Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From June 6 to 29, 2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study conducted in compliance with OECD Guideline No. 423 without any deviation.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2008
Report date:
2007

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
adopted 17 December 2001
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. QA statement)
Remarks:
Swiss legislation on Good Laboratory Practice (inspected in April-May, 2005 / signed in November, 2005)
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Appearance: liquid yellow pale
- Stability under storage conditions: stable
Specific details on test material used for the study:
Storage conditions: In the refrigerator (range of 5 +/- 3 °C), light protected

Test animals

Species:
rat
Strain:
Wistar
Remarks:
HanRcc:WIST (SPF)
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: RCC Ltd, Laboratory Animal Services, CH-4414 Füllinsdorf, Switzerland
- Age at study initiation: 11 weeks
- Weight at study initiation: 166.1-197.1 g
- Fasting period before study: Animals were fasted for approximately 18 hours (access to water was permitted) period before administration of test material and for approximately 3 h after dosing.
- Housing: Animals were housed in groups of 3 in Makrolon type-4 cages with wire mesh tops and standard softwood bedding.
- Diet: Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, bact number 16/07 (Provimi Kliba AG, CH-4303, Kaiseraugst, Switzerland), ad libitum.
- Water: Community tap water from Füllinsdorf, ad libitum.
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22 +/- 3 °C
- Humidity: 30-70 %
- Air changes: 10-15 changes/h
- Photoperiod: 12 h dark / 12 h light

IN-LIFE DATES: From: June 06, 2007 To: June 29, 2007

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
polyethylene glycol
Remarks:
PEG 300
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 0.2 g/mL
- Justification for choice of vehicle: PEH 30 was chosen after a non-GLP solubility trial.
- Lot/batch no. (if required): 1310049

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION: Test material was prepared shortly before each dosing occasion using a magnetic stirrer as homogenizer. The test item was weighed into a tared glass beaker on a suitable precision balance and the
vehicle added (weightvolume). 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: Using available information on the toxicity of the test material, 2000 mg/kg bw was chosen as the starting dose.
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
- Sighting study: 3 females / dose
- Main study: 3 females / dose
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of weighting: On test days 1 (prior to administration), 8 and 15.
- Frequency of observations:
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.
Clinical observations: 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. All abnormalities were recorded.
- 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.
Statistics:
None

Results and discussion

Preliminary study:
See below
Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: estimated from the flow chart from OECD Guideline 423 (Appendix 2d)
Mortality:
No deaths occurred during the study.
Clinical signs:
All animals showed a hunched posture from the 2-hour up to the 5-hour reading. A slightly ruffled fur was noted in three animals on test day 1 from the 2 hours up to the 3 hours post-dose and in the three further animals at the 3-hour observation. Light beige feces were recorded in all animals from test day 2 to 4. Three animals were sedated from the 1- or 2-hour up to the 5-hour reading.
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
Other findings:
None

Any other information on results incl. tables

None

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The acute oral median lethal dose (LD50) of the test material in the female Sprague-Dawley CD strain rat can be estimated from the flow chart from OECD Guideline 423 (Appendix 2d) as being greater than 5000 mg/kg bodyweight.. Under the test conditions, the substance is not classified according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
Executive summary:

In an acute oral toxicity study (limit test) performed according to OECD Guideline No. 423 and in compliance with GLP, female Wistar rats were administered a single oral dose of test material diluted in PEG-300 by gavage.

Following a sighting study using one animal at a dose level of 2000 mg/kg bw, additional 4 animals were administered a single oral dose of test item at 2000 mg/kg bw (main study). Animals were then observed for mortality, clinical signs and bodyweights for 14 days and at the end of the study the surviving animals were sacrificed for macroscopic examination.

The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 30 minutes, 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.

All animals showed a hunched posture from the 2-hour up to the 5-hour reading. A slightly ruffled fur was noted in three animals on test day 1 from the 2 hours up to the 3 hours post-dose and in the three further animals at the 3-hour observation. Light beige feces were recorded in all animals from test day 2 to 4. Three animals were sedated from the 1- or 2-hour up to the 5-hour reading.

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 acute oral median lethal dose (LD50) of the test material in the female Sprague-Dawley Wistar strain rat can be estimated from the flow chart from OECD Guideline 423 (Appendix 2d) as being greater than 5000 mg/kg bodyweight.

Under the test conditions, the substance is not classified according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

This study is considered as acceptable and satisfies the requirement for acute oral toxicity endpoint.