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 and mice for the test chemical. The LD50 value is2800 mg/kg bw. The study 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 the low vapour pressure of the test chemical, which is reported to be 1.056E-4 Pa (7.92E-07 mm Hg). 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 experimental study conducted on rats for the test chemical. The study concluded that 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
Study period:
25-03-1986 to 08-04-1986
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 401 (Acute Oral Toxicity)
Principles of method if other than guideline:
Acute oral toxicity study of the given test chemical in rats.
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Remarks:
SPF
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Iffa-Credo, Rrussels, Belgium
- Age at study initiation: 8 weeks, 6 days
- Weight at study initiation: Male: 360-402 g, Female: 205-251 g
- Fasting period before study: Animals were fasted overnight before dosing till 3-4 hours after administration of test substance.
- Housing: Animals were housed in Macrolon cage
- Diet (e.g. ad libitum): Standard laboratory animal diet (RMH-B, pellet diameter 10 mm)
- Water (e.g. ad libitum): Tap water, ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-22 °C
- Humidity (%):30-75 %
- Photoperiod (hrs dark / hrs light): Artificial light sequence was 12 hrs dark, 12 hrs light

IN-LIFE DATES: From: 25-03-1986 to 08-04-1986
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
Milli-RO
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 1800, 2400 and 3200 mg/kg body weight
- Amount of vehicle (if gavage): 10 ml/kg body weight
- Justification for choice of vehicle: Milli-RO water
DOSAGE PREPARATION (if unusual): test substance was formulated in Milli-RO water at 1800, 2400 and 3200 mg/kg body weight.
Doses:
1800, 2400 and 3200 mg/kg body weight
No. of animals per sex per dose:
Total: 30
1800 mg/kg bw: 5 male, 5 female
2400 mg/kg bw: 5 male, 5 female
3200 mg/kg bw: 5 male, 5 female
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical sign observed daily and body weight on day 0,7 and 14
- Necropsy of survivors performed: yes
- Other examinations performed: survival,clinical signs, body weight, gross pathology and histopathology were examined.
Statistics:
LD50 was calculated with U-distribution and maximum likelihood (Finney).
Preliminary study:
Dose range finding investigation: In order to establish an appropriate dose range, eleven groups of Sprague-Dawley rats, each comprising 1 male and 1 female, were dosed with a single oral dose of the test substance at 75, 100, 130, 180, 240, 320, 420, 750, 1300, 2400 and 4200 mg/kg bw, respectively. Both animals of the 4200 mg/kg bw and the male of the 2400 mg/kg bw group were found dead within a few minutes after dosing. Surviving animals revealed no signs of evident systemic toxicity during the 7 days observation period. Macroscopic examination at autopsy of all animals revealed no gross abnormalities.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
2 800 mg/kg bw
Based on:
test mat.
95% CL:
2 500 - 3 400
Remarks on result:
other: 50 % mortality observed
Sex:
male
Dose descriptor:
LD50
Effect level:
2 500 mg/kg bw
Based on:
test mat.
95% CL:
2 300 - 3 100
Remarks on result:
other: 50 % mortality observed
Sex:
female
Dose descriptor:
LD50
Effect level:
2 800 mg/kg bw
Based on:
test mat.
95% CL:
2 200 - 16 500
Remarks on result:
other: 50 % mortality observed
Mortality:
When treated with 3200 mg/kg bw, 5 male and 3 female died.
When treated with 2400 mg/kg bw, 1 male and 2 female died.
When treated with 1800 mg/kg bw, No mortality were observed in treated male and female rats.
Clinical signs:
Lethargy, tremors, convulsions, coma, laboured breathing and diarrhoea sign of toxicity were observed.Other findings were observed unbalanced gait and bloody nose or eye encrustation.
In surviving animals, these signs were reversible since generally as of day 3 no more abnormalities were observed during the 14 day observation period
Body weight:
No effect on body weight gain was observed in treated male and female rats.
Gross pathology:
Hyperemia, petechiae or erosion of stomach frequently followed by bloody gastrointestinal content were observed in dead animals. Other gross internal findings were renal hyperemia, motteled kidney, aqueous gastrointestinal content (possibly test substance) and autolysis.At the end of study in surviving animals no test substance related gross abnormalities were observed
Other findings:
not specified

Table 1: Day of death and Number of Mortalities after oral administration of TEAB in the rats

Day of Death

Mortalities (Male/ Female)

 

Dosage mg/kg

 

1800

2400

3200

0

0/0

1/1

5/3

1

0/0

0/1

0/0

Sum

 

 

 

Male / Female

0/0

1/2

5/3

Sexes combined

0

3

8

Note: the dose groups comprised 5 male and 5 female rats. Indicated are the number of animals (Male / Female) found dead during the 14 day observation period.

