Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: 

 

The acute oral toxicity dose (LD50) for test chemical was considered based on experimental study, the value considered to be >2000 mg/kg bw in rats and mice. Thus, comparing this value with the criteria of CLP regulation, test chemical cannot be classified for acute oral toxicity.

 

Acute Inhalation toxicity:

The test substance has very low vapor pressure (0.00000256 Pa at 25 degC). Also, the particle size distribution of the substance was found to vary in the size of 55-250 µm, so the potential for the generation of inhalable dust forms is low. Moreover 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 unlikely to occur, and therefore acute toxicity by inhalation route was considered to be waived.

 

Acute Dermal toxicity:

 

The acute dermal toxicity dose (LD50) for test chemical was considered based on experimental study, the value considered to be >2000 mg/kg bw in rabbits. Thus, comparing this value with the criteria of CLP regulation, test chemical cannot be classified for acute dermal toxicity.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from study report
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Principles of method if other than guideline:
The aim of this study was to assess the toxicity potential of test chemical after single oral administration in rats.
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:Bharat Serum and Vaccines Limited.
- Age at study initiation:8- 11 weeks at the time of dosing.
- Health Status :Healthy young adult animals were used for the study. Females were nulliparous and non pregnant.
- Weight at study initiation:Minimum: 183 g and Maximum: 216 g (Individual body weights were within ± 5% prior to treatment after overnight fasting).
- Fasting period:The animals were fasted for minimum 16-18 hours prior to dosing and for 3-4 hours post dosing.
- Housing:The animals were housed individually in polycarbonate cages.
- Bedding:All cages were provided with corn cobs.
- Room Sanitation:The experimental room floor and work tops were swept and mopped with disinfectant solution every day.
- Cages and water bottle:All the cages and water bottles were changed at least twice every week.
- Diet (e.g. ad libitum):All animals were provided conventional laboratory rodent diet (Nutrivet Life Sciences, Pune) ad libitum.
- Water (e.g. ad libitum):Aqua guard filtered tap water was provided ad libitum via drinking bottles.
- Acclimation period:Animal nos. 1-3 were acclimatized for five days, 4-6 for seven days, 7-9 for nine days and 10-12 for six days prior to administration of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Minimum: 20.00 °C and Maximum: 23.60 °C.
- Humidity (%):Minimum: 37.40% and Maximum: 61.80%
- Air changes (per hr):More than 12 changes per hour.
- Photoperiod (hrs dark / hrs light):12:12

Route of administration:
oral: unspecified
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Amount of vehicle:10 ml
- Justification for choice of vehicle:Corn oil was selected because test item was not soluble in distilled water.
- Lot/batch no. (if required):MKBD4650
- Purity:N/A

MAXIMUM DOSE VOLUME APPLIED:10 ml/kg body weight.

Doses:
G1 = 300 mg/kg bw
G2 = 2000 mg/kg bw
No. of animals per sex per dose:
Twelve female rats
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:Daily
- Necropsy of survivors performed: yes
At the end of 14 day observation period, all the survived rats were euthanised by overdose of CO2 for external and internal observations.
- Other examinations performed:
Clinical Observation
After test item administration, individual animals were frequently observed at 30 minutes, 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Additionally, on day 0 animal nos. 7-9 were observed at 6 hours post dosing. Subsequently, all the animals were observed once a day during the 14 day observation period.

