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Acute Toxicity: dermal

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Administrative data

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:
Experimental result performed using standard test methods

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Principles of method if other than guideline:
To assess the dermal toxicity of test chemical after dose application by dermal route in rats and an observation period of 14 days.
GLP compliance:
yes
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Details on test material:
Identification : STYRENATED PHENOL
Appearance : Clear yellowish viscous liquid
Batch number : Lot 2/04
CAS No. : 61788-44-1
AI Content : Monostyryl Phenol – 11.20%
Distyryl Phenol– 44.65%
Tristyryl Phenol– 43.59%
Manufactured date : May, 2013
Expiry Date : September, 2014
Storage conditions : Room temperature (20 - 30 °C)

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Animals and Husbandry
Test System Specifications
Species :Rat (Rattus norvegicus)
Strain :Wistar
Sex :Male and Female
Number of Animals :10 (Five per sex)
Supplier/Source :In-House Bred at Sa-Ford, Animal Facility
(CPCSEA Registration No. 1256/bc/09/CPCSEA)
Health Status : Healthy young adult animals were used for the study. Females were nulliparous and non pregnant
Body weight of animals :Male: Minimum: 228 g and Maximum: 255 g
(Prior to Treatment) Female: Minimum: 213 g and Maximum: 238 g
Acclimatisation :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 labeled with study no., study type, test system, sex, dose, group, animal number, experimental start and completion date.
Randomization: Animals were selected manually. No computer generated randomization program was used.

Husbandry Conditions
Diet : All animals were provided conventional laboratory rodent diet (Nutrivet Life Sciences, Pune) ad libitum. Batch No 400004.
Bedding : All cages were provided with corn cobs (Sparconn Life Sciences Bangalore) Batch No.: SPAR – 25/2014
Water : Aqua guard filtered tap water was provided ad libitum via drinking bottles.
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.

Experimental Room Condition
Temperature : Minimum: 19.80 °C Maximum: 23.20 °C
Relative humidity: Minimum: 50.60% Maximum: 61.10%
Light-dark-rhythm: 12:12
Air Changes : More than 12 changes per hour

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
8.2 Preparation of Application Site
Approximately 24 h prior to treatment, the fur of dorsal area of the trunk (approximately 10% body surface area) of rats was clipped by using clipper.
8.3 Test Item Application Procedure
The test item was applied uniformly over clipped dorsal area of rat skin. Individual rat was applied with an amount of test item, calculated based on the density (1.0898) and latest body weight. Test item was held in contact with the skin with a porous gauze dressing (Approx. 10% of body surface area of rat) and non-irritating tape throughout a 24-hour exposure period. It was ensured that the animals cannot ingest the test item. At the end of the exposure period, residual test item was removed by using distilled water. The animals were dosed between 11:19 to 11:32 a.m.
8.4 Limit Test
Five male and five female wistar rats were treated with test item by a single dermal application at the dose level of 2000 mg/kg body weight.
Since no test item related mortality was observed, the study was terminated with limit test only.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 per sex (male and female)
Control animals:
not required
Details on study design:
Preparation of Application Site
Approximately 24 h prior to treatment, the fur of dorsal area of the trunk (approximately 10% body surface area) of rats was clipped by using clipper.

Test Item Application Procedure
The test item was applied uniformly over clipped dorsal area of rat skin. Individual rat was applied with an amount of test item, calculated based on the density (1.0898) and latest body weight. Test item was held in contact with the skin with a porous gauze dressing (Approx. 10% of body surface area of rat) and non-irritating tape throughout a 24-hour exposure period. It was ensured that the animals cannot ingest the test item. At the end of the exposure period, residual test item was removed by using distilled water. The animals were dosed between 11:19 to 11:32 a.m.

Limit Test
Five male and five female wistar rats were treated with test item by a single dermal application at the dose level of 2000 mg/kg body weight.
Since no test item related mortality was observed, the study was terminated with limit test only.


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).

Statistical Analysis
No statistical analysis was performed since the study was terminated with limit test.

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.

Results and discussion

Preliminary study:
No details available
Effect levels
Key result
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no other details available
Mortality:
No mortality was observed at limit dose of 2000 mg/kg body weight of test item during the 14 day observation period (refer table 3).
Clinical signs:
All the animals were observed with normal clinical signs throughout the experimental period (refer table 2).
Body weight:
Mean body weight of both males and females were observed with increase on day 7 and 14, as compared to day 0. ( refer table 1 and 4).
Gross pathology:
The external and internal gross pathological observation of all terminally sacrificed animals did not show any pathological abnormality (refer table 5).

Any other information on results incl. tables

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

 

Dose:2000 mg/ kg bodyweight                                                                              Density:1.0898

Animal No.

Sex

Dose Volume*

(ml)

Body Weight (gram)

Body Weight Change (%)

Day 0

Day 7

Day 14

Day 0-7

Day 0-14

1

Male

0.46

251

262

276

4.38

9.96

2

0.47

254

257

236

1.18

-7.09

3

0.43

236

256

283

8.47

19.92

4

0.47

255

284

306

11.37

20.00

5

0.42

228

247

279

8.33

22.37

6

Female

0.39

213

208

210

-2.35

-1.41

7

0.42

231

236

237

2.16

2.60

8

0.41

224

224

233

0.00

4.02

9

0.44

238

237

245

-0.42

2.94

10

0.40

217

223

232

2.76

6.91

Key:* = based on density and day 0 body weight


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

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

Keys: 1 = Normal

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

244.80

261.20

276.00

6.75

13.03

SD

12.11

13.85

25.29

3.98

12.22

n

5

5

5

5

5

Female

Mean

224.60

225.60

231.40

0.43

3.01

SD

10.16

11.80

13.01

2.07

3.00

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


Applicant's summary and conclusion

Interpretation of results:
other: Not classified
Conclusions:
Under the conditions of study in acute dermal toxicity study,the acute dermal median lethal dose of test chemical was determined to be >2000 mg/kg bw when tested in rats.
Executive summary:

Acute Dermal Toxicity was studied for test chemical in Wistar Rats,This study was performed as per OECD No.402.

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. Approximately 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 based on the density (1.0898) and latest body weight was applied by single dermal application and observed for 14 days after treatment.

On test day 0, calculated amount of test item 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.

Under the conditions of this; acute dermal toxicity study of test chemical in rats is as given below: The acute dermal median lethal dose of test chemical was  > 2000 mg/kg body weight. and the substance is not classified as per the CLP regulation.