Registration Dossier

Administrative data

Description of key information

Single oral application of 2000 mg submission substance per kg body weight via gavage did not cause lethality in male and female Wistar-rats during the 14 day observation period, resulting in an oral LD50 > 2000 mg/kg bw. No data on acute toxicity after inhalation are available. Reliable data from one guideline study on acute toxicity after dermal application is available for the submission substance and a surrogate substance. The LD50 values from these studies are above 2000 mg/kg bw as well.

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
Study period:
From 11 APR 1996 to 02 MAY 1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
Deviations:
no
GLP compliance:
yes
Remarks:
according to Principles of Good Laboratory Practice, annex of paragraph 19a, section 1 of the chemical law of July 25, 1994
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hoechst Aktiengesellschaft, Kastengrund, SPF breeding colony, Germany
- Age at study initiation: male animals approximately 7 weeks; female animals approximately 8 weeks
- Weight at study initiation: males mean: 182 g; females mean: 172 g
- Fasting period before study: from about 16 hours before to 3 - 4 hours after treatment
- Housing: in fully air-conditioned rooms in macrolon cages (type 4) on soft wood granulate in groups of 5 animals
- Diet: ssniff R/M-H (V 1534), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least one day (breeding at identical conditions)

ENVIRONMENTAL CONDITIONS
- Temperature (°C):22 +/- 3
- Humidity (%): 50 +/- 20
- Air changes (per hr): fully air conditioned rooms
- Photoperiod (hrs dark / hrs light):12/12
Route of administration:
oral: gavage
Vehicle:
other: starch mucilage (potato starch in deionised water)
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 20% suspension in 2% starch mucilage
- Amount of vehicle (if gavage): 10 mL/kg bw
Doses:
2000 mg/kg body weight
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations: symptoms were recorded twice every day (in the morning and in the afternoon), on weekends and public holidays only once.
- Frequency of and weighing: weekly
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no animal died during the 14-day observation period
Mortality:
No deaths occurred during the whole study.
Clinical signs:
The following clinical signs were observed after the application of the test material:
sunken flanks, squatting posture, decreased spontaneous activity, irregular respiration, stilted and uncoordinated gait. The clinical symptoms had reversed 6 to 8 hours after application.
Body weight:
Development of body weight was not impaired.
Gross pathology:
The animals killed at the end of the observation period showed no macroscopically visible changes.
Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Single oral application of 2000 mg test substance per kg body weight via gavage did not cause lethality in male and female Wistar-rats during the 14 day observation period, resulting in a LD50 > 2000 mg/kg bw.
Executive summary:

10 Wistar-rats (5 males and 5 females) were subjected to test acute oral toxicity. The test substance was administered by gavage at a dose of 2000 mg/kg body weight (vehicle starch mucilage). Body weight development was not impaired, general clinical signs observed were reversible within 8 hours after substance application and there were no macroscopically visible changes found. No animal died during the 14 day observation period, resulting in a LD50 > 2000 mg/kg bw.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
2 000 mg/kg bw
Quality of whole database:
The study is GLP compliant and has Klimisch score 1. Another reliable study (Klimisch 1 and GLP) performed with a surrogate of the submission substance supports the reported findings for the submission substance.