Body weight
All rats were weighed on days 0 (prior to dosing), 7 and 14. Animals were weighed immediately after found dead.

other:
Mortality
All the animals were observed twice daily (morning and evening) for morbidity and mortality, throughout the acclimatization and study period.
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: Non toxic to rat.
Mortality:
No mortality was observed in the animals treated with 300 and 2000 mg/kg dose throught out the 14 days observation period post dosing.
Clinical signs:
At 300 mg/kg, all the six animals (animal nos. 1-6) were observed normal throughout experimental period. At 2000mg/kg, animal no. 7 was observed normal at 30 minutes and 1 hour post dosing, mild to moderate lethargy at 2, 3, 4 and 6 hours and mild ataxia at 3, 4 and 6 hours followed by normal clinical sign till day 14. Animal no. 8 was observed normal at 30 minutes and 1 hour post dosing, mild to severe lethargy at 2, 3, 4 and 6 hours and mild to moderate ataxia at 3, 4 and 6 hours followed by normal clinical sign till day 14. Animal no. 9 was observed normal at 30 minutes and 1 hour post dosing, moderate to severe lethargy and mild to moderate ataxia at 2, 3, 4 and 6 hours followed by normal clinical sign till day 14. Animal nos. 10-12 were observed normal at 30 minutes and 1 hour post dosing, mild to moderate lethargy at 2, 3 and 4 hours and mild ataxia at 3 and 4 hours followed by normal clinical sign till day 14.
Body weight:
Mean body weight was observed with gain was observed in the animals treated with 300 and 2000 mg/kg body weight, during the 14 day observation period, as compared to day 0.
Gross pathology:
No external and internal gross pathological changes were seen in all the six animals treated with 300 and 2000 mg/kg body weight during terminal sacrifice

Table 1: Individual Animal Body Weight (g) andBody Weight Changes(%)

Sex:Female

Animal No.

Group/ Dose (mg/kg)

Body Weight (gram)

Body Weight Change (%)

Day 0

Day 7

Day 14

Day

0-7

Day

0-14

1

G1/ 300

210

223

234

6.19

11.43

2

204

229

234

12.25

14.71

3

200

230

237

15.00

18.50

4

216

240

253

11.11

17.13

5

204

240

246

17.65

20.59

6

198

237

218

19.70

10.10

7

G2/ 2000

210

219

229

4.29

9.05

8

197

228

224

15.74

13.71

9

195

220

235

12.82

20.51

10

187

217

229

16.04

22.46

11

183

219

228

19.67

24.59

12

185

215

229

16.22

23.78

 

Table 2: Summary of Animal Body Weight (g) and Body Weight Changes (%)

 

Sex:Female

Group/ Dose (mg/kg)

Rats Body Weight (g)

Body Weight Changes (%)

Day 0

Day 7

Day 14

0-7

0-14

G1/ 300

Mean

205.33

233.17

237.00

13.65

15.41

SD

6.65

6.91

11.97

4.87

4.10

n

6

6

6

6

6

G2/ 2000

Mean

192.83

219.67

229.00

14.13

19.02

SD

10.09

4.46

3.52

5.29

6.25

n

6

6

6

6

6

Keys:SD = Standard Deviation, n = Number of Animals


Table 3: Individual Animal Clinical Signs and Symptoms

 

Sex:Female

Animal No.

Group/ Dose (mg/kg)

Hours (Day 0)

1/2

1

2

3

4

6

1

G1/ 300

1

1

1

1

1

-

2

1

1

1

1

1

-

3

1

1

1

1

1

-

4

1

1

1

1

1

-

5

1

1

1

1

1

-

6

1

1

1

1

1

-

7

G2/ 2000

1

1

99+

99++

21+

99++

21+

99++

21+

8

1

1

99+

99++

21+

99+++

21+

99+++

21++

9

1

1

99++

21+

99+++

21++

99+++

21++

99+++

21++

10

1

1

99+

99++

21+

99++

21+

-

11

1

1

99+

99++

21+

99++

21+

-

12

1

1

99+

99++

21+

99++

21++

-

Interpretation of results:
other: Not classified
Conclusions:
Under the conditions of this; acute oral toxicity study of test chemical in female rats is as given below:
The acute oral LD50 value of test chemical was 5000 mg/kg body weight.Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that test chemical does not exhibit acute oral toxicity i.e it is acutely non toxic to animals.
Executive summary:

This study was performed as per OECD No. 423. Twelve female Wistar rats were selected for acute oral toxicity study.

