on this two-year experiment.
3. no effects observed - Statistically significant increase in uterine polyps and vaginal fibromas were observed in 1260 mg/kg bw/day treated female rats. The observed effects were typical of this strain and occur in ageing rats.
Other effects:
not specified
Details on results:
No effect on No. of litters, number of pups per litter at day 18 and pup weight at day 18 were observed in treated rats as compared to control.
Dose descriptor:
NOAEL
Remarks:
2
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical signs
histopathology: neoplastic
histopathology: non-neoplastic
mortality
organ weights and organ / body weight ratios
Dose descriptor:
NOAEL
Remarks:
3
Effect level:
1 250 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
clinical biochemistry
clinical signs
food consumption and compound intake
food efficiency
haematology
histopathology: neoplastic
histopathology: non-neoplastic
mortality
ophthalmological examination
organ weights and organ / body weight ratios
urinalysis
water consumption and compound intake
Critical effects observed:
not specified

2.

Lesions observed in the 2 year feeding study

Parameter

Control

1%

2%

Number of rats started

200

100

100

Number of survivors at 80 weeks

122

66

54

Number received by pathology

147

83

74

Number of males with tumors

14

5

4

Number of females with tumors

24

11

10

Total number of rats with tumors

38

16

14

Tumor type in each rat

 

 

 

Fibroadenoma, mammary                          

21

9

9

Adenocarcinoma

1

1

0

Lymphosarcoma

5

4

4

Lymphoblastoma

4

0

0

Fibrosacroma

3

0

0

Interstitial cell tumor

0

0

0

Endometrial sarcoma

0

0

0

Other tumors

5

2

4

Other pathology

 

 

 

Chronic pneumonia

30

13

13

Granular kidneys

31

9

18

Splenic enlargement

12

9

8

Splenic atrophy

12

3

4

Testicular atrophy

13

4

4

Adrenal enlargement

7

2

7

Chronic arteritis

9

2

5

3.

Mean body weights of rats fed test chemical at dose levels of 0-1250 mg/kg/day

 

 

Mean body weight (g)

 

 

Males

Females

Wk on test

Dose level (mg/kg/day)

0

50

250

1250

0

50

250

1250

0

 

91

88

89

87

83

81

79*

80

4

 

278

274

278

270*

186

185

188

189

8

 

377

378

382

363**

228

229

233

234*

12

 

432

432

439

414"*

255

253

258

258

25

 

515

514

520

488***

286

286

290

283

51

 

593

600

599

558***

352

346

351

327***

73

 

603

617

605

564***

390

394

391

354***

97

 

562

588

576

546

380

399

375

358

109

 

543

538

548

509

370

386

351

337*

The results are expressed as the means for all survivors in each group. Those marked with asterisks differ significantly (Student's t test) from the control value: *P < 0.05; **P < 0.01; ***P < 0.001.

Mean food and water consumption and calculated intake of colouring by rats fed test chemical in the diet at dose levels of 0-1250 mg/kg/day

Nominal dose level (mg/kg/day)

Food intake (g/rat/day)

Water intake (ml/rat/day)

Calculated intake of test chemical (mg/kg/day)

Males

0

20.5

32.4

--

50

20.4

31.4

47

250

20.8

33.2

242

1250

21.8"**

36.3***

1260

Females

0

17.4

33.5

--

50

17.4

33.4

49

250

32.0

32.0

246

1250

18.1"**

36.9

1260

The figures marked with asterisks differ significantly (Student's t test) from that of the controls: ***P < 0.001.

Organ weights and relative organ weights of rats fed test chemical in the diet at dose levels of 0—1250 mg/kg/day for 2 yr

Dose level (mg/kg/day)

No. of rats examined

Absolute and relative organ weights

Terminal body weight (g)

 

 

 

 

 

 

Caecum

 

 

Brain

Heart

Liver

Spleen

Kidneys

Stomach

Full

Empty

Adrenalst

Gonads$

 

 

Organ weight (g)

 

Males

 

 

 

 

 

 

 

 

 

 

 

 

0

19

2.35

1.15

13.99

1.58

4.22

2.29

2.82

1.10

66.3

3.21

522

50

21

2.29

1.44

13.13

1.35*

3.75

2.39

2.78

1.10

70.0

3.28

501

250

16

2.37

1.47

13.58

1.64

3.69

2.20

4.09**

1.20

78.1

2.98

500

1250

18

2.34

1.47

12.92*

1.45

3.79

2.20

8.98***

1.73***

74.1

3.20

468*

Females

 

