Registration Dossier

Administrative data

Description of key information

Acute oral toxicity: 

The acute oral toxicity dose (LD50) was considered based on different studies conducted on rats for the given test chemical. The 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, the given test chemical cannot be classified for acute oral toxicity.

Acute Inhalation Toxicity:

The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to low vapour pressure of the test chemical, which is reported to be 4.81E-9 hPa. Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver. 

 

Acute Dermal toxicity:

The acute dermal toxicity dose (LD50) was considered based on different studies conducted on rats and rabbits for the given test chemical. The studies concluded that LD50 value is >2000 mg/kg bw, for acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given 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 purpose of this study was to assess the Toxicological profile of test item to a single administration via oral route to Sprague Dawley rats. This study was designed to determine the acute toxicity at fixed dose levels by oral route of the test item.
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: National institute of biosciences Pune
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8-12 weeks
- Weight at study initiation: Mean:202.98g (=100%)
Minimum:196.1 g (-3.39%)
Maximum: 207.1 g (+2.03%)
- Fasting period before study: Approx. 16 hours or more
- Housing:The rats were housed in polycarbonate cages.
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences. Pune, was provided ad libimm from individual feeders.
- Water (e.g. ad libitum):Rodent feed supplied by the Nutrivet Life Sciences. Pune, was provided ad libimm from individual feeders
- Acclimation period: acclimatized 5 days prior to study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.5-23.0 degree celcius
- Humidity (%): 56-58.9%
- Air changes (per hr): 10-15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light and dark period

IN-LIFE DATES: From: 05-03-2018 To: 04-04-2018
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 300, and 2000 mg/kg bw
- Amount of vehicle (if gavage): 10 ml/kg body weight
Doses:
Group I: Step I and Step II: 300 mg/kg bw
Group II: Step I and Step II: 2000 mg/kg bw
No. of animals per sex per dose:
Three females were used at each step.
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical Observations and General Appearance: Animals were observed for clinical signs, mortality and morbidity, until sacrifice. Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at immediately (0 to 5 minutes), 5, 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily thereafter for 14 day. Daily observation was done as far as possible at the same time. The observations were included general clinical signs, observations of eyes, mucous membranes, respiratory, circulatory system and behavior pattern.
Body weights: Individual animal body weights were recorded, before fasting, prior to administration of the test item (fasting body weights), weekly thereafter and at termination on day 14. Weight changes were calculated and recorded.
- Necropsy of survivors performed: yes, necropsy was performed on all animals at the end of the study period on day 15. Macroscopic examination of all the orifices, cavities and tissues were made and the findings were recorded. All animals surviving the study period were sacrificed by the carbon dioxide asphyxiation technique.
- Other examinations performed: Histopathology: No gross abnormalities were observed in animals sacrificed terminally hence, no histopathology was performed.
Statistics:
Not specified
Preliminary study:
Not specified
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No Mortality observed
Mortality:
All animals survived through the study period of 14 days.
Clinical signs:
Group I Step I : Animals treated at the dose level of 300 mg/kg body weight resulted in polyurea with onset at 1 to 4 hours after the dosing. All animals were free of signs of toxicity on day 1 after the dosing.
Group I Step II : Animals treated at the dose level of 300 mg/kg body weight resulted in polyurea with onset at 4 to 6 hours after the dosing. All animals were free of signs of toxicity on day 1 after the dosing.
Group II Step I : Animals treated at the dose level of 2000 mg/kg body weight resulted in polyurea with onset at 1 to 2 hours after the dosing. All animals were free of signs of toxicity on day 1 to day 2 after the dosing.
Group II Step II : Animals treated at the dose level of 2000 mg/kg body weight resulted in polyurea with onset at 1 to 2 hours after the dosing. All animals were free of signs of toxicity on day 2 after the dosing.
Body weight:
Group I Step l (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 8.6496 and 15.90% respectively.
Group l Step ll (300 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 6.71% and 14.46% respectively.
Group ll Step 1 (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 5.4691% and 12.33% respectively.
Group II Step II (2000 mg/kg) – Percent body weight gain after 7 days and 14 days was found to be 6.4892% and 12.77% respectively.
Gross pathology:
Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups.
Other findings:
Not specified

Table 1: Summary of clinical signs of toxicity ad mortality

Group No.

