Registration Dossier

Administrative data

Description of key information

Oral LD50 (OECD guideline 401), rat 1200 mg/kg bw
Dermal LD50 (OECD guideline 402), rabbit > 2000 mg/kg bw (limit test)
Acute toxicity by inhalation was not tested according to REGULATION (EC) No 1907/2006, Annex VIII, Section 8.5, Column 2.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Reason / purpose:
read-across source
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS- Details on strain: Wistar / Crl:WI (Han)- Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany- Age at study initiation: approx. 10 weeks- Weight at study initiation: mean (1st group): 169.7 ± 4.16g; mean (2nd group): 179.3 ± 10.02g- Fasting period before study: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum.- Housing: single housing in Makrolon cage, type III- Diet (ad libitum): VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany- Water (ad libitum): Tap water- Acclimation period: at least 5 daysENVIRONMENTAL CONDITIONS- Temperature: 22°C ± 3°C- Humidity: 30 – 70%- Air changes (per hr): approx. 10- Photoperiod (hrs dark / hrs light): 12 / 12
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
deionized
Details on oral exposure:
VEHICLE- Concentration in vehicle: 20 g/100 mL- Amount of vehicle (if gavage): 10 mL/kg bw- Justification for choice of vehicle: Solution in deionized water.MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bwCLASS METHOD- Rationale for the selection of the starting dose:By request of the sponsor a starting dose of 2000 mg/kg bw was chosen in the first step with 3 female animals. As only one animal died, 2000 mg/kg bw were administered to 3 female rats in the second step.
Doses:
Starting dose: 2000 mg/kg bwSecond step dose: 2000 mg/kg bw
No. of animals per sex per dose:
3 female rats per test group
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days- Frequency of observations: A check for any dead or moribund animals was made at least once each workday.- Necropsy of survivors performed: yes Necropsy with gross-pathology examination on the last day of the observation period after sacrifice by CO2-inhalation in a chamber with increasing concentrations over time. Necropsy of the animal that died during the study as early as possible after death.- Other examinations performed: Clinical signs: Recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals. Body weight: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation. Histopathology: No histological examinations were performed.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
In the first 2000 mg/kg bw test group one animal was found dead at hour 5 after administration.No mortality occurred in the second test group.
Clinical signs:
1st group: Clinical signs in the two surviving animals of the first 2000 mg/kg bw test group revealed impaired general state, dyspnoea and piloerection from hour 1 until hour 5 after administration. In the animal that died in this test group poor general state, dyspnoea, piloerection and abdominal position were observed at hour 4 only.2nd group:In the second test group impaired general state, dyspnoea and piloerection were noticed from hour 1, 3, or 4 until hour 5 after administration.
Body weight:
The mean body weight of the surviving animals increased within the normal range throughout the study period, with exception of one animal of the first test group, which did not adequately gain weight during the second post-exposure week.
Gross pathology:
Deceased animal:In the animal that died in the first test group the following macroscopic pathologic findings were observed: Liquid, light discolored content in the stomach, red discoloration of the glandular stomach and small intestine.Sacrificed animals:There were no macroscopic pathological findings in the surviving animals sacrificed at the end of the observation period.

Body weight changes:

Individual body weight changes

Dose [mg/kg bw]

2000

2000

Group

1

2

Animal No.

1

2

3

mean

sd.

1

2

3

mean

sd.

Body weight at study day [g]

 

 

 

 

 

 

 

 

 

 

