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, mice and rabbits for the given test chemical. The LD50 value is 1000 mg/kg bw in rats. The value concluded that the LD50 value is between 300-2000 mg /kg bw, for acute oral toxicity. Thus, comparing this value and range with the criteria of CLP regulation, the given test chemical can be classified in “Category 4” for acute oral toxicity.

Acute Inhalation Toxicity:

The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to low vapour pressure of the test chemical, which is reported to be 4.9E-9 Pa (3.68E-11 mmHg). 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 given test chemical. The study concluded that LD50 value is >2000 mg/kg bw, for acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Justification for type of information:
data is from experimental report.
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Principles of method if other than guideline:
Acute Oral toxicity test was carried out to study the effects of test chemical on rats.
GLP compliance:
no
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Age:7 to 9 weeks
Sex: Female, nulliparous and non pregnant. It has been observed that females are generally more sensitive than males to toxic effects
Body weight range:200±20g
Identification:By cage tag and corresponding colour body marking
Acclimation:One week in experimental room after veterinary examination.
Randomization:After acclimation and veterinary examination randomly selected in groups of three females.
Nutritional conditions:Fasted overnight prior to treatment. Food was offered three hours after dosing.
Husbandry
Environmental conditions:Air conditioned rooms with 10-15 air changes per hour, temperature between 22-25 0C, relative humidity 40-60% and illumination cycle set to 12 hours artificial fluorescent light and 12 hours dark.
Accommodation:Groups of three animals of similar sex in polypropylene cages with stainless steel grill top, facilities for food and water bottle, and bedding of clean paddy husk.
Diet:Pelleted feed
Water: Aqua Guard filter water was kept in PVC bottles, ad libitum
Route of administration:
oral: gavage
Vehicle:
other: distilled water
Details on oral exposure:
- Concentration in vehicle: 2000 mg/kg b. wt and 300 mg/kg b. wt
- Amount of vehicle (if gavage):10 ml/kg b.wt
Doses:
No. of dose groups:Three + one vehicle control
Group I: Dist. water, 10ml/kg body wt.
Group II : 2000 mg/kg body wt.
Group III : 300 mg/kg body wt.
Group IV : 300 mg/kg body wt.
No. of animals per sex per dose:
No. of animals per dose group:Three (3 females)/step
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: BODY WEIGHT: The body weight of all animals was observed weekly on day 0 (pre treatment),7th and 14th (post treatment).
CLINICAL SIGNS: The treated animals were closely observed for clinical signs of intoxication ,first 4 hours and every 1 hours interval for 24 hrs after dosing and thereafter twice a day for 14 days.All rats were observed at least twice daily to observe any clinical signs or behavioral changes.
- Necropsy of survivors performed: yes,necropsy was carried out on all animals which died during the study or surviving animals were sacrified at the end of the study to observe any gross pathological changes.
- Other examinations performed: clinical signs, body weight,organ weights, histopathology study:The organ which showed gross pathological change during necropsy subjected for histopathological study.
Statistics:
no data available
Preliminary study:
no data available
Sex:
female
Dose descriptor:
LD50 cut-off
Effect level:
1 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: 50% mortality was observed
Mortality:
Dose level 300 mg/kg b.wt
Step-I: The test compound did not produce any mortality throughout observation period.
Step-II: The test compound did not produce any mortality throughout the period of observation.
Dose level 2000 mg/kg b.wt.
All the Wistar albino rats died within the 4 hrs. of administration of test compound.
Clinical signs:
Dose level 300 mg/kg b.wt- Test compound did not produce any clinical signs of intoxication throughout the observation period of 14 days.
Dose level 2000 mg/kg b.wt- Test compound produced high to sever signs of intoxication viz; abdominal respiration, convulsion, tremor and hind leg paralysis and finally death after 4 hrs.
Body weight:
All the animals treated with test chemical at the dose level of 300 mg/kg b.wt showed decrease in normal gain in body weight on day 7th as compared to control group. Whereas, on day 14th all the rats showed normal gain weight as compared to control group
Gross pathology:
Necropsy finding:
A.EXTRENAL:
i:skin: skin and hair coat was observed wet
ii:all external orifices: normal
B.INTERNAL:
i:subcutaneous: no change was observed
ii:superficial and deep lymph node: no change in mesenteric lymph node
ABDOMINAL CAVITY:
i. opening and general examination: in the abdominal cavity all the organs were present in normal position
ii. spleen: normal upto highest tested dose level 2000 mg/kg bw
iii. digestive sysytem: no gross changes were observed in stomach and intestine upto highest tested dose level 2000 mg/kg bw
iv. liver and biliary ducts: no gross pathological changes were observed
v. excretory sysytem: no gross pathological changes were observed upto highest tested dose level 2000 mg/kg bw
vi. adrenal: observed normal
vii. male/female genital organs: showed normal color,consistancy and no inflammartory changes upto highest tested dose level 2000 mg/kg bw
2. THORACIC CAVITY:
i. opening and general examination: thoracic cavity was found to be normal withut any fluid,mucous or blood etc.
ii. lungs: severe congestion was observed at the dose level of 2000 mg/kg bw
iii. heart: no changes were observed in color and consistancy. heart found normal upto highest tested dose level 2000 mg/kg bw
iv. thyroid: normal in shape,size and surface upto highest tested dose level 2000 mg/kg bw
3. CRANIAL CAVITY:
brain: normal in shape and size
Other findings:
no data available

