Registration Dossier

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 range with the criteria of CLP regulation, the given test chemical can be classified into the “Category 3” for acute oral toxicity.

Acute Inhalation Toxicity:

The acute toxicity inhalation study need not be conducted because exposure to humans via inhalation route is not likely taking into account the low vapour pressure of the substance, which is reported as 3.36E-13 mm Hg. Thus, exposure to inhalable dust, mist and vapour of the test chemical is highly unlikely. Therefore this study is considered for waiver.

 

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:
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 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 193.9 to 206.9 grams.
Body weights at the start : Female Mean: 199.69 g (= 100 %); Minimum : 193.9 g (- 2.90 %); Maximum : 206.9 g (+ 3.61 %)
- 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 (%): 56.1% to 58.4%.
- 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: 28-05-2018 to 15-06-2018
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: The lesion observed in the gross pathological examination are the local irritation effects due to the administration of test item and were considered haemodynamic in nature and hence the organs/tissues were not processed for histopathology.
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 - Two 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:
Group I Step I : 300 mg/kg - Animals treated at the dose level of 300 mg/kg resulted in nasal discharge, salivation, reduced locomotor activity, ataxic gait, convulsions and test item coloured faeces and urine with onset at 5 minutes to 1 hour after the dosing. The only surviving animal was free of signs of toxicity on day 2 after the dosing.
Group II Step I : 50 mg/kg - Animals treated at the dose level of 50 mg/kg resulted in ataxic gait with onset at 30 minutes to 1 hour after the dosing after the dosing. All animals were free of signs of toxicity on day 1 after the dosing.
Group II Step II : 50 mg/kg - Animals treated at the dose level of 50 mg/kg resulted in ataxic gait with onset at 1 hour after the dosing after the dosing. All animals were free of signs of toxicity from 6 hours to day 1 after the dosing.
Body weight:
Group I Step I (300 mg/kg) – Percent body weight gain after 7 days and 14 days was found to be 6.29% and 13.15% respectively.
Group II Step I (50 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 6.48% and 13.47% respectively.
Group II Step II (50 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 7.72% and 16.16% respectively.
Gross pathology:
Gross pathological examination did not reveal any abnormalities in terminally sacrificed animals from 300 mg/kg and 50 mg/kg dose groups.
Gross pathological examination revealed stomach with test item coloured liquid ingesta and mucosa. The observed colouration of mucosa could be removed by cleaning it with normal saline. The external examination revealed salivation from mouth in found dead animals 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/122/084

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

Nasal discharge

3

1,2

3

30 min.

30 min. - 2 hrs.

2/3

Salivation

3

1,2

3

30 min.

30 min. - 2 hrs.

Reduced locomotor activity

3

1,2

3

30 min.

30 min. - 6 hrs.

Ataxic gait

3

1,2

3

5 min. - 30 min.

10 min. - day 1

Convulsions

2

1,2

30 min.

Test item coloured faeces and urine

1

3

1 hr. - day 1

 

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

Ataxic gait

3

4

5,6

1 hr. - 6 hrs.

30 min. - 6 hrs.

0/3

 

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

Ataxic gait

3

7,9

8

1 hr. - 6 hrs.

1 hr. - 4 hrs.

0/3

  

Table No.II 

Mean Body Weight and Percent Body Weight Gain (g)

 

Laboratory Test Item Code :TAS/122/084

Test System : Sprague Dawley Rat

Sex : Female

Group I :

Step

No.

Dose

(mg/kg body weight)

 

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

300

Mean

195.67

206.10

6.29

219.40

6.45

13.15

± SD

2.48

-

-

-

-

-

  

Group II :

Step

No.

Dose

(mg/kg body weight)

 

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

50

Mean

203.23

216.40

6.48

230.60

6.56

13.47

± SD

4.23

4.40

1.09

4.81

0.39

1.57

 

Group II :

Step

No.