 

 

 

 

 

 

 

 

 

 

 

0

26

2.17

1.17

10.54

1.13

2.98

1.82

2.71

0.95

49.0

82.5

350

50

20

2.12

1.15

11.04

1.03

2.80

1.84

2.70

0.93

53.4

81.1

361

250

20

2.12

1.10

10.62

1.03

2.76

1.78

2.77

0.96

49.4

72.0

330

1250

13

2.12

1.11

9.85

0.99

2.59

1.61

6.76***

1.31***

55.5

83.3

325

 

 

Relative organ weight (g/100 g bedy weight)

 

Males

 

 

 

 

 

 

 

 

 

 

 

 

0

19

0.46

0.29

2.72

0.31

0.83

0.45

0.55

0.21

12.9

0.62

 

50

21

0.46

0.29

2.62

0.27

0.75

0.48

0.55

0.22

14.2

0.70

 

250

16

0.46

0.30

2.75

0.33

0.75

0.45

0.84**

0.25

15.8

0.60

 

1250

18

0.51*

0.32

2.81

0.31

0.84

0.48

1.91***

0.37***

16.8

0.70

 

Females

 

 

 

 

 

 

 

 

 

 

 

 

0

26

0.63

0.34

3.05

0.33

0.88

0.53

0.80

0.28

14.4

23.8

 

50

20

0.60

0.33

3.12

0.29

0.81

0.53

0.78

0.27

15.1

22.6

 

250

20

0.65

0.34

3.25

0.32

0.86

0.55

0.87

0.30

15.1

21.8

 

1250

13

0.66

0.35

3.06

0.31

0.81

0.50

2.07***

0.41***

17.3

25.8

 

ɫ Absolute and relative weights of this organ are expressed in mg and mg/100 g body weight, respectively.

‡ Absolute and relative weights of female gonads are expressed in mg and mg/100 g body weight, respectively.

Values are the results expressed as mans for the number of rats shown and those marked with asterisks differ significantly the control values: *P < 0.05; **P < 0.01; ***P < 0.001.

Conclusions:
The No Observed Adverse Effect Level (NOAEL) was considered to be 1000 mg/kg/day, when rodents were treated with the given test chemical during repeated dose toxicity study.
Executive summary:

Data available from the various sources was reviewed to determine the toxic nature of the given test chemical. The studies are as mentioned below:

 

Combined Repeated Dose and Carcinogenicity was performed to determine the toxic nature of the test chemical in Sprague Dawley rats. Two-year tests were run in which 400 male and female Sprague Dawley rats were distributed into various treatment groups. The treatment groups were - 100 rats/dose (50/sex) and 100 rats/sex for controls. The test chemical (500 mg/kg bw/day, 1000 mg/kg bw/day) was mixed into a commercial laboratory chow diet for each feeding study. Rats were individually caged. Each animal received in the Pathology Laboratory was given a careful gross examination, and all lesions were recorded. Of those not received, most were discarded because of advanced postmortem autolysis. So far as can be determined, none of the discarded animals had tumors. Lesions which could not clearly be identified as neoplastic or non-neoplastic, and tumors, the type of which was uncertain or uncommon, were also sectioned and examined microscopically. Tissues sectioned include the following: liver, lung, heart, spleen, pancreas, stomach, small intestine, colon, kidney, adrenal, thyroid, testis (or ovary and uterus), leg muscles, leg bones with included marrow, and any tumor or other unusual condition. Sections of the mammary glands were also observed for tumors. The ingestion of the test chemical had no effect on survival, growth, or weights at autopsy of heart, liver, kidney, spleen, or testes in the test animals. No effect due to ingestion of the test chemical on either tumor incidence or tumor type was noted in Sprague Dawley strain on this two-year experiment. Hence, the No Observed Adverse Effect level was considered to be 1000 mg/kg bw/day in both male and female Sprague Dawley rats.