SUMMARY OF BODY WEIGHT (GM)

Group

Day 0

Day 7

% Gain/loss

Day 14

% Gain/loss

Group-I distilled water 10ml/kg

211.10

215.91

3.76

219.5

6.28

Group-II

2000 mg/kg b. wt

205.11

-

-

-

-

Group-III

300 mg/kg b. wt

211.91

215.51

1.69

218.26

5.99

Group-IV

300 mg/kg b. wt

204.98

209.65

2.27

211.1

6.44

CLINICAL SIGNS AND MORTALITY

Group: I (Vehicle Control)                                                           Dose: 10 ml/kg b.wt

                                                                                                                                 

                                                           

FEMALE RATS

 

 

 

Parameters

Incidence of clinical signs observed after dosing

Mortality

Day 0

DAY

Min

Hour

30

1

2

4

6

1

2

3

4

5

6

7

8

9

10

11

12

13

14

Total*

Mortality (total)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

 

0/3

 

Clinical Signs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0         =   No clinical sign (Normal)

+         =   Clinical Sign

CLINICAL SIGNS AND MORTALITY

Group: II                                                                                           Dose: 2000 mg/kg b. wt.

 

FEMALE RATS

 

 

Parameters

Incidence of clinical signs observed after dosing

Mortality

Day 0

DAY

Min

Hour

30

1

2

4

6

1

2

3

4

5

6

7

8

9

10

11

12

13

14

Total*

Mortality (total)

0

0

0

3

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

 

 

3/3

Clinical Signs

3

3

3

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Convulsion

++

+++

+++

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Tremor

++

+++

++++

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Abdominal respiration

+++

+++

+++

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

Paralysis

++

+++

+++

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

 0           =   Normal

+            =    Mild

++           =   Moderate

+++         =    High

++++      =   Severe

CLINICAL SIGNS AND MORTALITY

Group: III                                                                                          Dose: 300 mg/kg b.wt

                                            

 

FEMALE RATS

 

 

Parameters

Incidence of clinical signs observed after dosing

Mortality

Day 0

DAY

Min

Hour

30

1

2

4

6

1

2

3

4

5

6

7

8

9

10

11

12

13

14

Total

Mortality (total)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0/3

Clinical Signs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0            =   Normal

+            =    Mild

++           =   Moderate

+++         =   High

 ++++=        Severe

CLINICAL SIGNS AND MORTALITY

Group: IV                                                                                         Dose: 300 mg/kg b.wt.

