Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: The acute oral toxicity dose (LD50) for target chemical was considered based on different experimental studies conducted on rats and mice, the LD50 values were considered in between 5000-10000 mg/kg bw in mice, and 6800 mg/kg bw in rats. These studies concluded that the LD50 value is >2000 mg/kg bw, for acute oral toxicity. Thus, comparing this value with the criteria of CLP regulation, test chemical cannot be classified for acute oral toxicity.

Acute Inhalation Toxicity:The acute inhalation toxicity dose (LC50) for test chemical was considered based on experimental study conducted on rats, the LC50 value was considered to be >5 mg/L (>5000 mg/m3) in rats. Thus, comparing this value with the criteria of CLP regulation, test chemical cannot be classified for acute inhalation toxicity.

Acute Dermal toxicity:The acute dermal toxicity dose (LD50) for test chemical was considered based on experimental study conducted on rats and rabbits, the LD50 value was considered to be >2000 mg/kg bw in rats, and 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:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from study report
Qualifier:
according to
Guideline:
other: As mentioned below
Principles of method if other than guideline:
Acute oral toxicity of test chemical in mice.
GLP compliance:
not specified
Test type:
other: not specified
Limit test:
no
Species:
mouse
Strain:
not specified
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 4-5 week old
- Fasting period before study: 4 hours
- Housing: The animals were housed individually in cages.
- Diet (e.g. ad libitum): pelleted diet, ad libitum
- Water (e.g. ad libitum): water, ad libitum
Route of administration:
oral: gavage
Vehicle:
other: groundnut oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 20% w/v solution
Doses:
2000, 5000 and 10000 mg/kg
No. of animals per sex per dose:
Total = 10
2000 mg/kg - one male and one female
5000 mg/kg - three male and three female
10000 mg/kg - one male and one female
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 7 days
- Frequency of observations and weighing: The animals were observed for signs of toxicity for 7 days. Survivors were weighed before killing for post-mortem examination at the end of the one week observation period.
- Necropsy of survivors performed: yes, all animals dying were autopsied. All survivors were killed and examined post mortem after 7 days.
Statistics:
The approximate LD50 value is estimated from the results and the test substance is given a toxicity rating according to Hodge and Sterner's classification.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 - < 5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no further details available
Mortality:
At 2000 mg/kg – No mortality was observed.
At 5000 mg/kg – 1 mouse died within 24 hours and 1 mouse died within 96 hours after treatment.
At 10000 mg/kg – Both the mice were died within 48 and 66 hours.
Clinical signs:
At 2000 mg/kg – The male mouse was somnolent, showing signs of stress and exhibiting labored breathing within 18 hours after treatment but recovered within 42 hours. The female mouse appeared unaffected by the treatment.
At 5000 mg/kg – The mice were showing signs of stress and exhibiting lacrimation within 30 mins after treatment and within 2 hours somnolence and laboured breathing was also observed. These mice were comatose within 18-42 hours after treatment. The surviving mice continued to show signs of stress throughout the 7 day observation period.
At 10000 mg/kg – The mice were showing signs of stress and exhibiting lacrimation within 30 mins after treatment and within 2 hours somnolence and laboured breathing was also observed. These mice were comatose within 18-42 hours after treatment.
Body weight:
Surviving animals gained weight during the 7 day observation period and presented a normal appearance at autopsy.
Gross pathology:
Autopsy of the animals that died revealed gaseous distension and irritation of the stomach and intestines. The mice dying after 48 hours and later had pale soft rose-pink livers, pale spleens and mottled kidneys. Hemorrhaging was also present in the ileum of these mice.
Other findings:
No data
Interpretation of results:
other: Not classified
Conclusions:
The acute oral toxicity dose LD50 value was considered in between 5000-10000 mg/kg bw, when 10 male and female White mice were treated with test chemical via oral gavage route.
Executive summary:

The acute oral toxicity of test chemical was tested in 10 male and female White mice at thedose concentration of 2000, 5000 and 10000 mg/kgbw. The given test chemical was dissolved as 20% w/v solutionin groundnut oil and administered via oral gavage route. The animals were observed for signs of toxicity for 7 days. Survivors were weighed before killing for post-mortem examination at the end of the one week observation period. All animals dying were autopsied. All survivors were killed and examined post mortem after 7 days.The approximate LD50 value is estimated from the results and the test substance is given a toxicity rating according to Hodge and Sterner's classification. Mortality was observed as - At 2000 mg/kg – No mortality was observed. At 5000 mg/kg – 1 mouse died within 24 hours and 1 mouse died within 96 hours after treatment. At 10000 mg/kg – Both the mice were died within 48 and 66 hours. Clinical signs were observed such as - At 2000 mg/kg – The male mouse was somnolent, showing signs of stress and exhibiting labored breathing within 18 hours after treatment but recovered within 42 hours. The female mouse appeared unaffected by the treatment. At 5000 mg/kg – The mice were showing signs of stress and exhibiting lacrimation within 30 mins after treatment and within 2 hours somnolence and laboured breathing was also observed. These mice were comatose within 18-42 hours after treatment. The surviving mice continued to show signs of stress throughout the 7 day observation period. At 10000 mg/kg – The mice were showing signs of stress and exhibiting lacrimation within 30 mins after treatment and within 2 hours somnolence and laboured breathing was also observed. These mice were comatose within 18-42 hours after treatment. Surviving animals gained weight during the 7 day observation period and presented a normal appearance at autopsy. Autopsy of the animals that died revealed gaseous distension and irritation of the stomach and intestines. The mice dying after 48 hours and later had pale soft rose-pink livers, pale spleens and mottled kidneys. Hemorrhaging was also present in the ileum of these mice. Therefore, LD50 value was considered in between 5000-10000 mg/kg bw, when 10 male and female White mice were treated with test chemical via oral gavage route.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
5 000 mg/kg bw
Quality of whole database:
Data is Klimisch 2 and from study report

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
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 403 (Acute Inhalation Toxicity)
Principles of method if other than guideline:
Acute inhalation toxicity of test chemical in Wistar rats.
GLP compliance:
yes
Test type:
standard acute method
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Age : 7 to 9 weeks
Sex : Male and Female
Body weight range : 200 ±20g
Identification : By cage tag and corresponding colour body marking
Acclimatization : Twenty healthy albino rats were selected and acclimatized for standard laboratory condition for period of one week in experimental room under veterinary examination.
Randomization :After acclimatization and veterinary examination all the selected rats randomly divided into two groups of five females and five males.
Nutritional conditions :For feeding conventional Laboratory diets may be used with an unlimited supply of drinking water.

Route of administration:
inhalation: aerosol
Type of inhalation exposure:
nose only
Vehicle:
other: Distilled water
Details on inhalation exposure:
All exposure was conducted in a dynamic nose-only cylindrical chamber built from stainless steel and glass. The chamber had a volume of 8 liters with inner and outer chamber to minimize the fluctuation in concentration and temperature. All the animals uniformly exposed to aerosol drug concentration.
For the inhalation purpose the rats were placed in polycarbonate holder tubes positioned radically around exposure chamber, so that only the snouts and nostrils of the animals were exposed to the aerosol. The chamber was maintained at a slightly negative pressure to prevent leakage of the test atmosphere from the system, as well as its dilution with outside air. The exhaust air was decontaminated by subsequent passage through 1% NaOH solution, silica gel and activated charcoal filters.
Duration of exposure:
6 h
No. of animals per sex per dose:
10 (5 males and 5 females)
Control animals:
not specified
Details on study design:
Ten healthy wistar albino rats of both sex body weight 200±20 gm were selected for study after acclimatization. The test groups of animals were exposed to aerosol at the concentration of 5 mg/L for period of 4 hrs. After exposure all the animals were closely observed for clinical signs of toxicity at various internals such as 1 hr, 2 hrs, 4 hrs, and 6 hrs on the day of test compound aerosol exposure and later on twice a day throughout the experimentation period of 14 days. The necropsy was performed on all the animals at termination of experiment.
All the albino rats exposed to aerosol at the concentration of 5 mg/L did not show any clinical signs of intoxication. Furthermore, no mortality was observed throughout the period of observation.
Sex:
male/female
Dose descriptor:
LC50
Effect level:
> 5 mg/L air
Based on:
dissolved
Exp. duration:
6 h
Mortality:
The test compound did not elicit any mortality at test concentration viz, 5.0 mg/L throughout the period of observation after exposure.
Clinical signs:
The test compound did not elicit any clinical signs of intoxication at the tested aerosol concentration of 5 mg/L observed for the period of 14 days.
Body weight:
The body weight of animals exposed to test compound, observed on day 0th (pre treatment) and day 7th (post treatment) did not differ significantly as compared to day 0th. Whereas, body weight of animals observed on day 14th showed slight increase as compared to day 0 and 7th.
Gross pathology:
Necropsy-:
Necropsy was carried out on all the animals that died during the study or surviving animals were sacrificed at the end of the study to observe any gross pathological changes.
Interpretation of results:
other: Not classified
Conclusions:
The acute lethal Concentration (LC50) of test chemical in wister albino rats was found to be more than 5 mg/L (>5 mg/L).
Acute toxicity of test chemical to rat by inhalation route indicates that the substance exhibits acute toxicity by the inhalation route in the Category 4 which is relatively non toxic.
Executive summary:

The acute inhalation study of test compound was conducted in albino rat under the OECD-Guideline -403 for testing of chemicals. In limit test, ten healthy wistar albino rats of body weight 200±20 gm were selected for study after acclimatization. The test group of animals was exposed to aerosol at the concentration of 5 mg/L for period of 4 hrs. After exposure all the animals were closely observed for clinical signs of toxicity at various internals such as 1 hr, 2 hrs, 4 hrs, and 6 hrs on the day of test compound aerosol exposure and later on twice a day throughout the experimentation period of 14 days. The necropsy was performed on all the animals at termination of experiment. All the albino rats exposed to aerosol at the concentration of 5 mg/L did not show any clinical signs of intoxication. Furthermore, no mortality was observed throughout the period of observation. The necropsy was performed on all the animals at the termination of study did not show any gross pathological changes. In confirmatory test, after 72 hrs, the result obtained from limit test was confirmed in another 10 animal of both sex at similar concentration following same guideline. Ten healthy wistar albino rats of both sex body weight 200±20 gm were selected for study after acclimatization. The test groups of animals were exposed to aerosol at the concentration of 5 mg/L for period of 4 hrs. After exposure all the animals were closely observed for clinical signs of toxicity at various internals such as 1 hr, 2 hrs, 4 hrs, and 6 hrs on the day of test compound aerosol exposure and later on twice a day throughout the experimentation period of 14 days. The necropsy was performed on all the animals at termination of experiment. All the albino rats exposed to aerosol at the concentration of 5 mg/L did not show any clinical signs of intoxication. Furthermore, no mortality was observed throughout the period of observation. The body weight of animals exposed to test compound, observed on day 0th (pre treatment) and day 7th (post treatment) did not differ significantly as compared to day 0th. Whereas, body weight of animals observed on day 14th showed slight increase as compared to day 0 and 7th. The necropsy was performed on all the animals at the termination of study did not show any gross pathological changes. Hence, the acute lethal Concentration (LC50) of test compound in wister albino rats was considered to be >5 mg/L. The result obtained from present investigation can be concluded that test compound test chemical is acutely non toxic to wistar albino rats.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LC50
5 mg/m³
Quality of whole database:
Data is Klimisch 2 and from study report

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:
Acute Dermal Toxicity Study of test chemical in rats
GLP compliance:
yes
Test type:
fixed dose procedure
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 (CPCSEA Registration No. 1256/bc/09/CPCSEA)
- Females (if applicable) nulliparous and non-pregnant:yes
- Weight at study initiation:
Male:Minimum: 223 g and Maximum: 276 g
Female:Minimum: 212 g and Maximum: 232 g

- Fasting period before study:
- Housing:All cages were provided with corn cobs (Sparconn Life Sciences Bangalore) Batch No.: SPAR – 24/2013
- Diet (e.g. ad libitum): All animals were provided conventional laboratory rodent diet (Nutrivet Life Sciences, Pune) ad libitum. Batch No 400012
- 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 6 days prior to administration of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Minimum: 20.50 °C Maximum: 24.40 °C
- Humidity (%): Minimum: 36.60% Maximum: 57.30%
- Air changes (per hr): More than 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12:12

IN-LIFE DATES: From: To:30-01-2014 to 22-02-2014
Type of coverage:
semiocclusive
Vehicle:
water
Details on dermal exposure:
TEST SITE
- Area of exposure: fur of dorsal area of the trunk
- % coverage: 10%
- Type of wrap if used: porous gauze dressing and non-irritating tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes
- Time after start of exposure: 24 hrs

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- Constant volume or concentration used: yes
- For solids, paste formed: yes

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.2 ml
Duration of exposure:
24 hrs
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
10 (5/sex)
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: observed at 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing)
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: Yes
Statistics:
Not specified
Preliminary study:
Not specified
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other:
Remarks:
No mortality observed
Mortality:
No mortality was observed at limit dose of 2000 mg/kg body weight of test item during the 14 day observation period (refer table 3).
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 (refer table 2).
Body weight:
The male and female animals were observed with body weight gain compared to day 0 throughout the experiment (refer table 1 and 4).
Gross pathology:
The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality (refer table 5).
Other findings:
No data

