Reaction products of 2-ethyl-2-[[(1-oxoallyl)oxy]methyl]-1,3-propanediyl diacrylate and N-methylaniline

Acute Toxicity: dermal

Administrative data

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14 February 2020 - 14 May 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 402 (Acute Dermal Toxicity: Fixed Dose Procedure)

Version / remarks:

2017

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Reaction products of 2-ethyl-2-[[(1-oxoallyl)oxy]methyl]-1,3-propanediyl diacrylate and N-methylaniline

EC Number:

951-458-1

Molecular formula:

Not applicable - UVCB

IUPAC Name:

Reaction products of 2-ethyl-2-[[(1-oxoallyl)oxy]methyl]-1,3-propanediyl diacrylate and N-methylaniline

Test material form:

liquid

Details on test material:

Substance name: Reaction products of 2-ethyl-2-[[(1-oxoallyl)oxy]methyl]-1,3-propanediyl diacrylate and N-methylaniline
Test item handling: Use amber glassware or wrap container in aluminum foil
EC Number: 951-458-1
Physical Description: Light orange liquid
Purity/Composition: UVCB
Storage Conditions: At room temperature protected from light

Specific details on test material used for the study:

Physical Description: Light orange liquid
Purity/Composition: UVCB
Storage Conditions: At room temperature protected from light
Purity/Composition correction factor: No correction factor required
Substance Name: Reaction products of 2-ethyl-2-[[(1-oxoallyl)oxy]methyl]-1,3-propanediyl diacrylate and N-methylaniline
Chemical name (IUPAC): 2,2-bis[({3-[methyl(phenyl)amino]propanoyl}oxy)methyl]butyl 3-[methyl(phenyl)amino]propanoate
EC number: 951-458-1
Molecular formula: EtC(CH2OCOCH2CH2NMePh)3
Molecular weight: 617.79 g/mol
Specific gravity / density: 1.08 at 25°C

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
Source: Charles River Deutschland, Sulzfeld, Germany
Females: Nulliparous and non-pregnant
Age at study initiation: Young adult animals (approximately 9 weeks old) were selected
Weight at study initiation: 185 to 197 g
Fasting period before study: Yes

Housing
On arrival, animals were group housed (up to 5 animals of the same sex together) in polycarbonate cages (Makrolon MIV type; height 18 cm.) and following assignment to the study, animals were individually housed in polycarbonate cages (Makrolon MIII 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 was documented in the study records. Animals were separated during designated procedures/activities. Each cage was clearly labeled.

Diet
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.

Acclimation period
The animals were allowed to acclimate to the Test Facility toxicology accommodation for at least 5 days before the commencement of dosing.

Method of randomisation in assigning animals to test and control groups
Animals were assigned to the study at the discretion of the coordinating biotechnician, with all animals within ± 20% of the sex mean body weights. Animals in poor health or at extremes of body weight range were not assigned to 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.

ENVIRONMENTAL CONDITIONS
Temperature: The actual daily mean temperature during the study period was 20 to 21°C
Humidity: The actual daily mean relative humidity during the study period 36 to 53%
Air changes: Ten or greater air changes per hour with 100% fresh air (no air recirculation) were maintained in the animal rooms
Photoperiod: A 12-hour light/12-hour dark cycle was maintained

IN-LIFE DATES:
Study Initiation Date: 14 Feb 2020
Initiation of Dosing: 04 Mar 2020
Completion of In-life: 14 Apr 2020
Experimental Start Date: 03 Mar 2020
Experimental Completion Date: 14 Apr 2020

Administration / exposure

Type of coverage:

semiocclusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
Area of exposure: Approximately 18 cm²
% coverage: Approximately 10% of the total body surface

Type of wrap
The test item was held in contact with the skin with a dressing, consisting of a surgical gauze patch (Surgy 1D), successively covered with Coban elastic bandage. A piece of Micropore tape was additionally used for fixation of the bandages in females only.

REMOVAL OF TEST SUBSTANCE
Washing: The skin was cleaned of residual test item using water
Time after start of exposure: The application period was 24 hours

TEST MATERIAL
Amount applied
The dose volume for each animal was based on the body weight measurement prior to dosing. Dose volume (mL/kg body weight) was calculated as follows: Dose level (g/kg) / spec.gravity or density (g/mL) * purity correction factor.