FEMALE RATS

 

 

Parameters

Incidence of clinical signs observed after dosing

Mortality

Day 0

DAY

Min

Hour

30

1

2

4

6

1

2

3

4

5

6

7

8

9

10

11

12

13

14

Total*

Mortality (total)

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0/3

Clinical Signs

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

 

0            =   Normal

+            =    Mild

++           =   Moderate

+++         =   High

++++      =   Severe

SUMMARY OF NECROPSY FINDINGS

 

S. No.

 

Fate

 

Wistar albino rats

Dose (mg/kg b. wt)

Distilled water (10 ml/kg)

 

2000

 

 

300

 

300

1

Terminal sacrifice

3/3

0/3

3/3

3/3

2

Found Dead

0/3

3/3

0/3

0/3

3

Abnormalities detected

NAD

Severe congestion

In lung

NAD

NAD

 

 

NAD - No abnormality recorded

INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS

Group: I (Vehicle Control)                                                           Dose: 10ml/kg b.wt

                                                                                                                    

FEMALE RATS

 

Animal ID

Fate

Time

Gross Findings

20161-1

TS

Day 14

NAD

20161-2

TS

Day 14

NAD

20161-3

TS

Day 14

NAD

 

Day 0 is the day of dose administration.

TS- Terminal Sacrifice

NAD- No abnormality Detected

INDIVIDUAL ANIMAL FATE & NECROPSY FINDINGS

Group: II                                                                                            Dose: 2000 mg/kg b.wt.                  

FEMALE RATS

 

Animal ID

Fate

Time

Gross Findings

20161-4

FD

4 hrs.

 

Severe congestion in lungs

20161-5

FD

4 hrs.

20161-6

FD

4 hrs.

 

Day 0 is the day of dose administration.

TS- Terminal Sacrifice

NAD- No abnormality Detected

FD- Found Dead

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The lethal concentration (LD50) cut-off value for acute oral toxicity test was considered to be 1000 mg/kg bw, when female wistar albino rats were treated with test chemical orally via gavage according to OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).
Executive summary:

The acute oral toxicity study of the given test chemical was conducted in wistar albino rats under the OECD Guideline-423 for testing of chemicals. The healthy wistar albino rats of body weight 200±20 gm were selected for study after acclimatization to standard laboratory condition and divided into test group and vehicle control group each having three animals. The study was conducted stepwise as follow: Starting dose 2000 mg/kg body weight: The test chemical was mixed with distilled water and administered orally at the dose level of 2000 mg/kg body weight (dose volume 10ml/kg) to three female rats. However; vehicle control group treated with the distilled water at the dose level of 10 ml/kg b.wt.  The treated animals were closely observed continuously for clinical signs of intoxication during first four hours of test compound administration. Thereafter, all the animals were observed periodically at one hour interval for 24 hrs and once daily for a period of 14 days.  The necropsy was performed on all animals died during the study. The Wistar albino rats treated with the test chemical showed convulsion, tremor, abdominal respiration, hind leg paralysis condition and death within the 4 hrs of administration of test compound. No clinical signs and mortality were observed in vehicle control group. The necropsy finding showed severe congestion in lungs of the test group. Furthermore, no gross pathological change was observed in vehicle control group. Final dose 300 mg/kg body weight: Step -I: The test chemical was mixed with distilled water and administered orally at the dose level of 300 mg/kg body wt. (dose volume 10 ml/kg) to three female rats. All the animals were closely observed individually for clinical sign of toxicity and mortality once in 30 minutes, 1, 2, 4 and 6 hrs intervals during the first 24 hrs and thereafter, once daily for a period of 14 days. Necropsy was conducted on all the animals at the termination of study. The test chemical administered at the dose level of 300 mg/kg b.wt did not produce any mortality throughout the observation period of 14 days. Furthermore, no clinical signs toxicity was recorded in any of the treated rats throughout the period of observation. The necropsy finding did not show any gross pathological changes. Step -II: After 72 hrs, the result of step-I was confirmed by administration of same dose level (300 mg/kg. b.wt) of test chemical in additional three animals of same sex (OECD-423 guidelines). The test chemical administered at the dose level of 300 mg/kg b.wt did not produce any mortality and clinical sign of intoxication throughout the observation period of 14 days. The test chemical did not elicit any gross pathological changes in animals sacrificed at the end of experimentation. The lethal concentration (LD50) cut-off value for acute oral toxicity test was considered to be 1000 mg/kg bw, when female wistar albino rats were treated with test chemical orally via gavage according to OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
1 000 mg/kg bw
Quality of whole database:
Data is Klimisch 2 and from experimental report.

