Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From October 20, 1999 to November 3, 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report Date:
2000

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
Feb 1987
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Purity: approximately 81%
- Physical state: off white paste
- Lot/batch No.: 876 Tk
- Storage condition of test material: room temperature, in the dark
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source:
- Batch No.of test material: 876 TK
- Date received: 10 May 1999
- Purity: 92%
- Physical appearance: white paste

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in the dark

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test material was used as supplied. It was weighed out according to the animals individual body weight.

Test animals

Species:
rat
Strain:
Sprague-Dawley
Remarks:
Crl: CD (SD) IGS BR
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River (UK)
- Age at study initiation: approximately eight to twelve weeks old
- Weight at study initiation: males 206 to 232 g, and the females 214 to 225 g.
- Housing: in suspended polypropylene cages furnished with wood flakes. The animals were housed individually during the 24-hour exposure period and in groups of five, by sex, for the remainder of the study
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25 ºC
- Humidity (%): 30-70 %
- Air changes (per hr): approximately fifteen changes per hour
- Photoperiod (hrs dark / hrs light): twelve hours continuous light and twelve hours darkness

Administration / exposure

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): with cotton wool moistened with distilled water to remove any residual test material.
- Time after start of exposure: After the 24-hour contact period



TEST SITE
- Area of exposure: Back and flank of animals
- % coverage: approximating to 10% of the total body surface area.
- Type of wrap if used: Surgical gauze and self-adhesive bandage

REMOVAL OF TEST SUBSTANCE
- Washing: Residual test substance was wiped off with cotton wool moistened with distilled water
- Time after start of exposure: After a 24-hour contact period

TEST MATERIAL
- Amount applied: not specified
- Constant volume or concentration used: yes
- For solids, paste formed: yes

VEHICLE
- No vehicle used. The undiluted test substance was used.
Duration of exposure:
24-hours
Doses:
2174 mg/kg (equivalent to 2000 mg/kg bodyweight of pure test material).
No. of animals per sex per dose:
Ten animals (five males and five females)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: The animals were observed for deaths or overt signs of toxicity ½, 1, 2 and 4 hours after dosing and subsequently once daily for fourteen days. Individual bodyweights were recorded prior to application of the test material on Day 0 and on Days 7 and 14.
- Necropsy of survivors performed: yes
- Scoring system: Scale from Draize J H (1977)
Statistics:
Mortality data was used to estimate the acute dermal median lethal dose (LD50) of the test substance

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 174 mg/kg bw
Based on:
test mat.
Remarks on result:
other: The dose 2174 mg/kg is equivalent to 2000 mg/kg bodyweight of pure test material
Mortality:
There were no deaths.
Clinical signs:
No signs of systemic toxicity were noted during the study. Very slight to well-defined erythema was noted at the treatment sites of all animals one and two days after dosing, in nine animals three days after dosing and in four animals four days after dosing. Very slight oedema was noted at the treatment sites of eight animals one day after dosing, in six animals two days and three days after dosing and in two animals four days after dosing. On the treated site of some animals a slight brown discoloration of the epidermis during days one to seven (in four animals), bleeding four days after dosing (in one animal), desquamation was noted three days (in four animals) and four days (in five animals), and crust formation after five days (in five female animals), six and seven days (in four animals each), and after eight and nine days (in three animals) was noted.
Body weight:
All animals showed expected gain in bodyweight during the study except for one female animal which showed bodyweight loss during the first week and expected gain in bodyweight during the second week.
Gross pathology:
No abnormalities were noted at necropsy.
Other findings:
Residual test material was noted at the treatment sites of all animals during the study and prevented accurate evaluation of oedema in two males one to five days after dosing.
One female showed body weight loss during the first week andd expected gain in body weight during the second week. All other animlas showed normal body weights.

Any other information on results incl. tables

Table 1: Individual clinical observations and mortality data

Dose level [mg/kg]

Animal number and sex

Effects noted after dosing (hours)

Effects noted after dosing (days)

1/2

1

2

4

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2174

1-0

male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-1

male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-2

male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-3

male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-4

male

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-0

female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-1

female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-2

female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-3

female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-4

female

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0 = no signs of systemic toxicity

Table 2.1: Individual dermal reactions - males

Dose level [mg/kg]

Animal number and sex

Observation

Effects noted after dosing (days)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2174