0

165

171

173

169.7

4.16

183

187

168

179.3

10.02

7

182

182

-

182.0

0.00

195

203

187

195.0

8.00

14

184

192

-

188.0

5.66

206

212

201

206.3

5.51

sd. = standard deviation

Interpretation of results:
Category 5 based on GHS criteria
Endpoint:
acute toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Reason / purpose:
read-across source
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
According to guideline.
Route of administration:
oral: gavage
Vehicle:
water
Doses:
no data
No. of animals per sex per dose:
5-10
Control animals:
not specified
Sex:
male/female
Dose descriptor:
LD50
Effect level:
1 200 mg/kg bw
Based on:
test mat.
Sex:
female
Dose descriptor:
LD50
Effect level:
977 mg/kg bw
Based on:
test mat.
Sex:
male
Dose descriptor:
LD50
Effect level:
1 427 mg/kg bw
Based on:
test mat.
Mortality:
Occurred, but no detailed data.
Clinical signs:
Diarrhoea, spastic gait, decreased activity, lateral position, hunched posture, laboured respiration, coma, death.
Body weight:
Not stated.
Gross pathology:
Necropsy of died animals: haemorrhages in gastro-intestinal tract and vascular congestion in the liverNecropsy of survivors: no adverse effects
Other findings:
Not stated.
Interpretation of results:
Category 4 based on GHS criteria
Endpoint:
acute toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Reason / purpose:
read-across source
Species:
rat
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 1 500 mg/kg bw
Based on:
act. ingr.
Interpretation of results:
Category 4 based on GHS criteria
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
01 June 2012 - 30 August 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS- Details on strain: Wistar / Crl:WI (Han)- Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany- Age at study initiation: approx. 10 weeks- Weight at study initiation: mean (1st group): 169.7 ± 4.16g; mean (2nd group): 179.3 ± 10.02g- Fasting period before study: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum.- Housing: single housing in Makrolon cage, type III- Diet (ad libitum): VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany- Water (ad libitum): Tap water- Acclimation period: at least 5 daysENVIRONMENTAL CONDITIONS- Temperature: 22°C ± 3°C- Humidity: 30 – 70%- Air changes (per hr): approx. 10- Photoperiod (hrs dark / hrs light): 12 / 12
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
deionized
Details on oral exposure:
VEHICLE- Concentration in vehicle: 20 g/100 mL- Amount of vehicle (if gavage): 10 mL/kg bw- Justification for choice of vehicle: Solution in deionized water.MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bwCLASS METHOD- Rationale for the selection of the starting dose:By request of the sponsor a starting dose of 2000 mg/kg bw was chosen in the first step with 3 female animals. As only one animal died, 2000 mg/kg bw were administered to 3 female rats in the second step.
Doses:
Starting dose: 2000 mg/kg bwSecond step dose: 2000 mg/kg bw
No. of animals per sex per dose:
3 female rats per test group
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days- Frequency of observations: A check for any dead or moribund animals was made at least once each workday.- Necropsy of survivors performed: yes Necropsy with gross-pathology examination on the last day of the observation period after sacrifice by CO2-inhalation in a chamber with increasing concentrations over time. Necropsy of the animal that died during the study as early as possible after death.- Other examinations performed: Clinical signs: Recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals. Body weight: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation. Histopathology: No histological examinations were performed.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
In the first 2000 mg/kg bw test group one animal was found dead at hour 5 after administration.No mortality occurred in the second test group.
Clinical signs:
1st group: Clinical signs in the two surviving animals of the first 2000 mg/kg bw test group revealed impaired general state, dyspnoea and piloerection from hour 1 until hour 5 after administration. In the animal that died in this test group poor general state, dyspnoea, piloerection and abdominal position were observed at hour 4 only.2nd group:In the second test group impaired general state, dyspnoea and piloerection were noticed from hour 1, 3, or 4 until hour 5 after administration.
Body weight:
The mean body weight of the surviving animals increased within the normal range throughout the study period, with exception of one animal of the first test group, which did not adequately gain weight during the second post-exposure week.
Gross pathology:
Deceased animal:In the animal that died in the first test group the following macroscopic pathologic findings were observed: Liquid, light discolored content in the stomach, red discoloration of the glandular stomach and small intestine.Sacrificed animals:There were no macroscopic pathological findings in the surviving animals sacrificed at the end of the observation period.

Body weight changes:

Individual body weight changes

Dose [mg/kg bw]

2000

2000

Group

1

2

Animal No.

1

2

3

mean

sd.

1

2

3

mean

sd.

Body weight at study day [g]

 

 

 

 

 

 

 

 

 

 