Duration of exposure:

24 hours

Doses:

Range Finding Study: 2000 mg/kg
Main Study: 2000 mg/kg

No. of animals per sex per dose:

Range Finding Study: One female at 2000 mg/kg bw
Main Study: Two females at 2000 mg/kg bw

Control animals:

no

Remarks:

Adjacent areas of untreated skin of each animal served as controls

Details on study design:

Duration of observation period following administration: 14 days
Frequency of observations and weighing: The skin reactions were assessed approximately 24, 48 and 72 hours after the removal of the dressing and test item.
Necropsy of survivors performed: Yes

CLINICAL OBSERVATIONS

Post-dose Observations
Post-dose observations were performed at periodic intervals on the day of dosing (at least three times) and once daily thereafter. The observation period was 14 days.
All the animals were examined for reaction to dosing. The onset, intensity and duration of these signs was recorded, particular attention being paid to the animals during and for the first hour after dosing.

Body Weights
Animals were weighed individually on Day 1 (pre-dose), 8 and 15.

Irritation
The skin reactions were assessed approximately 24, 48 and 72 hours after the removal of the dressing and test item. Adjacent areas of untreated skin of each animal served as controls.
The following numerical scoring system was used in the scoring of skin reactions:

Erythema and eschar formation:
No erythema 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beef redness) * 4
*. Where signs of necrosis or corrosion (injuries in depth) prevent erythema scoring, the maximum grade for erythema (= 4) is given.

Oedema formation:
No oedema 0
Very slight oedema (barely perceptible) 1
Slight oedema (edges of area well-defined by definite raising) 2
Moderate oedema (raised approximately 1 millimeter) 3
Severe oedema (raised more than 1 millimeter and extending beyond the area of exposure) 4

Terminal Procedures
All animals were sacrificed by oxygen/carbon dioxide procedure at the end of the observation period. All animals assigned to the study were subjected to necropsy and descriptions of all internal macroscopic abnormalities were recorded.

Statistics:

All results presented in the tables of the report are calculated using values as per the raw data rounding procedure and may not be exactly reproduced from the individual data presented.

The dermal LD50 value of the test item was ranked within the following ranges: 0-50, 50-200, 200-1000 or 1000-2000 mg/kg b.w. or as exceeding 2000 mg/kg b.w.

The results can be evaluated according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of the United Nations (including all amendments) and the Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008 on classification, labelling and packaging of items and mixtures (including all amendments).

Results and discussion

Preliminary study:

Range Finder Study: No mortality occurred

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

no indication of skin irritation up to the relevant limit dose level

Mortality:

No mortality occurred.

Clinical signs:

other: No clinical signs of systemic toxicity were noted.

Gross pathology:

Abnormalities of the liver (right medial lobe diaphragmatic hernia and left median lobe accessory liver) was found in one animal at macroscopic post mortem examination. Both abnormalities are the result of a birth defect and therefore not considered test item-related. The other animals showed no abnormalities.

Other findings:

No skin irritation was noted for any of the animals at any time point.
The dermal LD50 value of the test item in Wistar Han rats was established to exceed 2000 mg/kg body weight.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The dermal LD50 value of the test item in Wistar Han rats was established to exceed 2000 mg/kg body weight.

Based on these results, the test item does not have to be classified and has no obligatory labelling requirement for acute dermal 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:

The objective of this study was to determine the potential toxicity of C894, when given by a single dermal dose.  

The study was carried out based on the guideline described in:  

OECD No. 402 (2017) "Acute Dermal Toxicity".

Initially, the test item was administered to a single female Wistar Han rat by a single dermal application at 2000 mg/kg body weight for 24 hours in a range finder study. Based on the results, the main study was performed by dosing two additional females at 2000 mg/kg. All animals were subjected to daily observations and weekly determination of body weight.

Macroscopic examination was performed after terminal sacrifice (Day 15).

No mortality occurred and no clinical signs of systemic toxicity were noted. Furthermore, no skin irritation was noted for any of the animals at any time point.

The body weight gain shown by the surviving animals during the observation period was within the range expected for rats used in this type of study.

Macroscopic post mortem examination of the animals at termination revealed no test item-related abnormalities.  

The dermal LD50 value of the test item in Wistar Han rats was established to exceed 2000 mg/kg body weight.  

Based on these results, the test item does not have to be classified and has no obligatory labelling requirement for acute dermal 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).