The animals were fasted for minimum 16-18 hours prior to dosing and for 3-4 hours post dosing, with food withheld but drinking water providedad libitum. The time intervals between dosing were determined by the onset, duration and severity of toxic signs.

Three rats of group G1 were dosed with starting dose of 300 mg/kg body weight and the animals did not show any mortality so another three animals of the group G1 were dosed with 300 mg/kg body weight and no mortality was observed. Three animals of the group G2 were dosed with 2000 mg/kg body weight and no mortality was observed, therefore another three animals of same group G2 were dose with 2000 mg/kg and no mortality was observed. Hence, further dosing was stopped.

Body weights were re­corded on day 0 (prior to dosing) 7 and 14. Mean body weight was observed with gain was observed in the animals treated with 300 and 2000 mg/kg body weight, during the 14 day observation period, as compared to day 0.

At 300 mg/kg, all the six animals (animal nos. 1-6) were observed normal throughout experimental period. At 2000mg/kg, animal no. 7 was observed normal at 30 minutes and 1 hour post dosing, mild to moderate lethargy at 2, 3, 4 and 6 hours and mild ataxia at 3, 4 and 6 hours followed by normal clinical sign till day 14. Animal no. 8 was observed normal at 30 minutes and 1 hour post dosing, mild to severe lethargy at 2, 3, 4 and 6 hours and mild to moderate ataxia at 3, 4 and 6 hours followed by normal clinical sign till day 14. Animal no. 9 was observed normal at 30 minutes and 1 hour post dosing, moderate to severe lethargy and mild to moderate ataxia at 2, 3, 4 and 6 hours followed by normal clinical sign till day 14.Animal nos. 10-12 were observed normal at 30 minutes and 1 hour post dosing, mild to moderate lethargy at 2, 3 and 4 hours and mild ataxia at 3 and 4 hours followed by normal clinical sign till day 14.

No external and internal gross pathological changes were seen in all the six animals treated with 2000 mg/kg body weight during terminal sacrifice.

The acute oralL D50 value of test chemical was 5000 mg/kg body weight.Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that test chemical does not exhibit acute oral toxicity i.e it is acutely non toxic to animals.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The data is K1 level.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
Waiver

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Data is from study report
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Principles of method if other than guideline:
The objective of the study was to assess the dermal toxicity of test chemical after single dose application by dermal route in rats.
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:In-House Bred at sa-Ford, Animal Facility
- Age at study initiation:N/A
- Health Status:Healthy young adult animals were used for the study. Females were nulliparous and non pregnant.
- Weight (Prior to Treatment):Male:Minimum: 238 g and Maximum: 274 g ,Female:Minimum: 234 g and Maximum: 250 g
- Fasting period before study:N/A
- Housing:The animals were housed individually in polycarbonate cages.
- Bedding : All cages were provided with corn cobs.
- Room Sanitation : The experimental room floor and work tops were swept and mopped with disinfectant solution every day.
- Cages and water bottle : All the cages and water bottles were changed at least twice every week.
- Diet (e.g. ad libitum):All animals were provided conventional laboratory rodent diet (Nutrivet Life Sciences, Pune) ad libitum.
- Water (e.g. ad libitum):Aqua guard filtered tap water was provided ad libitum via drinking bottles.
- Acclimation period:All animals were acclimatized to the test conditions for 5 days prior to administration of the test item.
- Randomization : Animals were selected manually. No computer generated randomization program was used.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):Minimum: 20.40 °C and Maximum: 23.10 °C
- Humidity (%):Minimum: Minimum: 38.40% and Maximum: 58.70%
- Air changes (per hr):More than 12 changes per hour
- Photoperiod (hrs dark / hrs light):12:12