 

The above study is supported with another combined repeated dose & carcinogenicity study conducted in Wistar male and female rats treated with test chemical in the concentration of 0, 50, 250, 1250 mg/kg/day (Nominal dose level) 47, 242 and 1260 mg/kg bw/day (calculated intake) for male and 49, 246 and 1260 mg/kg bw/day (calculated intake) for female orally in fed. No significant effect on survival of treated rats were observed as compared to control. Significant decrease in body weight in male rats from week 4 to 73 and in female rat from 51 and 73 and increase in food consumption was observed in 1260 mg/kg bw/day treated rats as compared to control. Slight effect on body weight and food consumption was due to a reduction in the absorption or utilization of the nutrient. Average compound intake of male rats were 47, 242 and 1260 mg/kg bw/day and for female rats 49, 246 and 1260 mg/kg bw/day. Increase in water consumption was observed in 1260 mg/kg bw/day treated male rats as compared to control. Increased water loss was observed due to production of softer, moister faeces and a compensatory increase in water intake. Similarly, decreased in packed cell volume was observed at month 6 and 12 in male and at month 18 in female rats and slightly increased hemoglobin concentrations in female rats at termination of study and decrease in glutamic-oxalacetic transaminase activities in male and female rats but not significant in female rats were observed at 1260 mg/kg bw/day. Observed effect were not dose related or consistent between the sexes. Increase level of protein in urine at 12 months in female was observed at 1260 mg/kg bw/day and Semi-quantitative analysis of urine for bilirubin and ketones was hindered at months 18 and 24 due to the contamination of the urine with test chemical which interfered with the colour reaction. Statistically significant increase in absolute and relative full and empty caecum weight were observed in male and female at 1260mg/kg bw/day and increase in absolute and relative full caecum weight in male rats at 242 mg/kg bw/day were observed as compared to control. The observed effect was not statistically significant as compared to control. In addition, Non-neoplastic transitional-cell hyperplasia of bladder, inflammatory cell infiltrate of the seminal vesicles and testicular interstitial-cell hyperplasia were observed in 47 mg/kg bw/day male and 1260 mg/kg bw/day treated male and female rats. Statistically significant increase in renal calcification and renal pelvic epithelial hyperplasia, lung oedema and hemorrhage, lymph-node hemorrhage and degenerative changes in the brain and nerve, degenerative and inflammatory changes in heart, inflammatory changes in thymus, aortic calcification and atrial thrombi were observed in 1260 mg/kg bw/day treated female rats. Statistically significant increase in neoplastic uterine polyps and vaginal fibromas were observed in 1260 mg/kg bw/day treated female rats. The observed effects were typical of this strain and occur in ageing rats. No effect on number of litters, number of pups per litter at day 18 and pup weight at day 18 were observed in treated rats as compared to control. Therefore, NOAEL was considered to be 1250 mg/kg body weight /day when Wistar male and female rats were treated with test chemical orally in fed for 2 years.

 

Thus based on the above studies, the No Observed Adverse Effect Level (NOAEL) was considered to be 1000 mg/kg/day, when rodents were treated with the given test chemical during repeated dose toxicity study.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
Data is Klimicsh 2 and from handbook or collection of data.

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: inhalation
Data waiving:
exposure considerations
Justification for data waiving:
a short-term toxicity study does not need to be conducted because exposure of humans via inhalation in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment
Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
Waiver

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: dermal
Data waiving:
exposure considerations
Justification for data waiving:
a short-term toxicity study does not need to be conducted because exposure of humans via the dermal route in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment
Endpoint conclusion
Endpoint conclusion:
no study available
Quality of whole database:
Waiver

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Repeated dose toxicity: Oral

Data available from the various sources was reviewed to determine the toxic nature of the given test chemical. The studies are as mentioned below:

 

Combined Repeated Dose and Carcinogenicity was performed to determine the toxic nature of the test chemical in Sprague Dawley rats. Two-year tests were run in which 400 male and female Sprague Dawley rats were distributed into various treatment groups. The treatment groups were - 100 rats/dose (50/sex) and 100 rats/sex for controls. The test chemical (500 mg/kg bw/day, 1000 mg/kg bw/day) was mixed into a commercial laboratory chow diet for each feeding study. Rats were individually caged. Each animal received in the Pathology Laboratory was given a careful gross examination, and all lesions were recorded. Of those not received, most were discarded because of advanced postmortem autolysis. So far as can be determined, none of the discarded animals had tumors. Lesions which could not clearly be identified as neoplastic or non-neoplastic, and tumors, the type of which was uncertain or uncommon, were also sectioned and examined microscopically. Tissues sectioned include the following: liver, lung, heart, spleen, pancreas, stomach, small intestine, colon, kidney, adrenal, thyroid, testis (or ovary and uterus), leg muscles, leg bones with included marrow, and any tumor or other unusual condition. Sections of the mammary glands were also observed for tumors. The ingestion of the test chemical had no effect on survival, growth, or weights at autopsy of heart, liver, kidney, spleen, or testes in the test animals. No effect due to ingestion of the test chemical on either tumor incidence or tumor type was noted in Sprague Dawley strain on this two-year experiment. Hence, the No Observed Adverse Effect level was considered to be 1000 mg/kg bw/day in both male and female Sprague Dawley rats.