0

165

171

173

169.7

4.16

183

187

168

179.3

10.02

7

182

182

-

182.0

0.00

195

203

187

195.0

8.00

14

184

192

-

188.0

5.66

206

212

201

206.3

5.51

sd. = standard deviation

Interpretation of results:
Category 5 based on GHS criteria
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
yes
Remarks:
Only limited information are available.
GLP compliance:
no
Test type:
standard acute method
Limit test:
yes
Species:
rat
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 1 500 mg/kg bw
Based on:
act. ingr.
Interpretation of results:
Category 4 based on GHS criteria
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Remarks:
Only basic data given.
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
yes
Remarks:
Limited information concerning test conditions and/or experimental methods.
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
According to guideline.
Route of administration:
oral: gavage
Vehicle:
water
Doses:
no data
No. of animals per sex per dose:
5-10
Control animals:
not specified
Sex:
male/female
Dose descriptor:
LD50
Effect level:
1 200 mg/kg bw
Based on:
test mat.
Sex:
female
Dose descriptor:
LD50
Effect level:
977 mg/kg bw
Based on:
test mat.
Sex:
male
Dose descriptor:
LD50
Effect level:
1 427 mg/kg bw
Based on:
test mat.
Mortality:
Occurred, but no detailed data.
Clinical signs:
Diarrhoea, spastic gait, decreased activity, lateral position, hunched posture, laboured respiration, coma, death.
Body weight:
Not stated.
Gross pathology:
Necropsy of died animals: haemorrhages in gastro-intestinal tract and vascular congestion in the liverNecropsy of survivors: no adverse effects
Other findings:
Not stated.
Interpretation of results:
Category 4 based on GHS criteria
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
1 200 mg/kg bw
Quality of whole database:
The whole data base is reliable and of high quality.

Acute toxicity: via inhalation route

Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: dermal
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Reason / purpose:
read-across source
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS- Details on strain: Wistar / Crl:WI (Han) SPF- Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany- Age at study initiation: approx. 9 weeks (males) and 13 weeks (females), respectively- Weight at study initiation: mean (males): 273.0 ± 6.63g; mean (females): 225.8 ± 3.19g- Fasting period before study: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum.- Housing: single housing in Makrolon cage, type III- Diet (ad libitum): VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany- Water (ad libitum): Tap water- Acclimation period: at least 5 daysENVIRONMENTAL CONDITIONS- Temperature: 22°C ± 3°C- Humidity: 30 – 70%- Air changes (per hr): approx. 10- Photoperiod (hrs dark / hrs light): 12 / 12
Type of coverage:
semiocclusive
Vehicle:
water
Remarks:
deionized water
Details on dermal exposure:
TEST SITE- Area of exposure: clipped epidermis (dorsal and dorsolateral parts of the trunk)- % coverage: about 40 cm² (corresponds to at least 10% of the body surface)REMOVAL OF TEST SUBSTANCE- Washing: rinsing of the application site with warm water- Time after start of exposure: 24 hoursTEST MATERIAL- Amount(s) applied (volume or weight with unit): 5.71 mL/kg bw- Concentration (if solution): 35 g/100 mL- Constant volume or concentration used: yes- For solids, paste formed: no- Form of application: solution in deionized water- Test item preparation and homogenization: For better handling the test item was ground with mortar and pestle. The test item preparation was produced shortly before application by stirring with a magnetic stirrer.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 animals/sex/dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days- Frequency of observations: A check for any dead or moribund animals was made at least once each workday.- Necropsy of survivors performed: yes- Other examinations performed: Clinical signs: Recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals. Scoring of skin findings: Individual readings 30 – 60 minutes after removal of the semi-occlusive dressing (day 1), weekly thereafter and on the last day of observation. The evaluation of skin reactions was performed according to Draize, J.H. (1959): Appraisal of the safety of chemicals in foods, drugs and cosmetics. The association of food and drug officials of the United States Austin, Texas. Body weight: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation. Pathology: Necropsy with gross-pathology examination on the last day of the observation period after sacrifice with CO2 in a chamber with increasing concentrations over time.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
No systemic clinical signs were observed during clinical examination.
Body weight:
Males:The mean body weight of the male animals increased within the normal range throughout the study period.Females:The mean body weight of the female animals did not significantly change during the first post-exposure observation week, probably due to the bandage procedure, but increased during the second week within the normal range.
Gross pathology:
There were no macroscopic pathological findings in the animals sacrificed at the end of observation period.
Other findings:
Local effects: No local effects were observed.

Body weight changes:

Individual body weight changes

Dose [mg/kg bw]

2000

Sex

male

Animal No.

1

2

3

4

5

mean

sd.

Body weight at study day [g]

 

 

 

 

 

 

 

0

274

267

282

266

276

273.0

6.63

7

278

284

301

281

285

285.8

8.93

14

307

304

330

299

308

309.6

11.93

 

Individual body weight changes

Dose [mg/kg bw]

2000

Sex

female

Animal No.

1

2

3

4

5

mean

sd.