Table 2: Daily incidence of Cage side observation for male and female rats after a single oral dose of TEAB: 1800 mg/kg bw

 

Day of observation

observation

Sex

0a)

0b)

0c)

0d)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

No abnormalities

M

2/5

3/5

3/5

4/5

2/5

4/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

 

5/5

5/5

5/5

 

F

2/5

2/5

1/5

 

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

5/5

Lethargy

M

3/5

2/5

2/5

1/5

1/5

 

 

 

 

 

 

 

 

 

 

 

 

 

 

F

3/5

3/5

4/5

5/5

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Tremors

M

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

F

 

 

 

2/5

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Bloody nose/eye encrustation

M

 

 

 

 

2/5

1/5

 

 

 

 

 

 

 

 

2/5

 

 

 

 

F

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Diarrhoea

M

 

 

 

 

1/5

 

 

 

 

 

 

 

 

 

 

 

 

 

 

F

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Table 3: Daily incidence of Cage side observation for male and female rats after a single oral dose of TEAB: 2400 mg/kg bw

 

Day of observation

observation

Sex

0a)

0b)

0c)

0d)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

No abnormalities

M

 

 

 

 

2/4

4/4

4/4

4/4

4/4

4/4

4/4

4/4

4/4

4/4

4/4

4/4

4/4

4/4

 

F

 

 

1/5

 

2/3

3/3

3/3

3/3

3/3

3/3

3/3

3/3

3/3

3/3

3/3

3/3

3/3

3/3

Lethargy

M

4/4

4/4

4/4

4/4

1/4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

F

5/5

5/5

4/5

5/5

1/3

 

 

 

 

 

 

 

 

 

 

 

 

 

Tremors

M

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

F

 

 

 

3/5

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Laboured breathing

M

 

1/4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

F

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Unbalanced gait

M

 

 

 

1/5

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

F

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Bloody nose/eye encrustation

M

 

 

 

 

1/4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

F

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Table 4: Daily incidence of Cage side observation for male and female rats after a single oral dose of TEAB: 3200 mg/kg bw

 

Day of observation

observation

Sex

0a)

0b)

0c)

0d)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

No abnormalities

M

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

F

1/4

 

 

 

1/2

2/2

2/2

2/2

2/2

2/2

2/2

2/2

2/2

2/2

2/2

2/2

2/2

2/2

Lethargy

M

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

F

1/4

2/2

2/2

2/2

1/2

 

 

 

 

 

 

 

 

 

 

 

 

 

Tremors

M

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

F

 

 

 

2/2

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Convulsions

M

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

F

 

1/4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Comatous

M

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

F

 

1/4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

a)     Approx. 1/4 - 1 1/4   hours postdosing

b)     Approx. 1 3/4- 2 3/4 hours postdosing

c) Approx. 3 1/4 – 4 1/4 hours postdosing

d) Approx. 4 1/2 – 5 1/2 hours hours postdosing

Interpretation of results:
other: Not classified
Conclusions:
The acute oral LD50 value was considered to be 2800 mg/kg bw, with 95% confidence limit of 2500-3400 mg/kg bw for both male and female rats, 2500 mg/kg bw, with 95% confidence limit of 2300-3100 mg/kg bw for male rats and 2800 mg/kg bw, with 95% confidence limit of 1650- 2200 mg/kg bw for female rats, when groups of 5 male and 5 female Sprague-Dawley rats were treated with the given test chemical orally by gavage in Milli-RO water.
Executive summary:

In a acute oral toxicity study, groups of 5 male and 5 female Sprague-Dawley rats were treated with the given test chemical at the dose concentrations of 1800, 2400 and 3200 mg/kg body weight as per OECD 401.

