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Description of key information

Both the dermal and oral LD50 were found to be higher than the limit dose of 2000 mg/kg

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 September 19, 2008 to October 7, 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
2001
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
2008
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
2002
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Strain as stated in the report: Wistar rat strain Crl:Wl (Han) outbred
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: approximately 11-12 weeks
- Weight at study initiation: group 1: 183-208 g / group 2: 181-199 (body weight variation did not exceed +/-20% of the sex mean)
- Sex: female (nulliparous and non-pregnant)
- Fasting period before study: food was withheld overnight (for a maximum of 20 hours) prior to dosing until 3-4 hours after administration of the test substance
- Housing: Group housing of 3 animals per cage in labeled Macrolon cages (MIV type; height 18 cm), containing sterilized sawdust as bedding material (Litalabo, SPPS, Argenteuil, France) and paper as cage-enrichment (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom)
- Diet ad libitum: pelleted rodent diet (SM R/M-Z from SSNIFF Spezialdiäten GmbH, Soest, Germany)
- Water ad libitum: tap water
- Acclimation period: At least 5 days before start of treatment under laboratory conditions
- Number of animal per dose group: 3

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18.9-21.2°C
- Humidity (%): 43-75%
- Air changes (per hr): 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12h artificial fluorescent light and 12h darkness per day
Route of administration:
oral: gavage
Vehicle:
propylene glycol
Details on oral exposure:
VEHICLE
- Volume applied: 10 ml/kg bw (2000 mg/kg bw)
- Justification for choice of vehicle: selection based on trial formulations performed at Notox and on test substance data supplied
- Specific gravity: 1.036
- Formulations (w/w, 4 grams substance with 17 grams vehicle) were prepared within 4h prior to dosing. Homogeneity was accomplished to a visually acceptable level.

ACUTE TOXIC CLASS METHOD:
- Method: Oral gavage, using plastic feeding tubes
- Frequency: single dosage, on day 1
- Dose level (volume): 2000 mg/kg bw (10 mL/kg) bw
Doses:
2000 mg/kg (10 mL/kg) bw
No. of animals per sex per dose:
3 females per dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 15 days

OBSERVATIONS:
- Frequency of observation for mortality/viability: twice daily
- Frequency of weighing: Days 1 (pre-administration), 8 and 15
- Frequency of observation for clinical signs: At periodic intervals on the day of dosing (day 1) and once daily thereafter, until day 15.
- Necropsy: At the end of the observation period (day 15), all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal abnormalities were recorded.

Statistics:
Electronic data capture:
REES Centron Environmental Monitoring System version SQL 2.0 (REES scientific, Trenton, NJ, USA) and
TOXDATA version 8.0 (NOTOX BV, 's-Hertogenbosch, The Netherlands)
No statistical analysis was performed.
Preliminary study:
Dose: 2000 mg/kg bw
No mortality occured
Hunched posture was noted in all animals between days 1 and 8 and piloerection was noted in all animals between days 1 and 4. Uncoordinated movements were observed among the animals on day 1.
The body weight gain shown by the animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.
No abnormalities were found at macroscopic post mortem examination of the animals.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: none of the animals died
Mortality:
No mortality occured.
Clinical signs:
other: Hunched posture was noted in all animals between days 1 and 8 and piloerection was noted in all animals between days 1 and 4. Uncoordinated movements were observed among the animals on day 1.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.

Table 1: Mean body weights in gram (Dose level: 2000 mg/kg bw)

Test day

1

8

15

 

 

 

 

Group 1 females

197 +/- 13

222 +/- 15

230 +/- 22

Group 2 females

188 +/- 10

206 +/- 15

215 +/- 15

Body weights on day one were assessed before application of test article.