Step No.

Dose

Mg/kg

Observed signs

Total no. of animals

Animal no.

Period of signs in days

From-To

Mortality

I

1

300

Polyurea

3

1

1 hrs-6hrs

0/3

 

 

 

 

 

2

4 hrs-6hrs

 

 

 

 

 

 

3

2 hrs-6hrs

 

I

2

300

Polyurea

3

4,6

4 hrs-6hrs

0/3

 

 

 

 

 

5

6 hrs

 

II

1

2000

Polyurea

3

7

1 hrs-6hrs

0/3

 

 

 

 

 

8

2 hrs-day 1

 

 

 

 

 

 

9

1 hrs- day 1

 

II

2

2000

Polyurea

3

10,12

2 hrs-day 1

0/3

 

 

 

 

 

11

1 hrs- day 1

 

 

Table 2: Mean body weight and percent body weight gain (g)

Group

Step no.

 

Before fasting body weight

Day 0

Body weight

Day 7

% body weight gain

Day 0-7

Body weight

Day 14

% body weight gain

Day 7-14

% body weight gain

Day 0-14

I

1

Mean

201.13

218.50

8.64

233.07

6.67

15.90

 

 

± SD

4.78

3.97

0.68

3.81

1.31

2.08

I

2

Mean

202.30

215.87

6.71

231.57

7.26

14.46

 

 

± SD

3.56

2.86

0.47

6.16

1.45

1.07

II

1

Mean

204.97

216.17

5.46

230.23

6.52

12.33

 

 

± SD

2.53

3.95

0.65

2.45

0.90

0.47

II

2

Mean

203.50

216.70

6.48

229.50

5.91

12.77

 

 

± SD

4.19

5.52

0.66

5.32

0.26

0.56

 

 

Table 3: Summary of gross pathological findings

Group

Step no.

Dose

Mg/kg

Animal numbers

Animal fate

Gross Pathological Findings

I

1

300

1-3

TS

No abnormality detected

I

2

300

4-6

TS

No abnormality detected

II

1

2000

7-9

TS

No abnormality detected

II

2

2000

10-12

TS

No abnormality detected

TS= Terminal Sacrifice

Table 4: Individual animal-Body weight and Percent body weight gain (g)

Group I                       Step 1                                          Dose: 300 mg/kg bw

Animal no.

Before fasting body weight

Day 0

Body weight

Day 7

% body weight gain

Day 0-7

Body weight Day 14

% body weight gain

Day 7-14

% body weight gain

Day 0-14

1

205.6

221.8

7.88

233.4

5.23

13.52

2

196.1

214.1

9.18

229.1

7.01

16.83

3

201.7

219.6

8.87

236.7

7.79

17.35

 

Group I                       Step 2                                          Dose: 300 mg/kg bw

Animal no.

Before fasting body weight

Day 0

Body weight

Day 7

% body weight gain

Day 0-7

Body weight Day 14

% body weight gain

Day 7-14

% body weight gain

Day 0-14

4

202.6

216.1

6.66

231.2

6.99

14.12

5

198.6

212.9

7.20

225.6

5.97

13.60

6

205.7

218.6

6.27

237.9

8.83

15.65

 

Group II                       Step 1                                          Dose: 2000 mg/kg bw

Animal no.

Before fasting body weight

Day 0

Body weight

Day 7

% body weight gain

Day 0-7

Body weight Day 14

% body weight gain

Day 7-14

% body weight gain

Day 0-14

7

205.9

217.2

5.49

230.2

5.99

11.80

8

202.1

211.8

4.80

227.8

7.55

12.72

9

206.9

219.5

6.09

232.7

6.01

12.47

 

Group II                       Step 2                                          Dose: 2000 mg/kg bw

Animal no.