The given test substance was formulated in Milli-RO water and administered as 10 ml/kg body weight via oral gavage route.

In order to establish an appropriate dose range, eleven groups of Sprague-Dawley rats, each comprising 1 male and 1 female, were dosed with a single oral dose of the test substance at 75, 100, 130, 180, 240, 320, 420, 750, 1300, 2400 and 4200 mg/kg bw, respectively. Both animals of the 4200 mg/kg bw and the male of the 2400 mg/kg bw group were found dead within a few minutes after dosing. Surviving animals revealed no signs of evident systemic toxicity during the 7 days observation period. Macroscopic examination at autopsy of all animals revealed no gross abnormalities.

Based on the toxicity observed in the dose range finding investigation three groups of animals, each comprising 5 males and 5 females, were dosed with a single oral dose of the test substance at 1800, 2400 and 3200 mg/kg body weight, respectively.

Animals were observed for mortality and clinical signs daily and body weight on day 0,7 and 14 day. Necropsy of survivors performed. Gross pathology and histopathology were examined. LD50 was calculated with U-distribution and maximum likelihood (Finney).

Mortality was observed as, when treated with 3200 mg/kg bw, 5 male and 3 female died; when treated with 2400 mg/kg bw, 1 male and 2 female died; when treated with 1800 mg/kg bw, no mortality were observed in male and female rats.

Lethargy, tremors, convulsions, coma, laboured breathing and diarrhoea sign of toxicity were observed. Other findings were observed unbalanced gait and bloody nose or eye encrustation.

In surviving animals, these signs were reversible since generally as of day 3 no more abnormalities were observed during the 14 day observation period. No effect on body weight gain was observed in treated male and female rats. Hyperemia, petechiae or erosion of stomach frequently followed by bloody gastrointestinal content were observed in dead animals. Other gross internal findings were renal hyperemia, motteled kidney, aqueous gastrointestinal content (possibly test substance) and autolysis.At the end of study in surviving animals no test substance related gross abnormalities were observed

Under the condition of the study, the acute oral LD50 value was considered to be 2800 mg/kg bw, with 95% confidence limit of 2500-3400 mg/kg bw for both male and female rats, 2500 mg/kg bw, with 95% confidence limit of 2300-3100 mg/kg bw for male rats and 2800 mg/kg bw, with 95% confidence limit of 1650- 2200 mg/kg bw for female rats, when groups of 5 male and 5 female Sprague-Dawley rats were treated with the given test chemical orally by gavage in Milli-RO water.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 800 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:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: National Institute of Biosciences, Pune.
- Age at study initiation: Young adult male and female rats aged between 8 – 12 weeks were used.
- Weight at study initiation: The weight range of approximately 216.8 to 249.7 grams at initiation of dosing.
Body weights at the start : Male Mean : 240.96 g (= 100 %) Minimum : 233.9 g (- 2.93 %) Maximum : 249.7 g (+ 3.63 %)
Female Mean: 220.24 g (= 100 %)Minimum : 216.8 g (- 1.56 %) Maximum : 223.9 g (+ 1.66 %)
- 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 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): 19.7 to 21.8 degree centigrade.
- Humidity (%): 56.2% to 60.1%
- Air changes (per hr): Ten to fifteen air changes per hour.
- Photoperiod (hrs dark / hrs light): An artificial light and dark cycle of 12 hours each was provided to the room.

IN-LIFE DATES: 11-07-2017 to 26-07-2017
Type of coverage:
occlusive
Vehicle:
water
Remarks:
(Distilled water)
Details on dermal exposure:
TEST SITE
- Area of exposure: Trunk (dorsal surface and sides from scapular to pelvic area)
- % coverage: Approximately 10% of the body surface area.
- Type of wrap if used: Porous gauze dressing and non-irritating tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Distilled water was used to remove residual test item.

