Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Acute oral toxicity: 

The acute oral toxicity dose (LD50) was considered based on experimental study conducted on rats for the test chemical. The LD50 value is between 50 - 300 mg/kg bw, for acute oral toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical can be classified in “Category 3” for acute oral toxicity.

Acute Inhalation Toxicity:

The acute inhalation toxicity dose (LC50) was considered based on different studies conducted on rats for the test chemical. The LC50 value considered to be 1300 mg/m3. The study concluded that LC50 is between 1 to ≤ 5 mg/L, for acute inhalation toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical can be classified in “Category 4” for acute inhalation toxicity.

Acute Dermal toxicity:

The acute dermal toxicity dose (LD50) was considered based on experimental study conducted on rats for the test chemical. The 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 experimental study report.
Qualifier:
according to guideline
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 or test system and environmental conditions:
TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: Female rats of the age of approximately 8 to 12 weeks old were used at the commencement of its dosing.
- Weight at study initiation: Body weight range was 193.7 to 201.8 grams.
Body weights at the start : Female Mean: 198.47 g (= 100 %); Minimum : 193.7 g (- 2.40 %); Maximum : 201.8 g (+ 1.68 %)
- Identification: Each female rat was individually identified by the picric acid marking.
- Fasting period before study: Approximately 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 libitum from individual feeders.
- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period: 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.0 to 22.0 degree centigrade.
- Humidity (%): 54.4% to 58.9%.
- Air changes (per hr): at least ten to fifteen air changes per hour of 100% fresh air that had been passed through the HEPA filters
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room

IN-LIFE DATES: 10-10-2017 to 28-10-2017
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 50 mg/kg , 50 mg/kg and 300 mg/kg
MAXIMUM DOSE VOLUME APPLIED: 10 ml per kg of body weight
Doses:
Group I Step I : 300 mg/kg
Group II Step I : 50 mg/kg
Group II Step II : 50 mg/kg
No. of animals per sex per dose:
Total No. of animals : 9
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Twice daily
- Necropsy of survivors performed: yes, necropsy was performed on all animals, found dead and sacrificed 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:
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.
Histopathology:
Gross pathological examination revealed unabsorbed residual test item in stomach in found dead animals from 300 mg/kg dose group. No gross pathological changes attributable to the treatment were observed in stomach and hence no histopathology was considered.
Statistics:
not specified
Preliminary study:
not specified
Sex:
female
Dose descriptor:
LD50
Effect level:
> 50 - <= 300 mg/kg bw
Based on:
test mat.
Mortality:
Group I Step I : 300 mg/kg - Three animals died at 1 hour after the dosing.
Group II Step I : 50 mg/kg - All animals survived through the study period of 14 days.
Group II Step II : 50 mg/kg - All animals survived through the study period of 14 days.
Clinical signs:
other: Group I Step I : 300 mg/kg - Animals treated at the dose level of 300 mg/kg resulted in reduced locomotor activity, loss of righting reflex and convulsions with onset at 30 minutes after the dosing. Group II Step I : 50 mg/kg - Animals treated at the d
Gross pathology:
Gross pathological examination did not reveal any abnormalities in all animals sacrificed terminally from 50 mg/kg dose group.
Gross pathological examination revealed unabsorbed residual test item in the stomach in found dead animal from 300 mg/kg dose group.
Other findings:
not specified

Table No. I 

Summary of Clinical Signs of Toxicity and Mortality

 

Laboratory Test Item Code :TAS/122/058

Test System : Sprague Dawley Rat

Sex : Female

Group I :

Step

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal Nos.

Period of signs in days

From - to

Mortality

I

300

Reduced locomotor activity

3

1,2,3

30 min.

3/3

Loss of righting reflex   

3

1,2,3

30 min.

Convulsions

3

1,2,3

30 min.

 

Group II :

Step

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal Nos.

Period of signs in days

From - to

Mortality

I

50

Reduced locomotor activity

3

4,5,6

2 hrs.

0/3

Ataxic gait    

3

4,5,6

2 hrs. - 4 hrs.

 

Group II :

Step

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal Nos.

Period of signs in days

From - to

Mortality

II

50

Reduced locomotor activity

3

7,8,9

2 hrs.

0/3

Ataxic gait    

3

7,9

8

2 hrs. - 4 hrs.

