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

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 February 2020 to 22 October 2020

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.1300 (Acute inhalation toxicity)

Version / remarks:

August 1998

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

September 2009

Deviations:

no

GLP compliance:

yes

Test type:

concentration x time method

Limit test:

no

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:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Rationale for alternative/additional species to rat (if applicable)
- Source: Charles River Deutschland, Sulzfeld, Germany
- Females (if applicable) nulliparous and non-pregnant: [yes]
- Rationale for use of males (if applicable)
- Age at study initiation: approximately 9-10 weeks old
- Weight at study initiation: Males: 241 to 334 g, Females: 168 to 231 g.
- Fasting period before study: unspecified
- Housing: polycarbonate cages (Makrolon MIV type; height 18 cm.) containing sterilized wooden fibers as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG, Rosenberg, Germany) equipped with water bottles.
- Historical data: unspecified
- Diet (e.g. ad libitum): Pelleted rodent diet (SM R/M-Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany) was provided ad libitum throughout the study, except during designated procedures.
- Water (e.g. ad libitum): Municipal tap-water was freely available to each animal via water bottles.
- Acclimation period: 5 days
- Microbiological status when known: unspecified
- Method of randomisation in assigning animals to test and control groups: Animals were assigned to the study at the discretion of the coordinating biotechnician.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 20 °C
- Humidity (%): 48 to 55%
- Air changes (per hr): Ten or greater air changes per hour with 100% fresh air (no air recirculation)
- Photoperiod (hrs dark / hrs light): 12-hour light/12-hour dark cycle

IN-LIFE DATES: From: 24 February 2020 To: 10 April 2020

Administration / exposure

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

air

Mass median aerodynamic diameter (MMAD):

>= 1 - <= 4 µm

Geometric standard deviation (GSD):

>= 1.5 - <= 3

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: unspecified
- Exposure chamber volume: unspecified
- Method of holding animals in test chamber: polycarbonate restraining tubes
- Source and rate of air (airflow): 22, 36 and 14 L/min for the exposures at 0.5, 1 and 5 mg/L, respectively
- Method of conditioning air: unspecified
- System of generating particulates/aerosols: aerosol was generated by means of a Collison nebulizer and pressurized air.
- Method of particle size determination: The samples were drawn with a flow of 2, 3 or 10 L/min from the test atmosphere through a tube mounted in one of the free animal ports of the exposure chamber. The samples were collected with an 8 stage Marple personal cascade impactor containing fiber glass filters (TE-290-GF. Tisch Environmental, Cleves, Ohio, USA) and a fiber glass back-up filter (SEC-290-F1, Westech, Upper Stondon, Bedfordshire, England). Amounts of test item collected were measured gravimetrically. Subsequently the Mass Median Aerodynamic Diameter (MMAD) and the Geometric Standard Deviation (GSD) were determined based on OECD guidance document No 39.
- Treatment of exhaust air: unspecified
- Temperature, humidity, pressure in air chamber: The temperature of the atmosphere during the exposures was between 20.9 and 23.1 °C. The relative humidity was between 13 and 27%. Pressure in air chamber is unspecified.

TEST ATMOSPHERE
- Brief description of analytical method and equipment used: aerosol was generated by means of a Collison nebulizer and pressurized air.
- Samples taken from breathing zone: yes
- Time needed for equilibrium of exposure concentration before animal exposure: Due to the small volume of the exposure chamber the equilibrium time was negligible

VEHICLE
- Composition of vehicle (if applicable):
- Concentration of test material in vehicle (if applicable):
- Justification of choice of vehicle:
- Lot/batch no. (if required):
- Purity:

TEST ATMOSPHERE (if not tabulated)
- Particle size distribution:
- MMAD (Mass median aerodynamic diameter) / GSD (Geometric st. dev.):

CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: unspecified

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

0.5 mg/L, 1 mg/L and 5 mg/L

No. of animals per sex per dose:

5

Control animals:

yes

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: twice daily
- Necropsy of survivors performed: yes
- Clinical signs including body weight: body weights were determined on days 1, 2, 4, 8 and 15.
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:

Statistics:

No statistical analysis will be performed (the method used is not intended to allow the calculation of a precise LC50 value).

Results and discussion

Preliminary study:

Not reported.

Effect levels

Effect levels 1

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

5 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

not determinable due to absence of adverse toxic effects

Remarks:

At 5 mg/L, hunched posture and quick breathing were seen for all animals between Days 1 and 3.

Effect levels 2

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

1 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

not determinable due to absence of adverse toxic effects

Remarks:

At 1 mg/L, no clinical signs of systemic toxicity were seen for the animals.

Effect levels 3

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

0.5 mg/L air

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

not determinable due to absence of adverse toxic effects

Remarks:

At 0.5 mg/L, piloerection was noted for one female on Days 6 and 7 post-exposure.

Mortality:

No mortality occurred at any of the exposure levels.

Clinical signs:

other:

Remarks:

At 0.5 mg/L, piloerection was noted for one female on Days 6 and 7 post-exposure. At 1 mg/L, no clinical signs of systemic toxicity were seen for the animals. At 5 mg/L, hunched posture and quick breathing were seen for all animals between Days 1 and 3.

