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Description of key information

Acute oral toxicity: 

The acute oral toxicity dose (LD50) was considered based on different studies conducted on rats for the test chemical. 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 0.0446 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 study conducted on rats for the test chemical. The studies concluded that LD50 value is >2000 mg/kg bw, for acute dermal toxicity. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is ffrom publication
Qualifier:
according to guideline
Guideline:
other: As mentioned below
Principles of method if other than guideline:
Acute oral toxicity of test chemical in rats
GLP compliance:
not specified
Test type:
other: not specified
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals or test system and environmental conditions:
not specified
Route of administration:
oral: unspecified
Vehicle:
not specified
Details on oral exposure:
not specified
Doses:
5000 mg/kg bw
No. of animals per sex per dose:
10(5/sex)
Control animals:
not specified
Details on study design:
Not specified
Statistics:
Not specified
Preliminary study:
Not specified
Sex:
male/female
Dose descriptor:
LD50
Effect level:
5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other:
Remarks:
Mortality observed
Mortality:
One female died on day 2 of experement
Clinical signs:
other: Not specified
Gross pathology:
Necropsy findings of this animal were dark and mottled lungs and liver, reddened pylorus, and gas-filled GI tract.
surviving animals showed signs of decreased activity, ataxia, diarrhea, gasping, and urinary incontinence.
Interpretation of results:
other: Not classified
Conclusions:
The acute oral LD50 of test chemical in 5 male and 5 female rats was estimated to be 5000 mg/kg .
Executive summary:

Acute oral toxicity study of test chemical was conducted on 5 male and 5 female Sprague- Dawley rats at the dose concentration of 5000 mg/kg bw. The test chemical was administered via oral route. All animals were maintained under close observation for recording toxic signs and time of death for 14 days. One female died on day 2.No clinical signs were observed during 14 days observation period. Necropsy findings were dark and mottled lungs and liver, reddened pylorus, and gas-filled GI tract. surviving animals showed signs of decreased activity, ataxia, diarrhea, gasping, and urinary incontinence. Therefore, LD50 value was considered to be 5000 mg/kg bw, when rats were treated with test chemical via oral route.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
5 000 mg/kg bw
Quality of whole database:
Data is Klimisch 2 and from publication

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:
Input of 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
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Principles of method if other than guideline:
The objective of the study was to assess the dermal toxicity of test chemical after single dose application by dermal route in rats.
GLP compliance:
yes
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: In-House Bred at sa-Ford, Animal Facility
- Health Status : Healthy young adult animals were used for the study.
- Females were nulliparous and non-pregnant: yes
- Age at study initiation: No data available
- Weight at study initiation: Male: Minimum: 234 g and Maximum: 258 g Female: Minimum: 228 g and Maximum: 246 g
- Fasting period before study: No data available
- Housing: The animals were housed individually in polycarbonate cages.
- Bedding: All cages were provided with corn cobs (Sparconn Life Sciences Bangalore) Batch No.: SPAR – 24/2013
- 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): All animals were provided conventional laboratory rodent diet (Nutrivet Life Sciences, Pune) ad libitum. Batch No. 400012.
- Water (e.g. ad libitum): Aqua guard filtered tap water was provided ad libitum via drinking bottles.
- Acclimation period: All animals were acclimatized to the test conditions for 5 days prior to administration of the test item.
- Identification: During Acclimatization, animals were temporarily marked by permanent marker, on their tails. After acclimatization, the animals were marked by toe pad micro tattooing and cage cards. Individual cage cards were labelled with project /Study No., species, strain, sex, animal ID. and No. of animal per cage, experiment start and end date.
- Randomization: Animals were selected manually. No computer generated randomization program was used.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Minimum: 20.40 °C Maximum: 23.10 °C
- Humidity (%): Minimum: 38.40% Maximum: 56.00%
- Air changes (per hr): More than 12 changes per hour
- Photoperiod (hrs dark / hrs light): 12:12

IN-LIFE DATES: From: February 10, 2014 To: March 01, 2014
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: Dorsal area of the trunk
- % coverage: Approximately 10% body surface area
- 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

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2000 mg/kg body weight.
- Concentration (if solution): No data available
- Constant volume or concentration used: yes
- For solids, paste formed: no (The substance is liquid)
Duration of exposure:
24-hour
Doses:
2000 mg/kg body weight
No. of animals per sex per dose:
10 (Five per sex)
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Daily
- Necropsy of survivors performed: yes
- Other examinations performed:
Clinical Observation
After test item administration, 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.

Mortality
Animals were observed twice daily for any mortality during the experimental period.