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
other justification
Justification for data waiving:
the study does not need to be conducted because exposure of humans via inhalation is not likely taking into account the vapour pressure of the substance and/or the possibility of exposure to aerosols, particles or droplets of an inhalable size
Endpoint conclusion
Quality of whole database:
waiver

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
data is from study report.
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Principles of method if other than guideline:
Acute dermal toxicity test was carried out to study the effects of test chemical on rats.
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: In-House Bred
- Age at study initiation: Healthy young adult were used.Females were nulliparous and non pregnant.
- Weight at study initiation: Male: Minimum: 218 g and Maximum: 261 g
(Prior to Treatment) Female:Minimum: 210 g and Maximum: 221 g
- Housing: Bedding : All cages were provided with corn cobs (Sparconn Life Sciences Bangalore) Batch No.: SPAR – 25/2014
Husbandry : The animals were housed individually in polycarbonate cages.
Room Sanitation : The experimental room floor and work tops were swept and mopped with disinfectant solution every day.
Cages and water bottle : All the cages and water bottles were changed at least twice every week.
- Diet (e.g. ad libitum): animals were provided conventional laboratory rodent diet ad libitum
- Water : Aqua guard filtered tap water was provided ad libitum via drinking bottles.
- Acclimation period: All animals were acclimatized to the test conditions for 6 days prior to administration of the test item.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Minimum: 19.90 °C Maximum: 23.30 °C
- Humidity (%): Minimum: 52.30% Maximum: 66.50%
- Air changes (per hr): More than 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12:12
Type of coverage:
semiocclusive
Vehicle:
other: distilled water
Details on dermal exposure:
TEST SITE
- Area of exposure: dorsal area of the trunk
- % coverage: Approx. 10% of body surface area of rat
- Type of wrap if used: porous gauze dressing and non-irritating tape

REMOVAL OF TEST SUBSTANCE
- Washing (if done): residual test item was removed by using distilled water
- Time after start of exposure: 24-hour exposure period

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg body weight
- Concentration (if solution): Individual rat was applied with an amount of test item moistened with 0.2 ml distilled water
- Constant volume or concentration used: yes
- For solids, paste formed: yes

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.2 ml
Duration of exposure:
24 hrs
Doses:
2000 mg/kg body weight
No. of animals per sex per dose:
total:10 animals
2000 mg/kg bw:male 5 and female:5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: individual animals were frequently observed at 1, 2, 3 and 4 hours post dosing on day 0 (day of dosing). Subsequently, all animals were observed once a day during the 14 day observation period for clinical signs, teice daily for mortality
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology
Statistics:
no data available
Preliminary study:
no data available
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:
No mortality was observed at limit dose of 2000 mg/kg body weight of test item during the 14 day observation period
Clinical signs:
All the animals were observed with normal clinical signs throughout the experimental period
Body weight:
In males and females, mean body weight was observed with gain on day 7 and 14, as compared to day 0
Gross pathology:
The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality
Other findings:
no data available

Individual Animal Body Weight (g) andBody Weight Changes(%)

Dose:2000 mg/ kg bodyweight                                                                                                         

Animal No.

Sex

Body Weight (gram)

Body Weight Change (%)

Day 0

Day 7

Day 14

Day 0-7

Day 0-14

1

Male

222

232

248

4.50

11.71

2

237

243

256

2.53

8.02

3

261

279

314

6.90

20.31

4

255

276

297

8.24

16.47

5

218

229

232

5.05

6.42

6

Female

214

231

244

7.94

14.02

7

212

222

228

4.72

7.55

8

221

237

253

7.24

14.48

9

210

222

241

5.71

14.76

10

213

220

232

3.29

8.92

Individual Animal Clinical Signs and Symptoms

Dose:2000 mg/kg body weight

Animal

No.

Sex

Hour(s) - Day 0

Day

1

2

3

4

1

2

3

4

5

6

7

1

Male

1

1

1

1

1

1

1

1

1

1

1

2

1

1

1

1

1

1

1

1

1

1

1

3

1

1

1

1

1

1

1

1

1

1

1

4

1

1

1

1

1

1

1

1

1

1

1

5

1

1

1

1

1

1

1

1

1

1

1

6

Female

1

1

1

1

1

1

1

1

1

1

1

7

1

1

1

1

1

1

1

1

1

1

1

8

1

1

1

1

1

1

1

1

1

1

1

9

1

1

1

1

1

1

1

1

1

1

1

10

1

1

1

1

1

1

1

1

1

1

1

 

Animal

No.