Dose

(mg/kg body weight)

 

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

II

50

Mean

200.17

215.60

7.72

232.50

7.84

16.16

± SD

2.27

1.47

0.84

2.86

0.60

1.10

 

Table No.III

Summary of Gross Pathological Findings

 

Laboratory Test Item Code :TAS/122/084

Test System : Sprague Dawley Rat

Sex : Female Group I :

Step

No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

300

1, 2

FD

Stomach with test item coloured liquid ingesta and mucosa. The observed colouration of mucosa could be removed by cleaning it with normal saline. The external examination revealed salivation from mouth.

3

TS

No abnormality detected

 

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 nasal discharge, salivation, reduced locomotor activity, ataxic gait, convulsions and test item coloured faeces and urine with onset at 5 minutes to 1 hour after the dosing.

Two 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 ataxic gait with onset at 30 minutes to 1 hour 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 ataxic gait with onset at 1 hour 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 terminally sacrificed animals from 300 mg/kg and 50 mg/kg dose groups. Gross pathological examination revealed stomach with test item coloured liquid ingesta and mucosa. The external examination revealed salivation from mouth in found dead animals 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:
300 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:
other justification
Justification for data waiving:
other:
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 experimental 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) or to establish a non-lethal dose level of 2000 milligram of test item per kilogram of body weight.
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
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: Young adult (8 to 10 weeks old) female rats were used.
- Weight at study initiation: The weight range of approximately 221.5 to 227.4 grams at initiation of dosing.
Body weights at the start : Female Mean: 225.23 g (= 100 %); Minimum : 221.5 g (- 1.66 %); Maximum : 227.4 g (+ 0.96 %)
- 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 (%): 50.4% to 60.3%.
- 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: 07-05-2018 To: 17-06-2018
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 : 2000 mg/kg
Main Study: Group II : 2000 mg/kg
No. of animals per sex per dose:
Dose Range Finding Study: 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: Based on the available literature a single dose of 2000 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity were observed during first 48 hours.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Dose Range Finding Study: Group I : Animal treated at the dose level of 2000 mg/kg body weight survived through the study period of 14 days.
Main Study: Group II : Animals treated at the dose level of 2000 mg/kg body weight survived through the study period of 14 days.
Clinical signs:
Dose Range Finding Study: Group I : Animal 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.
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 I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 8.35% and 15.85% respectively.
Main Study: Group II (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 5.88% and 12.57% respectively.
Gross pathology:
Gross pathological examination did not reveal any abnormalities in animals from 2000 mg/kg dose group 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 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/122/084

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group

No.

Dose mg/kg

Observed Signs

Total Number of

Animals

Animal No.

Period of signs in days

 From - to

 

Mortality

I

2000

No clinical signs observed

1

1

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

2, 3

Day 0 - Day 14

0/2

 

Table No. II 

Summary of Evaluation of Dermal Reaction

 

Laboratory Test Item Code :TAS/122/084

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group

No.

Dose mg/kg

Dermal Reaction

Total Number of

Animals

Animal

No.

Period of signs

in days

 From – to

I

2000

No dermal reaction observed

1

1

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

2, 3

Day 0 - Day 14

  

Table No.III

Mean Body Weight and Percent Body Weight Gain (g)

 

Laboratory Test Item Code :TAS/122/084

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

2000

Mean

221.50

240.00

8.35

256.60

6.92

15.85

± 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

227.10

240.45

5.88

255.65

6.32

12.57

± SD

0.42

0.07

0.23

0.49

0.17

0.43

 

 

Table No.IV

Summary of Gross Pathological Findings

 

Laboratory Test Item Code :TAS/122/084

Test System : Sprague Dawley Rat

Sex : Female

Dose Finding Study:

Group No.

Dose

mg/kg

Animal Number

Animal Fate

Gross Pathological Findings

I

2000

1

TS

No abnormality detected

 

                    Main Study:

Group No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

II

2000

2, 3

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 2000 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity were observed during first 48 hours.