Interpretation of results:
GHS criteria not met
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

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 August 27, 2009 to October 14, 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
1987
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Japan MAFF Testing Guideline of 12 Nosan No. 8147
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
standard acute method
Limit test:
yes
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Strain as stated in the report: Rat/Wistar/CrI:WI (Han) SPF
- Source: Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: males: approximately 8 weeks, females: approximately 12 weeks
- Weight at study initiation: males: mean weight: 251.4 g (Sd: 1.52), females: mean weight: 203.8 g (Sd: 5.93)
- 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 ad libitum
- Acclimation period: at least 5 days before the beginning of the experimental pahse
- Other information: The female animals were nulliparous and non-pregnant

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-26°C
- Humidity (%): 20-80%
- Air changes (per hr): air-conditioned rooms
- Photoperiod (hrs dark / hrs light): 12h/12h
Type of coverage:
semiocclusive
Vehicle:
water
Remarks:
Double distilled
Details on dermal exposure:
TEST SITE
- Area of exposure: dorsal and dorsolateral parts of the trunk
- % coverage: about 40 cm2 (corresponds to at least 10% of the body surface)
- Type of wrap if used: semi occlusive dressing (the bandage consists of four layers absorbent gauze, Beiersdorf AG)

REMOVAL OF TEST SUBSTANCE
- Washing: with warm water
- Time after start of exposure: 24h

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2.86 g/kg
- Concentration (if solution): 2000 mg/kg bw

VEHICLE
- Double distilled water

TEST SUBSTANCE PREPARATION:
The test-item preparation was produced for the application group shortly before application by stirring with a high speed homogenizer (Ultra-Turrax) and a magnetic stirrer.
The homogeneity of the test-item preparation during application was provided by stirring with a magnetic stirrer.
Duration of exposure:
24 h
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5 males, 5 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations for mortality and clinical symptoms: several times on the day of administration and at least once daily thereafter each working day for the individual animals.
- Frequency of weighing: shortly before administration (day 0) , weekly thereafter and on the last day of observation.
- Necropsy: all animals were sacrificed and subjected to gros-pathology on the last day of the observation period.
Statistics:
Calculations were performed using Microsoft Excel 2003 and checked with a calculator.
No statistical analysis was performed.
Preliminary study:
not performed
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: None of the animals died. No clinical signs were observed.
Mortality:
No mortality occurred.
Clinical signs:
other: No systemic clinical signs and no local effects were observed during clinical examination.
Gross pathology:
No macroscopic findings were noted at necropsy.

Table 1: Mean body weights in gram (Dose level: 2000 mg/kg bw)

Test day

0

7

14

 

 

 

 

Treated males

251 +/- 2

275 +/- 3

303 +/- 3

Treated females

204 +/- 6

208 +/- 5

216 +/-8

Body weights on day one were assessed before application of test article.

Interpretation of results:
GHS criteria not met
Conclusions:
Under the conditions of the study, the LD50 after dermal application was found to be greater than 2000 mg/kg bw in male and female rats.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
2 000 mg/kg bw

Additional information

Acute oral toxicity

The acute oral toxicity of the test item was assessed in the rat following OECD guideline 423. The test material was administered by oral gavage to two subsequent groups of three female Wistar rats at 2000 mg/kg body weight. Animals were subjected to daily observations and weekly determination of body weight. Macroscopic examination was performed after terminal sacrifice (Day 15). No mortality occurred. Hunched posture was noted in all animals between Days 1 and 8 and piloerection was noted in all animals between Days 1 and 4. Uncoordinated movements were observed among the animals on Day 1. The mean body weight gain shown by the animals over the study period was considered to be normal. No abnormalities were found at macroscopic post mortem examination of the animals. The oral LD50 value in Wistar rats was established to exceed 2000 mg/kg body weight.

Acute dermal toxicity

In an acute dermal toxicity study (Limit Test), young adult Wistar rats (5 males and 5 females) were dermally exposed to a single dose of 2000 mg/kg bw test item (as suspension in doubly distilled water) to the clipped skin (dorsal

and dorso-lateral parts of the trunk) and covered by semi occlusive dressing for 24 hours. The application area comprised at least 10% of body surface area. The animals were observed for 14 days. No mortality occurred. Accordingly, the acute dermal median lethal dose (LD50) was determined to be LD50, dermal, rat > 2000 mg/kg bw. No signs of systemic toxicity or skin effects were observed in the animals. The mean body weight of the animals increased within the normal range throughout the study period. No macroscopic pathologic abnormalities were noted in the animals examined at the end of the study.

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for acute toxicity under Regulation (EC) No. 1272/2008.