Registration Dossier

Toxicological information

Acute Toxicity: oral

Currently viewing:

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from 2003-09-17 to 2003-10-10
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study, although OECD 423 version of 1996 rather than the updated version of 2001.

Data source

Reference
Reference Type:
study report
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
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Janvier, Le Genest-Saint-Isle, France
- Age at study initiation: 8 weeks old
- Weight at study initiation: 270 ± 10 g for the males, and 222 ± 3 g for the females
- Fasting period before study: overnight pre-testing fasting
- Housing: in polycarbonate cages with stainless steel lids. Each cage contained 1 to 7 rats of the same sex during acclimation, and 3 rats of the same sex and group during the treatment period.
- Diet : ad libitum
- Water: ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±2°C
- Humidity (%): 30 to 70%
- Air changes (per hr): 12 hour light/dark cycle
- Photoperiod (hrs dark / hrs light): 12 air changes per hour.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentrations in vehicle: 20 and 200 mg/ml
- Amount of vehicle (if gavage): 10 ml/kg
- Justification for choice of vehicle: solubility

MAXIMUM DOSE VOLUME APPLIED: 10 ml/kg

CLASS METHOD
- Rationale for the selection of the starting dose:
the dose-level used as the starting dose was selected from one of the 3 fixed levels: 25, 200 or 2000 mg/kg bw. As no information on the toxic potential of the test item was available, for animal welfare reasons, the starting dose of 200 mg/kg bw was chosen
Doses:
200 and 2000 mg/kg/bw
No. of animals per sex per dose:
3 males and females given 200 mg/kg bw, 3 males given 2000 mg/kg bw
Control animals:
other: Historical data of control animals dosed by oral route
Details on study design:
- Duration of observation period following administration: 14 days after first day of dosing (D1)

- Frequency of observations and weighing: the animals were observed frequently during the hours following administration of the test item, for detection of possible treatment-related clinical signs. Thereafter, observation of the animals was made at least once a day. Type, time of onset and duration of clinical signs were recorded for each animal individually. The animals were weighed individually just before administration of the test item on day 1 and then on days 8 and 15.

- Necropsy of survivors performed: yes

Results and discussion

Effect levels
Sex:
male
Dose descriptor:
LD50
Effect level:
200 - 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: toxicity is comparable in females
Mortality:
No mortality occurred at the 200 mg/kg dose in both sexes. At the 2000 mg/kg dose-level, 2out of 3 males were found dead on day 2 or 3.
Clinical signs:
At 200 mg/kg:
Hypoactivity, piloerection and dyspnea, together with hypersalivation on day 1, were observed in all males on days 1 and 2. In all females, only hypoactivity and piloerection were noted on day 1.
At 2000 mg/kg:
Hypoactivity, piloerection and dyspnea were recorded prior to death. In the surviving male, the same clinical signs, together with swollen abdomen and loud breathing on day 3, were observed from day 1 up to day 3; loud breathing persisted until day 12.
Body weight:
The overall body weight gain of the animals given 200 mg/kg was similar to that of CIT historical control animals.
The body weight gain of the surviving male given 2000 mg/kg was reduced during the first week of the study when compared to CIT historical control animals.
Gross pathology:
Macroscopic examination of the main organs of the animals revealed no apparent abnormalities.

Applicant's summary and conclusion

Interpretation of results:
other: Toxicity category 4 / Harmful
Remarks:
Criteria used for interpretation of results: other: CLP (Reg. 1272/2008/EC) + expert judgement and Directive 67/548/EEC
Conclusions:
The oral LD50 of the test item was comprised between 200 and 2000 mg/kg in rats, with a LD50 cut-off of 1000 mg/kg.
Executive summary:

The acute oral toxicity of the test item was evaluated in rats according to OECD guideline 423 (1996) and EU method B.1tris. The study was conducted in compliance with the principles of Good Laboratory Practice Regulations.  

The test item was prepared in corn oil and was administered by oral route (gavage), under a volume of 10 mL/kg, to groups of three fasted Sprague-Dawley rats. Males were used in the initial step starting with a dose-level of 200 mg/kg bw. As no deaths occured, another assay was

carried out on 3 other males at 2000 mg/kg bw . After this second assay as 2/3 animals died, another assay was carried out on 3 females at the next lower dose-level of 200 mg/kg bw.

At each step, clinical signs, mortality and body weight gain were checked for a period of up to 14 days following the single administration of the test item. All animals were subjected to necropsy.

At the 200 mg/kg bw dose-level, no mortality occured. Hypoactivity, piloerection and dyspnea, together with hypersalivation on day 1, were observed in all males on days 1 and 2. In all females, only hypoactivity and piloerection were noted on day 1. When compared to historical control data, the body weight gain of the animals was not affected by treatment with the test item. At necropsy, no apparent abnormalities were observed in any animal.

At the 2000 mg/kg bw dose-level, 2out of 3 males were found dead on day 2 or 3. Hypoactivity, piloerection and dyspnea were recorded prior to death. In the surviving male, the same clinical signs, together with swollen abdomen and loud breathing on day 3, were observed from day 1 up to day 3; loud breathing persisted until day 12. The body weight gain of the surviving male given 2000 mg/kg was reduced during the first week of the study when compared to historical control animals. At necropsy, no apparent abnormalities were observed.

Under these experimental conditions, the oral LD50of the test item was comprised between 200 and 2000 mg/kg in male rats. Toxicity was comparable in females. The LD50 cut-off is established at 1000 mg/kg.