Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report Date:
2010

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (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
Test material form:
solid: particulate/powder

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
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: From 180 to 220g (± 20%)
- Fasting period before study: the animals were fasted during the night before treatment but had free access to water. Fodd was given back approximately 4 hours after administration of the test item.
- Housing: in polycarbonate cages with a stainless steel lid (43 x 22 x 20 cm).
- Diet (e.g. ad libitum): ad libitum (SSNIFF R/M-H pellet diet)
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2°C
- Humidity (%): 30 to 70%
- Air changes (per hr): approximately 12 cycles/hour of filtered, non-recycled air
- Photoperiod (hrs dark / hrs light): 12h/12h

IN-LIFE DATES: no data

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Details on oral exposure:
VEHICLE
The solubility assay first started at the concentration of 200 mg/mL, and the first choice vehicle was drinking water treated by reverse osmosis.
Since a heterogeneous suspension at 200 mg/mL was obtained with drinking water treated by reverse osmosis, the test substance was tested with a 0.5% methylcellulose aqueous solution.
A homogenous suspension was obtained at the concentration of 200 mg/mL in a 0.5% methylcellulose aqueous solution.

DOSAGE PREPARATION (if unusual): The test substance was administered as a homogeneous suspension in the vehicle. The test substance was ground to a fine powder, using a mortar and pestle, and then mixed with the required quantity of vehicle.
Dose formulations preparations were prepared extemporaneously on the day of each administration.
The dose formulations were stored at room temperature and delivered to the study room in brown flasks.

CLASS METHOD (if applicable):
- Rationale for the selection of the starting dose:
Since no relevant toxicity data were available for the estimation of a lethal dose-level and any existing data were taken into account by the Sponsor, the starting dose level was 300 mg/kg for ethical reasons.
After treatment at the starting dose-level, the next higher dose-level of 2,000 mg/kg was tested.

ADMINISTRATION:
The dose formulations were administered once by gavage, using a plastic syringe fitted with a plastic gavage tube. The quantity of dose formulation administered to each animal was adjusted according to the bodyweight recorded on the day of the dose administration.
A constant dosage-volume of 10 mL/kg was used.
Doses:
300 and 2000 mg/kg bw
No. of animals per sex per dose:
300 mg/kg bw: 3 females
2000 mg/kg bw: 3+3 females
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Clinical observations: At least once during the first 30 minutes, periodically during the first 4 hours, then once a day, at approximately the same time.
- Morbidity and mortality: Frequently during the hours following administration, then at least once a day until the end of the observation period, including weekends and public holidays.
- Bodyweight: Just before treatment on Day 1; then on Days 8 and 15.
- Necropsy of survivors performed: Yes (macroscopic). The main organs included the digestive tract, heart, kidneys, liver, lungs, pancreas, spleen and any other organs with obvious abnormalities. All gross observations were recorded individually for each animal.
Statistics:
None

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD0
Effect level:
>= 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occured in any animals at any tested dose.
Clinical signs:
No clinical signs were observed.
Body weight:
When compared to laboratory historical control data, a lower body weight gain was noted between day 1 and day 8 in 1/6 females given 2000 mg/kg bw (30 g vs 41 +/- 9g in control data base). The body weight gain returned to normal thereafter. The body weight gain of the other animals given 300 or 2000 mg/kg bw was not affected by test-item treatment.
Gross pathology:
At necrospsy, in one female given 2000 mg/kg bw, both horns of uterus were dilated and had translucent content. No apparent abnormalities were observed in the other females given 300 or 2000 mg/kg bw.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The oral LD0 of the test substance was equal or higher than 2000 mg/kg bw.
Executive summary:

In an acute oral toxicity study, performed according to the OECD 423 test guidelines and in compliance with GLP, groups of fasted Sprague-Dawley female rats were administered a single oral dose of the test substance in 0.5 % methylcellulose by gavage. The animals were observed for mortality, clinical signs and body weight for 14 days and then necropsied for macroscopic observations.

Three animals were first given an oral dose of 300 mg/kg bw. As no mortality nor clinical signs were observed, a higher dose was tested. Therefore two independent and successive groups of 3 female rats each were given the maximum dose of 2000 mg/kg bw. Again, no mortality nor clinical signs were observed at 2000 mg/kg bw. A lower body weight gain was noted in 1/6 tested females between day 1 and 8 but the body gain returned to normal thereafter. At necropsy, one female treated with 2000 mg/kg bw, presented a dilatation of both horns of uterus accompagnied of a translucent content. Since no apparent abnormalities were observed in the other animals at the both given doses, these effects were considered not significant.

The acute oral LD0 was found equal or greater than 2000 mg/kg bw.