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

Endpoint summary

Description of key information

Two Acute oral studies have been undertaken. The results of both studies classify the substance as harmful if swallowed. The key study was chosen as it follows an OECD guideline and was conducted to GLP.

In an acute inhalation study the 4 hr LC50 in rats was greater than 5.82 mg/L.

In an acute dermal study the LD50 was greater than 2000 mg/kg bw.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Reference 1

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between: 10th September 2003 and 07th November 2003

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.

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Date of inspection: November 2002 Date of Signature: March 2003

Test type:

acute toxic class method

Limit test:

yes

Species:

rat

Strain:

other: HanBrl: Wist (SPF)

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:RCC Ltd, Laboratory Animal Services, CH-4414 Fullinsdorf, Switzerland

- Age at study initiation: 12 weeks

- Fasting period before study: Approximately between 17 and 18 hours.

- Housing: In groups of three in Makrolon type-4 cages with wire mesh tops and standard softwood bedding ('Lignocel' Schill AG, CH-4132 Muttenz,Switzerland).

- Diet (e.g. ad libitum):Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch No. 4/03 and 54/03 (Provimi Kliba AG. CH-4303, Kaiseraugst, Switzerland, ad libitum

- Water (e.g. ad libitum): Community tap water from Fullinsdorf in Switzerland ad libitum.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): Set to achieve limits of 19 deg C to 25 deg C.

- Humidity (%): Set to achieve limits of 30 to 70%.

- Air changes (per hr): The rate of air exchanges was 10 to 15 changes per hour.

- Photoperiod (hrs dark / hrs light): automatically controlled light cycle of 12 hours light and 12 hours dark.

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

Route of administration:

oral: gavage

Vehicle:

polyethylene glycol

Details on oral exposure:

VEHICLE
- Concentration in vehicle: The test item was diluted in vehicle (PEG 300) at concentrations of 0.2 g/mL and 0.03 g/ml, respectively and administered at a volume dosage of 10 mL/kg.

- Amount of vehicle (if gavage): 10ml/kg

- Justification for choice of vehicle: The vehicle was chosen after a non-GLP solubility trial which was performed before the study initiation date. This trial formulation is excluded from the GLP statement of compliance. PEG 300 was found to be a suitable vehicle.

- Lot/batch no. (if required): 448174/1 21203148

MAXIMUM DOSE VOLUME APPLIED:
2000 mg/kg

The test material was administered orally undiluted at a dose level of 2000 mg/kg, and as a solution in arachis oil BP at a dose level of 300 mg/kg.

Dosing was performed sequentially.

DOSAGE PREPARATION (if unusual):
In the absence of data regarding the toxicity of the test material, 300 mg/kg was chosen as the starting dose.

The volume administered to each animal was calculated according to the fasted bodyweight at the time of dosing. Treatment of animals was sequential. Sufficient time was allowed between each group and each dose level to confirm the survival of the previously dosed animals.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: Dose levels are in terms of the test item as supplied by the sponsor. In the absence of data regarding the toxicity of the test material, 300 mg/kg was chosen as the starting dose.

Doses:

300 (with vehicle) and 2000 (undiluted) mg/kg.

No. of animals per sex per dose:

3 females at 2000 mg/kg bw
6 females at 300 mg/kg bw

Control animals:

no

Details on study design:

- Duration of observation period following administration: 18 days


- Frequency of observations and weighing: Observations were monitored during acclimitisation and at approximately 1, 2, 3 and 5 hours after dosing and daily from day 2 to 18.


- Necropsy of survivors performed: yes

- Other examinations performed: clinical signs, body weight, necropsy.

Statistics:

None recorded.

Preliminary study:

Not applicable.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 - < 2 000 mg/kg bw

Based on:

test mat.

Mortality:

All 2000 mg/kg treated animals were found dead 5 hours after treatment.

Clinical signs:

other: Slightly ruffled fur with hunched posture was noted in all 2000 mg/kg treated animals 3 hours after administration, No clinical signs were observed in the 300 mg/kg treated animals during the course of the study.

Gross pathology:

All animals which died spontaneously during the observation period were necropsied as soon as they were found dead. All surviving animals were killed at the end of the observation period by an intraperitoneal injection of Vetanarcol at a dose of at least 2.0 mUkg body weight (equivalent to at least 324 mg sodium pentobarbitone/kg body weight) and discarded after macroscopic examinations were performed. No organs or tissues were retained. No macroscopic findings were recorded at necropsy.

Other findings:

- Organ weights: not retained

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The median lethal dose of 2-(2'-Vinyloxy ethoxy) ethyl acrylate after single oral administration to female rats, observed over a period of 14 days is:
300 mg/kg body weight < LD50 (female rat) < 2000 mg/kg body weight.

Executive summary:

Three groups, each of three female HanBrl: WIST (SPF) rats were treated with 2-(2'Vinyloxy ethoxy) ethyl acrylate by oral gavage administration at a dosage of 2000 mg/kg or 300 mg/kg body weight. The test item was diluted in vehicle (PEG 300) at concentrations of 0.2 g/mL and 0.03 g/ml, respectively and administered at a volume dosage of 10 mL/kg. The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Mortality/viability was recorded twice daily during test days 1-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically. The following females were treated and percentage of mortality was observed: 2000 mg/kg: 100% and 300 mg/kg: 0%.