1 hr. - 4 hrs.

  

Table No.II

Mean Body Weight and Percent Body Weight Gain (g)

 

Laboratory Test Item Code :TAS/122/058

Test System : Sprague Dawley Rat

Sex : Female

Group I :

Step

No.

Dose

(mg/kg body weight)

 

Before Fasting Body weight

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

300

Mean

197.60

-

-

-

-

-

± SD

3.46

-

-

-

-

-

Group II :

Step

No.

Dose

(mg/kg body weight)

 

Before Fasting Body weight

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

50

Mean

197.57

211.13

6.87

228.10

8.04

15.45

± SD

1.82

2.39

0.22

2.36

0.45

0.45

 Group II :

Step

No.

Dose

(mg/kg body weight)

 

Before Fasting Body weight

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

II

50

Mean

200.23

214.80

7.27

232.67

8.31

16.19

± SD

1.78

1.87

0.10

3.76

0.93

0.92

Table No.III

Summary of Gross Pathological Findings

 

Laboratory Test Item Code :TAS/122/058

Test System : Sprague Dawley Rat

Sex : Female

                    Group I :

Step

No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

300

1 - 3

FD

Stomach : Unabsorbed residual test item

 

                     Group II : 

Step

No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

50

4 - 6

TS

No abnormality detected

 

                    Group II : 

Step

No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

II

50

7 - 9

TS

No abnormality detected

 

                    FD = Found dead

         TS = Terminal sacrifice

 

Interpretation of results:
Category 3 based on GHS criteria
Conclusions:
Under the condition of the study, the acute oral LD50 value of the given test chemical was considered in between 50 – 300 mg/kg body weight. Thus, it was concluded that the acute toxicity study of test chemical, when administered via oral route in Sprague Dawley rats falls into the “Category 3” according to criteria of CLP.
Executive summary:

The reported study was designed and conducted to determine the acute oral toxicity profile of the given test chemical 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 reduced locomotor activity, loss of righting reflex and convulsions with onset at 30 minutes after the dosing. Three animals died at 1 hour after the dosing. As mortality was observed at 300 mg/kg dose group, additional three female animals were treated with the lower dose of 50 mg/kg of the test item (Step - I).

Administration of the test item at 50 mg/kg resulted in reduced locomotor activity and ataxic gait with onset at 2 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 additional female animals were added to the study and treated with the dose of 50 mg/kg of the test item (Step - II). Administration of the test item at 50 mg/kg resulted in reduced locomotor activity and ataxic gait with onset at 1 to 2 hours after the dosing and no mortality at 24 hours after the dosing.

All animals from 50 mg/kg dose group survived through the study period of 14 days. Gross pathological examination did not reveal any abnormalities in all animals sacrificed terminally from 50 mg/kg dose group. Gross pathological examination revealed unabsorbed residual test item in the stomach in found dead animal from 300 mg/kg dose group.

Under the condition of the study, the acute oral LD50 value of the given test chemical was considered in between 50 – 300 mg/kg body weight. Thus, it was concluded that the acute toxicity study of test chemical, when administered via oral route in Sprague Dawley rats falls into the “Category 3” according to criteria of CLP.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
200 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
Type of information:
experimental study
Adequacy of study:
weight of evidence
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 authoritative database.
Qualifier:
according to guideline
Guideline:
other: As mentioned below
Principles of method if other than guideline:
Acute inhalation toxicity of the given test chemical in rat.
GLP compliance:
not specified
Test type:
other: not specified
Limit test:
no
Species:
rat
Strain:
not specified
Sex:
not specified
Details on test animals or test system and environmental conditions:
not specified
Route of administration:
inhalation: aerosol
Type of inhalation exposure:
not specified
Vehicle:
other: aqueous solutions
Remark on MMAD/GSD:
not specified
Details on inhalation exposure:
VEHICLE
- Concentration in vehicle: 550 mg/m³ from a 20% solution while the 2600 mg/m³ and 5600 mg/m³ from a 60% solution.
Analytical verification of test atmosphere concentrations:
not specified
Duration of exposure:
4 h
Concentrations:
550 mg/m³ from a 20% solution
2600 mg/m³ and 5600 mg/m³ from a 60% solution
No. of animals per sex per dose:
not specified
Control animals:
not specified
Details on study design:
- Necropsy of survivors performed: yes
- Other examinations performed: Animals were observed for mortality and clinical signs.
Statistics:
not specified
Preliminary study:
not specified
Sex:
not specified
Dose descriptor:
LC0
Effect level:
550 mg/m³ air
Based on:
test mat.
Exp. duration:
4 h
Sex:
not specified
Dose descriptor:
LC50
Effect level:
1 300 mg/m³ air
Based on:
test mat.
Sex:
not specified
Dose descriptor:
LC100
Effect level:
2 600 mg/m³ air
Based on:
test mat.
Exp. duration:
2 h
Sex:
not specified
Dose descriptor:
LC100
Effect level:
5 600 mg/m³ air
Based on:
test mat.
Exp. duration:
1 h
Mortality:
550 mg/m3 - Animals were survived.
2600 mg/m³ and 5600 mg/m³ - Caused the death of all the animals.
Clinical signs:
other: The symptoms were attempts to escape, respiratory distress, seizures.
Body weight:
not specified
Gross pathology:
The findings from the necropsy were unspecific (hyperemia of inner organs). Nothing conspicuous was found in the animals which survived.
Other findings:
not specified
Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The acute inhalation toxicity dose (LC50) was assumed to be 1300 mg/m3, when rats were treated with the given test chemical via inhalation route by aerosols.
Executive summary:

Acute inhalation toxicity study of the given test chemical was conducted in rats at the dose concentration of 550 mg/m³, 2600 mg/m³ and 5600 mg/m³. Rats were exposed to aerosols from aqueous solutions of test chemical.

The given test chemical was exposed as a 20% solution to form 550 mg/m³ and 60% solution to form 2600 mg/m³ and 5600 mg/m³.

The inhalation of a concentration of 550 mg/m³ from a 20% solution was exposed for 4 hours; the inhalation of 2600 mg/m³ exposed for 2 hours and 5600 mg/m³ for 1 hour from a 60% solution.

Animals were observed for mortality and clinical signs.Necropsy of survivors performed. Rats exposed to a concentration of 550 mg/m³ from a 20% solution exposed for 4 hours was survived. While the inhalation of 2600 mg/m³ exposed for 2 hours and 5600 mg/m³ exposed for 1 hour from a 60% solution caused the death of all the animals.

The symptoms were attempts to escape, respiratory distress, seizures. The findings from the necropsy were unspecific (hyperemia of inner organs). Nothing conspicuous was found in the animals which survived.

Under the condition of this, the LC100 value was considered to be 2600 mg/m³ with the exposure time of 2 hours.

Considering the above signs and mortality, the LC50 value was assumed to be 1300 mg/m3, when rats were treated with the given test chemical via inhalation route by aerosols.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LC50
Value:
1 300 mg/m³ air
Quality of whole database:
Data is Klimisch 2 and from authoritative database.

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 experimental study report.
Qualifier:
according to guideline
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) or to establish a non-lethal dose level of 2000 milligram of test item per kilogram of body weight.
GLP compliance:
yes
Test type:
other: Acute Dermal Toxicity
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: National Institute of Biosciences, Pune
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Young adult (8 to 10 weeks old) female rats were used.
- Weight at study initiation: The weight range of approximately 228.0 to 247.7 grams at initiation of dosing.
Body weights at the start : Female Mean: 235.60 g (= 100 %); Minimum : 228.0 g (- 3.23 %); Maximum : 247.7 g (+ 5.14 %)
- Identification: Each rat was individually identified by the cage number.
- Housing: The rats were individually housed in polycarbonate cages with paddy husk as bedding.
- Diet (e.g. ad libitum): Rodent feed was provided ad libitum from individual feeders.
- Water (e.g. ad libitum): Water was provided ad libitum from individual bottles attached to the cages. All water was from a local source and passed through the reverse osmosis membrane before use.
- Acclimation period:at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.2 to 21.8 degree centigrade
- Humidity (%): 56.0% to 59.1%.
- Air changes (per hr): The animal room was independently provided with at least ten to fifteen air changes per hour of 100% fresh air that had been passed through the HEPA filters.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: From: 31-10-2017 To: 24-11-2017
Type of coverage:
occlusive
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. Elizabethan collar was placed on each animal for first 24 hours after application of the test item. These collars prevent ingestion of test item.