Body weight:

Overall body weight gain in males and females was within the range expected for rats of this strain and age used in this type of study and were therefore considered not indicative of toxicity.

Gross pathology:

No test item related abnormalities were found at macroscopic post mortem examination of the animals.
One male exposed to 5 mg/L showed seminal vesicles that were reduced in size. This finding is occasionally seen for rats of this age and strain and therefore considered not related to treatment with the test item.

Any other information on results incl. tables

Concentration
At 0.5 mg/L, the time-weighted mean actual concentration was 0.52 ± 0.02 mg/L. The nominal concentration (amount of test item used divided by the volume of pressurized air used) was 2.1 mg/L. This resulted in a generation efficiency (ratio of actual and nominal concentration) of 25%. At 1 mg/L, the time-weighted mean actual concentration was 1.2 ± 0.03 mg/L. The nominal concentration was 2.0 mg/L. This resulted in a generation efficiency of 57%. At 5 mg/L, the time-weighted mean actual concentration was 5.1 ± 0.1 mg/L. The nominal concentration was 7.9 mg/L. This resulted in a generation efficiency of 65%. The concentrations were measured at several time points that were equally distributed over the exposure periods, the results of which demonstrated that the item was sufficiently stable. The variation in concentrations was caused by adjustments to the generation equipment. By calculating the time-weighted mean concentration, effects of the variations were taken into account resulting in an actual reflection of the mean exposure concentration over time.
Particle Size
At 0.5 mg/L, the MMAD was 1.6 µm (gsd 2.1) and 1.8 µm (gsd 1.8). At 1 mg/L, the MMAD was 1.6 µm (gsd 2.1) and 2.0 µm (gsd 2.0). At 5 mg/L, the MMAD was 2.8 µm (gsd 1.9) and 2.3 µm (gsd 1.8). All values were within the recommended range of 1 - 4 µm.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The inhalation LC50, 4h value of C894 in Wistar Han rats was established to exceed 5 mg/L. Based on these results the test item does not have to be classified and has no obligatory labelling requirement for acute inhalation 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 assess the acute inhalation toxicity of the test item in rats of both sexes in rats following a single 4 hour nose-only exposure to one or more defined concentrations. Animals were retained for a 14 day post exposure observation period.

The study was carried out based on the guidelines described in:
• OECD Guideline 403. Acute Inhalation Toxicity, September 2009.
• EPA Health Effects Test Guideline OPPTS 870.1300. Acute Inhalation Toxicity, August 1998.
• EC No 440/2008, part B. Acute Toxicity (inhalation), May 2008, amended by COMMISSION REGULATION (EU) No 260/2014 (24 January 2014).
• Appendix to Director General Notification, No. 12-Nousan-8147. Agricultural Production Bureau, Ministry of Agriculture, Forestry and Fisheries of Japan (JMAFF), November 2000.

The test item was administered as an aerosol by nose only inhalation for 4 hours to three groups of five male and five female Wistar Han rats at target concentrations of 0.5, 1 and 5 mg/L. Mortality and clinical signs were observed at several days during the observation period and body weights were determined on Days 1, 2, 4, 8 and 15. For the animals exposed to 1 mg/L, body weights were determined on Days 17 and 19. Macroscopic examination was performed after terminal sacrifice on Day 15 for the animals exposed to 0.5 and 5 mg/L and on Day 19 for the animals exposed to 5 mg/L.

At 0.5 mg/L, the time-weighted mean actual concentration was 0.52 ± 0.02 mg/L. The nominal concentration (amount of test item used divided by the volume of pressurized air used) was 2.1 mg/L. This resulted in a generation efficiency (ratio of actual and nominal concentration) of 25%. At 1 mg/L, the time-weighted mean actual concentration was 1.2 ± 0.03 mg/L. The nominal concentration was 2.0 mg/L. This resulted in a generation efficiency of 57%. At 5 mg/L, the time-weighted mean actual concentration was 5.1 ± 0.1 mg/L. The nominal concentration was 7.9 mg/L. This resulted in a generation efficiency of 65%.

The concentrations were measured at several time points that were equally distributed over the exposure periods, the results of which demonstrated that the item was sufficiently stable. The variation in concentrations was caused by adjustments to the generation equipment. By calculating the time-weighted mean concentration, effects of the variations were taken into account resulting in an actual reflection of the mean exposure concentration over time.

At 0.5 mg/L, the MMAD was 1.6 µm (gsd 2.1) and 1.8 µm (gsd 1.8). At 1 mg/L, the MMAD was 1.6 µm (gsd 2.1) and 2.0 µm (gsd 2.0). At 5 mg/L, the MMAD was 2.8 µm (gsd 1.9) and 2.3 µm (gsd 1.8). All values were within the recommended range of 1 - 4 µm.

No mortality occurred at any of the exposure levels.

No clinical signs of systemic toxicity were seen in any of the animals during exposure. At 0.5 mg/L, piloerection was noted for one female on Days 6 and 7 post-exposure. At 1 mg/L, no clinical signs of systemic toxicity were seen for the animals. At 5 mg/L, hunched posture and quick breathing were seen for all animals. The animals had completely recovered from these signs on Day 4.

Overall body weight gain in males and females was within the range expected for rats of this strain and age used in this type of study and were therefore considered not indicative of toxicity.