As the dose range finding study revealed no mortality or clinical signs of toxicity 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 and 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.

Under the condition of the study, 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 and mice 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 nasal discharge, salivation, reduced locomotor activity, ataxic gait, convulsions and test item coloured faeces and urine with onset at 5 minutes to 1 hour after the dosing. Two 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 ataxic gait with onset at 30 minutes to 1 hour 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 ataxic gait with onset at 1 hour 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 terminally sacrificed animals from 300 mg/kg and 50 mg/kg dose groups. Gross pathological examination revealed stomach with test item coloured liquid ingesta and mucosa. The external examination revealed salivation from mouth in found dead animals 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 mentioned in peer-reviewed journal for the test chemical and was conducted in group 5 males and 5 females CFY rats.

In preliminary study, the rats (N = 4; 2 males, 2 females) were fasted, orally administered a single dose of the given test chemical (0, 100, 400, and 1600 mg/kg; 10% suspension in aqueous gum tragacanth [0.5%]), and observed for 14 days. No rats died in the control and low-dose groups. Three rats died in the middle-dose group within 19 hours of dosing, and all died in the high-dose group within 2 hours of dosing.

Hence, the experiment was repeated with the dose concentration of 0, 100, 160, 250, 400, or 640 mg/kg bw. Animals were observed for mortality. No rats died in the control and low-dose groups. 8/ 10 died in the 160 mg/kg group; 7 died in the 250 mg/kg group; and 9 died in both the 400 and 640 mg/kg groups. The rats that died within 19 hours.

Therefore, the LD50 value was considered to be 110 mg/kg bw, with 95% confidence limit of 63-176 mg/kg bw, when group 5 males and 5 females CFY rats were treated with the given test chemical via oral route.

 

Both the above studies are contradicted with the study mentioned in publication, handbook and database for the test chemical. The acute oral toxicity study of the given test chemical was conducted in 36 male HA/ICR mice at the dose concentration of 378 mg/kg bw. Animals were observed for mortality. The LD50 value was determined by probit analysis. 50% mortality was observed at 378 mg/kg bw. Therefore, LD50 value was considered to be 378 mg/kg bw, when 36 male HA/ICR mice were treated with the given test chemical via oral route.

 

Even though from the above contradicting study, the LD50 value is mentioned as 378 mg/kg bw in mice, the study will not be considered for the classification of the test chemical. As the test animal “rat” is most preferable species for the acute toxicity study.

Thus, from rest of the experimental studies conducted on rats, the LD50 value was considered to be 300 mg/kg bw, as 50% mortality was observed near to this dose. Therefore, comparing this value with the criteria of CLP regulation, the given test chemical can be classified into the “Category 3”for acute oral toxicity.

                                                               

Acute Inhalation Toxicity:

The acute toxicity inhalation study need not be conducted because exposure to humans via inhalation route is not likely taking into account the low vapour pressure of the substance, which is reported as 3.36E-13 mm Hg. Thus, exposure to inhalable dust, mist and vapour of the test chemical is highly unlikely. Therefore this study is considered for waiver.

 

Acute Dermal Toxicity:

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 2000 mg/kg body weight of the test item was administered to 1 female animal. No death or clinical signs of toxicity were observed during first 48 hours.

As the dose range finding study revealed no mortality or clinical signs of toxicity 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 and 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.

Under the condition of the study, 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”.

 

Justification for classification or non-classification

Based on the above studies on test chemical, it can be concluded that the LD50 value is between 50 - 300 mg/kg bw, for acute oral toxicity and LD50 value is >2000 mg/kg bw, for acute dermal toxicity. Thus, comparing this range and value with the criteria of CLP regulation, the given test chemical can be classified into the “Category 3” for acute oral toxicity and cannot be classified for acute dermal toxicity. For acute inhalation toxicity wavier was added so, not possible to classify.