1-0

male

Erythema

Oedema

Other

2

1

Rt

2

1

Rt

1

1

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-1

male

Erythema

Oedema

Other

2

od

RtBr

2

od

RtBr

1

od

RtBr

0

od

RtBr

0

od

RtBr

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

G

0

0

G

0

0

G

1-2

male

Erythema

Oedema

Other

2

1

Rt

1

1

Rt

1

1

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1-3

male

Erythema

Oedema

Other

2

1

Rt

2

1

Rt

2

1

Rt

1

1

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

G

0

0

G

1-4

male

Erythema

Oedema

Other

2

od

Rt

2

od

Rt

1

od

Rt

0

od

RtBs

0

od

RtBr

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

Rt

0

0

G

0

0

G

0

0

G

0 = no signs of dermal irritation; G = glossy skin; Bs = Bleeding; Rt = residual test material; Br = light brown discoloration of the epidermis; od = adverse reactions prevent accurate evaluation of oedema

Table 2.2: Individual dermal reactions - females

Dose level [mg/kg]

Animal number and sex

Observation

Effects noted after dosing (days)

1

2

3

4

5

6

7

8

9

10

11

12

13

14

2174

2-0

female

Erythema

Oedema

Other

2

1

Rt

1

1

Rt

1

1

RtD

1

1

RtD

0

0

RtCf

0

0

Cf

0

0

Cf

0

0

Cf

0

0

Cf

0

0

G

0

0

G

0

0

G

0

0

G

0

0

G

2-1

female

Erythema

Oedema

Other

2

1

RtBr

2

1

RtBr

2

1

RtBrD

1

0

RtBrD

0

0

RtBrCf

0

0

Cf

0

0

Cf

0

0

Cf

0

0

Cf

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-2

female

Erythema

Oedema

Other

2

1

Rt

2

0

Rt

1

0

RtD

0

0

RtD

0

0

RtCf

0

0

Cf

0

0

Cf

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

2-3

female

Erythema

Oedema

Other

2

1

RtBr

2

1

RtBr

2

1

RtBrD

2

0

RtBrD

0

0

RtBrCf

0

0

BrCf

0

0

BrCf

0

0

Cf

0

0

Cf

0

0

G

0

0

G

0

0

G

0

0

G

0

0

G

2-4

female

Erythema

Oedema

Other

2

1

Rt

1

0

Rt

0

0

Rt

0

0

D

0

0

Cf

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0 = no signs of dermal irritation; D = desquamation; Br = light brown discoloration of the epidermis; G = glossy skin; Cf = crust formation; Rt = residual test material

Table 3: Individual bodyweight and weekly changes

 

Doselevel

mg/kg

Animal Number and Sex

Bodyweight

(g) atDay

Bodyweight Gain (g)

During Week

 

 

0

7

14

1

2

2174

1-0 Male

206

238

286

32

48

1-1 Male

228

285

336

57

51

1-2 Male

232

280

345

48

65

1-3Male

220

257

312

37

55

1-4 Male

215

201

322

46

61

2-0 Female

214

215

234

1

19

2-1 Female

220

225

236

5

11

2-2 Female

225

220

245

-5

25

2-3 Female

224

235

265

11

30

2-4 Female

225

226

250

1

24

 

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The acute dermal median lethal dose (LD50) of the test substance in the Sprague-Dawley CD strain rat was found to be greater than 2174 mg/kg bodyweight (equivalent to 2000 mg/kg bodyweight of pure test material).
Executive summary:

The Acute Dermal Toxicity assay for the test substance was performed in Sprague-Dawley CD rats. One female group and one male group, of 5 animals each one, were treated with a single dose of 2174 mg/kg bodyweight, equivalent to 2000 mg/kg bodyweight of pure test material (Limit test). The test material was applied uniformly to an area of shorn skin (approximating to 10% of the total body surface area). A piece of surgical gauze was placed over the treatment area and semi-occluded with a piece of self-adhesive bandage. After the 24-hour contact period the bandage was carefully removed and the treated skin and surrounding hair wiped with cotton wool moistened with distilled water. Clinical observation was conducted at 1/2,1, 2 and 4 hours after dosing and subsequently once daily for fourteen days. Body weight was controlled before the administration, on day 0 and on days 7 and 14 after the administration. All animals were subjected to gross necropsy. No test substance related effects were noted from clinical observations or post-mortem examination. The acute dermal median lethal dose (LD50) of the test substance in the Sprague-Dawley CD strain rat was found to be greater than 2174 mg/kg bodyweight (equivalent to 2000 mg/kg bodyweight of pure test material).