Before fasting body weight

Day 0

Body weight

Day 7

% body weight gain

Day 0-7

Body weight Day 14

% body weight gain

Day 7-14

% body weight gain

Day 0-14

10

198.9

210.4

5.78

223.4

6.18

12.32

11

204.5

219.0

7.09

231.9

5.89

13.40

12

207.1

220.7

6.57

233.2

5.66

12.60

Interpretation of results:
other: Not classified
Conclusions:
It was concluded that the acute oral toxicity dose (LD50) value was considered to be >2000 mg/kg bw, when Sprague Dawley rats were treated with the given test chemical via oral route.
Executive summary:

The reported study was designed and conducted to determine the acute oral toxicity profile as per OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method) in Sprague Dawley rats.

Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Step - I). Administration of the test item at 300 mg/kg resulted in polyurea with onset at 6 hours after the dosing and no mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, three female animals were added to the study and treated with the same dose of 300 mg/kg of the test item (Step - II). Administration of the test item at 300 mg/kg resulted in polyurea with onset at 4 to 6 hours after the dosing and no mortality at 24 hours after the dosing. No mortality was observed at 300 mg/kg dose group, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - I). Administration of the test item at 2000 mg/kg resulted in polyurea with onset at 1 to 2 hours after the dosing and no mortality at 24 hours after the dosing. As no mortality were observed at 24 hours after the dosing, additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - II). Administration of the test item at 2000 mg/kg resulted in polyurea with onset at 1 to 2 hours after the dosing and no mortality at 24 hours after the dosing. Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups.

It was concluded that the acute oral toxicity dose (LD50) value was considered to be >2000 mg/kg bw, when Sprague Dawley rats were treated with the given test chemical via oral route.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Data is Klimisch 1 and from study report.

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:
the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size
Endpoint conclusion
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:
This study was designed to determine the dermal LD50 of the test item (up to 2000 mg/kg bw) or to establish a non-lethal dose level of 2000 mg/kg bw.
GLP compliance:
yes
Test type:
other: Acute Dermal Toxicity
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
- Source: National institute of biosciences Pune
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 8-10 weeks
- Weight at study initiation: Mean:227.60g (=100%)
Minimum:221.6 g (-2.64%)
Maximum: 233.3 g (+2.50%)
- Housing:The rats were housed in polycarbonate cages with paddy husk and bedding
- Diet (e.g. ad libitum): Rodent feed supplied by the Nutrivet Life Sciences. Pune, was provided ad libimm from individual feeders.
- Water (e.g. ad libitum):Water was provied ad libitum
- Acclimation period: acclimatized 5 days prior to study

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-22.5 degree celcius
- Humidity (%): 55.3-58.8%
- Air changes (per hr): 10-15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light and dark period

IN-LIFE DATES: From: 06-03-2018 To: 04-04-2018
Type of coverage:
semiocclusive
Vehicle:
water
Details on dermal exposure:
TEST SITE
- Area of exposure: the trunk (dorsal surface and sides from scapular to pelvic area)
- % coverage: approximately 10% of the total body surface area
- Type of wrap if used: The test item was held in contact with the skin using a porous gauze dressing and non irritating tape around the animal to cover the exposure site for first 24 hours exposure period. The test item was held in contact with the skin using a porous gauze dressing and non irritating tape around the animal to cover the exposure site for first 24 hours exposure period.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): The wrapping was removed and the test site wiped free of excess test item. Distilled water was used to remove residual test item.
- Time after start of exposure:24 hrs