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
Study period:
From 27 APR 2011 to 11 MAY 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
according to Hungarian GLP Regulations: 9/2001 (III.30.) which comply with OECD/European community requirements
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Janvier S.A.S.
- Age at study initiation:young adult rats
- Weight at study initiation: between 211 and 245 g
- Housing: individual caging in type II cages (polypropylene/polycarbonate) with Lignocel Hygienic Animal Bedding produced by J. Rettenmaier & Söhne GmbH+Co.KG (Holzmühle 1, 73494 Rosenberger, Germany);
- Diet: ssniff® SM R/M-Z+H "Autoclavable complete feed for rats and mice – breeding and maintenance" produced by ssniff Spezialdiäten GmbH, D-59494
Soest Germany; ad libitum
- Water: tap water from municipal supply; ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.4 to 24.8
- Humidity (%): 31 to 68
- Air changes (per hr): 15 to 20
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: back
- % coverage: 10% of total body surface
- Type of wrap if used: Sterile gauze pads were placed on the skin of rats to cover the test item. These gauze pads were kept in contact with the skin by a patch
with adhesive hypoallergenic plaster. The entire trunk of the animal was then wrapped with semi occlusive plastic wrap for 24 hours.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): with water of body temperature
- Time after start of exposure: 24 hours
Duration of exposure:
24 hours
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: once weekly (recorded on Day 0 (before test item administration) and on Days 7 and 14)
- Necropsy of survivors performed: yes (All animals were anaesthetised with Euthasol® 40% and exsanguinated. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed. All macroscopic changes were recorded.)
- Other examinations performed: clinical signs (Clinical observations were performed on the day of treatment at 1 and 5 hours after application of the test item and once each day for 14 days thereafter. Observations included the skin and fur, eyes and mucous membranes, the respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.)
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no animal died during the 14-day observation period
Mortality:
No mortality occurred after a 24-hour dermal exposure of test material administered at 2000 mg/kg bw to RjHan:(WI) Wistar rats followed by a 14-day observation period.
Clinical signs:
No clinical signs were observed after the treatment with the test item or during the 14-day observation period.
Body weight:
Body weight development was not impaired.
Gross pathology:
There was no evidence of the test item-related observations at a dose level of 2000 mg/kg bw at necropsy.
Other findings:
After treatment with the test material no local signs were observed after the treatment with the test item or during the 14-day observation period.

INDIVIDUAL CLINICAL OBSERVATIONS

DOSE LEVEL: 200 mg/kg bw                                                                                               SEX: MALE

Cage No.

Animal No.

Observations

Observation days

Frequency

0

1

2

3

4

5

6

7

8

9

10

11

12

13

14

1h

5h

1

6235

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

2

6236

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

3

6237

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

4

6238

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

5

6239

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

DOSE LEVEL: 2000 mg/kg bw                                                                                                                         SEX: FEMALE

6

6240

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

7

6241

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

8

6242

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

9

6243

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

10

6244

Symptom Free

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

16/16

Remarks:            +: present

                               h=hour (s)                          Treatment day=Day 0

                               Frequency of observation = number of occurrence of observation / total number of observations

INDIVIDUAL BODY WEIGHT AND BODY WEIGHT GAIN

DOSE LEVEL: 2000 mg/kg bw                                                 SEX: MALE

Cage No.

Animal No.

Body Weight (g) Days

Body Weight Gain (g)

0

7

14

0-7

7-14

0-14

1

6235

234

275

336

41

61

102

2

6236

242

294

357

52

63

115

3

6237

236

290

352

54

62

116

4

6238

245

292

351

47

59

106

5

6239

230

284

341

54

57

111

Mean:

237.4

287.0

347.4

49.6

60.4

110.0

Standard deviation:

6.1

7.7

8.6

5.6

2.4

6.0

DOSE LEVEL: 2000 mg/kg bw                                             SEX: FEMALE

6

6240

224

227

253

3

26

29

7

6241

211

231

243

20

12

32

8

6242

217

220

237

3

17

20

9

6243

218

226

244

8

18

26

10

6244

221

231

245

10

14

24

Mean:

218.2

227.0

244.4

8.8

17.4

26.2

Standard deviation:

4.9

4.5

5.7

7.0

5.4

4.6

Remark:              Treatment day = Day 0

 

INDIVIDUAL INTERNAL AND EXTERNAL MACROSCOPIC OBSERVATIONS

DOSE LEVEL: 2000 mg/kg bw                                                                                                                         SEX: MALE

Cage No.

Animal No.

