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:
other: Guideline and GLP-Study without deviations

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2005

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
yes

Test material

Constituent 1
Reference substance name:
BIBR 1048 Oxa-Ester
IUPAC Name:
BIBR 1048 Oxa-Ester
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder

Test animals

Species:
rat
Strain:
other: Wistar Han / Crl:WI(Han)
Sex:
male/female
Details on test animals or test system and environmental conditions:
Strain / Stock Supplier: Wistar Han / Crl:WI(Han), Charles River Deutschland GmbH Sandhofer Weg 7 D-97633 Sulzfeld
Body weight (at administration): males: 189-209 g
females: 171 - 183 g
Age (at administration): males: approx. 7 weeks
females: approx. 8 weeks
Selection of species: international guidelines
Identification of animals: by coloured marks and cage label
Number of animals: 6 male and 6 female animals
Groups: 2 groups of 3 male and 3 female animals, each
Duration of experiment: at least 5 adaptation days
1 test day
2 recovery weeks

Diet
ssniff® R/M-H V1534 (ssniff Spezialdiaten GmbH, D-59494 Soest; composition: see Appendix 2) served as food. Feeding was discontinued approx. 16 hours before administration; only tap water was then available ad libitum. Three hours after administration feeding was continued.
Periodic analysis of the food for contaminants based on EPA/USA2 is conducted at least twice a year by LUFA-ITL3.

Housing
The animals were transferred from the animal housing room to the laboratory 2 hours before administration to allow the animals to acclimatize to the laboratory conditions. The animals remained in the laboratory until the 3 h p.a. observations were completed. They were returned to the animal housing room where the 6 and 24 h observations were recorded.
Granulated textured wood (Granulat A2, J. Brandenburg, D-49424 Goldenstedt) was used as bedding material for the cages. The cages were changed and cleaned twice a week.
Periodic analysis of the bedding material for contaminants based on EPA/USA is conducted at least once a year by LUFA-ITL.
During the 14-day observation period the animals were kept in groups of 3 animals in MAKROLON cages (type III) at a room temperature of 22°C ± 3°C (maximum range) and a relative humidity of 55% + 15% (maximum range). Deviations from the maximum range caused for example during cleaning procedures are dealt with in SOPs.
The rooms were lit (150 lux at approx. 1.50 m room height) and darkened for periods of 12 hours each.

Drinking water
Drinking water in bottles was offered ad libitum. Drinking water is examined according to the 'Deutsche Trinkwasserverordnung 2001' [German Regulations on drinking water 2001] by the Hamburger Wasserwerke, D-20539 Hamburg, at least four times a year.
In addition, drinking water samples taken at LPT are analysed by LUFA-ITL once a year for means of bacteriological investigations according to the 'Deutsche Trinkwasserverordnung 2001, Anlage 1' [German Regulations on drinking water 2001, Addendum 1]. Certificates of analysis of diet, drinking water and bedding material are QAU archived.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: 0.5% aqueous hydroxyethylcellulose suspension
Details on oral exposure:
The test item was suspended to the appropriate concentrations in the above-mentioned vehicle.
Doses:
300, 2000 mg/kg bw
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
Principle of the ATC test method
This procedure permits the identification of the 'acute-toxic-class' (ATC), a measurement of the acute toxicity by the oral route.
The test item is administered orally by gavage at a single dose level to a group of experimental animals. The dose used is selected from a series of defined dose levels. Due to the small number of animals used with this method, there is no need to perform a range finding test.
The test item is tested using a stepwise procedure, each step uses three animals of one sex.
The results of each step determine if:
o no further testing is needed,
o the next step will be performed with the same dose, but with animals of the other sex,
o the next step will be performed at the next higher or next lower dose level.
If available information suggested that mortality was unlikely at 2000 mg/kg body weight, a limit test was conducted. If test item related mortality
was produced, further testing at the next lower level was carried out according to Annex 2d of OECD guideline 423. If there was no information on the test item and for animal welfare reasons, dosing was started at 300 mg/kg body weight and testing continued as described in Annex 2c of
OECD guideline 423. As suggested by the sponsor, this study was started with the recommended dose of 300 mg/kg body weight.

Starting at 300 mg/kg body weight

o Testing at 300 mg/kg body weight:

Three animals of sex 1 are treated at 300 mg/kg body weight (first step).
If two or three animals die, testing at 50 mg/kg body weight should be performed. If fewer than two animals die, the substance should be re-tested (second step) with 300 mg/kg body weight, using three animals of the other sex.
If, in this second step, two or three animals die, testing at 50 mg/kg body weight should be performed. If, in this second step, no to one animal
dies, testing at 2000 mg/kg body weight should be performed.

o Testing at 2000 mg/kg body weight:

If the results of the test at 300 mg/kg body weight indicate the need for further testing at a higher dose level. Three animals of one sex ('sex V)
are treated at 2000 mg/kg body weight (first step).
If two to three animals die, no further testing is necessary. If no to one animal dies, the substance should be retested (second step) with
2000 mg/kg body weight, using three animals of the other sex ('sex 2').
Statistics:
No statistical analysis was performed (The method used is not intended to allow the calculation of a precise LD50 value).

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality observed during the test.
Clinical signs:
No clinical signs observed.
Body weight:
No inhibition of body weight gain observed.
Gross pathology:
No necropsy findings.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the present test conditions, single oral administrations of 300 or 2000 mg test substance/kg b.w. to rats did not reveal any toxic
symptoms. The animals gained the expected body weight throughout the whole study period. No findings were noted at necropsy.

LD50 (24 hours, 14 days): exceeding 2000 mg/kg b.w.