Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
5 Apr - 6 May 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2016
Report Date:
2016

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
adopted 17 December 2001
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rat
Strain:
other: Crl:CD(SD), SPF
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 8 weeks
- Weight at study initiation: 172.8 - 195.7 g
- Fasting period before study: overnight, approx. 16 h prior and 4 h after dosing
- Housing: 1 animal per cage in stainless wire mech cages (260W x 350D x 210H mm)
- Diet: Teklad Certified Irradiated Global 18% Protein Rodent Diet 2918C (Envigo RMS. Ltd., USA), ad libitum
- Water: public tap water filtered and irradiated by ultraviolet light, ad libitum
- Acclimation period: 4 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.2 - 23.1
- Humidity (%): 45.1 - 54.5
- Air changes (per hr): 10 - 15
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 5 mL/kg bw

CLASS METHOD
- Rationale for the selection of the starting dose: Due to the low expected toxicity of the test substance 2000 mg/kg bw was selected as the starting dose.
Doses:
2000 mg/kg bw (step 1)
300 mg/kg bw (step 2 and 3)
No. of animals per sex per dose:
6 (300 mg/kg bw); 3 (2000 mg/kg bw)
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Animals were observed 30 min and 1, 2, 4 and 6 h after dosing and thereafter once daily for 14 days. Individual body weights were recorded prior to dosing on Day 0 and on Days 1, 3, 7 and on the day of necropsy, Day 14. The body weights of animals found dead during the observation period were recorded prior to necropsy.
- Necropsy of survivors performed: yes
- Other examinations performed: Histopathological examination was performed on animals with gross findings in the thoracic cavity, stomach and/or liver at 2000 mg/kg bw.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
500 mg/kg bw
Based on:
test mat.
Remarks on result:
other: LD50 cut-off according to OECD 423
Mortality:
All animals dosed at 2000 mg/kg were found dead on Days 1, 2 and 3 after dosing, respectively.
No mortality was observed at 300 mg/kg bw.
Clinical signs:
Salivation and/or lacrimation were observed in two animals at 2000 mg/kg bw from 0.5 and 1 h after dosing. Abnormal gait, salivation, staining around mouth, decrease in locomotor activity, decrease of fecal volume and/or lacrimation were observed on Days 1, 2 and 3 after dosing. Three animals were found dead in a state of lying on side, dorsal position, prone position or staining around mouth on Days 1, 2 and 3 after dosing, respectively. These clinical signs were considered to be test substance-related.
During the observation period, no clinical abnormalities were observed in any animal at 300 mg/kg bw.
Body weight:
A tendency to suppressed body weight gain was observed in two animals at 2000 mg/kg bw on Day 1. These changes were considered to be test substance-related.
Normal body weight gains were observed in all animals at 300 mg/kg bw.
Gross pathology:
Macroscopic examination of dead animals revealed gross findings in the thoracic cavity, stomach and/or liver.
No grossly visible evidence of morphological abnormalities was observed in any animal at 300 mg/kg bw.
Other findings:
- Histopathology: Histopathological examination of the liver revealed granular and eosinophilic hepatocellular degeneration, single cell necrosis of hepatocytes and perivenular mononuclear cell infiltration. These findings showed centrilobular distribution. In the forestomach, epithelial necrosis with submucosal hemorrhage and edema were observed. The hepatic and gastric findings were considered to be associated with gross findings in the liver and forestomach, respectively. These necropsy findings were considered to be test substance-related.

Applicant's summary and conclusion

Interpretation of results:
other: Acute Tox. Cat. 4
Remarks:
according to Regulation (EC) 1272/2008
Conclusions:
In this acute oral toxicity study in rats a LD50 cut-off value of 500 mg/kg bw was found.
Executive summary:

Three animals dosed at 2000 mg/kg died on Days 1, 2 and 3 after dosing.

There were no deaths of any of the six animals at 300 mg/kg. No test substance-related effects were observed in clinical signs, body weight data or necropsy findings in any animal at 300 mg/kg.

Based on the result of the acute oral toxicity study in Sprague-Dawley rats the test item was classified to be Category 4

The LD50 cut-off is 500 mg/kg bw.