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):200, 1000 and 2000 mg/kg bw
- Constant volume or concentration used: yes
- For solids, paste formed: yes
Duration of exposure:
24 hrs
Doses:
Dase range finding study: Group I: 200, 1000 and 2000 mg/kg bw
Main study: Group II: 2000 mg/kg bw
No. of animals per sex per dose:
Dase range finding study: Group I: 200, 1000 and 2000 mg/kg bw : 1/dose
Main study: Group II: 2000 mg/kg bw: 2/dose
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical Observations and General Appearance: Animals were observed for clinical signs, mortality, until sacrifice. Onset, duration and severity of any sign were recorded. The clinical signs and mortality observations were conducted at 10, 30, 60 minutes, 2, 4 and 6 hours on the day of dosing and once daily thereafter for 14 day. Daily observation was done as far as possible at the same time. The observations were included general clinical signs, observations of eyes, mucous membranes, respiratory, circulatory system and behavior pattern.
Evaluation of Dermal Reaction: Dermal reaction was observed daily for study period of 14 days.
Body weights: Individual animal body weights were recorded pre-test (prior to administration of the test item), day 7 and at termination on day 14.
- Necropsy of survivors performed: yes, necropsy was performed on animals surviving at the end of the study. Macroscopic examination of all the orifices, cavities and tissues were made and the findings were recorded. All animals surviving the study period were sacrificed by the carbon dioxide asphyxiation technique (day 15).
- Other examinations performed: Histopathology: No gross abnormalities were observed in animals sacrificed terminally hence, no histopathology was performed.
Statistics:
not specified
Preliminary study:
Dose range finding study: A single dose of 200 mg/kg bw of the test item was administered to 1 female animal. No death or clinical signs of toxicity was observed during first 48 hours, hence, addtional 1 female animal was administered at the dosr 1000 mg/kg bw. Administration of 1000 mg/kg bw did not reveal any clinical signs of toxicity or death during first 48 hours, hence, additional 1 female animal was administered at the dose of 2000 mg/kg bw. Administration of 2000 mg/kg bw did not reveal any clinical signs of toxicity or death during first 48 hours.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality observed
Mortality:
Dose Range Finding Study: All animals survived through the study period of 14 days at 200 mg/kg, 1000 mg/kg and 2000 mg/kg body weight .
Main Study: Group II : All animals survived through the study period of 14 days.
Clinical signs:
Dose Range Finding Study: Animals treated at the dose level of 200 mg/kg, 1000 mg/kg and 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days.
Main Study: Group II : Animals treated at the dose level of 2000 mg/kg body weight did not result in any signs of toxicity during the study period of 14 days.
Body weight:
Dose Range Finding Study:
Group 1 (200 mg/kg) - Percent body weight gain after 7 days and I4 days was found to be 4.61% and 8.15% respectively.
Group I (1000 mg/kg) - percent body weight gain utter 7 days and 14 days was found to be 5.4296 and 9.70% respectively.
Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.939% and 8.99% respectively.
Main Study:
Group II (2000 mg/kg) - Percent body weight gain alter 7 days and 14 days was found to be 3.58% and 9.0394% respectively.
Gross pathology:
Gross pathological examination did not reveal any abnormalities in animals from 200 mg/kg, 1000 mg/kg and 2000 mg/kg dose groups from dose range finding study and main study sacrificed terminally.
Other findings:
Evaluation of Dermal Reaction -
Dose Range Finding Study: Group I : Animal treated at the dose level of 200 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Group I : Animal treated at the dose level of 1000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Group I : Animal treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.
Main Study: Group II : Animals treated at the dose level of 2000 mg/kg body weight did not result in any skin reaction during the study period of 14 days.

TABLE 1:SUMMARY OF BODY WEIGHT (GM)

Group

Animal ID

Day 0

Day 7

%

Gain/loss

Day 14

%

Gain/loss

Group-I

2000 mg/kg bw

201320-1

202.5

211.1

4.24

218.7

8.00

 

201320-2

200.9

210.7

4.87

215.1

7.07

 

201320-3

203.2

212.9

4.78

217.8

7.19

 

201320-4

203.2

210.2

3.09

216.9

6.38

 

201320-5

201.2

209.7

4.23

216.1

7.41

 

201320-6

200.4

211.9

5.82

218.5

9.10

 

201320-7

205.7

213.9

4.00

219.9

6.90

 

201320-8

204.9

212.4

3.70

220.1

7.50

 

201320-9

208.4

214.9

3.20

221.8

6.50

 

201320-10

206.9

213.9

3.40

219.9

6.30

Group-II

2000 mg/kg bw

201320-11

208.2

214.5

3.10

222.1

6.70

 

201320-12

206.8

213.9

3.50

220.8

6.78

 

201320-13

209.6

215.9

3.00

219.1

4.60

 

201320-14

203.4

213.6

5.10

218.4

7.40

 

201320-15

205.9

214.7

4.40

220.0

6.90

 

201320-16

208.7

217.9

4.40

221.8

6.30

 

201320-17

205.9

214.1

3.99

219.8

6.75

 

201320-18

207.8

215.5

4.00

219.8

6.20

 

201320-19

202.6

212.9

5.10

217.3

7.30

 

201320-20

201.6

212.6

5.50

218.6

8.50

TABLE 3: CLINICAL SIGNS AND MORTALITY

Group-I Limit test                            Dose :2000 mg/kg b.wt.