Sex

Day

8

9

10

11

12

13

14

1

Male

1

1

1

1

1

1

1

2

1

1

1

1

1

1

1

3

1

1

1

1

1

1

1

4

1

1

1

1

1

1

1

5

1

1

1

1

1

1

1

6

Female

1

1

1

1

1

1

1

7

1

1

1

1

1

1

1

8

1

1

1

1

1

1

1

9

1

1

1

1

1

1

1

10

1

1

1

1

1

1

1

Key: 1 = Normal

Individual Animal Mortality Record

 Dose:2000 mg/kg body weight

       Animal No.

Sex

Days of Observation (0 to 14)

Morning Observations

Evening Observations

1

Male

No mortality and morbidity

No mortality and morbidity

2

No mortality and morbidity

No mortality and morbidity

3

No mortality and morbidity

No mortality and morbidity

4

No mortality and morbidity

No mortality and morbidity

5

No mortality and morbidity

No mortality and morbidity

6

Female

No mortality and morbidity

No mortality and morbidity

7

No mortality and morbidity

No mortality and morbidity

8

No mortality and morbidity

No mortality and morbidity

9

No mortality and morbidity

No mortality and morbidity

10

No mortality and morbidity

No mortality and morbidity

Summaryof Animal Body Weight (g) and Body Weight Changes (%)

Dose:2000 mg/kg body weight

Sex

Body Weight (gram)

Body Weight Changes (%)

Day 0

Day 7

Day 14

Day 0-7

Day 0-14

Male

Mean

238.60

251.80

269.40

5.44

12.59

SD

19.19

24.06

34.59

2.20

5.80

n

5

5

5

5

5

Female

Mean

214.00

226.40

239.60

5.78

11.95

SD

4.18

7.30

9.91

1.88

3.43

n

5

5

5

5

5

Keys:SD= Standard deviation, n = Number of animals

GrossNecropsyObservation

 Dose:2000 mg/kg body weight                                               Mode of Death:Terminal Sacrifice

Animal No.

Sex

Gross Observation

External

Internal

1

Male

No abnormality detected

No abnormality detected

2

No abnormality detected

No abnormality detected

3

No abnormality detected

No abnormality detected

4

No abnormality detected

No abnormality detected

5

No abnormality detected

No abnormality detected

6

Female

No abnormality detected

No abnormality detected

7

No abnormality detected

No abnormality detected

8

No abnormality detected

No abnormality detected

9

No abnormality detected

No abnormality detected

10

No abnormality detected

No abnormality detected

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

Acute dermal toxicity study was conducted as per OECD No.402 by using the given test chemical in Wistar Rats. Five male and five female healthy young adult rats were randomly selected and used for conducting acute dermal toxicity study. Rats free from injury and irritation of skin were selected for the study. Twenty four hours prior to dermal application of test item, approximately 10% of body surface area of each rat was clipped. A limit dose of 2000 mg/ kg body weight of test item moistened with 0.2 ml distilled water was applied by single dermal application and observed for 14 days after treatment. On test day 0, an amount of test chemical moistened with 0.2 ml distilled water was applied directly on the intact skin of clipped area of rats; the porous gauze dressing was put on to the intact skin of clipped area. This porous gauze dressing was covered with a non-irritating tape. After the 24-hour application period, the dressings were removed and the skin was gently wiped with distilled water. The skin reactions were assessed. The animals were observed daily for mortality and clinical signs, during the acclimatization period. All animals were observed for clinical signs at approximately 1, 2, 3 and 4 hours after treatment on day 0 and once daily during test days 1 - 14. Mortality was recorded after application on test day 0 and twice daily during days 1-14 (at least once on the day of sacrifice). Local signs / Skin reactions were observed daily from test days 1-14 (in common with clinical signs). Body weights were recorded on day 0 (prior to application) and on day 7 and 14. All animals were necropsied and examined macroscopically. No mortality was observed in any animal till the end of the experimental period. All the animals were observed with normal clinical signs throughout the experimental period. In males and females, mean body weight was observed with gain on day 7 and 14, as compared to day 0. The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality. Under the conditions of this, the LD50 value was considered to be >2000 mg/kg bw, when male and female wistar rats were 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, mice and rabbits for the given test chemical. The studies are summarized as below -

 