Body weight
All rats were weighed on days 0 (prior to dosing), 7 and 14.

Local Signs/Skin Reactions
All animals were observed once daily during days 1-14 (in common with clinical signs).

Pathology
At the end of 14 day observation period, all the surviving rats were euthanised by overdose of CO2 and subjected to gross pathology examination, for external and internal observations.
Statistics:
No statistical analysis was performed since the study was terminated with limit test.
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other:
Remarks:
No mortality 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:
other: No systemic or local signs of toxicity were observed at limit dose of 2000 mg/kg body weight of test item during the experimental period, except female animal nos. 8 and 10 were observed with erythema from day 2 to 8 and scab from day 9 to 12.
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

Table 1: Individual Animal Body Weight (g) and Body Weight Changes(%)

Dose:2000 mg/ kg bodyweight                                                                              Density:0.9464

Animal No.

Sex

Dose (ml) Applied*

Body Weight (gram)

Body Weight Change (%)

Day 0

Day 7

Day 14

Day 0-7

Day 0-14

1

Male

0.52

247

245

265

-0.81

7.29

2

0.49

234

215

243

-8.12

3.85

3

0.52

245

240

256

-2.04

4.49

4

0.55

258

250

261

-3.10

1.16

5

0.52

244

238

264

-2.46

8.20

6

Female

0.52

244

242

251

-0.82

2.87

7

0.52

246

246

249

0.00

1.22

8

0.49

231

235

231

1.73

0.00

9

0.48

228

229

226

0.44

-0.88

10

0.52

246

250

254

1.63

3.25

Key:* = Based on density of test item and day 0 body weight taken prior to dose application.


Table 2: 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

8

9

10

11

12

13

14

1

Male

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

2

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

3

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

4

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

5

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

6

Female

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

7

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

8

1

1

1

1

1

65+

65+

65+

65+

65+

65+

65+

146

146

146

146

1

1

9

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

10

1

1

1

1

1

65+

65+

65+

65+

65+

65+

65+

146

146

146

146

1

1

Key: 1 = Normal, 65 = Erythema, 146 = Scab, + = Mild


Table 3: 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


Table 4: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

245.60

237.60

257.80

-3.31

5.00

SD

8.56

13.46

8.98

2.82

2.82

n

5

5

5

5

5

Female

Mean

239.00

240.40

242.20

0.60

1.29

SD

8.77

8.44

12.76

1.09

1.78

n

5

5

5

5

5

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


Table 5: GrossNecropsyObservation

 

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

Animal No.

Sex

Gross Observation

External

Internal

1

Male

No abnormalities detected

No abnormalities detected

2

No abnormalities detected

No abnormalities detected

3

No abnormalities detected

No abnormalities detected

4

No abnormalities detected

No abnormalities detected

5

No abnormalities detected

No abnormalities detected

6

Female

No abnormalities detected

No abnormalities detected

7

No abnormalities detected

No abnormalities detected

8

No abnormalities detected

No abnormalities detected

9

No abnormalities detected

No abnormalities detected

10

No abnormalities detected

No abnormalities detected


Interpretation of results:
other: Not classified
Conclusions:
The acute dermal median lethal dose of test chemical was >2000 mg/kg body weight.It can be classified under CLP Classification: “Category- Not classified”.
Executive summary:

 Acute Dermal Toxicity Study of test chemical in Wistar Rats, was performed as per OECD No.402.

Five male and five female healthy young adult rats were selected and used for conducting acute dermal toxicity 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 was applied by single dermal application and observed for 14 days after treatment.

On test day 0 test item was applied directly on the intact skin of clipped area of rats; the surgical gauze patch was put on to the intact skin of clipped area. This porous gauze dressing was covered with a non-irritating adhesive 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. No systemic or local signs of toxicity were observed at limit dose of 2000 mg/kg body weight of test item during the experimental period, except female animal nos. 8 and 10 were observed with erythema from day 2 to 8 and scab from day 9 to 12.

Body weight gain was observed in male and female animals on day 7 and 14 as compared to day 0, except decline in mean body weight gain was observed in males on day 7. External and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality.

Thus, acute dermal median lethal dose of test chemical was >2000 mg/kgbody weight and can be classified as CLP Classification: “Category- Not classified”.