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 hours
Duration of exposure:
24 hours
Doses:
Dose Range Finding Study:
Group I : 200 mg/kg
Group I : 1000 mg/kg
Group I : 2000 mg/kg
Main Study: Group II : 2000 mg/kg
No. of animals per sex per dose:
Dose Range Finding Study:
Group I : 200 mg/kg - 1
Group I : 1000 mg/kg - 1
Group I : 2000 mg/kg - 1
Main Study: Group II : 2000 mg/kg - 2
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Twice daily
- 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: 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 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.
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 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 at 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.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality was observed
Mortality:
Dose Range Finding Study: The 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:
other: 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 m
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: Animals treated at the dose level of 200 mg/kg, 1000 mg/kg and 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 No. I

Summary of Clinical Signs of Toxicity and Mortality

 

Laboratory Test Item Code :TAS/122/058

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group

 No.

Dose mg/kg

                            Observed Signs

Total Number of

Animals

 

Animal Nos.

Period of signs in days

 From - to

 

Mortality

I

200

No clinical signs observed

1

1

Day 0 - Day 14

0/1

 

Group

 No.

Dose mg/kg

                            Observed Signs

Total Number of

Animals

 

Animal Nos.

Period of signs in days

 From - to

 

Mortality

I

1000

No clinical signs observed

1

2

Day 0 - Day 14

0/1

 

Group

 No.

Dose mg/kg

                            Observed Signs

Total Number of

Animals

 

Animal Nos.

Period of signs in days

 From - to

 

Mortality

I

2000

No clinical signs observed

1

3

Day 0 - Day 14

0/1

 

Main Study:

Group

 No.

Dose mg/kg

                            Observed Signs

Total Number of

Animals

 

Animal Nos.

Period of signs in days

 From - to

 

Mortality

II

2000

No clinical signs observed

2

4, 5

Day 0 - Day 14

0/2

 

Table No. II 

Summary of Evaluation of Dermal Reaction

 

Laboratory Test Item Code :TAS/122/058

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group

 No.

Dose mg/kg

                          

Dermal Reaction

Total Number of

Animals

Animal Nos.

Period of signs

in days

 From - to

I

200

No dermal reaction observed

1

1

Day 0 - Day 14

 

Group

 No.

Dose mg/kg

                          

Dermal Reaction

Total Number of

Animals

Animal Nos.

Period of signs

in days

 From - to

I

1000

No dermal reaction observed

1

2

Day 0 - Day 14

 

Group

 No.

Dose mg/kg

                          

Dermal Reaction

Total Number of

Animals

Animal Nos.

Period of signs

in days

 From - to

I

2000

No dermal reaction observed

1

3

Day 0 - Day 14

 

Main Study:

Group

 No.

Dose mg/kg

                          

Dermal Reaction

Total Number of

Animals

Animal Nos.

Period of signs

in days

 From - to

II

2000

No dermal reaction observed

2

4, 5

Day 0 - Day 14

 

Table No.III

Mean Body Weight and Percent Body Weight Gain (g)

 

Laboratory Test Item Code :TAS/122/058

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group No.

Dose

(mg/kg body weight)

 

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

200

Mean

231.4

247.0

6.74

260.6

5.51

12.62

± SD

-

-

-

-

-

-

 

Group No.

Dose

(mg/kg body weight)

 

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

1000

Mean

228.3

239.3

4.82

250.4

4.64

9.68

± SD

-

-

-

-

-

-

 

Group No.

Dose

(mg/kg body weight)

 

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

2000

Mean

228.0

239.6

5.09

252.5

5.38

10.75

± SD

-

-

-

-

-

-

 

Main Study:

Group No.

Dose

(mg/kg body weight)

 

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

II

2000

Mean

245.15

256.25

4.53

267.95

4.57

9.31

± SD

3.61

2.47

0.53

0.64

0.76

1.35

Table No.IV 

Summary of Gross Pathological Findings

 

Laboratory Test Item Code :TAS/122/058

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

200

1

TS

No abnormality detected

 

Group No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

1000

2

TS

No abnormality detected

 

Group No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

2000

3

TS

No abnormality detected

 

                    Main Study:

Group No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

II

2000

4, 5

TS

No abnormality detected

 

                     TS = Terminal Sacrifice 

 

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 test chemical. The studies are summarized as below –

The reported study was designed and conducted to determine the acute oral toxicity profile of the given test chemical 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 reduced locomotor activity, loss of righting reflex and convulsions with onset at 30 minutes after the dosing. Three animals died at 1 hour after the dosing. As mortality was observed at 300 mg/kg dose group, additional three female animals were treated with the lower dose of 50 mg/kg of the test item (Step - I).

