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Description of key information

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 Feb 2019 - 19 March 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
2001
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1100 (Acute Oral Toxicity)
Version / remarks:
2002
Deviations:
no
Qualifier:
according to
Guideline:
other: EC No 440/2008 Part B. Acute Oral Toxicity, Acute Toxic Class Method
Version / remarks:
May 2008
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes
Specific details on test material used for the study:
Identification: Esacure 3644
Physical Description: Light yellow solid
Purity/Composition: UVCB
Storage Conditions: At room temperature protected from light
Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
ANIMALS
Species: Rat
Strain: Crl: WI(Han)
Condition: Outbred, SPF-Quality
Source: Charles River Deutschland, Sulzfeld, Germany
Number of Animals: 6 Females (nulliparous and non-pregnant). Each dose group consisted of 3 animals.
Age at the Initiation of Dosing: Young adult animals (approximately 8-10 weeks old) were selected.
Weight at the Initiation of Dosing: 159 to 193 g.

SELECTION
Animals were assigned to the study at the discretion of the coordinating bio technician according to body weights, with all animals within ± 20% of the sex mean. Animals in poor health or at extremes of body weight range were not assigned to the study.
Before the initiation of dosing, a health inspection was performed and any assigned animal considered unsuitable for use in the study were replaced by alternate animals obtained from the same shipment and maintained under the same environmental conditions.
The disposition of all animals was documented in the study records.

HOUSING
On arrival and following assignment to the study, animals were group housed (up to 3 animals together) in polycarbonate cages (Makrolon MIV type; height 18 cm.) containing sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) equipped with water bottles. The room in which the animals were kept were documented in the study records. Animals were separated during designated procedures/activities. Each cage was clearly labeled.

ENVIRNOMENT
Target temperatures of 18 to 24°C with a relative target humidity of 40 to 70% were maintained. The actual daily mean temperature during the study period was 20 to 21°C with an actual daily mean relative humidity of 41 to 52%. A 12-hour light/12-hour dark cycle was maintained. Ten or greater air changes per hour with 100% fresh air (no air recirculation) were maintained in the animal rooms.

FOOD
Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) was provided ad libitum throughout the study, except during designated procedures. The feed was analyzed by the supplier for nutritional components and environmental contaminants. Results of the analysis were provided by the supplier and are on file at the Test Facility. It is considered that there were no known contaminants in the feed that would interfere with the objectives of the study.

WATER
Municipal tap-water was freely available to each animal via water bottles. Periodic analysis of the water was performed, and results of these analyses are on file at the Test Facility. It is considered that there were no known contaminants in the water that would interfere with the objectives of the study.

ANIMAL ENRICHMENT
For psychological/environmental enrichment, animals were provided with paper (Enviro-dri, Wm. Lillico & Son (Wonham Mill Ltd), Surrey, United Kingdom), except when interrupted by study procedures/activities.

VETERINARY CARE
Veterinary care was available throughout the course of the study; however, no examinations or treatments were required.
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE

Supplier: Sigma-Aldrich, Steinheim, Germany
Specific gravity: 0.92

The dosing formulations were stirred continuously during dose administration.

A dose volume of 10 mL/kg body weight was used for each dose. The dose volume for each animal was based on the body weight measurement prior to dosing.

Trial preparations were performed to select the suitable vehicle and to establish a suitable formulation procedure. These trials were not performed as part of this study and these preparations were not used for dosing. Raw Data of these trials will be retained by the Test Facility


MAXIMUM DOSE

2000 mg/kg
A single dose of test item was administered to the appropriate animals by oral gavage on Day 1, using a syringe with a plastic gavage cannula attached.
Doses:
The first group was treated at a dose level of 2000 mg/kg.
Based on the results, one additional group was dosed at 2000 mg/kg.
No. of animals per sex per dose:
Each dose group consisted of 3 animals females (nulliparous and non-pregnant)
(2 test groups in total)
Control animals:
no
Details on study design:
POST-DOSE OBSERVATIONS
The observation period was 14 days.
Frequency: At periodic intervals on the day of dosing (at least three times) and at least once daily thereafter.
Procedure: All the animals will be examined for reaction to dosing. The onset, intensity and duration of these signs will be recorded (if appropriate), particular attention being paid to the animals during and for the first hour after dosing.

BODY WEIGHTS
Frequency: On Days 1 (predose), 8 and 15.
Procedure: Animals will be individually weighed. A fasted weight will be recorded on the day of dosing. Terminal body weights will also be collected from animals if found dead or euthanized moribund after Day 1.

TERMINAL PROCEDURES
Necropsy of survivors performed: yes
All moribund animals and animals surviving to the end of the observation period will be sacrificed by oxygen/carbon dioxide procedure.
All animals assigned to the study are subjected to necropsy and descriptions of all internal macroscopic abnormalities will be recorded.
Sex:
female
Dose descriptor:
LD50
Effect level:
> 5 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred.
Clinical signs:
Hunched posture and/or piloerection were noted for all animals on Day 1.
Body weight:
The mean body weight gain shown by the animals over the study period was considered to be similar to that expected for normal untreated animals of the same age and strain.
The incidence of slight body weight loss between days 8 and 15 in one individual animal was considered not indicative of toxicity, based on the absence of any corroborative findings in this animal.
Gross pathology:
No abnormalities were found at macroscopic post mortem examination of the animals.
Other findings:
No other observations

Full details of results are in appendix 1 attached to backround material

Interpretation of results:
GHS criteria not met
Conclusions:
The oral LD50 value of Esacure 3644 in Wistar Han rats was established to exceed 2000 mg/kg body weight.

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

Based on these results, Esacure 3644 does not have to be classified and has no obligatory labelling requirement for acute oral toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2017) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).
Executive summary:

Esacure 3644 does not have to be classified and has no obligatory labelling requirement for acute oral toxicity according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (2017) (including all amendments) and Regulation (EC) No 1272/2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
LD50 > 5000 mg/kg bw

Additional information

Justification for classification or non-classification