Body weight at study day [g]

 

 

 

 

 

 

 

0

224

222

230

225

228

225.8

3.19

7

231

220

234

226

230

228.2

5.40

14

237

238

240

239

238

238.4

1.14

sd. = standard deviation

Interpretation of results:
Category 5 based on GHS criteria
Endpoint:
acute toxicity: dermal
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Reason / purpose:
read-across source
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS- Weight at study initiation: 2.4 - 2.9 kgNo further data are available.
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE- Area of exposure: approx. 25% (abraded and shaved)- Type of wrap if used: occlusive (Elastoplast)REMOVAL OF TEST SUBSTANCE- Time after start of exposure: 24 hTEST MATERIAL- Amount(s) applied (volume or weight with unit): 2 g/kg
Duration of exposure:
24 h
Doses:
2 g/kg bw (corresponding to 500 mg a.s./kg bw)
No. of animals per sex per dose:
3
Control animals:
not required
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 500 mg/kg bw
Based on:
act. ingr.
Mortality:
None
Clinical signs:
Moderate to severe erythema, edema, and atonia were observed in all animals during this study. By Day 6 of the study, all animals showed signs of desquamation and fissuring. Five animals were observed to have moderate to marked desquamation and one animal had slight desquamation. Moderate to marked fissuring was noted in all animals in this study. In addition, all animals had eschar formation and exfoliation. Signs of dermal irritation were evident in all six anmals at the termination of this study.
Body weight:
No weight losses.
Gross pathology:
No data
Interpretation of results:
GHS criteria not met
Endpoint:
acute toxicity: dermal
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Reason / purpose:
read-across source
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
No data.
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE- Area of exposure: approx. 10%- Type of wrap if used: occlusive (Elastoplast)REMOVAL OF TEST SUBSTANCE- Time after start of exposure: 24 hTEST MATERIAL- Amount(s) applied (volume or weight with unit): 2 g/kg
Duration of exposure:
24 h
Doses:
2 g/kg bw (corresponding to 500 mg a.s./kg bw)
No. of animals per sex per dose:
3
Control animals:
not required
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 500 mg/kg bw
Based on:
act. ingr.
Mortality:
None
Clinical signs:
Severe erythema and slight eschar formation at 24 h; necrosis by Day 2–14 with sloughing of the skin by Day 8–14; hyper-pigmentation of new skin by Day 14; no signs of systemic toxicity.
Body weight:
Decrease in body weight (220 g) during 14 d observation period in one rabbit with intact skin.
Gross pathology:
No data
Interpretation of results:
GHS criteria not met
Endpoint:
acute toxicity: dermal
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Reason / purpose:
read-across source
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
No data.
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE- Area of exposure: approx. 10%- Type of wrap if used: occlusive (Elastoplast)REMOVAL OF TEST SUBSTANCE- Time after start of exposure: 24 hTEST MATERIAL- Amount(s) applied (volume or weight with unit): 2 g/kg
Duration of exposure:
24 h
Doses:
2 g/kg bw (corresponding to 500 mg a.s./kg bw)
No. of animals per sex per dose:
3
Control animals:
not required
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 500 mg/kg bw
Based on:
act. ingr.
Mortality:
None
Clinical signs:
Severe erythema and eschar formation at 24 h; necrosis by Day 5–21; necrotic tissues sloughed and leaving skin hyper-pigmented at Day 21; no further signs of systemic toxicity than body weight changes.
Body weight:
Weight loss in all except one rabbit.
Gross pathology:
No data
Interpretation of results:
GHS criteria not met
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
01 June 2012 - 30 August 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS- Details on strain: Wistar / Crl:WI (Han) SPF- Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany- Age at study initiation: approx. 9 weeks (males) and 13 weeks (females), respectively- Weight at study initiation: mean (males): 273.0 ± 6.63g; mean (females): 225.8 ± 3.19g- Fasting period before study: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum.- Housing: single housing in Makrolon cage, type III- Diet (ad libitum): VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany- Water (ad libitum): Tap water- Acclimation period: at least 5 daysENVIRONMENTAL CONDITIONS- Temperature: 22°C ± 3°C- Humidity: 30 – 70%- Air changes (per hr): approx. 10- Photoperiod (hrs dark / hrs light): 12 / 12
Type of coverage:
semiocclusive
Vehicle:
water
Remarks:
deionized water
Details on dermal exposure:
TEST SITE- Area of exposure: clipped epidermis (dorsal and dorsolateral parts of the trunk)- % coverage: about 40 cm² (corresponds to at least 10% of the body surface)REMOVAL OF TEST SUBSTANCE- Washing: rinsing of the application site with warm water- Time after start of exposure: 24 hoursTEST MATERIAL- Amount(s) applied (volume or weight with unit): 5.71 mL/kg bw- Concentration (if solution): 35 g/100 mL- Constant volume or concentration used: yes- For solids, paste formed: no- Form of application: solution in deionized water- Test item preparation and homogenization: For better handling the test item was ground with mortar and pestle. The test item preparation was produced shortly before application by stirring with a magnetic stirrer.
Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 animals/sex/dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days- Frequency of observations: A check for any dead or moribund animals was made at least once each workday.- Necropsy of survivors performed: yes- Other examinations performed: Clinical signs: Recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals. Scoring of skin findings: Individual readings 30 – 60 minutes after removal of the semi-occlusive dressing (day 1), weekly thereafter and on the last day of observation. The evaluation of skin reactions was performed according to Draize, J.H. (1959): Appraisal of the safety of chemicals in foods, drugs and cosmetics. The association of food and drug officials of the United States Austin, Texas. Body weight: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation. Pathology: Necropsy with gross-pathology examination on the last day of the observation period after sacrifice with CO2 in a chamber with increasing concentrations over time.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
No systemic clinical signs were observed during clinical examination.
Body weight:
Males:The mean body weight of the male animals increased within the normal range throughout the study period.Females:The mean body weight of the female animals did not significantly change during the first post-exposure observation week, probably due to the bandage procedure, but increased during the second week within the normal range.
Gross pathology:
There were no macroscopic pathological findings in the animals sacrificed at the end of observation period.
Other findings:
Local effects: No local effects were observed.

