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Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 May 2020 - 11 September 2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2020
Report date:
2020

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Reaction mass of 2- (palmitoylamino)ethyl acrylate and 2-(stearoylamino)ethyl acrylate
EC Number:
950-492-4
Cas Number:
2361378-62-1
Molecular formula:
C21-23H39-43NO3
IUPAC Name:
Reaction mass of 2- (palmitoylamino)ethyl acrylate and 2-(stearoylamino)ethyl acrylate
impurity 1
Reference substance name:
Amides, C16-18, N-(hydroxyethyl)
EC Number:
309-819-0
EC Name:
Amides, C16-18, N-(hydroxyethyl)
Cas Number:
101226-97-5
IUPAC Name:
Amides, C16-18, N-(hydroxyethyl)
impurity 2
Reference substance name:
Zirconium compounds
IUPAC Name:
Zirconium compounds
Test material form:
solid: granular
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material: test material provided by sponsor: 91112Y
- Expiration date of the lot/batch: 12 March 2021
- Purity test date: 92.4%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Controlled room temperature (15-25℃, ≤70% relative humidity)
- Solubility and stability of the test substance in the solvent/dispersant/vehicle/test medium: 1% methylcellulose was appropriate for treatment

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item was freshly formulated at concentrations of 30 and 200 mg/mL, stirred with a magnetic stirrer during an overnight period before the treatment.

Test animals

Species:
rat
Strain:
other: Crl:WI (Wistar)
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH (Address: Sandhofer Weg 7, D-97633 Sulzfeld, Germany)
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: Young healthy adult rats, 8-11 weeks old
- Weight at study initiation: 196 - 246 g
- Fasting period before study: At least 5 days
- Housing: Type II. or III. polycarbonate cages (3 animals/cage), Certified laboratory wood bedding and nest building material.
- Diet: ssniff SM R/M "Autoclavable complete diet ad libitum
- Water: tap water from the municipal supply ad libitum
- Acclimation period: During the acclimation period of at least 5 days, the animals were kept under the same controlled environment conditions as during the experimental period.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.1 – 23.9°C
- Humidity (%): 31 - 68%
- Air changes (per hr): 15-20 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12 hours daily, from 6.00 a.m. to 6.00 p.m.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
methylcellulose
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 1 % Methyl cellulose
- Justification for choice of vehicle: The selection of the vehicle was made during trial formulations with the test item. In order of preference, recommended vehicles were: distilled water, 0.5 or 1% methylcellulose, PEG 400, oil (corn) and dimethyl sulfoxide. 1% methylcellulose was appropriate for treatment; therefore, it was found to be a suitable vehicle for formulations.
- Lot/batch no. (if required): S11183

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Per the guideline, a starting dose of 300 mg/kg bw was selected as there was no information available on the substance.
Doses:
300 mg/kg bw (Group 1 and 2), 2000 mg/kg bw (Group 3 and 4)
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days

- Frequency of observations and weighing:
Morbidity and mortality: twice daily (at the beginning and end of each working day)
Clinical observations: after dosing at least once during the first 30 minutes, then 1, 2, 3, 4 and 6 hours after the treatment and once each day for 14 consecutive days thereafter or until death. Individual observations were performed on the skin and fur, eyes and mucous membranes and also respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma. The time of death was recorded as precisely as possible.
Body weights: Days -1 (prior to removal of food), 0 (prior to administration), 7 and 14 with a precision of 1 g.

- Necropsy of survivors performed: Yes; All animals were subjected to a necropsy and a macroscopic examination. exsanguinated after verification of narcosis following an injection of sodium pentobarbital. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs were observed. All gross macroscopic changes were recorded for each animal.

Statistics:
No statistical evaluation was performed in this study

Results and discussion

Preliminary study:
Not applicable
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortality occurred in the study during the 14-day observation period at the dose levels of 300 or 2000 mg/kg bw.
Clinical signs:
All animals were symptom-free during the 14-day observation period.
Body weight:
There were no test item related body weight changes. Body weights were within the range commonly recorded for this strain and age.
Gross pathology:
There was no evidence of the macroscopic changes at dose levels of 300 or 2000 mg/kg bw at necropsy.
Other findings:
Initially, three females (Group 1) were treated at a dose level of 300 mg/kg bw. As no mortality was observed, a confirmatory group (Group 2) was treated at the same dose level. No mortality was observed in the confirmatory group; therefore, one further group (Group 3) was treated at the dose level of 2000 mg/kg bw. As no mortality was observed, a confirmatory group (Group 4) was treated at the same dose level. No mortality was observed in this group and the study was terminated.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The LD50 in female Crl:WI Wistar rats is > 2000 mg/kg bw.
Executive summary:

In an acute oral toxicity test (20/071-001P), 4 groups of female Crl:WI Wistar rats (3/group) were administered the test item (92.4%) in 1% Methyl cellulose by oral gavage at 300 mg/kg bw (2 steps) and 2000 mg/kg bw (2 steps). Animals were observed for 14 days.

The LD50 was > 2000 mg/kg bw.

No mortality occurred in the study during the 14-day observation period at the dose levels of 300 or 2000 mg/kg bw. All animals were symptom-free during the 14-day observation period. There were no test item related body weight changes. Body weights were within the range commonly recorded for this strain and age. There was no evidence of the macroscopic changes at dose levels of 300 or 2000 mg/kg bw at necropsy.