Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
May 09, 2012 - June 13, 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
- Storage condition of test material: At room temperature in the dark
Specific details on test material used for the study:
pH (1% in water, indicative range): 6.0 - 5.8

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. 8-12 weeks old)
- Weight at study initiation: 185-200 g (1st group), 158-172 g (2nd group) and 149-151 g (3rd group)
- Fasting period before study: Animals were deprived of food overnight prior to dosing
- 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. maximum relative humidity of 71 or 72% 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: May 22, 2012 to June 13, 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
- The vehicle was selected based on trial formulations performed at WIL Research Europe and on test substance data supplied by the sponsor

DOSE VOLUME APPLIED
2000 mg/kg (10 mL/kg) body weight
300 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 (1 group)
- 300 mg/kg body weight (2 groups)
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, a second and third group were also tested at 300 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.

Animals were deprived of food until 3-4 hours after administration of the test substance.

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:
Mortality/Viability: Twice daily. Animals showing pain, distress or discomfort, which was considered not transient in nature or was likely to become more severe, were sacrificed for humane reasons based on OECD guidance document on humane endpoints (ENV/JM/MONO/ 2000/7). The time of death was recorded as precisely as possible.
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: All animals were subjected to necropsy and descriptions of all internal macroscopic abnormalities 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:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Mortality:
- 2000 mg/kg bw: all females were found dead or were sacrificed for ethical reasons at approx. one and a half hours after dosing on Day 1
- 300 mg/kg bw (both groups): no mortality occurred
Clinical signs:
- 2000 mg/kg bw: no clinical signs were scored in two animals which were found dead within 2 hours after dosing. One animal showed severe lethargy, cramped posture, flat posture and slow breathing on Day 1.
- 300 mg/kg bw: hunched posture, uncoordinated movements and/or piloerection were noted in all animals on Days 1 and/or 2.
Body weight:
The body weight gain shown by the surviving 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 500 mg/kg body weight.

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Conclusions:
In an acute oral toxicity study with the substance in rats, performed according to OECD/EC test guidelines, an LD50 of > 300 - < 2000 mg/kg bw was determined. Based on this result, the substance is classified for acute toxicity by the oral route (Category 4) with Harmful if swallowed (H302).
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 three subsequent groups of three female Wistar rats at 2000 (1 group) and 300 mg/kg body weight (2 groups). At 2000 mg/kg bw, all females were found dead or were sacrificed for ethical reasons at approx. one and a half hours after dosing on Day 1. At 300 mg/kg bw, no mortality occurred. At 2000 mg/kg bw, no clinical signs were scored in two animals which were found dead within 2 hours after dosing. One animal showed severe lethargy, cramped posture, flat posture and slow breathing on Day 1. At 300 mg/kg bw, hunched posture, uncoordinated movements and/or piloerection were noted in all 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 > 300 - < 2000 mg/kg bw. Based on this result, the substance is classified for acute toxicity by the oral route (Category 4) with Harmful if swallowed (H302). According to the OECD 423 test guideline the LD50 cut-off value was considered to be 500 mg/kg body weight.