Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Experimental starting date: 12 February 2008 and Experimental completion date: 06 March 2008.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report Date:
2008

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid

Test animals

Species:
rat
Strain:
other: HanRcc: WIST (SPF)
Sex:
female
Details on test animals and environmental conditions:
- Animals: Rat, HanRcc: WIST(SPF)
- Rationale: Recognized by international guidelines as a recommended test system.
- Breeder: RCC Ltd
- Laboratory Animal Services Wölferstrasse 4, 4414 Füllinsdorf / Switzerland
- Age when treated: 11 weeks
- Identification: Unique cage number and corresponding color-coded spots on the tail. The animals were marked at acclimatization start.
- Randomization: Selected by hand at time of delivery. No computer generated randomization program.
- Acclimatization: Under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.

Husbandry

- Conditions: Standard Laboratory Conditions. Air-conditioned with 10-15 air changes per hour, and continuously monitored environment with ranges for room temperature 22 ± 3 °C and for relative humidity between 30-70 % (values above 70 % during cleaning process possible), automatically controlled light cycle of 12 hours light and 12 hours dark, music during the daytime light period.
- Accommodation: In groups of three in Makrolon type-4 cages with wire mesh tops and standard softwood bedding ('Lignocel' Schill AG, CH-4132 Muttenz/Switzerland).
- Diet: Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 77/07 (Provimi Kliba AG, CH-4303 Kaiseraugst/Switzerland) ad libitum.
- Water: Community tap water from Füllinsdorf ad libitum.


Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
Vehicle
- Identification: Corn oil
- Description: Yellowish oily liquid
- Batch number: 18787208
- Source: Carl Roth GmbH & Co. D-76185 Karlsruhe / Germany
- Stability of the Vehicle: Stable under storage conditions; expiration date: October, 2009

Dose Formulation
Dose levels are in terms of the test item as supplied by the sponsor.
The dose formulations were made shortly before each dosing occasion using a magnetic stirrer, a spatula and an Ultra-Turrax (Janke & Kunkel, D-79219 Staufen) as homogenizers.
The test item was first reduced into a fine powder using a mortar and a pestle. Thereafter, the test item was weighed into a tared glass beaker on a suitable precision balance and the vehicle was added (weight:volume).
Homogeneity of the test item in the vehicle was maintained during administration using a magnetic stirrer.

Treatment
The animals received a single dose of the test item by oral gavage administration at 2000 mg/kg body weight after being fasted for approximately 18 hours (access to water was permitted). Food was provided again approximately 3 hours after dosing.
The dosing volume was 10 mL/kg body weight.
Rationale: Oral administration was considered to be an appropriate application method as it is a possible route of human exposure during manufacture, handling and use of the test item.
Doses:
2000 mg/kg body weight
No. of animals per sex per dose:
3 females rats
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 15 days
- Frequency of observations and weighing:
- Mortality / Viability: Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15. Body Weights On test days 1 (prior to administration), 8 and 15. Clinical Signs Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1. Once daily during days 2-15.
- Necropsy of survivors performed: yes, All animals were killed at the end of the observation period by Carbon dioxide asphyxiation and discarded after macroscopic examinations were performed. No organs or tissues were retained.
- Other examinations performed: clinical signs, body weight.
Statistics:
No statistical analysis was used.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No deaths occurred during the study.
Clinical signs:
No clinical signs were observed during the course of the study.
Body weight:
The body weight of the animals was within the range commonly recorded for this strain and age.
Gross pathology:
No macroscopic findings were recorded at necropsy.

Any other information on results incl. tables

Mortality/Clinical signs

 Dose mg/kg bw  Animal N° Sex   Signs                      Test days                                  
                    1  2  3  4  5  6  8 10  11  12  13  14  15 
         0.5*  1*  2*  3* 5*                             
 2000  1  F No clinical signs   √  √ √  √   √ √   √ √  √  √  √  √  √   √ √  √  √  √  √ 
 2000  2  F   No clinical signs  √  √ √  √  √  √  √  √  √  √  √  √   √  √ √  √  √  √  √   
 2000  4  F   No clinical signs  √  √ √   √  √  √  √  √  √  √  √  √  √  √  √  √  √  √ √ 
 2000  5  F   No clinical signs  √  √  √  √  √  √  √  √  √  √  √  √  √  √  √  √  √ √  √ 
 2000  6  F   No clinical signs  √  √  √  √  √  √  √  √  √  √  √  √  √  √ √   √  √  √  √

Key:  √noted

* Examinations were performed within the first 30 minutes and 1, 2, 3 and 5 hours after treatment.

No clinical signs were evident in any animal during the acclimatization period.

Body weight

 Dose mg/kg Animal N°  Sex   Day 1 (treatment)   Day 8  Day 15
 2000  1  F  177.2 188.7  203.2 
 2000 2  F  195.5 201.8  216.9 
 2000  3  F  177.9 189.8  197.7 
 2000  1  F  187.0  203.9  205.9
 2000  2  F  188.1  206.5  215.1
 2000  3  178.0  203.1  210.9

Body weight are presented in grams

Macroscopic Findings

 Dose mg/kg bw  Animal N°  Sex  Mode of death  Findings
 2000  1  F  S  No macroscopic findings
 2000  2  F  S  No macroscopic findings
 2000  3  F  S  No macroscopic findings
 2000  1  S  No macroscopic findings
 2000  2  F  S  No macroscopic findings
2000  3  F  S  No macroscopic findings

S: Scheduled necropsy

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The median lethal dose of the test item after single oral administration to female rats, observed over a period of 14 days is:
LD50 (female rat): greater than 2000 mg/kg body weight.
Executive summary:

The purpose of this study was to assess the acute toxicity of the test substance when administered by a single oral gavage to rats, followed by an observation period of 14 days.

Two groups, each of three female HanRcc:WIST (SPF) rats, were treated with the test item by oral gavage administration at a dosage of 2000 mg/kg body weight. The

test item was diluted in vehicle (corn oil) at a concentration of 0.2 g/mL and administered at a dosing volume of 10 mL/kg.

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded.

All animals were examined for clinical signs within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Mortality/viability was recorded at approximately 30 minutes, 1, 2, 3 and 5 hours after administration on test day 1 (with the clinical signs) and twice daily during days 2-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically.

All animals survived until the end of the study period.

No clinical signs were observed during the course of the study.

The body weight of the animals was within the range commonly recorded for this strain and age.

No macroscopic findings were recorded at necropsy.

The median lethal dose of the test item after single oral administration to female rats, observed over a period of 14 days is:

LD50 (female rat): greater than 2000 mg/kg body weight