External Observations

Internal Observations

Organ/Tissue

1

6235

No external observations

No internal observations

Not applicable

2

6236

No external observations

No internal observations

Not applicable

3

6237

No external observations

No internal observations

Not applicable

4

6238

No external observations

No internal observations

Not applicable

5

6239

No external observations

No internal observations

Not applicable

DOSE LEVEL: 2000 mg/kg bw                                                                                                                      SEX: FEMALE

6

6240

No external observations

No internal observations

Not applicable

7

6241

No external observations

No internal observations

Not applicable

8

6242

No external observations

In estrus

Uterus

9

6243

No external observations

No internal observations

Not applicable

10

6244

No external observations

No internal observations

Not applicable

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this limit test the acute dermal median lethal dose (LD50) of the test item was found to be higher than 2000 mg/kg body weight in male and female RjHan:(WI) Wistar rats.
Executive summary:

An acute dermal toxicity study was performed with the test material in RjHan:(WI) Wistar rats, in compliance with OECD Guideline No.: 402. A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test item was applied as supplied as a single dermal 24-hour exposure followed by a 14-day observation period. Clinical observations were performed on all animals at 1 and 5 hours after dosing and daily for 14 days thereafter. Body weight was measured prior to dosing on Day 0 and on Days 7 and 14. Rats were euthanized and a gross macroscopic examination performed at the end of the 2-week observation period (Day 14). The results of the study were summarized as follows: No mortality occurred. No clinical signs were observed after the treatment with the test item or during the 14-day observation period and no local dermal signs were existing. The body weight and body weight gain of treated animals did not show any adverse effects. There was no evidence of the test item-related observations at a dose level of 2000 mg/kg bw at necropsy. Taken from this observations a LD50 is > 2000 mg/kg body weight.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
discriminating dose
2 000 mg/kg bw
Quality of whole database:
The study is GLP compliant and has Klimisch score 1.

Additional information

10 Wistar-rats (5 males and 5 females) were subjected to test acute oral toxicity of the submission substance (OECD 401, GLP compliant). Within this limit test the test substance was administered by gavage at a dose of 2000 mg/kg body weight (vehicle starch mucilage). Body weight development was not impaired, general clinical signs observed were reversible within 8 hours after substance application and there were no macroscopically visible changes found. No animal died during the 14 day observation period, resulting in a LD50 > 2000 mg/kg bw.

The results for the submission substance were verfied by another study performed with a surrogate of the submission substance using the same study design (OECD 401, Limit test, GLP). As no irreversible effects were seen and no animal died during the 14 day observation period the resulting LD50 is > 2000 mg/kg bw for rats.

An acute dermal toxicity study was performed with the the submission subtsance in RjHan:(WI) Wistar rats, in compliance with OECD Guideline No. 402. A limit test was carried out at 2000 mg/kg body weight (bw) in both sexes (5 rats/sex). The test item was applied as supplied as a single dermal 24-hour exposure followed by a 14-day observation period. Within this study the following results were obtained: No mortality occurred. No clinical signs were observed after the treatment with the test item or during the 14-day observation period and no local dermal signs were existing. The body weight and body weight gain of treated animals did not show any adverse effects. There was no evidence of the test item-related observations at a dose level of 2000 mg/kg bw at necropsy. Taken from this observations a LD50 is > 2000 mg/kg body weight.

The results for the submission substance were verfied by another study performed with a surrogate of the submission substance using the same study design (OECD 402, Limit test, GLP). As no irreversible effects were seen and no animal died during the 14 day observation period the resulting LD50 is > 2000 mg/kg bw for rats.


Justification for selection of acute toxicity – oral endpoint
Reliable study performed with the submission substance and a surrogate substance

Justification for selection of acute toxicity – dermal endpoint
Reliable study performed with the submission substance and a surrogate substance

Justification for classification or non-classification

The oral LD50 values for the submission substance as well as for a surrogate of the submission substance are > 2000 mg/kg bw, the limit for classification. No lethality was observed after single dermal application of ≥ 2000 mg submission substance and a surrogate per kg bw. No data on acute toxicity after inhalation exposure are available.

The submission substance does not have to be classified for acute toxicity according to Regulation (EC) No 1272/2008 or to Council Directive 67/548/EEC.

Furthermore, the submission substance does not have to be classified for specific target organ toxicity – single exposure according to Regulation (EC) No 1272/2008, as no specific toxic effects were observed after acute exposure.