Body weight changes:

Individual body weight changes

Dose [mg/kg bw]

2000

Sex

male

Animal No.

1

2

3

4

5

mean

sd.

Body weight at study day [g]

 

 

 

 

 

 

 

0

274

267

282

266

276

273.0

6.63

7

278

284

301

281

285

285.8

8.93

14

307

304

330

299

308

309.6

11.93

 

Individual body weight changes

Dose [mg/kg bw]

2000

Sex

female

Animal No.

1

2

3

4

5

mean

sd.

Body weight at study day [g]

 

 

 

 

 

 

 

0

224

222

230

225

228

225.8

3.19

7

231

220

234

226

230

228.2

5.40

14

237

238

240

239

238

238.4

1.14

sd. = standard deviation

Interpretation of results:
Category 5 based on GHS criteria
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
no
Remarks:
pre-GLP
Test type:
fixed dose procedure
Limit test:
yes
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS- Weight at study initiation: 2.4 - 2.9 kgNo further data are available.
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE- Area of exposure: approx. 25% (abraded and shaved)- Type of wrap if used: occlusive (Elastoplast)REMOVAL OF TEST SUBSTANCE- Time after start of exposure: 24 hTEST MATERIAL- Amount(s) applied (volume or weight with unit): 2 g/kg
Duration of exposure:
24 h
Doses:
2 g/kg bw (corresponding to 500 mg a.s./kg bw)
No. of animals per sex per dose:
3
Control animals:
not required
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 500 mg/kg bw
Based on:
act. ingr.
Mortality:
None
Clinical signs:
Moderate to severe erythema, edema, and atonia were observed in all animals during this study. By Day 6 of the study, all animals showed signs of desquamation and fissuring. Five animals were observed to have moderate to marked desquamation and one animal had slight desquamation. Moderate to marked fissuring was noted in all animals in this study. In addition, all animals had eschar formation and exfoliation. Signs of dermal irritation were evident in all six anmals at the termination of this study.
Body weight:
No weight losses.
Gross pathology:
No data
Interpretation of results:
GHS criteria not met
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Similar to Guideline with acceptable restrictions.
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Qualifier:
according to
Guideline:
other: Standard Procedure #10, Rabbit Acute Percutaneous Toxicity Studies
Deviations:
no
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
3 animals instead of 5 animals were used.The observation period was 72 h instead of 14 d. The test substance was applied under occlusive instead of semi-occlusive conditions.
Principles of method if other than guideline:
Three male and three female New Zealand Albino rabbits each in the weight range of 2200-2800 g constitute a test group. Animals are randomized into groups according to sex and weight. Areas on the animals' backs approximating 10% of the total body surface are clipped prior to treatment with an Oster small animal clipper. The backs of three animals (2 females, 1 male) are abraded and three (2 males, 1 female) are left intact. Abrading is accomplished with a clipper head which produces minor incisions through the stratum corneum which are not deep enough to produce bleeding.2 g/kg of the undiluted test material are applied to the test sites and spread with a tongue depressor over the clipped area. A material in dry form may be moistened, if necessary. The test material is covered with a layer of gauze and covered with a rubber dam and secured in place by several wrappings of 3 inch Elastoplast tape. The animals are placed in harnesses to prevent removal of the bandage and test material. The harnesses and wrappings are removed after 24 hours. At this time the skin is evaluated for degree of irritation. Daily observations are made and recorded for 14 days thereafter. On the fourteenth day the animals are counted, weighed and sacrificed. If deaths were to occur due to this test procedure, an LD50 value could be determined. In our experience, however, with the types of materials that we test, this does not occur so that the test provides information relative to primary irritation.
GLP compliance:
no
Remarks:
pre-GLP
Test type:
fixed dose procedure
Limit test:
yes
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
No data.
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE- Area of exposure: approx. 10%- Type of wrap if used: occlusive (Elastoplast)REMOVAL OF TEST SUBSTANCE- Time after start of exposure: 24 hTEST MATERIAL- Amount(s) applied (volume or weight with unit): 2 g/kg
Duration of exposure:
24 h
Doses:
2 g/kg bw (corresponding to 500 mg a.s./kg bw)
No. of animals per sex per dose:
3
Control animals:
not required
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 500 mg/kg bw
Based on:
act. ingr.
Mortality:
None
Clinical signs:
Severe erythema and slight eschar formation at 24 h; necrosis by Day 2–14 with sloughing of the skin by Day 8–14; hyper-pigmentation of new skin by Day 14; no signs of systemic toxicity.
Body weight:
Decrease in body weight (220 g) during 14 d observation period in one rabbit with intact skin.
Gross pathology:
No data
Interpretation of results:
GHS criteria not met
Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
Refer to the Category Approach Justification document provided in IUCLID6 Section 13.
Qualifier:
according to
Guideline:
other: Standard Procedure #10, Rabbit Acute Percutaneous Toxicity Studies
Deviations:
no
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
yes
Remarks:
3 animals instead of 5 animals were used. The observation period was 72 h instead of 14 d. The test substance was applied under occlusive instead of semi-occlusive conditions.
Principles of method if other than guideline:
Three male and three female New Zealand Albino rabbits each in the weight range of 2200-2800 g constitute a test group. Animals are randomized into groups according to sex and weight. Areas on the animals' backs approximating 10% of the total body surface are clipped prior to treatment with an Oster small animal clipper. The backs of three animals (2 females, 1 male) are abraded and three (2 males, 1 female) are left intact. Abrading is accomplished with a clipper head which produces minor incisions through the stratum corneum which are not deep enough to produce bleeding.2 g/kg of the undiluted test material are applied to the test sites and spread with a tongue depressor over the clipped area. A material in dry form may be moistened, if necessary. The test material is covered with a layer of gauze and covered with a rubber dam and secured in place by several wrappings of 3 inch Elastoplast tape. The animals are placed in harnesses to prevent removal of the bandage and test material. The harnesses and wrappings are removed after 24 hours. At this time the skin is evaluated for degree of irritation. Daily observations are made and recorded for 14 days thereafter. On the fourteenth day the animals are counted, weighed and sacrificed. If deaths were to occur due to this test procedure, an LD50 value could be determined. In our experience, however, with the types of materials that we test, this does not occur so that the test provides information relative to primary irritation.
GLP compliance:
no
Remarks:
pre-GLP
Test type:
fixed dose procedure
Limit test:
yes
Species:
rabbit
Strain:
New Zealand White
Sex:
male/female
Details on test animals and environmental conditions:
No data.
Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE- Area of exposure: approx. 10%- Type of wrap if used: occlusive (Elastoplast)REMOVAL OF TEST SUBSTANCE- Time after start of exposure: 24 hTEST MATERIAL- Amount(s) applied (volume or weight with unit): 2 g/kg
Duration of exposure:
24 h
Doses:
2 g/kg bw (corresponding to 500 mg a.s./kg bw)
No. of animals per sex per dose:
3
Control animals:
not required
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 500 mg/kg bw
Based on:
act. ingr.
Mortality:
None
Clinical signs:
Severe erythema and eschar formation at 24 h; necrosis by Day 5–21; necrotic tissues sloughed and leaving skin hyper-pigmented at Day 21; no further signs of systemic toxicity than body weight changes.
Body weight:
Weight loss in all except one rabbit.
Gross pathology:
No data
Interpretation of results:
GHS criteria not met
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw
Quality of whole database:
The whole data base is reliable and of high quality.