Type of coverage:
semiocclusive
Vehicle:
other: distilled water
Details on dermal exposure:
TEST SITE
- Area of exposure:The test item was applied uniformly over clipped dorsal area of rat skin.
- % coverage:Approximately 10% body surface area of rat.
- Type of wrap if used:The porous gauze dressing and non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done):The residual test item was removed by using distilled water.
- Time after start of exposure:24-hour.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):A limit dose of 2000 mg/ kg body weight of test item was applied.
- Constant volume or concentration used: yes
- For solids, paste formed: yes

VEHICLE
- Amount(s) applied (volume or weight with unit):0.2 ml distilled water.
- Concentration (if solution):N/A
- Lot/batch no. (if required):N/A
- Purity:N/A
Duration of exposure:
24 hrs
Doses:
2000 mg/kg body weight.
No. of animals per sex per dose:
10 (Five per sex)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:Daily
- Necropsy of survivors performed: yes
At the end of 14 day observation period, all the surviving rats were euthanised by overdose of CO2 and subjected to gross pathology examination, for external and internal observations.
- Other examinations performed:
- Clinical signs : After test item administration, individual animals were frequently observed at 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Subsequently, all animals were observed once a day during the 14 day observation period.

- Body weight: All rats were weighed on days 0 (prior to dosing), 7 and 14.

other:
- Local Signs/Skin Reactions
All animals were observed once daily during days 1-14 (in common with clinical signs).

- Mortality
Animals were observed twice daily for any mortality during the experimental period.
Statistics:
No statistical analysis was performed since the study was terminated with limit test.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: Non toxic to animals.
Mortality:
No mortality was observed at limit dose of 2000 mg/kg body weight of test item during the 14 day observation period.
Clinical signs:
No systemic or local signs of toxicity were observed at limit dose of 2000 mg/kg body weight of test item during the experimental period.
Body weight:
The male and female animals were observed with body weight gain compared to day 0 throughout the experiment.
Gross pathology:
The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality.

Table 1: Individual Animal Body Weight (g) andBody Weight Changes(%)

 

Dose:2000 mg/ kg bodyweight                                                                                                         

Animal No.

Sex

Body Weight (gram)

Body Weight Change (%)

Day 0

Day 7

Day 14

Day 0-7

Day 0-14

1

Male

245

247

229

0.82

-6.53

2

245

264

280

7.76

14.29

3

238

248

257

4.20

7.98

4

274

297

269

8.39

-1.82

5

251

244

272

-2.79

8.37

6

Female

249

249

265

0.00

6.43

7

238

254

262

6.72

10.08

8

246

250

267

1.63

8.54

9

234

227

244

-2.99

4.27

10

250

254

263

1.60

5.20

 

 

Table 2: Individual Animal Clinical Signs and Symptoms

 

Dose:2000 mg/kg body weight

Animal No.

Sex

Hour(s) - Day 0

Day

1

2

3

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

1

Male

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

2

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

3

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

4

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

5

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

6

Female

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

7

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

8

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

9

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

10

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

Key: 1 = Normal


Table 3: Individual Animal Mortality Record

 

Dose:2000 mg/kg body weight

       Animal No.

Sex

Days of Observation (0 to 14)

Morning Observations

Evening Observations

1

Male

No mortality and morbidity

No mortality and morbidity

2

No mortality and morbidity

No mortality and morbidity

3

No mortality and morbidity

No mortality and morbidity

4

No mortality and morbidity

No mortality and morbidity

5

No mortality and morbidity

No mortality and morbidity

6

Female

No mortality and morbidity

No mortality and morbidity

7

No mortality and morbidity

No mortality and morbidity

8

No mortality and morbidity

No mortality and morbidity

9

No mortality and morbidity

No mortality and morbidity

10

No mortality and morbidity

No mortality and morbidity


Interpretation of results:
other: Not classified
Conclusions:
The acute dermal median lethal dose of test chemical was >2000 mg/kg body weight.Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that test chemical does not exhibit acute dermal toxicity i.e it is acutely non toxic to animals.
Executive summary:

This study was performed as per OECD No.402.

