Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 20 March 2012 to 06 April 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014
Report Date:
2014

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
2001
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Version / remarks:
2008
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
2002
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nohsan, Notification No. 8147, April 2011; including the most recent partial revisions.
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Appearance: Off-white powder with lumps
- Storage condition of test material: At room temperature in the dark
Specific details on test material used for the study:
- pH: 4.4 - 4.5

Test animals

Species:
rat
Strain:
other: Crl:WI (Han)
Sex:
female
Details on test animals and environmental conditions:
- Source: Charles River Deutschland, Sulzfeld, Germany
- Age at study initiation: Young adult animals (approx. 8 weeks old)
- Weight at study initiation: 148-156 g (1st group); 152-168 g (2nd group)
- Fasting period before study: Animals were deprived of food overnight prior to dosing and until 3-4 hours after administration of the test substance.
- Housing: Group housing of 3 animals per cage in labeled Macrolon cages
- Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).
- Water: Free access to tap water.
- Acclimation period: At least 5 days

ENVIRONMENTAL CONDITIONS (set to maintain)
- Temperature (°C): 18 – 24
- Humidity (%): 40 - 70
- Air changes (per hr): approx 15
- Photoperiod (hrs dark / hrs light): 12/12

Variations to these conditions occurred (i.e. minimum relative humidity of 31, 32 or 39% on a single day, respectively). Based on the laboratory’s extensive experience with variations in these parameters and absence of any clinical signs among the animals that could be associated to these variations, these were considered to have no effect on the outcome of the study.

IN-LIFE DATES: From: 20 March 2012 to 06 April 2012

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
propylene glycol
Remarks:
(specific gravity: 1.036)
Details on oral exposure:
GAVAGE METHOD: plastic feeding tubes.

Frequency: single dosage, on Day 1.

VEHICLE
- Justification for choice of vehicle: The vehicle was selected based on trial formulations performed at WIL Research Europe

DOSE VOLUME APPLIED: 2000 mg/kg (10 mL/kg) body weight

DOSAGE PREPARATION: The formulations (w/w) were prepared within 4 hours prior to dosing. Homogeneity was accomplished to a visually acceptable level. Adjustment was made for specific gravity of the vehicle. No correction was made for purity of the test substance.
Doses:
2000 mg/kg body weight
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
The toxicity of the test substance was assessed by stepwise treatment of groups of 3 females. The first group was treated at a dose level of 2000 mg/kg bw. Thereafter the second group was also tested at 2000 mg/kg bw. The onset, duration and severity of the signs of toxicity were taken into account for determination of the time interval between the dose groups.

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Twice daily
Body weights: Days 1 (pre-administration), 8 and 15
Clinical signs: At periodic intervals on the day of dosing (Day 1) and once daily thereafter, until Day 15.
- Necropsy of survivors performed: At the end of the observation period, all animals were sacrificed by oxygen/carbon dioxide procedure and subjected to necropsy. Descriptions of all internal macroscopic abnormalities were recorded.
- Other examinations performed: none.
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
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Hunched posture was noted in all animals on Days 1 and/or 2. In addition, lethargy, flat posture, uncoordinated movements, restless behavior, slow or shallow breathing, salivation and/or ptosis were noted in three animals on Days 1 and/or 2.
Body weight:
The body weight gain shown by the animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.

Any other information on results incl. tables

According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 5000 mg/kg body weight.

Applicant's summary and conclusion

Interpretation of results:
other: Category 5 based on GHS criteria and not classified according to Regulation (EC) No 1272/2008 (CLP Regulation)
Conclusions:
In an acute oral toxicity study with the substance in rats, performed according to OECD/EC test guidelines, an LD50 of >2000 mg/kg bw was determined. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 5000 mg/kg body weight.
Executive summary:

The acute oral toxicity of the substance was determined in accordance with OECD 423 (2001) and according to GLP principles. The substance was administered by oral gavage to two subsequent groups of three female Wistar rats at 2000 mg/kg body weight. No mortality occurred. Hunched posture was noted in all animals on Days 1 and/or 2. In addition, lethargy, flat posture, uncoordinated movements, restless behavior, slow or shallow breathing, salivation and/or ptosis were noted in three animals on Days 1 and/or 2. No abnormalities were found at macroscopic post mortem examination of the animals. The acute oral toxicity (LD50) was determined to be >2000 mg/kg bw. According to the OECD 423 test guideline, the LD50 cut-off value was considered to be 5000 mg/kg body weight. Based on these results, the substance does not need to be classified for acute toxicity by the oral route according to Regulation (EC) No 1272/2008 (CLP Regulation) and should be classified as may be harmful if swallowed (Category 5) according to the GHS.