The acute oral toxicity study of the given test chemical was conducted in wistar albino rats under the OECD Guideline-423 for testing of chemicals. The healthy wistar albino rats of body weight 200±20 gm were selected for study after acclimatization to standard laboratory condition and divided into test group and vehicle control group each having three animals. The study was conducted stepwise as follow: Starting dose 2000 mg/kg body weight: The test chemical was mixed with distilled water and administered orally at the dose level of 2000 mg/kg body weight (dose volume 10ml/kg) to three female rats. However; vehicle control group treated with the distilled water at the dose level of 10 ml/kg b.wt.  The treated animals were closely observed continuously for clinical signs of intoxication during first four hours of test compound administration. Thereafter, all the animals were observed periodically at one hour interval for 24 hrs and once daily for a period of 14 days.  The necropsy was performed on all animals died during the study. The Wistar albino rats treated with the test chemical showed convulsion, tremor, abdominal respiration, hind leg paralysis condition and death within the 4 hrs of administration of test compound. No clinical signs and mortality were observed in vehicle control group. The necropsy finding showed severe congestion in lungs of the test group. Furthermore, no gross pathological change was observed in vehicle control group. Final dose 300 mg/kg body weight: Step -I: The test chemical was mixed with distilled water and administered orally at the dose level of 300 mg/kg body wt. (dose volume 10 ml/kg) to three female rats. All the animals were closely observed individually for clinical sign of toxicity and mortality once in 30 minutes, 1, 2, 4 and 6 hrs intervals during the first 24 hrs and thereafter, once daily for a period of 14 days. Necropsy was conducted on all the animals at the termination of study. The test chemical administered at the dose level of 300 mg/kg b.wt did not produce any mortality throughout the observation period of 14 days. Furthermore, no clinical signs toxicity was recorded in any of the treated rats throughout the period of observation. The necropsy finding did not show any gross pathological changes. Step -II: After 72 hrs, the result of step-I was confirmed by administration of same dose level (300 mg/kg. b.wt) of test chemical in additional three animals of same sex (OECD-423 guidelines). The test chemical administered at the dose level of 300 mg/kg b.wt did not produce any mortality and clinical sign of intoxication throughout the observation period of 14 days. The test chemical did not elicit any gross pathological changes in animals sacrificed at the end of experimentation. The lethal concentration (LD50) cut-off value for acute oral toxicity test was considered to be 1000 mg/kg bw, when female wistar albino rats were treated with test chemical orally via gavage according to OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method).

 

The above result is supported with the study mentioned in another experimental report for the given test chemical. The acute oral toxicity study was carried out to determine mean lethal dose (MLD) in male and female wistar rats as per OECD Guideline 401 (Acute Oral Toxicity). Animals were given doses as follows: 3 male/female (200 mg/kg) and 3 male(2000 mg/kg). All animals were observed for mortality. All male rats died at dose 2000 mg/kg bw. Under the condition of this, the mean lethal dose (MLD) of test chemical in male and female wistar rat was considered to be > 200<= 2000 mg/kg of body weight. Acute oral toxicity of test chemical to rat by oral route indicates that test chemical exhibits acute toxicity by the oral route in the "Category 4" as per CLP criteria.

 

These studies are further supported with the data mentioned in peer-reviewed journal and the acute oral toxicity study was performed in groups of 10 fasted wistar rats using test chemical. 50% mortality was observed at dose 1050 and 930 mg/kg bw. Clinical signs like behavioral muscle contraction or spasticity and convulsions or effect on seizure threshold were observed.Hence,LD50 value was considered to be 930 mg/kg bw in females and 1050 mg/kg bw in males, when groups of 10 fasted wistar rats were treated with test chemical orally.

 

This result is supported by the data available in peer-reviewed journal for the given test chemical. The acute oral toxicity study was performed in male Fischer 344 rats. 50% mortality was observed at dose 1126 mg/kg bw. Hence, LD50 value was considered to be 1126 mg/kg bw, when male Fischer 344 rats were treated with test chemical orally.

 

Along with the above studies one more study was conducted to determine acute oral toxicity dose for the given test chemical mentioned in peer-reviewed journal. The acute oral toxicity study was performed in rats. Animals were observed for mortality. 50% mortality was observed at dose 1600 mg/kg bw. Hence, LD50 value was considered to be 1600 mg/kg bw, when rat were treated with test chemical orally.