Five male and five female healthy young adult rats were randomly selected and used for conducting acute dermal toxicity study. Rats free from injury and irritation of skin were selected for the study. Twenty four hours prior to dermal application of test item, approximately 10% of body surface area of each rat was clipped. A limit dose of 2000 mg/ kg body weight of test item moistened with 0.2 ml distilled water was applied by single dermal application and observed for 14 days after treatment.

On test day 0, an amount of test item moistened with 0.2 ml distilled water was applied directly on the intact skin of clipped area of rats; the surgical gauze patch was put on to the intact skin of clipped area.This gauze patch was covered with a semi-occlusive dressing.The dressing was wrapped around the abdomen and anchored with non-irritating adhesive tape.After the 24-hour application period, the dressings were removed and theskin was gently wiped with distilled water.The skin reactions were assessed.

The animals were observed daily for mortality and clinical signs, during the acclimatization period. All animals were observed for clinical signs at approximately 1, 2, 3 and 4 hours after treatment on day 0 and once daily during test days 1‑14. Mortality was recorded after application on test day 0 and twice daily during days 1-14 (at least once on the day of sacrifice). Local signs / Skin reactions were observed daily from test days 1-14 (in common with clinical signs). Body weights were re­corded on day 0 (prior to application) and on day 7 and 14. All animals were necropsied and examined macroscopically.

No mortality was observed in any animal till the end of the experimental period.

No clinical signs and any skin reaction were observed throughout the experimental period in all treated animals.

The male and female animals were observed with body weight gain compared to day 0 throughout the experiment.

The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality.


Under the conditions of this; acute dermal toxicity study of test chemical in rats is as given below:

The acute dermal median lethal dose of test chemical was >2000 mg/kg body weight.Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that test chemical does not exhibit acute dermal toxicity i.e it is acutely non toxic to animals.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The data is K1 level.

Additional information

Acute oral toxicity:

In different studies, the given test chemical has been investigated for acute oral toxicity to a greater or lesser extent. Often are the studies based on in-vivo experiments in rodents, i.e. most commonly in rats for test chemical. The studies are summarized as below –

This study was performed as per OECD No. 423.Twelve female Wistar rats were selected for acute oral toxicity study.

The animals were fasted for minimum 16-18 hours prior to dosing and for 3-4 hours post dosing, with food withheld but drinking water providedad libitum. The time intervals between dosing were determined by the onset, duration and severity of toxic signs.

Three rats of group G1 were dosed with starting dose of 300 mg/kg body weight and the animals did not show any mortality so another three animals of the group G1 were dosed with 300 mg/kg body weight and no mortality was observed. Three animals of the group G2 were dosed with 2000 mg/kg body weight and no mortality was observed, therefore another three animals of same group G2 were dose with 2000 mg/kg and no mortality was observed. Hence, further dosing was stopped.

Body weights were re­corded on day 0 (prior to dosing) 7 and 14. Mean body weight was observed with gain was observed in the animals treated with 300 and 2000 mg/kg body weight, during the 14 day observation period, as compared to day 0.

At 300 mg/kg, all the six animals (animal nos. 1-6) were observed normal throughout experimental period. At 2000mg/kg, animal no. 7 was observed normal at 30 minutes and 1 hour post dosing, mild to moderate lethargy at 2, 3, 4 and 6 hours and mild ataxia at 3, 4 and 6 hours followed by normal clinical sign till day 14. Animal no. 8 was observed normal at 30 minutes and 1 hour post dosing, mild to severe lethargy at 2, 3, 4 and 6 hours and mild to moderate ataxia at 3, 4 and 6 hours followed by normal clinical sign till day 14. Animal no. 9 was observed normal at 30 minutes and 1 hour post dosing, moderate to severe lethargy and mild to moderate ataxia at 2, 3, 4 and 6 hours followed by normal clinical sign till day 14.Animal nos. 10-12 were observed normal at 30 minutes and 1 hour post dosing, mild to moderate lethargy at 2, 3 and 4 hours and mild ataxia at 3 and 4 hours followed by normal clinical sign till day 14.