 

The above study is supported with another combined repeated dose & carcinogenicity study conducted in Wistar male and female rats treated with test chemical in the concentration of 0, 50, 250, 1250 mg/kg/day (Nominal dose level) 47, 242 and 1260 mg/kg bw/day (calculated intake) for male and 49, 246 and 1260 mg/kg bw/day (calculated intake) for female orally in fed. No significant effect on survival of treated rats were observed as compared to control. Significant decrease in body weight in male rats from week 4 to 73 and in female rat from 51 and 73 and increase in food consumption was observed in 1260 mg/kg bw/day treated rats as compared to control. Slight effect on body weight and food consumption was due to a reduction in the absorption or utilization of the nutrient. Average compound intake of male rats were 47, 242 and 1260 mg/kg bw/day and for female rats 49, 246 and 1260 mg/kg bw/day. Increase in water consumption was observed in 1260 mg/kg bw/day treated male rats as compared to control. Increased water loss was observed due to production of softer, moister faeces and a compensatory increase in water intake. Similarly, decreased in packed cell volume was observed at month 6 and 12 in male and at month 18 in female rats and slightly increased hemoglobin concentrations in female rats at termination of study and decrease in glutamic-oxalacetic transaminase activities in male and female rats but not significant in female rats were observed at 1260 mg/kg bw/day. Observed effect were not dose related or consistent between the sexes. Increase level of protein in urine at 12 months in female was observed at 1260 mg/kg bw/day and Semi-quantitative analysis of urine for bilirubin and ketones was hindered at months 18 and 24 due to the contamination of the urine with test chemical which interfered with the colour reaction. Statistically significant increase in absolute and relative full and empty caecum weight were observed in male and female at 1260mg/kg bw/day and increase in absolute and relative full caecum weight in male rats at 242 mg/kg bw/day were observed as compared to control. The observed effect was not statistically significant as compared to control. In addition, Non-neoplastic transitional-cell hyperplasia of bladder, inflammatory cell infiltrate of the seminal vesicles and testicular interstitial-cell hyperplasia were observed in 47 mg/kg bw/day male and 1260 mg/kg bw/day treated male and female rats. Statistically significant increase in renal calcification and renal pelvic epithelial hyperplasia, lung oedema and hemorrhage, lymph-node hemorrhage and degenerative changes in the brain and nerve, degenerative and inflammatory changes in heart, inflammatory changes in thymus, aortic calcification and atrial thrombi were observed in 1260 mg/kg bw/day treated female rats. Statistically significant increase in neoplastic uterine polyps and vaginal fibromas were observed in 1260 mg/kg bw/day treated female rats. The observed effects were typical of this strain and occur in ageing rats. No effect on number of litters, number of pups per litter at day 18 and pup weight at day 18 were observed in treated rats as compared to control. Therefore, NOAEL was considered to be 1250 mg/kg body weight /day when Wistar male and female rats were treated with test chemical orally in fed for 2 years.

 

Thus based on the above studies, the No Observed Adverse Effect Level (NOAEL) was considered to be 1000 mg/kg/day, when rodents were treated with the given test chemical during repeated dose toxicity study.

 

Repeated dose toxicity: Inhalation

A short-term toxicity study does not need to be conducted because exposure of humans via inhalation in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment. Also, the given test chemical has very low vapor pressure 6.79E-42 mm Hg, so the potential for the generation of inhalable vapors is very low. The normal conditions of use of this substance will not result in aerosols, particles or droplets of an inhalable size, so exposure to humans via the inhalatory route will be highly unlikely and therefore this end point for repeated inhalation toxicity was considered for waiver.

 

Repeated dose toxicity: Dermal

A short-term toxicity study does not need to be conducted because exposure of humans via dermal in production and/or use is not likely as based on the provided thorough and rigorous exposure assessment. Also, the acute dermal toxicity value for 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 test chemical shall not exhibit 28 day repeated dose toxicity by the dermal route. In addition, there is no data available that suggests that test chemical shall exhibit repeated dose toxicity by the dermal route. Hence this end point was considered for waiver.

 

Justification for classification or non-classification

Based on the data available, the given test chemical does not exhibit toxic nature upon repeated exposure by oral route of exposure. Hence, it is not likely to classify as toxic as per the criteria mentioned in CLP regulation. For repeated inhalation and repeated dermal toxicity waiver was added so, not possible to classify.