Additional information

There are no data on acute toxicity for C9-12AS Na (CAS n.a.) available. Therefore this endpoint is covered by read across to structurally related alkyl sulfates, i.e. C8AS Na (CAS 142-31-4) and C12AS Na (CAS 151-21-3) for acute oral and dermal toxicity as well as C10-16AS NH4 (CAS 68081-96-9), C10-16AS Mg (CAS 68081-97-0) and C12-13AS K (CAS 91783-22-1) for acute dermal toxicity. During evaluation of the human health hazards of the alkyl sulfates within the AS category it turned out that alkyl sulfates with a carbon chain length of C12 bear the most concern for potential adverse effects on human health. Alkyl sulfates of this chain length have to be classified as harmful if swallowed while alkyl sulfates having a chain length of C8 and higher than C14 does not have to be classified for acute toxicity after oral ingestion. The same trend is also seen for the irritating properties of alkyl sulfates (see endpoint summary of IUCLID section 7.3.1 for details). The possibility of a read-across to other alkyl sulfates in accordance with Regulation (EC) No 1907/2006 Annex XI 1.5. Grouping of substances and read-across approach was assessed. In Annex XI 1.5 it is given that a read-across approach is possible for substances, whose physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity. The AS reported within the AS category show structural similarity. The most important common structural feature of the category members is the presence of a predominantly linear aliphatic hydrocarbon chain with a polar sulfate group, neutralized with a counter ion. This structural feature confers the surfactant properties of the alkyl sulfates. The surfactant property of the members of the AS category in turn represent the predominant attribute in mediating effects on mammalian health. Therefore, the AS of the AS category have similar physico-chemical, environmental and toxicological properties, validating the read across approach within the category. The approach of grouping different AS for the evaluation of their effects on human health and the environment was also made by the OECD in the SIDS initial assessment profile [1] and by a voluntary industry programme carrying out Human and Environmental Risk Assessments (HERA [2]), further supporting the read across approach between structurally related AS.

Acute oral toxicity

The study conducted with C12AS Na (CAS 151-21-3) was performed according to OECD Guideline 401 with acceptable restrictions on Wistar rats (Potokar, 1983). Both sexes were dosed with the test substance at unknown doses via gavage. Mortalities occurred but no details were available. Clinical signs of toxicity comprised of diarrhoea, spastic gait, decreased activity, lateral position, hunched posture, laboured respiration and coma. Necropsy of dead animals revealed haemorrhages in gastro-intestinal tract and vascular congestion in the liver. No findings were observed in animals sacrificed upon study termination. The LD50 was given as 1200 mg/kg bw for males and females, 977 mg/kg bw for females and 1427 mg/kg for males.

The second study conducted with C12AS Na (CAS 151-21-3, analytical purity 30%) was performed similar to OECD Guideline 401 (Esposito, 1976). 5 Wistar rats of each sex were dosed with the test substance at the limit dose of 5000 mg/kg bw via gavage and observed for mortalities and behaviour for 14 days. Mortalities occurred 24 hours (2/5 males) and 3 days after treatment (1/5 females). No depression of animals was observed during the post exposure period. The LD50 was determined to be greater than 5000 mg/kg bw based on the test substance and greater than 1500 mg/kg bw based on the active ingredient.

The study conducted with C8AS Na (CAS 142-31-4) was performed according to OECD Guideline 423 with Wistar rats. Two groups of three female rats were treated with 2000 mg/kg bw via gavage according to the Acute Toxic Class method. The observation period was 14 days. One animal was found dead 5 h after administration in application group 1. No mortality occurred in the second test group. Clinical signs of toxicity comprised impaired general state, dyspnoea and piloerection. All animals gained weight throughout the whole study period. Upon pathology no findings were observed in the surviving animals of both groups.

The available data of the alkyl sulphate category suggest alkyl sulfates with a carbon chain length of C12 to bear the most concern for potential adverse effects on human health. As C9-12AS Na (CAS n.a.) contains carbon chain lengths of C12 up to 30% the LD50 is very likely to be greater than the LD50 of pure C12AS Na. Therefore the LD50 of C12 ASO4 is used as a worst case scenario for C9-12AS Na (CAS n.a.). Thus, the oral LD50 is established at 1200 mg/kg bw.

Acute dermal toxicity 

Regarding the acute dermal toxicity four studies are available for the read-across substances C10-16AS NH4 (CAS 68081-96-9), C10-16AS Mg (CAS 68081-97-0), C12-13AS K (CAS 91783-22-1) and C8AS Na (CAS 142-31-4).