Parameters

Incidence of clinical signs observed after dosing on

Mortality

 

Day 0

 

DAY

 

Min

Hour

 

 

30

1

2

4

6

1

2

3

4

5

6

7

8

9

10

11

12

13

14

Total

%

%

Mortality (total)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0/10

 

00

Clinical signs-local

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

 

 

Redness

-

-

-

-

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

Pain

-

-

-

-

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

Swelling

-

-

-

-

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

Systemic signs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

- = OBSERVED AFTER 24 HOURS

0 = NO CLINICAL SIGNS

+ =MILD

++ = MODERATE

+++ =HIGH

++++=SEVERE

 

 

TABLE 3 CONTINUED

GROUP :II CONFIRMATORY TEST        Dose:2000 mg/kg b.wt

Parameters

Incidence of clinical signs observed after dosing on

Mortality

Day 0

 

DAY

Min

Hour

 

30

1

2

4

6

1

2

3

4

5

6

7

8

9

10

11

12

13

14

Total

%

Mortality (total)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0/10

0

Clinical signs-local

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

Redness

-

-

-

-

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Pain

-

-

-

-

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Swelling

-

-

-

-

-

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Systemic signs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

- = OBSERVED AFTER 24 HOURS

0 = NO CLINICAL SIGNS

+ =MILD

++ = MODERATE

+++ =HIGH

++++=SEVERE

  

 

Table 4 :SUMMARY OF NECROPSY FINDINGS

S. NO.

FATE

WISTAR ALBINO RATS

DOSE (mg/kg b.wt)

2000

(limit test)

2000

(confirmatory test)

1.

Terminal sacrifice

10/10

10/10

2.

Found dead

0/10

0/10

3.

Abnormalities detected

0/10

0/10

Interpretation of results:
other: Not classified
Conclusions:
It was concluded that the acute dermal median lethal dose (LD50) of the given test chemical, when administered to female Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.
Executive summary:

The reported study was designed and conducted to determine the acute dermal toxicity profile of the given test chemical as per OECD Guideline 402 (Acute Dermal Toxicity) in Sprague Dawley rats.

In the dose range finding study a single dose of 200 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity was observed during first 48 hours, hence, additional 1 female animal was administered with the dose of 1000 mg/kg body weight. Administration of 1000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours; hence, additional 1 female animal was administered at the dose of 2000 mg/kg body weight. Administration of 2000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours.

As the dose range finding study revealed no mortality or clinical signs at the maximum dose of 2000 mg/kg, the main study was initiated with two additional animals. The animals were administered with a dose of 2000 mg/kg body weight in sequential manner at 48 hours intervals.

Animals from dose range finding study treated at the dose levels of 200 mg/kg, 1000 mg/kg and 2000 mg/kg and animals from main study treated at the dose level of 2000 mg/kg exhibited normal body weight gain and revealed no clinical signs of toxicity or mortality during the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

It was concluded that the acute dermal median lethal dose (LD50) of the given test chemical, when administered to female Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
Data is Klimisch 1 and from study report.

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 the given test chemical. The studies are summarized as below –

 

The reported study was mentioned in study report and conducted to determine the acute oral toxicity profile as per OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method) by using the given test chemical in Sprague Dawley rats.

