2-Propenoic acid, 2-[2-(ethenyloxy)ethoxy]ethyl ester

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

between 19th January 2009 and the 10th March 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with agreed protocols, with no/or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.2 (Acute Toxicity (Inhalation))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Date of inspection: 19/08/08 Date of Signature: 04/03/09

Test type:

fixed concentration procedure

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

451-690-9

EC Name:

-

Cas Number:

86273-46-3

Molecular formula:

C9H14O4

IUPAC Name:

2-[2-(ethenyloxy)ethoxy]ethyl prop-2-enoate

Test material form:

other: liquid

Details on test material:

- Name of test material (as cited in study report): not applicable as same as test material
- Molecular formula (if other than submission substance): not applicable as same as test material
- Molecular weight (if other than submission substance): not applicable as same as test material
- Smiles notation (if other than submission substance): not applicable as same as test material
- InChl (if other than submission substance): not applicable as same as test material
- Structural formula attached as image file (if other than submission substance): not applicable as same as test material
- Substance type: clear colourless liquid
- Physical state: liquid
- Analytical purity: 99.5%
- Impurities (identity and concentrations): see section 1.2
- Composition of test material, percentage of components: see section 1.2
- Isomers composition: not stated
- Purity test date: not stated
- Lot/batch No.: 7D23-3323
- Expiration date of the lot/batch: not stated
- Stability under test conditions: not stated
- Storage condition of test material: Store cold at 4 deg C, in the dark

Test animals

Species:

rat

Strain:

other: Hsd RccHan:WIST

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS

- Source:
Harlan Ltd, Oxon, UK

- Age at study initiation:
eight to twelve weeks old

- Weight at study initiation:
200g to 350g.

- Fasting period before study:
Not applicable

- Housing:
solid-floor polypropylene cages with stainless steel lids, furnished with softwood flakes (Datesand Ltd., Cheshire, UK) and provided with environmental enrichment items: wooden chew blocks (B & K Universal Ltd, Hull, UK) and cardboard “fun tunnels” (Datesand Ltd., Cheshire, UK). The animals were housed in groups of 5 by sex.

- Diet:
ad libitum

- Water:
ad libitum

- Acclimation period:
at least five days


ENVIRONMENTAL CONDITIONS

- Temperature:
19 - 25 deg C

- Humidity:
30% - 70%

- Air changes (per hr):
at least fifteen changes per hour

- Photoperiod (hrs dark / hrs light):
twelve hours continuous light and twelve hours darkness


IN-LIFE DATES:
From: Day 0 To: End of study

Administration / exposure

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

other: unchanged (no vehicle)

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus:
a glass concentric jet nebuliser (Radleys, Saffron Walden, Essex, UK) located at the top of the exposure chamber. The nebuliser was connected to a glass syringe attached to an infusion pump, which provided a continuous supply of test material under pressure, and to a metered compressed air supply.

- Exposure chamber volume:
approximately 30 litres

- Method of holding animals in test chamber:
Each rat was individually held in a tapered, polycarbonate restraining tube fitted onto a single tier of the exposure chamber and sealed by means of a rubber ‘O’ ring. Only the nose of each animal was exposed to the test atmosphere.

- Source and rate of air:
Compressed air was supplied by means of an oil free compressor and passed through a water trap and respiratory quality filters before it was introduced to the nebuliser.

- Method of conditioning air:
water trap and respiratory quality filters

- System of generating particulates:
a glass concentric jet nebuliser (Radleys, Saffron Walden, Essex, UK)

- Method of particle size determination:
The particle size of the generated atmosphere inside the exposure chamber was determined three times during the exposure period usinf a Marple Personal Cascade Impactor (Schaefer instrument Ltd, Oxon, UK)

- Treatment of exhaust air:
filtered

- Temperature, humidity, pressure in air chamber: temperature and relative humidity inside the exposure chamber were measured by an electronic thermometer/humidity meter (Hanna Instruments Ltd, Beds., UK) located in a vacant port in the animals’ breathing zone of the chamber and recorded every thirty minutes throughout the four-hour exposure period.


TEST ATMOSPHERE
- Brief description of analytical method used:
glass fibre filters (Gelman type A/E 25 mm) placed in a filter holder. The holder was temporarily sealed in a vacant port in the exposure chamber in the animals’ breathing zone and a suitable, known volume of exposure chamber air was drawn through the filter using a vacuum pump (Gravimetric).

- Samples taken from breathing zone:
yes


VEHICLE
- Composition of vehicle (if applicable):
Not applicable

- Concentration of test material in vehicle:
Not applicable

- Justification of choice of vehicle:
Not applicable

- Lot/batch no. (if required):
Not applicable

- Purity: Not applicable


TEST ATMOSPHERE (if not tabulated)

- Particle size distribution:
tabulated
- MMAD (Mass median aerodynamic diameter:
2.66 µm


CLASS METHOD (if applicable)
- Rationale for the selection of the starting concentration: Not applicable.

