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Administrative data

Description of key information

acute oral toxicity (female rat, OECD 423): LD50 = 5000 mg/kg bw (cut-off value)
acute dermal toxicity (male and female rat, OECD 402): LD50 > 2000 mg/kg bw

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:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-guideline study
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
adopted in 2001
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, Landesinstitut für Arbeitsschutz und Produktsicherheit, München, Germany
Test type:
acute toxic class method
Limit test:
yes
Species:
rat
Strain:
other: Wistar Crl: WI(Han)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River, Sulzfeld, Germany
- Age at study initiation: 8 - 10 weeks
- Weight at study initiation: 154 - 167 g
- Fasting period before study: Food was withheld for 16 to 19 h before until 4 h after dosing.
- Housing: The animals were housed in groups in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding.
- Diet: Altromin 1324 maintenance diet for rats and mice, ad libitum
- Water: tap water, sulphur acidified to a pH value of approximately 2.8, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES:
- Step 1
From: 27 Jul 2012
To: 10 Aug 2012
- Step 2
From: 16 Aug 2012
To: 30 Aug 2012
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 0.2 g/mL
- Justification for choice of vehicle: The vehicle was chosen due to its non-toxic characteristics.

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
6 females (3/step)
Control animals:
other: not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were made several times on the day of dosing (especially during the first 30 min until 4 h after application) and once daily thereafter until the end of the study period. Animals were weighed on Days 1, 8, and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight
Sex:
female
Dose descriptor:
LD50
Effect level:
5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: cut-off value
Mortality:
All animals survived until the end of the observation period.
Clinical signs:
After application of the test substance, slightly reduced spontaneous activity and slight piloerection were noticed in all animals. The effects were reversible within 4 h. Moreover, the first set of animals showed bradykinesia, kyphosis and half eyelid-closure, that reversed within 3 h after application.
Body weight:
No effects on body weight and body weight gain were observed (Table 1).
Gross pathology:
Necropsy revealed no substance-related findings.

Table 1: Absolute Body Weights and Body Weight Gain

Animals

Body weight (g)

Increase (%)

Step

Number

Day 1

Day 8

Day 15

Day 1 - 15

1

1

156

187

198

27

2

154

176

185

20

3

165

175

200

21

2

4

164

191

208

27

5

167

189

202

23

6

164

185

197

20

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
CLP: not classified
DSD: not classified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The available information comprises an adequate and reliable study (Klimisch score 1), and is thus sufficient to fulfil the standard information requirements set out in Annex VII, 8.5, of Regulation (EC) No 1907/2006.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

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:
other: GLP-guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
adopted in 1987
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, Landesinstitut für Arbeitsschutz und Produktsicherheit, München, Germany
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
other: WISTAR Crl: WI(Han)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River, Sulzfeld, Germany
- Age at study initiation: 8 - 9 weeks (males), 12 - 13 weeks (females)
- Weight at study initiation: 256 - 278 g (males), 211 - 224 g (females)
- Housing: The animals were housed in groups in IVC cages, type III H, polysulphone cages on Altromin saw fibre bedding.
- Diet: Altromin 1324 maintenance diet for rats and mice, ad libitum
- Water: tap water, sulphur acidified to a pH value of approximately 2.8, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES:
From: 16 Aug 2012
To: 30 Aug 2012
Type of coverage:
semiocclusive
Vehicle:
corn oil
Details on dermal exposure:
TEST SITE
- Area of exposure: clipped, healthy skin of the dorsal area of the trunk
- % coverage: approximately 10%
- Type of wrap if used: gauze and non-irritant tape, fixed with a semiocclusive dressing

REMOVAL OF TEST SUBSTANCE
- Washing (if done): After an exposure period of 24 h, the residual test substance was removed using the vehicle.
Duration of exposure:
24 h
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Clinical observations were made several times on the day of dosing (especially during the first 30 min until 4 h after application) and once daily thereafter until the end of the study period. The animals were weighed on Day 1 prior to application and on Days 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: primary skin irritation (Draize scoring system)
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
All animals survived until the end of the study period.
Clinical signs:
No signs of sytemic toxicity were observed.
Body weight:
A slight weight loss was recorded for 1/5 males and 3/5 females during the first week, but all animals showed weight gain during the second week (Table 1).
Gross pathology:
The necropsy revealed no test substance-related findings.
Other findings:
Skin irritation:
No erythema and oedema formation was noticed during the observation period. In 1/5 males, eschar formation was observed on Day 8, that resolved within 24 h (Table 2). In 4/5 females, scratches, eschar formation or wounds were noticed which were fully reversible within the observation period in 3/5 females (Table 3).