Initially, three female animals were treated at the dose level of 300 mg/kg body weight of the test item (Step - I). Administration of the test item at 300 mg/kg resulted in polyurea with onset at 6 hours after the dosing and no mortality at 24 hours after the dosing. As no mortality was observed at 24 hours after the dosing, three female animals were added to the study and treated with the same dose of 300 mg/kg of the test item (Step - II). Administration of the test item at 300 mg/kg resulted in polyurea with onset at 4 to 6 hours after the dosing and no mortality at 24 hours after the dosing. No mortality was observed at 300 mg/kg dose group, hence additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - I). Administration of the test item at 2000 mg/kg resulted in polyurea with onset at 1 to 2 hours after the dosing and no mortality at 24 hours after the dosing. As no mortality were observed at 24 hours after the dosing, additional three female animals were treated with the higher dose of 2000 mg/kg of the test item (Step - II). Administration of the test item at 2000 mg/kg resulted in polyurea with onset at 1 to 2 hours after the dosing and no mortality at 24 hours after the dosing. Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups.

It was concluded that the acute oral toxicity dose (LD50) value was considered to be >2000 mg/kg bw, when Sprague Dawley rats were treated with the given test chemical via oral route.

 

The above study is supported with the data mentioned in study report and the study was conducted by using the given test chemical under the OECD Guideline-423 for testing of chemicals in Wistar albino rats.

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. The study was conducted stepwise as follow: Starting dose 2000 mg/kg body weight: Step-I - The test compound was mixed with distilled water and administered orally at the dose level of 2000 mg/kg body weight (dose volume 10ml/kg body weight) to three female rats. However; vehicle control group treated with the distilled water at the dose level of 10 ml/kg b.wt.  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 once daily for a period of 14 days.  The necropsy was performed on all animals at termination of the study. The wistar albino rats treated with the test compound did not produce any mortality and clinical signs of toxicity throughout the observation period. Furthermore, No clinical signs and mortality were observed in vehicle control group. The necropsy finding did not reveal any gross pathological changes. Furthermore, no gross pathological change was observed in vehicle control group.

Step -II: After 72 hrs, the result of step-I was confirmed by administration of same dose level (2000 mg/kg. b.wt) of test compound in additional three animals of same sex (OECD-423 guidelines). The test compound administered at the dose level of 2000 mg/kg b.wt did not produce any mortality and clinical sign of intoxication throughout the observation period of 14 days. The test compound did not elicit any gross pathological changes in animals sacrificed at the end of experimentation.

The acute oral LD50 value was considered to be >2000 mg/kg bw, when rats were treated with test chemical via oral route. Thus, it was concluded that the test compound following the guideline OECD-423 is non toxic to wistar albino rats.

 

These studies are supported with the data available in peer-reviewed journal, authoritative database and different secondary reports for the given test chemical. The acute oral toxicity study was conducted on rats at the dose concentration of 34600 mg/kg bw administered via oral route. All animals were maintained under close observation for recording toxic signs and time of death for 14 days. No mortality observed at 34600 mg/kg bw dose level in treated animals. Therefore, the acute oral LD50 value was considered to be >34600 mg/kg bw, when rats were treated with the given test chemical via oral route.

 

This study is supported with the data available in authoritative database and different secondary sources for the given test chemical. The acute oral toxicity study was conducted on 5 male and 5 female rats at the dose concentration of 2000 mg/kg bw. The test chemical was administered via oral route with polyethylene glycol 400 as a vehicle. All animals were maintained under close observation for recording toxic signs and time of death for 14 days. No mortality observed within 14 days at 2000 mg/kg bw. Therefore, the acute oral LD50 value was considered to be >2000 mg/kg bw, when 10 male and female rats were treated with the given test chemical via oral route.

 

All the above studies are further supported with the data mentioned in secondary source for the given test chemical. Acute oral toxicity study was conducted on male and female Sprague-Dawley rats. Diluted compound was fed by stomach tube to Sprague-Dawley albino male and female rats in increasing doses at 0.3 and 0.2 fractional log intervals. Observations were made for toxic symptoms. No mortality observed at a dose of 50100 mg/kg bw. Therefore, the acute oral LD50 value was considered to be >50100 mg/kg bw, when male and female Sprague-Dawley rats were treated with the given 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, for acute oral toxicity. 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 acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to low vapour pressure of the test chemical, which is reported to be 4.81E-9 hPa. Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver. 