Analytical verification of test atmosphere concentrations:

no

Remarks:

Gravimetric only

Duration of exposure:

4 h

Concentrations:

Mean Achieved (mg/L) 5.82
Mean Mass Median Aerodynamic Diameter (µm) 2.66
Inhalable Fraction (% <4 µm) 68.8
Geometric Standard Deviation 3.14

No. of animals per sex per dose:

5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: All animals were observed for clinical signs at hourly intervals during exposure, immediately on removal from the restraining tubes at the end of exposure, one hour after termination of exposure and subsequently once daily for fourteen days. Individual bodyweights were recorded prior to treatment on the day of exposure and on Days 7 and 14 or at death.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs.

Statistics:

Data evaluations included the relationship, if any, between the animals’ exposure to the test material and the incidence and severity of all abnormalities including behavioural and clinical observations, necropsy findings, bodyweight changes, mortality and any other toxicological effects.
Using the mortality data obtained, an estimate of the acute inhalation median lethal concentration (LC50) of the test material was made.

Results and discussion

Preliminary study:

Not applicable

Effect levels

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 5.82 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Remarks on result:

other: CL not given

Mortality:

One male animal died during the study. One day after exposure, one male was found dead.
See Appendix 3 Mortality Data in attachment 1

Clinical signs:

other: Signs of hunched posture and pilo erection are commonly seen in animals for short periods on removal from the chamber following 4- hour inhalation studies. During exposure, increased respiratory rate was noted in all animals and there were frequent ins

Body weight:

Variations in body weight gain are frequently seen for female animals of this strain and age during this type of study and, in isolation, are considered not to be significant.
All male animals and four female animals exhibited a bodyweight loss or reduced bodyweight gain during week 1 but recovered to show normal development during week 2. Normal bodyweight development was noted for the other female during the study.
Appendix 6 Individual Bodyweights in the overall remarks section (attachment 1)

Gross pathology:

No macroscopic abnormalities were detected in animals that survived until Day 14 at necropsy.

The following macroscopic abnormalities were detected in the male animal that died during the course of the study at necropsy:
lungs - abnormally dark
liver - patchy pallor, accentuated lobular pattern
Spleen - pale
Stomach - gaseous distension

Other findings:

Not applicable.

Any other information on results incl. tables

The mortality data were summarised as follows:

Mean Achieved Atmosphere Concentration (mg/L)	Deaths
	Male	Female	Total
5.82	1/5	0/5	1/10

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Only one death occurred in a group of ten rats exposed to a mean achieved atmosphere concentration of 5.82 mg/L for four hours. It was therefore considered that the acute inhalation median lethal concentration (4 hr LC50) of T-FC35-Y, in the Sprague-Dawley Crl:CD® (SD) IGS BR strain rat, was greater than 5.82 mg/L.

Executive summary:

Introduction.

A study was performed to assess the acute inhalation toxicity of the test material.The method used followed that described in the OECD Guidelines for Testing of Chemicals (1981) No. 403 “Acute Inhalation Toxicity” referenced as Method B2 in Commission Directive 92/69/EEC “Acute Toxicity – Inhalation” (which constitutes Annex V of Council Directive 67/548/EEC).

Methods.

A group of ten HsdRccHan : WIST strain rats (five males and five females) was exposed to an aerosol atmosphere. The animals were exposed for four hours using a nose only exposure system, followed by a fourteen day observation period.

Results. The mean achieved atmosphere concentration was as follows:

Atmosphere Concentration
Mean Achieved (mg/L)	Standard Deviation	Nominal (mg/L)
5.82	0.18	24.2

The characteristics of the achieved atmosphere were as follows:

Mean Achieved Atmosphere Concentration (mg/L)	Mean Mass Median Aerodynamic Diameter (µm)	Inhalable Fraction (% <4 µm)	Geometric Standard Deviation
5.82	2.66	68.8	2.31

The mortality data were summarised as follows:

Mean Achieved Atmosphere Concentration (mg/L)	Deaths
	Male	Female	Total
5.82	1/5	0/5	1/10

Clinical Observations.

Common abnormalities noted during the study included increased respiratory rate, decreased respiratory rate, noisy respiration, hunched posture, pilo-erection and wet fur. There were frequent instances of laboured respiration and occasional instances of gasping respiration, isolated occurences of ataxia, sneezing and red/brown staining around the eyes or snout were also noted. Surviving male animals appeared normal from Day 8 post-exposure, all females appeared normal one day later.

Bodyweight.

All male animals and four female animals exhibited a bodyweight loss or reduced bodyweight gain during week 1 but recovered to show normal development during week 2. Normal bodyweight development was noted for the other female during the study.

Necropsy. No macroscopic abnormalities were detected amongst animals that survived until Day 14 at necropsy. The following macroscopic abnormality was detected in the animal that died during the course of the study: Lungs – abnormally dark. Liver - patchy pallor, accentuated lobular pattern Spleen - pale Stomach - gaseous distension.

Conclusion. Only one death occurred in a group of ten rats exposed to a mean achieved atmosphere concentration of 5.82mg/L for four hours. It was therefore considered that the acute inhalation median lethal concentration (4 hr LC₅₀) of the test material, in the HsdRccHan : WIST strain rat, was greater than 5.82mg/L.