Table 1: Absolute Body Weights and Body Weight Gain

Animals

Body weight (g)

Increase (%)

Sex

Number

Day 1

Day 8

Day 15

Day 1 - 15

male

21

274

305

339

24

22

264

285

310

17

23

256

255

273

7

24

278

291

319

15

25

270

281

313

16

female

26

213

213

219

3

27

224

221

221

-1

28

211

215

216

2

29

212

206

211

0

30

213

211

215

1

Table 2: Skin irritation – individual data – males

Days after Application

Animal No. 21

Animal No. 22

Animal No. 23

Animal No. 24

Animal No. 25

E/O

Comments

E/O

Comments

E/O

Comments

E/O

Comments

E/O

Comments

2

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

3

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

4

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

5

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

6

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

7

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

8

0/0

nsf

0/0

nsf

0/0

es

0/0

nsf

0/0

nsf

9

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

10

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

11

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

12

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

13

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

14

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

15

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

Comments:

E = erythema, O = oedema; 1, 2, 3, 4 = scoring system laid down in OECD Guideline 404 (Draize scoring system)

es = eschar, s = scratches, es* = eschar on large patches; w = small wounds; w (2) = wound (2 mm diameter) nsf = no specific finding

Table 3: Skin irritation – individual data – females

Days after Application

Animal No. 26

Animal No. 27

Animal No. 28

Animal No. 29

Animal No. 30

E/O

Comments

E/O

Comments

E/O

Comments

E/O

Comments

E/O

Comments

2

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

3

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

4

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

0/0

nsf

5

0/0

nsf

0/0

nsf

0/0

nsf

0/0

es*

0/0

es

6

0/0

nsf

0/0

nsf

0/0

nsf

0/0

es*

0/0

es

7

0/0

s

0/0

s

0/0

nsf

0/0

es*

0/0

nsf

8

0/0

w (2)

0/0

es

0/0

nsf

0/0

es*

0/0

nsf

9

0/0

w (2)

0/0

es

0/0

nsf

0/0

es*

0/0

nsf

10

0/0

w (2)

0/0

es

0/0

nsf

0/0

w

0/0

nsf

11

0/0

w (2)

0/0

es

0/0

nsf

0/0

w

0/0

nsf

12

0/0

w (2)

0/0

es

0/0

nsf

0/0

w

0/0

nsf

13

0/0

w

0/0

nsf

0/0

nsf

0/0

w

0/0

nsf

14

0/0

w

0/0

nsf

0/0

nsf

0/0

w

0/0

nsf

15

0/0

nsf

0/0

nsf

0/0

nsf

0/0

w

0/0

nsf

Comments:

E = erythema, O = oedema; 1, 2, 3, 4 = scoring system laid down in OECD Guideline 404 (Draize scoring system)

es = eschar, s = scratches, es* = eschar on large patches; w = small wounds; w (2) = wound (2 mm diameter) nsf = no specific finding

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
CLP: not classified
DSD: not classified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
The available information comprises an adequate and reliable study (Klimisch score 1), and is thus sufficient to fulfil the standard information requirements set out in Annex VII, 8.5, of Regulation (EC) No 1907/2006.

Additional information

Acute oral toxicity

In the oral toxicity study performed by Takawale (2003) according to OECD 423 and GLP, 3 female Wistar rats were treated with 2000 mg/kg bw of the test substance in corn oil by gavage. As no mortality occurred, another group of 3 female rats was treated with 2000 mg/kg bw of the test substance. Again, there was no mortality.

After application of the test substance, slightly reduced spontaneous activity and slight piloerection were noticed in all animals. The effects were reversible within 4 h. Moreover, the first set of animals showed bradykinesia, kyphosis and half eyelid-closure, that reversed within 3 h after application. During the 14 day-observation period, no further clinical signs and no effects on the body weight gain were observed in any animal. Macroscopic examinations at necropsy did not reveal any abnormalities.

According to the acute toxic class method described in the OECD guideline 423, if there is no mortality following administration of 2000 mg/kg bw in two separate steps, the LD50 cut-off limit is 5000 mg/kg bw. Therefore, the LD50 is considered to be 5000 mg/kg bw for female rats.

Acute dermal toxicity 

The acute dermal toxicity of the test substance was investigated in a study performed according to OECD 402 and GLP (Takawale, 2012). A single test substance dose of 2000 mg/kg bw was applied on the clipped, healthy skin of five male and five female Wistar Crl: WI(Han) rats. The treated skin site was covered with a semiocclusive dressing for 24 h. No mortality occurred during the 14 day-observation period. No clinical signs were observed and the necropsy revealed no test substance-related findings.

A slight weight loss was recorded for 1/5 males and 3/5 females during the first week, but all animals showed weight gain during the second week.

Scratches were observed in 2/5 females on Day 7. In these females, wounds or eschar formation, respectively, were observed by Day 8. The effects were reversible within 7 and 5 days, respectively. Eschar formation was further noticed in 1/5 males on Day 8 only and in 2/5 females on Day 5. The effect was reversible in one of these females by Day 6. The second female showed eschar formation on large patches that persisted from Day 5 to Day 9. In this female, small wounds were observed from Day 10 until the end of the observation period.

No erythema and oedema formation was observed during the observation period.

Therefore, the LD50 is > 2000 mg/kg bw for male and female rats.


Justification for selection of acute toxicity – oral endpoint
There is only one study available.

Justification for selection of acute toxicity – dermal endpoint
There is only one study available.

Justification for classification or non-classification

The available data on the acute oral and dermal toxicity of the test substance does not meet the criteria for classification according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and is therefore conclusive but not sufficient for classification.