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Toxicological information

Acute Toxicity: oral

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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from August 11th to September 8th, 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 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:
female
Details on test animals and environmental conditions:
Age: 7-8 weeks old
Supplier: Envigo RMS s.r.l., San Pietro al Natisone (UD), Italy
Weight range at arrival: 163-183 grams
Acclimatisation period: At least 5 days
Veterinary health check: During acclimatisation period
Animals per cage: 3 during the study; up to 5 during acclimatisation
Housing: Polisulphone solid bottomed cages measuring 59.5×38×20 cm with nesting material provided into suitable bedding bags
Cage control: Daily inspected and changed as necessary (at least 3 times/week)
Water: Drinking water supplied to each cage via a water bottle - Ad libitum
Diet: 4 RF 18 (Mucedola S.r.l., Via G. Galilei, 4, 20019, Settimo Milanese (MI) Italy)
Diet supply: Ad libitum throughout the study except for the dosing procedure
Room lighting: Artificial (fluorescent tubes), daily light/dark cycle of 12/12 hours
Air changes: Approximately 15 to 20 air changes per hour
Temperature range: 22 °C ± 2 °C
Relative humidity range: 55 % ± 15 %

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Remarks:
distelled
Details on oral exposure:
Concentrations: 200 and 30 mg/ml. The concentrations were calculated and expressed in terms of test item as supplied.
The formulations were administered in a range of approximately 10 to 60 minutes after preparation.
Fasting procedure: Food was removed from the cages overnight prior to dosing (Day -1) and was made available approximately 4 hours after dosing.
Dose calculation: On the day of dosing (Day 1), the amount of the formulated test item to be administered was calculated for each fasted animal according to body weight.
Dosing method: The formulated test item was administered, by gavage, at a dose volume of 10 ml/kg using a plastic feeding tube attached to a graded syringe.
Doses:
2000 mg/kg and 300 mg/kg
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
Duration of observation period following administration: 14 days
Mortality and morbidity: Throughout the study all animals were checked twice daily.
Clinical signs: Animals were observed for clinical signs as indicated below
– Day of dosing
· Session 1: on dosing
· Session 2: approximately 0.5 hour after dosing
· Session 3: approximately 2 hours after dosing
· Session 4: approximately 4 hours after dosing
– Daily thereafter for a total of 14 days (Session 1)
Body weight: All animals were weighed at allocation to the study (Day -1), on the day of dosing (Day 1) and on Days 2, 8 and 15.
Body weight change calculated for Days 2, 8 and 15 of the dosing phase was relevant to Day 1 of the phase.
Euthanasia method: Animals were sacrificed by carbon dioxide narcosis.
Necropsy procedure: Necropsy was carried out on all animals (gross necropsy examination for both external and internal abnormalities, with particular attention to the gastro-intestinal tract).

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 300 - < 2 000 mg/kg bw
Based on:
test mat.
Mortality:
All animals treated at 2000 mg/kg were found dead on Day 2.
No mortality occurred a in the second and third group of animals dosed at 300 mg/kg
Clinical signs:
Hunched posture, piloerection and decreased activity were observed on the day of dosing at 2000 mg/kg.
No clinical signs were observed in the second and third group of animals dosed at 300 mg/kg
Body weight:
Changes in body weight observed during the study were within the expected range for this strain and age of animals.
Gross pathology:
No abnormalities were observed at necropsy examination performed on early decedent animals dosed at 2000 mg/kg and in those sacrificed at the end of the observation period treated at 300 mg/kg.

Applicant's summary and conclusion

Interpretation of results:
other: Category 4 according to the CLP Regulation (EC) No. 1272/2008
Conclusions:
300 mg/kg < LD50 < 2000 mg/kg
Executive summary:

The acute toxicity of the test item was investigated following a single oral administration to the Sprague Dawley rat followed by a 14-day observation period. A first group of 3 female animals was initially dosed at 2000 mg/kg body weight (Step 1). Mortality occurred in all animals on Day 2. Hunched posture, piloerection and decreased activity were observed on the day of dosing. A second group of 3 female animals was then dosed at lower dose level of 300 mg/kg (Step 2). No death occurred and no signs of toxicity were seen. A third group of 3 female animals was then dosed at the same dose level (300 mg/kg, Step 3). No death occurred and no signs of toxicity were seen. No abnormalities were observed at necropsy examination in the early decedent animals and in those sacrificed at the end of the observation period. These results indicate that the test item induced effects of toxicological relevance (mortality) in the rat following oral administration of a single dose at 2000 mg/kg. No mortality nor signs of toxicity were observed following dosing at 300 mg/kg. These results indicate the acute toxicity estimate (ATE) to be greater than 300 but lower than 2000 mg/kg body weight.