Administration of the test item at 50 mg/kg resulted in reduced locomotor activity and ataxic gait with onset at 2 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 additional female animals were added to the study and treated with the dose of 50 mg/kg of the test item (Step - II). Administration of the test item at 50 mg/kg resulted in reduced locomotor activity and ataxic gait with onset at 1 to 2 hours after the dosing and no mortality at 24 hours after the dosing.

All animals from 50 mg/kg dose group survived through the study period of 14 days. Gross pathological examination did not reveal any abnormalities in all animals sacrificed terminally from 50 mg/kg dose group. Gross pathological examination revealed unabsorbed residual test item in the stomach in found dead animal from 300 mg/kg dose group.

Under the condition of the study, the acute oral LD50 value of the given test chemical was considered in between 50 – 300 mg/kg body weight. Thus, it was concluded that the acute toxicity study of test chemical, when administered via oral route in Sprague Dawley rats falls into the “Category 3” according to criteria of CLP.

The above study is supported with another study conducted on rats and mentioned in database for the test chemical.

Acute oral toxicity study of the given test chemical was conducted in rats at the dose concentration of 100 or 121 mg/kg bw. Necropsy of survivors performed. Animals were observed for mortality and clinical signs. 50% mortality was observed. Rats showed respiratory distress, apathy, staggering, shivering and clonic seizures. The necropsy did not reveal anything conspicuous except slight reddening of the mucous membranes in the intestine.

Therefore, the LD50 value was considered to be 121 mg/kg bw, when rats were treated with the given test chemical via oral route.

Thus, based on the above summarised studies on test chemical, all rats were died at 300 mg/kg bw, hence the LD100 was considered to be 300 mg/kg bw. Considering this value, the LD50 value can be expected to be 200 mg/kg bw or in between 100-200 mg/kg bw. Therefore, comparing this value / range with the criteria of CLP regulation, the given test chemical can be classified in “Category 3 (50 - ≤ 300)” for acute oral toxicity.

Acute inhalation toxicity:

In different studies, the given test chemical has been investigated for acute inhalation 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:

 

Acute inhalation toxicity study of the given test chemical was conducted in rats at the dose concentration of 550 mg/m³, 2600 mg/m³ and 5600 mg/m³. Rats were exposed to aerosols from aqueous solutions of test chemical.

The given test chemical was exposed as a 20% solution to form 550 mg/m³ and 60% solution to form 2600 mg/m³ and 5600 mg/m³.

The inhalation of a concentration of 550 mg/m³ from a 20% solution was exposed for 4 hours; the inhalation of 2600 mg/m³ exposed for 2 hours and 5600 mg/m³ for 1 hour from a 60% solution.

Animals were observed for mortality and clinical signs. Necropsy of survivors performed. Rats exposed to a concentration of 550 mg/m³ from a 20% solution exposed for 4 hours was survived. While the inhalation of 2600 mg/m³ exposed for 2 hours and 5600 mg/m³ exposed for 1 hour from a 60% solution caused the death of all the animals.

The symptoms were attempts to escape, respiratory distress, seizures. The findings from the necropsy were unspecific (hyperaemia of inner organs). Nothing conspicuous was found in the animals which survived.

Under the condition of this, the LC100 value was considered to be 2600 mg/m³ with the exposure time of 2 hours. Considering the above signs and mortality, the LC50 value was assumed to be 1300 mg/m3, when rats were treated with the given test chemical via inhalation route by aerosols.

 

The above study is supported by another study conducted on rats for the test chemical. The acute inhalation toxicity study of the given test chemical was conducted in 10 males/group Wistar rats at the dose concentrations of 0.190, 0.504, 0.708, 1.453, 1.980 mg/l air via inhalation route by nose only exposure for 4 hours.

Necropsy was performed on rats that died during exposure, during observation period and on rats that survived. The animals were observed for clinical signs and mortality. Post exposure observation time was 21 days. No statistical analysis mentioned.

