Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Jan 2004 - Feb 2004
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Title:
Unnamed
Year:
2004
Report Date:
2004

Materials and methods

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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Batch No.of test material: 2003 7026
- Expiration date of the lot/batch: 31-DEC-2013
- Purity test date: > 99%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature (20+/-3 °C) in closed containers in the dark. Not to be kept in a strong containment that allows the pressure to rise in case of a fire in the room.
- Stability under test conditions: Unknown in purified water; is exluded from the statement of compliance.

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: RCC Ltd, Laboratory Animal Services
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 10 wks
- Diet: Pelleted standard Provimi Kliba 3433 rat/ mouse maintenance diet, batch nos. 54/03 and 78/03 (Provimi Kliba AG, CH-4303 kaiseraugst/ Switzerland) ad libitum.
- Water: Community tap water from Fuellinsdorf ad libitum.
- Acclimation period: Under laboratory conditions, after health examination. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/- 3
- Humidity (%): 30-70%
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/ 12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
ADN by oral gavage administration at dosages of 1000 mg/ kg or 300 mg/ kg bw. The test item was diluted in vehicle at concentrations of 0.1 g/mL or 0.03 g/mL and administered at a volume dosage of 10 mL/kg.
Doses:
1000 mg/ kg or 300 mg/ kg bw
No. of animals per sex per dose:
3 females treated at 1000 mg/ kg
6 females treated at 300 mg/ kg
Control animals:
yes

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 1 000 mg/kg bw
Mortality:
1000 mg/ kg mortality: 3/3
300 mg/ kg mortality: 0/6
Clinical signs:
At the dose of 1000 mg/ kg, the fallowing observations were made:
All animals were observed with slightly to moderately ruffled fur, huntched posture from the 0- to the 1-hour reading and persisted in one animal up to the 2-hour reading. Sedation was noted in the same animals at the 0- and 1-hour reading and slight tremor at the 1-hour reading. One animal out of 3 showed moderate tremor as well as letearl recumbency at the 2-hour reading.
At the dose of 300 mg/ kg, the following observations were recorded:
All animals were observed with slightly to moderately ruffled fur, huntched posture from the 1- to 3-hour reading and persisted in two animals up to 5-hour reading. Hunched posture was noted from the 1-hour to the 2-hour reading and persisted in four animals up to the 3-hour and in another animal up to the 5-hour reading. Slight tremor was seen in one animal at the 1- and 2-hour reading and in three other animals at the 2- and 3- hour reading. Ventral recumbency was seen in one animal and tachypnea was observed in two animals at the 2- and 3- hour reading. Sedation was observed in one animal at the 3- hour reading.
Body weight:
The body weigh of the animals was within the range commonly recorded for this strain and age.
Gross pathology:
Necropsy:
The two 1000 mg/kg treated animals which died spontaneously approximately two hours after test item administration and the third animal of the same group which was killed in extremis approximately at the same time were necropsied as soon as they were found dead or killed.
All surviving animals were killed at the end of the observation period by an intraperitoneal injection of Vetanarcol at a dose of at least 2.0 mL/ kg bw (equivalent to at least 324 mg sodium pentobarbitone/ kg bw) and discarded after macroscopic examinations were performed. No organs or tissues were retained.
Other findings:
A stomach distended with gas was observed in all 1000 mg/ kg treated animals at necroskopy. No microscopic findings were recorded in the 300 mg/ kg treated animals.

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
The median lethal dose of ADN after single oral administartion to femal rats, observed over a period od 14 days is:
300 mg/ kg bw