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg bw
- For solids, paste formed: Paste
Duration of exposure:
24 hours
Doses:
A single dose of 2000 mg of the test item per kilogram of body weight was administered to ten rats (five males and five females).
No. of animals per sex per dose:
10 (5/sex).
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Twice daily
- Necropsy of survivors performed: Yes
- 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 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.
- Gross Pathology: 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).
- Histopathology:No gross abnormalities were observed in animals sacrificed terminally hence, no histopathology was performed.
Statistics:
not specified
Preliminary study:
not specified
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no mortality was observed
Mortality:
Sex : Male Group I - Animal treated at the dose level of 2000 mg/kg body weight: All animals survived through the study period of 14 days.
Sex : Female Group I - Animal treated at the dose level of 2000 mg/kg body weight: All animals survived through the study period of 14 days.
Clinical signs:
Sex : Male 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.
Sex : Female 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.
Body weight:
Sex : Male Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 9.02% and 18.32% respectively.
Sex : Female Group I (2000 mg/kg) - Percent body weight gain after 7 days and 14 days was found to be 4.58% and 9.42% respectively.
Gross pathology:
Gross pathological examination did not reveal any abnormalities in animals from 2000 mg/kg dose group.
Other findings:
- Other observations:Evaluation of Dermal Reaction
Sex : Male 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.
Sex : Female 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.

Table No. I

Summary of Clinical Signs of Toxicity and Mortality

Test System : Sprague Dawley Rat

Sex : Male

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

5

1 - 5

Day 0 - Day 14

0/5

 

 

Sex : Female 

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

5

6 - 10

Day 0 - Day 14

0/5

 

 

 Table No. II 

Summary of Evaluation of Dermal Reaction

Test System : Sprague Dawley Rat

Sex : Male 

Group

 No.

Dose mg/kg

                          

Dermal Reaction

Total Number of

Animals

 

Animal Nos.

Period of signs

in days

 From - to

 

Mortality

I

2000

No dermal reaction observed

5

1 - 5

Day 0 - Day 14

0/5

 

 

Sex : Female

Group

 No.

Dose mg/kg

                          

Dermal Reaction

Total Number of

Animals

 

Animal Nos.

Period of signs

in days

 From - to

 

Mortality

I

2000

No dermal reaction observed

5

6 - 10

Day 0 - Day 14

0/5

 

 

Table No.III

Mean Body Weight and Percent Body Weight Gain (g)

Test System : Sprague Dawley Rat

Sex : Male

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

240.96

262.66

9.02

285.08

8.53

18.32

± SD

5.85

5.01

1.04

6.02

0.66

1.50

 

  

Sex : Female

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

220.24

230.32

4.58

240.98

4.63

9.42

± SD

2.55

2.28

0.37

2.68

0.71

0.64

 

 

 

Table No.IV

Summary of Gross Pathological Findings

Test System : Sprague Dawley Rat

Sex : Male

Group No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

2000

1 - 5

TS

No abnormality detected

 

Sex : Female     

Group No.

Dose

mg/kg

Animal Numbers

Animal Fate

Gross Pathological Findings

I

2000

6 - 10

TS

No abnormality detected

           

TS = Terminal Sacrifice

 

Interpretation of results:
other: Not Classified
Conclusions:
The LD50 value was considered to be >2000 mg/kg bw, when male and female Sprague Dawley rats were occlusively treated with test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).
Executive summary:

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

The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days. Animals exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

Hence, the LD50 value was considered to be >2000 mg/kg bw, when male and female Sprague Dawley rats were occlusively treated with test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

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 key study was mentioned in study report to determine the acute oral toxicity profile of the given test chemical in groups of 5 male and 5 female Sprague-Dawley rats at the dose concentrations of 1800, 2400 and 3200 mg/kg body weight as per OECD 401.

The given test substance was formulated in Milli-RO water and administered as 10 ml/kg body weight via oral gavage route.

In order to establish an appropriate dose range, eleven groups of Sprague-Dawley rats, each comprising 1 male and 1 female, were dosed with a single oral dose of the test substance at 75, 100, 130, 180, 240, 320, 420, 750, 1300, 2400 and 4200 mg/kg bw, respectively. Both animals of the 4200 mg/kg bw and the male of the 2400 mg/kg bw group were found dead within a few minutes after dosing. Surviving animals revealed no signs of evident systemic toxicity during the 7 days observation period. Macroscopic examination at autopsy of all animals revealed no gross abnormalities.

Based on the toxicity observed in the dose range finding investigation three groups of animals, each comprising 5 males and 5 females, were dosed with a single oral dose of the test substance at 1800, 2400 and 3200 mg/kg body weight, respectively.