 

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 and rabbits for the given test chemical. The studies are summarized as below –

 

The reported study was mentioned in study report and conducted to determine the acute dermal toxicity profile as per OECD Guideline 402 (Acute Dermal Toxicity) by using the given test chemical in Sprague Dawley rats.

In the dose range finding study a single dose of 200 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity was observed during first 48 hours, hence, additional 1 female animal was administered with the dose of 1000 mg/kg body weight. Administration of 1000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours; hence, additional 1 female animal was administered at the dose of 2000 mg/kg body weight. Administration of 2000 mg/kg body weight did not reveal any clinical signs of toxicity or death during first 48 hours. As the dose range finding study revealed no mortality or clinical signs at the maximum dose of 2000 mg/kg, the main study was initiated with two additional animals. The animals were administered with a dose of 2000 mg/kg body weight in sequential manner at 48 hours intervals.

Animals from dose range finding study treated at the dose levels of 200 mg/kg, 1000 mg/kg and 2000 mg/kg and animals from main study treated at the dose level of 2000 mg/kg exhibited normal body weight gain and revealed no clinical signs of toxicity or mortality during the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

It was concluded that the acute dermal median lethal dose (LD50) of the given test chemical, when administered to female Sprague Dawley rats was considered to be >2000 mg/kg body weight. Thus, according to CLP criteria for acute toxicity rating for the chemicals, it infers that the given test chemical does not classify as an acute dermal toxicant. CLP Classification: “Not classified”.

 

This study is supported with the data mentioned in another study report and the acute dermal toxicity study was conducted by using the given test chemical according to OECD guideline 402 for testing of chemicals on Wistar albino rats. The summary of the study was as follows- LIMIT TEST (2000 mg/kg body weight): Ten healthy wistar albino rats of both sexes (ranging b.wt 200±20 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 drug 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 continuously for 4 hrs and thereafter at 1 hr interval for period of 24 hrs and twice a day for 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. 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 and mortality throughout the observation period of 14 days. Furthermore, test compound did not elicit any gross pathological changes at the test dose level.

CONFIRMATORY TEST: 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) 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 compound was applied dermally at the dose level of 2000 mg/kg b.wt to each animal. The treated animals were observed for clinical signs of intoxication and mortality continuously for 4 hrs and thereafter 1 hr interval for period of 24 hrs and twice a day for 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 died during the study or 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. Furthermore, the test compound did not elicit any clinical signs of toxicity in the entire observation period. No skin reaction was observed after 24 hrs of patch removal. The body weight of animals treated with test compound observed on days 7th 14th showed normal gain in body as compared to day 0 (pre-treatment)  

Result obtained from the present investigation it can be concluded that the test compound is acutely non toxic at the tested dose level of 2000 mg/kg b.wt in Wistar albino rats when applied by dermal route.

 

These studies are supported with the data available in authoritative database and different secondary sources for the test chemical. The acute dermal toxicity study was conducted on male and female New Zealand White rabbits at the dose concentration of 10000 mg/kg bw.

The diluted compound was applied in increasing doses at 0.2 fractional log intervals to the closely clipped, intact skin of New Zealand white male and female rabbits. The treated areas were covered with plastic strips and the animals placed in wooden stocks for periods up to 24 hr, after which time they were assigned to individual cages. Observations were made for toxic symptoms and, since there were no deaths, no autopsies were performed. No mortality was observed at 10000 mg/kg bw.

Hence, the acute dermal LD50 value was considered to be >10000 mg/kg bw, when male and female New Zealand White rabbits were treated with the given test chemical via dermal route occlusively.

 

All the above studies are further supported with the data mentioned in authoritative database and secondary source for the given test chemical. Acute dermal toxicity study was conducted on rabbits at the dose concentration of 7940 mg/kg bw. The test chemical was administered via dermal route. All animals were observed for mortality. No mortality was observed at 7940 mg/kg bw dose level. Hence, the acute dermal LD50 value was considered to be >7940 mg/kg bw, when rabbits were treated with test chemical via dermal route.

 

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