The key study was conducted with C8AS Na (CAS 142-31-4) according to OECD Guideline 402 (BASF, 2012). 5 Wistar rats per sex were treated with 2000 mg/kg bw under semi occlusive conditions for 24 h. No mortality occurred, no sign of systemic toxicity and of local irritation was observed.

The study conducted with C10-16AS NH4 (CAS 68081-96-9) was performed as limit test conducted similar to OECD Guideline 402 with 3 male and 3 female New Zealand White rabbits (Frank, 1975). The test substance (analytical purity 25.1%) was applied at 2000 mg/kg bw for 24 h under occlusive conditions. No mortalities occurred. Clinical signs of toxicity comprised of severe erythema and slight eschar formation at 24 h, necrosis by Day 2–14 with sloughing of the skin by Day 8–14 and hyper-pigmentation of new skin by day 14. No signs of systemic toxicity were observed. Hence, the LD50 value is greater than 2000 mg/kg bw based on the test material and greater than 500 mg/kg bw based on the active ingredient.

The study conducted with C10-16AS Mg (CAS 68081-97-0) was performed as limit test conducted similar to OECD Guideline 402 with 3 male and 3 female New Zealand White rabbits (Frank, 1975). The test substance (analytical purity 23.5%) was applied at 2000 mg/kg bw for 24 h under occlusive conditions. No mortalities occurred. Clinical signs of toxicity comprised of severe erythema and eschar formation at 24 h, necrosis by Day 5–21 and necrotic tissues sloughed and leaving skin hyper-pigmented at Day 21. Hence, the LD50 value is greater than 2000 mg/kg bw based on the test material and greater 500 mg/kg bw based on the active ingredient.

Another study similar to OECD Guideline 402 was performed with C12-13AS K (CAS 91783-22-1) on three male and three female New Zealand White rabbits (Benedict, 1978). Both sexes were dosed at 2000 mg/kg bw (analytical purity 25%) under occlusive conditions for 24 h. No mortalities occurred during the conduct of the study. Findings within this study comprised of moderate to severe erythema, edema, and atonia, desquamation and fissuring by Day 6 and eschar formation and exfoliation. Based on the above mentioned findings the LD50 is greater than 2000 mg/kg bw based on the test material and greater 500 mg/kg bw based on the active ingredient.

Within the studies mentioned above no mortalities or signs of systemic toxicity occurred after dermal application of alkyl sulfates with varying carbon chain length. Taken into account the generally low toxicity after oral application and the low dermal absorption rate of alkyl sulfates (approx. 1%) this result was expected. In agreement, no classification for acute dermal toxicity is required. For details on absorption please see section Toxicokinetics, metabolim and distribution.

Acute inhalation toxicity 

No studies for acute inhalation toxicity are available. However, testing the potential of acute toxicity via inhalation route of C9-12AS Na (CAS n.a.) is considered to be not justified. According to Regulation (EC) No 1907/2006, Annex VIII, Section 8.5, Column 2, in addition to the oral route (8.5.1), for substances other than gases, the information mentioned under 8.5.2 to 8.5.3 shall be provided for at least one other route. As information under 8.5.3 (dermal route) is provided, the requirement is fulfilled by using the most appropriate route of exposure.

AS is mainly used in liquid media and due to its very low vapour pressure [2] inhalation is not viewed as a significant route of exposure.Inhalation of AS may occur by inhalation of aerosols generated by spray cleaners or by inhalation of detergent dusts (e.g. washing powder). Taken into account that the acute toxicity of AS is generally low no further information on acute toxicity is expected by testing for acute inhalation toxicity as the predominant effect will comprise of local irritant effects rather than systemic toxicity. However, as the neat substance has to be classified as harmful if swallowed the substance will also be classified as harmful if inhaled (H332: Acute tox 4 and R20, respectively) in case the substance is available as neat powder.

[1] SIDS initial assessment profile, (2007);
http://www.aciscience.org/docs/Alkyl_Sulfates_Final_SIAP.pdf

[2] (HERA Draft report, 2002);
http://www.heraproject.com/files/3-HH-04-%20HERA%20AS%20HH%20web%20wd.pdf


Justification for classification or non-classification

The available data on acute oral toxicity meet the criteria for classification as Acute Toxic Category 4 (H302) according to Regulation (EC) 1272/2008 and as Xn (R22) according to Directive 67/548/EEC.

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