No rats died up to 0.708 mg/l; At 1.453 mg/l: 5/10 rats died within 1 to 19 days, At 1.980 mg/l: 6110 rats died within 4hrs to 2 days.

Signs of intoxication were: inactivity, piloerection, unkempt fur, and difficulties in breathing up to 19 days post exposure in all rats. Rats that died during the test showed dark red colored lungs always with emphysema, some rats showed lung edema. Rats that survived showed no pathologic findings up to 0.708 mg/L. At higher concentrations lungs with emphysema and mottled, some showed enlarged adrenals.

Therefore, the LC50 value was considered to be 1.45 mg/L (1450 mg/m3), when 10 males/group Wistar rats were treated with the given test chemical via inhalation route by nose only exposure for 4-hours.

 

These studies are supported with the study mentioned in different databases and conducted in 70 male and female Sprague Dawley rats at the dose concentrations of 0.01, 0.026, 0.30, 1.13, 1.48, 1.38, and 1.42 mg/L via inhalation route by concentrated vapors/aerosols for 4 hours exposure.

The average particle size (mass median diameter) ranged from 3.8 to 5.8 µm (standard deviation of 2.5 to 6.0). The animals were observed for mortality, clinical signs and body weight changes daily after exposure. Necropsy of survivors performed.

50% mortality was observed in animals between 1.13 - 1.48 mg/l. Daily observations indicated that body weight loss, nasal discharge, decreased activity, and possible slight respiratory difficulty occurred subsequent to exposure. No lesions were noted during post mortem examination.

Therefore, the LC50 value was considered in between 1.13 - 1.48 mg/l, when 70 male and female Sprague Dawley rats were treated with the given test chemical via inhalation route by concentrated vapours/aerosols for 4-hours exposure.

 

All the above studies are further supported with the study mentioned in databases and conducted in rats at the actual concentration of 1.97 mg/l via inhalation route for 4 hours.

Particle size was ~ 3.5 - 5.0 microns; 75-89% <10 microns. The animals were observed for clinical signs and mortality.

50% mortality was observed at 1.92 mg/l. Signs of toxicity such as ataxia and tremors were observed in animals. Hence, the LC50 value was considered to be 1.92 mg/l, when rats were treated with the given test chemical via inhalation route for 4-hours exposure.

 

Thus, based on the above summarised studies on test chemical, it can be concluded that the LC50 value is between 1 to ≤ 5 mg/L, for acute inhalation toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical can be classified in “Category 4” for acute inhalation toxicity.

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 test chemical. The studies are summarized as below –

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”.

 

The above study is contradicted with the study mentioned in database and conducted in rabbits for the test chemical.

Acute dermal toxicity study of the given test chemical was conducted in rabbits at the dose concentration of 309 mg/kg bw. The given test chemical was dissolved as a 50% aqueous preparation and applied to the skin of rabbits. Animals were observed for mortality and clinical signs. 50% mortality was observed. The animals showed distinct symptoms of systemic effects (salivation, cyanosis, respiratory distress, seizures). A local effect was slight reddening of the skin.

Therefore, the LD50 value was considered to be 309 mg/kg bw, when rabbits were treated with the given test chemical by dermal application.

The above study is supported with the study mentioned in secondary database for the test chemical. The acute dermal toxicity study of the given test chemical was conducted in rabbits at the dose concentration range of 205 - 375 mg/kg bw. 50% mortality was observed. Therefore, the LD50 value was considered in between 205 - 375 mg/kg bw, when rabbits were treated with the given test chemical by dermal application.

Under the condition of the above studies, the contradicting studies don’t have detailed data for classification of the test chemical. Therefore, the LD50 value was considered to be >2000 mg/kg bw, based on the reliable data mentioned in study report. 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 on test chemical, it can be concluded that LD50 value is between 50 - 300 mg/kg bw, for acute oral toxicity; LC50 value isbetween 1 to ≤ 5 mg/L, for acute inhalation toxicity and LD50 value is >2000 mg/kg bw, acute dermal toxicity. Thus, comparing these ranges / values with the criteria of CLP regulation, the given test chemical can be classified in “Category 3” for acute oral toxicity, “Category 4” for acute inhalation toxicity and cannot be classified for acute dermal toxicity.