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

223

245

256

9.87

14.80

2

276

291

312

5.43

13.04

3

238

253

257

6.30

7.98

4

242

254

261

4.96

7.85

5

242

250

283

3.31

16.94

6

Female

223

230

246

3.14

10.31

7

232

244

242

5.17

4.31

8

212

213

215

0.47

1.42

9

213

224

224

5.16

5.16

10

219

225

231

2.74

5.48

Key:* = Based on day 0 body weight taken prior to dose application.

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:
Result obtained from present investigation can be concluded that the test chemical is acutely non toxic at the tested dose level of 2000 mg/kg b.wt in wistar rats when applied by dermal route. The acute dermal LD50 of test chemical found to be more than 2000 mg/kg b.wt. (> 2000 mg/kg b.wt.).
Executive summary:

Acute dermal toxicity study of test chemical was conducted as per OECD No.402 in Wistar Rats. 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 the skin 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 114. 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). Bodyweights 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. Therefore, under the conditions of this; the acute dermal median lethal dose of test chemical was > 2000 mg/kg body weight. It can be concluded that test compound is acutely non toxic to wistar rats.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
200 mg/kg bw
Quality of whole database:
Data is Klimisch 1 and from experimental study report

Additional information

Acute oral toxicity:

In different experimental studies, 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 and mice for test chemical. The studies are summarized as below –

The acute oral toxicity of test chemical was tested in 10 male and female White mice at thedose concentration of 2000, 5000 and 10000 mg/kgbw. The given test chemical was dissolved as 20% w/v solutionin groundnut oil and administered via oral gavage route. The animals were observed for signs of toxicity for 7 days. Survivors were weighed before killing for post-mortem examination at the end of the one week observation period. All animals dying were autopsied. All survivors were killed and examined post mortem after 7 days.The approximate LD50 value is estimated from the results and the test substance is given a toxicity rating according to Hodge and Sterner's classification. Mortality was observed as - At 2000 mg/kg – No mortality was observed. At 5000 mg/kg – 1 mouse died within 24 hours and 1 mouse died within 96 hours after treatment. At 10000 mg/kg – Both the mice were died within 48 and 66 hours. Clinical signs were observed such as - At 2000 mg/kg – The male mouse was somnolent, showing signs of stress and exhibiting labored breathing within 18 hours after treatment but recovered within 42 hours. The female mouse appeared unaffected by the treatment. At 5000 mg/kg – The mice were showing signs of stress and exhibiting lacrimation within 30 mins after treatment and within 2 hours somnolence and laboured breathing was also observed. These mice were comatose within 18-42 hours after treatment. The surviving mice continued to show signs of stress throughout the 7 day observation period. At 10000 mg/kg – The mice were showing signs of stress and exhibiting lacrimation within 30 mins after treatment and within 2 hours somnolence and laboured breathing was also observed. These mice were comatose within 18-42 hours after treatment. Surviving animals gained weight during the 7 day observation period and presented a normal appearance at autopsy. Autopsy of the animals that died revealed gaseous distension and irritation of the stomach and intestines. The mice dying after 48 hours and later had pale soft rose-pink livers, pale spleens and mottled kidneys. Hemorrhaging was also present in the ileum of these mice. Therefore, LD50 value was considered in between 5000-10000 mg/kg bw, when 10 male and female White mice were treated with test chemical via oral gavage route.

The above experimental study is supported with the study mentioned in peer-reviewed journal, handbook and authoritative database, for the target chemical.Acute oral toxicity study of test chemical was conducted in rats at the dose concentration of 6800 mg/kg bw. 50% mortality was observed at 6800 mg/kg bw. Therefore, LD50 value was considered to be 6800 mg/kg bw, when rats were treated with test chemical via oral route.

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, test chemical cannot be classified for acute oral toxicity.