No external and internal gross pathological changes were seen in all the six animals treated with 2000 mg/kg body weight during terminal sacrifice.

The acute oralL D50value of test chemicalwas5000 mg/kg body weight.Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that test chemical does not exhibit acute oral toxicity i.e it is acutely non toxic to animals.

In another similar study, Acute oral toxicity study of test chemical was conducted in Wistar albino rats. The study was conducted under the OECD guideline 423 for testing of chemicals. The healthy wistar albino rats of body weight 200 ± 20 gm were selected for study after acclimatization to standard laboratory condition and divided into test compound and vehicle control group each having three animals.

Test compound was mixed with distilled water and administered orally at the dose level of 2000mg/kg bw (dose volume 10ml/l) to three female rats. However, vehicle control group treated with distilled water at a dose level of 10 ml/kg bw. The treated animals were closely observed for clinical signs of intoxication during first four hours of test compound administration. Thereafter, all the animals were observed periodically at one hour interval for 24 hrs and twice daily for a period of 14 days. The necropsy was performed on all animals at the termination of the study.

 The test compound did not elicit any clinical signs of intoxication throughout the period of observation at the tested dose level 2000 mg/kg bw. Furthermore, no mortality was recorded at the test dose level. Vehicle control group of animals were also free from any mortality and clinical signs.Necropsy results were: EXTERNAL : Skin- Skin and hair coat was observed wet., All external orifices- Normal   

B. INTERNAL : Subcutaneous- No change was observed, Superficial and deep lymph nodes- No change in mesenteric lymph node.

1.ABDOMINAL CAVITY: Opening and general examination- In the abdominal cavity all the organs were present in normal position, Spleen- Normal upto highest tested dose level 2000 mg/kg b.wt, Digestive system- No gross changes were observed in stomach and intestine upto highest tested dose level 2000 mg/kg b.wt, Liver and biliary ducts- No gross pathological changes were observed, Excretory system- No gross pathological changes were observed upto highest tested dose level 2000 mg/kg b.wt, Adrenal- Observed normal, Male/female genital organs – Showed normal colour, consistency and no inflammatory changes upto highest tested dose level 2000 mg/kg b.wt.

2. THORACIC CAVITY: Opening and general examination- Thoracic cavity was found to be normal without any fluid, mucous or blood etc, Lungs- observed normal, Heart- No changes were observed in color and consistency. Heart found normal upto highest tested dose level 2000 mg/kg b.wt, Thyroid- Normal in shape, size and surface upto highest tested dose level 2000 mg/kg b.wt.

3. CRANIAL CAVITY: Brain- Normal in shape and size.

Thus, based on the above summarised studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute oral toxicity.

Acute Inhalation toxicity:

The test substance has very low vapor pressure (0.00000256 Pa at 25 degC). Also, the particle size distribution of the substance was found to vary in the size of 55-250 µm, so the potential for the generation of inhalable dust forms is low. Moreover 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 unlikely to occur, and therefore acute toxicity by inhalation route was considered to be waived.

Acute Dermal Toxicity

In different studies, the given test chemical has been investigated for acute dermal toxicity to a greater or lesser extent. Often are the studies based on in-vivo experiments in rodents, i.e. most commonly in rats for test chemical. The studies are summarized as below –

This study was performed as per OECD No.402.

Five male and five female healthy young adult rats were randomly selected and used for conducting acute dermal toxicity study. Rats free from injury and irritation of skin were selected for the study. Twenty four hours prior to dermal application of test item, approximately 10% of body surface area of each rat was clipped. A limit dose of 2000 mg/ kg body weight of test item moistened with 0.2 ml distilled water was applied by single dermal application and observed for 14 days after treatment.