Animals were observed for mortality and clinical signs daily and body weight on day 0,7 and 14 day. Necropsy of survivors performed. Gross pathology and histopathology were examined. LD50 was calculated with U-distribution and maximum likelihood (Finney).

Mortality was observed as, when treated with 3200 mg/kg bw, 5 male and 3 female died; when treated with 2400 mg/kg bw, 1 male and 2 female died; when treated with 1800 mg/kg bw, no mortality were observed in male and female rats. Lethargy, tremors, convulsions, coma, laboured breathing and diarrhoea sign of toxicity were observed.Other findings were observed unbalanced gait and bloody nose or eye encrustation.

In surviving animals, these signs were reversible since generally as of day 3 no more abnormalities were observed during the 14 day observation period. No effect on body weight gain was observed in treated male and female rats. Hyperemia, petechiae or erosion of stomach frequently followed by bloody gastrointestinal content were observed in dead animals. Other gross internal findings were renal hyperemia, motteled kidney, aqueous gastrointestinal content (possibly test substance) and autolysis.At the end of study in surviving animals no test substance related gross abnormalities were observed

Under the condition of the study, the acute oral LD50 value was considered to be 2800 mg/kg bw, with 95% confidence limit of 2500-3400 mg/kg bw for both male and female rats, 2500 mg/kg bw, with 95% confidence limit of 2300-3100 mg/kg bw for male rats and 2800 mg/kg bw, with 95% confidence limit of 1650- 2200 mg/kg bw for female rats, when groups of 5 male and 5 female Sprague-Dawley rats were treated with the given test chemical orally by gavage in Milli-RO water.

 

The above study is supported with another study mentioned in study report for the test chemical. The acute oral toxicity profile of the given test chemical was conducted according to 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 did not result in any signs of toxicity and 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 did not result in any signs of toxicity and 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 diarrhoea, reduced locomotor activity and ataxic gait with onset at 4 hours after the dosing and no mortality after the dosing.

As no mortality was 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 diarrhoea, reduced locomotor activity and ataxic gait with onset at 2 to 4 hours after the dosing and no mortality after the dosing. Gross pathological examination did not reveal any abnormalities in animals from 300 mg/kg and 2000 mg/kg dose groups.

Hence, the lethal concentration (LD50) value for acute oral toxicity test was considered to be >2000 mg/kg bw, when female Sprague Dawley rats were treated with test chemical orally via gavage according to OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).

 

All the above studies are supported with the study mentioned in databases for the test chemical. The acute oral toxicity study was performed in mice by using test chemical at the dose concentration of 2000 mg/kg bw. Animals were observed for mortality and clinical signs. No mortality was observed at dose 2000 mg/kg bw. Clinical signs observed such as, changes in lungs and thorax. Hence, the lethal concentration (LD50) value for acute oral toxicity test was considered to be >2000 mg/kg bw, when mice were treated with test chemical orally.

 

Thus, based on the above summarised experimental studies on test chemical, the LD50 value is >2000 mg/kg bw. Therefore, 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 the low vapour pressure of the test chemical, which is reported to be 1.056E-4 Pa (7.92E-07 mm Hg). 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 study of the given test chemical was conducted as per OECD Guideline 402 (Acute Dermal Toxicity) in Sprague Dawley rats.

The test item was applied to shorn skin of 5 male and 5 female animals at 2000 mg/kg body weight. Administration of the test item at 2000 mg/kg did not result in any skin reaction at the site of application during the study period of 14 days. Administration of the test item did not result in any signs of toxicity and mortality during the study period of 14 days. Animals exhibited normal body weight gain through the study period of 14 days. Gross pathological examination did not reveal any abnormalities attributable to the treatment.

Hence, the LD50 value was considered to be >2000 mg/kg bw, when male and female Sprague Dawley rats were occlusively treated with test chemical by dermal application following 14 days of observation period according to OECD Guideline 402 (Acute Dermal Toxicity).

 

Justification for classification or non-classification

Based on the above studies on test chemical, it can be concluded that the LD50 value is >2000 mg/kg bw, for acute oral and acute dermal toxicity. Thus, comparing this value 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.