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

Additional information

Acute oral toxicity:

 

In different studies, the given test chemical has been investigated for acute oral toxicity to a greater or lesser extent. Often are the studies based on in-vivo experiments in rodents, i.e. most commonly in rats for test chemical. The studies are summarized as below –

 

1. Acute oral toxicity study of test chemical was conducted on 5 male and 5 female Sprague- Dawley rats at the dose concentration of 5000 mg/kg bw. The test chemical was administered via oral route. All animals were maintained under close observation for recording toxic signs and time of death for 14 days. One female died on day 2.No clinical signs were observed during 14 days observation period. Necropsy findings were dark and mottled lungs and liver, reddened pylorus, and gas-filled GI tract. surviving animals showed signs of decreased activity, ataxia, diarrhea, gasping, and urinary incontinence. Therefore, LD50 value was considered to be 5000 mg/kg bw, when rats were treated with test chemical via oral route.

 

2. Acute oral toxicity study of test chemical was conducted on 5 male and 5 female Sprague- Dawley rats at the dose concentration of 5000 mg/kg bw. The test chemical was administered via oral route. All animals were maintained under close observation for recording toxic signs and time of death for 14 days. No mortality and clinical signs were observed during 14 days of observation perid. Necropsy findings revealed three males and five females had decreased activity and ataxia. Therefore, LD50 value was considered to be 5000 mg/kg bw, when rats were treated with test chemical via oral route.

 

3. Acute oral toxicity study of test chemical was conducted on 5 male and 5 female wistar rats at the dose concentration upto 76800 mg/kg bw. The test chemical was administered via oral route. All animals were maintained under close observation for recording toxic signs and time of death for 14 days. One male died on day 4 with highest dose concentration of 76800 mg/kg bw. No clinical signs were observed during 14 days observation period. The necropsy findings included urinary staining of the abdomen, prominent serosal blood vessels in the stomach, cecum and intestines, and red fluid in the intestines. Therefore, LD50 value was considered to be 76800 mg/kg bw, when rats were treated with test chemical via oral route.

4. Acute oral toxicity study of test chemical was conducted 5 female Albino rats at the dose concentration of 5000 mg/kg bw. The test chemical was administered via oral route. All animals were maintained under close observation for recording toxic signs and time of death for 7 days. No mortality and clinical signs were observed during 7 days observation period. Therefore, LD50 value was considered to be 5000 mg/kg bw, when rats were treated with test chemical via oral route.

5. Acute oral toxicity study of test chemical was conducted 5 female rats at the dose concentration of 15000 mg/kg bw. The test chemical was administered via oral route. All animals were maintained under close observation for recording toxic signs and time of death for 7 days. No mortality and clinical signs were observed during 7 days observation period. Therefore, LD50 value was considered to be >15000 mg/kg bw, when rats were treated with test chemical via oral route.

6. Acute oral toxicity study of test chemical was conducted on 5 female Albino rats at the dose concentration of 1500 mg/kg bw. The test chemical was administered via oral route. All animals were maintained under close observation for recording toxic signs and time of death for 7 days.No mortality and clinical signs were observed during 7 days observation period. Therefore, LD50 value was considered to be >15000 mg/kg bw, when rats were treated with test chemical via oral route.

 

Thus, based on the above summarised studies on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw. 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 0.0446 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:

 

In study report, the given test chemical has been investigated for acute dermal toxicity to a greater or lesser extent. Often are the studies based on in-vivo experiments in rodents, i.e. most commonly in rats for test chemical. The study is summarized as below –

 

1.  Acute Dermal Toxicity Study of test chemical in Wistar Rats, was performed as per OECD No.402.

Five male and five female healthy young adult rats were selected and used for conducting acute dermal toxicity 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 was applied by single dermal application and observed for 14 days after treatment.

On test day 0 test item was applied directly on the intact skin of clipped area of rats; the surgical gauze patch was put on to the intact skin of clipped area. This porous gauze dressing was covered with a non-irritating adhesive 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. No systemic or local signs of toxicity were observed at limit dose of 2000 mg/kg body weight of test item during the experimental period, except female animal nos. 8 and 10 were observed with erythema from day 2 to 8 and scab from day 9 to 12.

Body weight gain was observed in male and female animals on day 7 and 14 as compared to day 0, except decline in mean body weight gain was observed in males on day 7. External and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality.

Thus, acute dermal median lethal dose of test chemical was >2000 mg/kgbody weight and can be classified as CLP Classification: “Category- Not classified”. 

 

Thus, based on the above summarised study on test chemical, it can be concluded that LD50 value is >2000 mg/kg bw. Thus, comparing this value with the criteria of CLP regulation, the given test chemical cannot be classified for acute dermal toxicity.

Justification for classification or non-classification

Based on the above studies on test chemical, it can be concluded that LD50 value is >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.