On test day 0, an amount of test item moistened with 0.2 ml distilled water was applied directly on the intact skin of clipped area of rats; the surgical gauze patch was put on to the intact skin of clipped area.This gauze patch was covered with a semi-occlusive dressing.The dressing was wrapped around the abdomen and anchored with non-irritating adhesive tape.After the 24-hour application period, the dressings were removed and theskin was gently wiped with distilled water.The skin reactions were assessed.

The animals were observed daily for mortality and clinical signs, during the acclimatization period. All animals were observed for clinical signs at approximately 1, 2, 3 and 4 hours after treatment on day 0 and once daily during test days 1‑14. Mortality was recorded after application on test day 0 and twice daily during days 1-14 (at least once on the day of sacrifice). Local signs / Skin reactions were observed daily from test days 1-14 (in common with clinical signs). Body weights were re­corded on day 0 (prior to application) and on day 7 and 14. All animals were necropsied and examined macroscopically.No mortality was observed in any animal till the end of the experimental period.No clinical signs and any skin reaction were observed throughout the experimental period in all treated animals.The male and female animals were observed with body weight gain compared to day 0 throughout the experiment.The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality.

Under the conditions of this; acute dermal toxicity study of test chemical in rats is as given below:

The acute dermal median lethal dose of test chemical was>2000 mg/kgbody weight.Thus by considering the CLP criteria for acute toxicity rating for the chemicals, it infers that test chemical does not exhibit acute dermal toxicity i.e it is acutely non toxic to animals.

In another study,Acute dermal toxicity test was performed on 10 wistar male and female rats to determine the LD50 value of test chemical. Ten healthy Wistar albino rats of both sex (ranging bw 200±30 gm) were selected for study after acclimatization. Approximate 10 percent back skin of total body surface area was prepared 24 hrs prior to application of test compound. The test chemical was applied dermally at the dose level of 2000 mg/kg b.wt for each animal. The treated animals were closely observed for clinical signs of intoxication during first four hours of test compound administration. Thereafter, all the animals were observed periodically at one hour interval for 24 hrs and twice daily for a period of 14 days. The body weight of each rat was observed on day 0 (pre treatment), 7th and 14th (post treatment). The necropsy was performed on all animals at the termination of the study. Rats were free from any systematic and local clinical signs when observed after 24 hours of patch removal. After 14 days observation no mortality was observed. No body weight change was observed.

Necropsy was conducted at the end of the observation on all the animals (day 15th), did not reveal any significant gross pathological changes related to compound toxicity. Skin- Skin and hair coat was observed wet, All external orifices- Normal  

INTERNAL: Subcutaneous- No changes were observed, Superficial and deep lymph nodes- No change in mesenteric lymph node.

1. ABDOMINAL CAVITY: Opening and general examination- In the abdominal cavity all the organs were present in normal position, Spleen- No changes were recorded, Digestive system- No gross changes were observed in stomach and intestine, Liver and biliary ducts

- No gross pathological changes were observed, Excretory system- No gross pathological changes were observed, Adrenal- Observed normal, Male/female genital organs – Showed normal colour, consistency and no inflammatory changes.

2. THORACIC CAVITY: Opening and general examination- Thoracic cavity was found to be normal without any fluid, mucous or blood etc, Lungs- No changes were recorded, Heart- No changes were observed in color and consistency, Heart found normal, Thyroid- Normal in shape, size and surface.

3. CRANIAL CAVITY: Brain- Normal in shape and size.

Thus, based on the above summarized studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.

Justification for classification or non-classification

Based on the above experimental studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw, for acute oral and dermal toxicity. Thus, comparing this value with the criteria of CLP regulation,test chemical cannot be classified for acute oral and dermal toxicity. For acute inhalation toxicity wavier were added so, not possible to classify.