 

Furthermore, the above study is supported with data mentioned in handbook for the given test chemical. The acute oral toxicity study was performed in rats. Animals were observed for mortality. 50% mortality was observed at dose 1200 mg/kg bw. Hence, LD50 value was considered to be 1200 mg/kg bw, when rats were treated with test chemical orally.

 

All the above rats studies are again supported with the study conducted on mice for the given test chemical. The acute oral toxicity study was performed in mouse. Animals were observed for mortality. 50% mortality was observed at dose 1070 mg/kg bw. Hence, LD50 value was considered to be 1070 mg/kg bw, when mouse were treated with test chemical orally.

 

Supporting with the above study one more study was conducted on mice for the given test chemical and mentioned in handbook. The acute oral toxicity study was performed in mouse. Animals were observed for mortality. 50% mortality was observed at dose 540 mg/kg bw. Hence, LD50 value was considered to be 540 mg/kg bw, when mouse were treated with test chemical orally.

 

Both the above mice studies are further supported with the data available in peer-reviewed journal and the acute oral toxicity study was performed in mouse using test chemical. Animals were observed for mortality. 50% mortality was observed at dose 900 mg/kg bw. Hence, LD50 value was considered to be 900 mg/kg bw, when mouse were treated with test chemical orally.

 

All the above studies on rats, mice are further supported with the study conducted on rabbits and mentioned in peer-reviewed journal. The acute oral toxicity study was performed in rabbits using test chemical. Animals were observed for mortality. 50% mortality was observed at dose 1700 mg/kg bw. Hence, LD50 value was considered to be 1700 mg/kg bw, when rabbit were treated with test chemical orally.

 

Thus, based on the above summarised studies on test chemical, it can be concluded that LD50 value is between 300-2000 mg /kg bw, for acute oral toxicity. Thus, comparing this value and range with the criteria of CLP regulation, the given test chemical can be classified in “Category 4” for acute oral toxicity

 

Acute Inhalation Toxicity:

The acute inhalation toxicity study need not be conducted because exposure to humans via inhalation route is not likely taking into account due to low vapour pressure of the test chemical, which is reported to be 4.9E-9 Pa (3.68E-11 mmHg). Thus, exposure to inhalable dust, mist and vapour of the chemical is highly unlikely. Therefore this study is considered for waiver. 

 

Acute Dermal Toxicity:

Acute dermal toxicity study was conducted as per OECD No.402 by using the given test chemical in Wistar Rats. Five male and five female healthy young adult rats were randomly selected and used for conducting acute dermal toxicity study. Rats free from injury and irritation of skin were selected for the study. Twenty four hours prior to dermal application of test item, approximately 10% of body surface area of each rat was clipped. A limit dose of 2000 mg/ kg body weight of test item moistened with 0.2 ml distilled water was applied by single dermal application and observed for 14 days after treatment. On test day 0, an amount of test chemical moistened with 0.2 ml distilled water was applied directly on the intact skin of clipped area of rats; the porous gauze dressing was put on to the intact skin of clipped area. This porous gauze dressing was covered with a non-irritating tape. After the 24-hour application period, the dressings were removed and the skin was gently wiped with distilled water. The skin reactions were assessed. The animals were observed daily for mortality and clinical signs, during the acclimatization period. All animals were observed for clinical signs at approximately 1, 2, 3 and 4 hours after treatment on day 0 and once daily during test days 1 - 14. Mortality was recorded after application on test day 0 and twice daily during days 1-14 (at least once on the day of sacrifice). Local signs / Skin reactions were observed daily from test days 1-14 (in common with clinical signs). Body weights were recorded on day 0 (prior to application) and on day 7 and 14. All animals were necropsied and examined macroscopically. No mortality was observed in any animal till the end of the experimental period. All the animals were observed with normal clinical signs throughout the experimental period. In males and females, mean body weight was observed with gain on day 7 and 14, as compared to day 0. The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality. Under the conditions of this, the LD50 value was considered to be >2000 mg/kg bw, when male and female wistar rats were 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 LD50 value is between 300-2000 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 in “Category 4” for acute oral toxicity and cannot be classified for acute dermal toxicity. For acute inhalation toxicity waiver was added so, not possible to classify.