Acute inhalation toxicity:

The acute inhalation study of test compound was conducted in albino rat under the OECD-Guideline -403 for testing of chemicals. In limit test, ten healthy wistar albino rats of body weight 200±20 gm were selected for study after acclimatization. The test group of animals was exposed to aerosol at the concentration of 5 mg/L for period of 4 hrs. After exposure all the animals were closely observed for clinical signs of toxicity at various internals such as 1 hr, 2 hrs, 4 hrs, and 6 hrs on the day of test compound aerosol exposure and later on twice a day throughout the experimentation period of 14 days. The necropsy was performed on all the animals at termination of experiment. All the albino rats exposed to aerosol at the concentration of 5 mg/L did not show any clinical signs of intoxication. Furthermore, no mortality was observed throughout the period of observation. The necropsy was performed on all the animals at the termination of study did not show any gross pathological changes. In confirmatory test, after 72 hrs, the result obtained from limit test was confirmed in another 10 animal of both sex at similar concentration following same guideline. Ten healthy wistar albino rats of both sex body weight 200±20 gm were selected for study after acclimatization. The test groups of animals were exposed to aerosol at the concentration of 5 mg/L for period of 4 hrs. After exposure all the animals were closely observed for clinical signs of toxicity at various internals such as 1 hr, 2 hrs, 4 hrs, and 6 hrs on the day of test compound aerosol exposure and later on twice a day throughout the experimentation period of 14 days. The necropsy was performed on all the animals at termination of experiment. All the albino rats exposed to aerosol at the concentration of 5 mg/L did not show any clinical signs of intoxication. Furthermore, no mortality was observed throughout the period of observation. The body weight of animals exposed to test compound, observed on day 0th (pre treatment) and day 7th (post treatment) did not differ significantly as compared to day 0th. Whereas, body weight of animals observed on day 14th showed slight increase as compared to day 0 and 7th. The necropsy was performed on all the animals at the termination of study did not show any gross pathological changes. Hence, the acute lethal Concentration (LC50) of test compound in wister albino rats was considered to be >5 mg/L. The result obtained from present investigation can be concluded that test compound test chemical is acutely non toxic to wistar albino rats.

Acute Dermal Toxicity:In different experimental studies, 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 and rabbits for test chemical. The studies are summarized as below –

Acute dermal toxicity study of test chemical was conducted as per OECD No.402 in Wistar Rats. 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 the skin 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). Bodyweights 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. Therefore, under the conditions of this; the acute dermal median lethal dose of test chemical was > 2000 mg/kg body weight. It can be concluded that test compound is acutely non toxic to wistar rats.

The above experimental study is supported with another study mentioned in study report for the target chemical.

The acute dermal toxicity study was conducted on wistar albino rats under OECD guideline-402 Guideline for Testing of Chemicals. In limit test, ten healthy wistar albino rats of both sex (ranging b.wt 200±20 gm) 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 article was applied dermally at the dose level of 2000 mg/kg b.wt for each animal. The treated animals were observed for clinical signs of intoxication and mortality at different time interval 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 which were sacrificed at termination of the study. The test compound applied at the dose level of 2000 mg/kg b.wt in Wistar albino rats did not show any clinical signs of toxicity throughout the observation period of 14 days.   However, after 24 hrs, side of application of test compound showed slight redness in skin. No mortality was observed throughout the period of observation (14 days). After 72 hrs a confirmatory test was conducted in same species of animals to confirm the limit test of test compound (OECD-402 guidelines). Ten healthy wistar albino rats of both sex (ranging b.wt 200±20 gm) 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 article was applied dermally at the dose level of 2000 mg/kg b.wt for each animal. The treated animals were observed for clinical signs of intoxication and mortality at different time interval 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 which were sacrificed at termination of the study. The test compound did not produce any mortality at the tested dose level of 2000 mg/kg b.wt in wistar albino rats throughout the period of observation. The test compound did not elicit any clinical signs of toxicity entire the observation period. However, slight redness of the skin was observed after 24th hrs of patch removal. The body weight of animals treated with test compound observed on days 7th & 14th showed normal gain in body weight as compared to day 0. Therefore, the acute lethal Concentration (LD50) of test chemical in wister albino rats was considered to be >2000 mg/kg bw. The result obtained from present investigation can be concluded that test chemical is acutely non toxic to wistar albino rats.

Both the above experimental studies are further supported with the study mentioned in peer-reviewed journal and authoritative database for the target chemical.Acute dermal toxicity study of test chemical was conducted in rabbits at the dose concentration of 2000 mg/kg bw. No mortality was observed at 2000 mg/kg bw. Therefore, LD50 value was considered to be >2000 mg/kg bw, when rabbits were treated with test chemical by dermal application.

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, 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 acute dermal toxicity; and LC50 value is >5 mg/L, for acute inhalation toxicity. Thus, comparing these values with the criteria of CLP regulation, test chemical cannot be classified for acute oral, acute dermal and acute inhalation toxicity.