Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2011

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
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Laboratories B.V.
- Age at study initiation: 10 weeks
- Weight at study initiation: 176.5g - 199.7g
- Accommodation: In groups of three in Makrolon type-4 cages with wire mesh tops and standard softwood bedding. Paper enrichment was included.
- Diet (e.g. ad libitum): Pelleted standard Teklad Rat-Mouse Diet 2914C, irradiated ad libitum (except for the overnight fasting period prior to intubation and approximately 3-4 hours post dose). Results of analyses for contaminants are archived at the laboratory.
- Water (e.g. ad libitum): Community tap water ad libitum. Results of bacteriological, chemical and contaminant analyses are archived at the laboratory.
- Acclimation period: Seven days under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.
- Identification: Unique cage number and corresponding color-coded spots on the tail. The animals were marked at acclimatisation start.
- Randomisation: Selected by hand at time of delivery. No computer generated randomisation program.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3°C
- Humidity (%): 30-70%
- Air changes (per hr): air conditioned with 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): automatically controlled light cycle of 12 hours light and 12 hours dark and music played during the daytime light period.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Remarks:
PEG 300
Details on oral exposure:
VEHICLE
- Concentration in vehicle: The test item was formulated in PEG 300 at a concentration of 0.2 g/mL
- Justification for choice of vehicle: The vehicle was chosen after a non-GLP solubility trial which was performed before the study initiation date. Therefore, solubility testing is excluded from the statement of compliance.

MAXIMUM DOSE VOLUME APPLIED: The dosing volume was 10 mL/kg body weight.

DOSAGE PREPARATION (if unusual): The dose formulations were prepared shortly before each dosing occasion. The test item was weighed into a tared glass beaker on a suitable precision balance and the vehicle was added (weight:volume). The formulations were prepared using a magnetic stirrer and a spatula as homogenisers. Homogeneity of the test item in the vehicle was maintained during administration using a magnetic stirrer.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: The starting dose could be selected from the fixed dose levels of 5, 50, 300, and 2000 mg/kg. Available information indicated test item is likely to be nontoxic considering to acute toxicity. A limit dose of 2000 mg/kg was used as a starting dose. One group of 3 females was dosed. Test itemrelated mortality was not observed during 24 hours and therefore in a second step another 3 females were treated at the same dose.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
2 groups of 3 animals per group
Control animals:
other: not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Viability / Mortality - daily during acclimatisation. Once before treatment and within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 (in common with the clinical signs). Twice daily during test days 2- 15.
Clinical Signs: daily during acclimatization and treatment. Additionally, within the first 30 minutes and approximately 1, 2, 3 and 5 hours after administration on test day 1.
Body Weights: on test days 1 (prior to administration), 8 and 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. An external examination and opening of the abdominal and thoracic cavities for examinations of major organs was performed. The appearance of any macroscopic abnormalities was recorded. No organs or tissues were retained.
- Other examinations performed: clinical signs, body weight
Statistics:
No statistical analysis was performed.

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 intercurrent deaths occurred during the course of the study.
Clinical signs:
Clinical signs, including slightly to moderately increased activity, hunched posture, slightly to moderately ruffled fur and slight to moderate salivation, were observed in all animals at various time points throughout test day I and on test day 2. No clinical signs were observed from test day 3 onwards until the end of the study.
Gross pathology:
The body weight of the animals was within the range commonly recorded for this strain and age.
Other findings:
Macroscopic findings: no macroscopic findings were recorded at necropsy.

Applicant's summary and conclusion

Interpretation of results:
other: not classified (EU-GHS)
Conclusions:
LD50 (female rat) > 2000 mg/kg bw
Executive summary:

The purpose of the present GLP study according to OECD Guideleine 423 and EU Method B.1 tris was to evaluate the potential toxic effect of the test item when administered as a single oral dose to Wistar rats. Two groups, each consisting of three female RccHan:WIST (SPF) rats, were treated with the test item by single oral gavage administration at a dose of 2000 mg/kg body weight. The test item was formulated in PEG 300 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 acclimatisation period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs before treatment, within the first 30 minutes and approximately 1, 2, 3 and 5 hours after treatment on test day 1 and once daily during test days 2-15. Mortality/viability was recorded before treatment, within the first 30 minutes and approximately 1, 2, 3 and 5 hours after administration on test day I (with the clinical signs) and twice daily during test days 2-15. Body weights were recorded on day 1 (prior to administration) and on test days 8 and 15. All animals were necropsied and examined macroscopically. No intercurrent deaths occurred during the course of the study.

Clinical signs, including slightly to moderately increased activity, hunched posture, slightly to moderately ruffled fur and slight to moderate salivation, were observed in all animals at various time points throughout test day 1 and on test day 2. No clinical signs were observed from test day 3 onwards until the end 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) > 2000 mg/kg bw

Based upon the referred classification criteria (Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008), the test item is not classified with respect to acute oral toxicity in the rat.