All 2000 mg/kg treated animals were found dead 5 hours after treatment. Slightly ruffled fur with hunched posture was noted in all 2000 mg/kg treated animals 3 hours after administration. No clinical signs were observed in the 300 mg/kg treated animals during the course of the study. The body weight of the animals was within the range commonly recorded for this strain and age. No macroscopic findings were recorded at necropsy.

Reference 2

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

Between February the 12th 2002 and March 15th 2002

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study conducted in accordance with generally accepted scientific principles, possibly with incomplete or methodological deficiencies, which do not affect the quality of relevant results.

Qualifier:

no guideline followed

GLP compliance:

no

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

Crj: CD(SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Japan Inc, Atsugi Breeding centre, 795 Shimofurusawa, Atsugi-shi, Kanagawa, Japan
- Age at study initiation: 6 weeks
- Weight at study initiation:
Male: 204g to 228g
Female: 141 to 166g
- Fasting period before study:
- Housing: Animals were housed individually in stainless steel bracket cages for rats (260W x 380D x 180Hmm)
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: acclimitised for 7 days prior to experiments


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3 deg C
- Humidity (%): 50 +/- 20%
- Air changes (per hr): 10 or more per hour
- Photoperiod (hrs dark / hrs light): 12 hours light followed by 12 hours of darkness.


IN-LIFE DATES: From: Day 1 Up to: Day 15

Route of administration:

oral: gavage

Vehicle:

corn oil

Details on oral exposure:

VEHICLE
- Concentration in vehicle: not stated
- Amount of vehicle (if gavage):
- Justification for choice of vehicle: not stated
- Lot/batch no. (if required): V9K2502
- Purity: not stated


MAXIMUM DOSE VOLUME APPLIED:
10 mL/kg suspensions

DOSAGE PREPARATION (if unusual):


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

Doses:

1024, 1280, 1600, 2000 and 2500 mg/kg suspensions

No. of animals per sex per dose:

1024 mg/kg Male: 5 Female: 5
1280 mg/kg Male: 5 Female: 5
1600 mg/kg Male: 5 Female: 5
2000 mg/kg Male: 5 Female: 5
2500 mg/kg Male: 5 Female: 5

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: days 1, 2, 3, 4, 8 and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs at 30 minutes, 1, 3, 6 hours and then daily up to day 15.

Statistics:

Mean values and standard deviations of body weights obtained in this study were calculated for each group. The LD50 and 95% confidence limit for each group were calculated by Probit method.

Sex:

female

Dose descriptor:

LD50

Effect level:

1 790 mg/kg bw

95% CL:

>= 1 494 - <= 2 160

Sex:

male

Dose descriptor:

LD50

Effect level:

2 026 mg/kg bw

95% CL:

>= 1 725 - <= 2 614

Mortality:

All of the deaths were observed on day 2. Deaths occurred in 1 male and 2 females at 1600 mg/kg, 1 male an 3 females at 2000 mg/kg and all males and females at 2500 mg/kg. Meanwhile, no death occurred in the 1024 and 1280 mg/kg group.
The Ld50 values were calculated by Probit method. As a result, the LD50 of HBM-AV values were 2026 mg/kg (95% confidence limit: 1725-2614 mg/kg) for males and 1790 mg/kg (95% confidence limit: 1494-2160 mg/kg) for females.

Clinical signs:

other: In the 1024 mg/kg group, no abnormal findings were noted through the observation period in both males and females. In the 1280 mg/kg group, at 3-6 hours after administration, decreased locomotor activity was observed in 3 males and 3 females. In the 1600

Gross pathology:

In all animals that died, stomach distension and dark redness of glandular stomach were observed, but in the animals that survived, no special changes were noted in males and females in any groups.

Other findings:

not reported

Discussion:

HBM-AV was administered by gavage to SD [Crj:CD(SD)] rats (five rats of each sex group) at doses of 1024, 1280, 1600, 2000 and 2,500 mg/kg. After administration, observations were performed for 15 days (including the day of treatment) and the potential acute toxicity and the LD₅₀ of HBM-AV were studied.

All of the deaths in any groups were observed on day 2. Deaths occurred in 1 males and 2 females at 1600 mg/kg, 1 male and 3 females at 2000 mg/kg and all males and females at 2500 mg/kg. Meanwhile, no death occurred in the 1024 and 1280 mg/kg group.

The LD50 values were calculated by Probit method. As a result, the LD50 values of HBM-AV were 2026 mg/kg (95% confidence limit: 1725 -2614 mg/kg) for males and 1790 mg/kg (95% confidence limit: 1494 -2160 mg/kg) for females.

At clinical signs, in 1280 mg/kg or higher groups decreased locomotor activity and abnormal gait were observed in both males and females at 3 -6 hours after administration. In 2000 mg/kg or higher groups, prone position, panting, sedation and hypothermia were observed in either males or females at 6 hours, and in severe cases of these symptoms, animals resulted in death. But in the surviving animals., these symptoms disappeared quickly. These symptoms were considered to be caused by administration of the test substance. Meanwhile in the 1024 mg/kg group, no abnormal findings were noted through the observation period in both males and females.

The body weights of all surviving animals in each group increased satisfactorily throughout the observation period.

At necropsy, in all animals that died, stomach distension and dark redness of glandular stomach considered congestion/hemorrhage were observed. But in the animals that survived, no special changes were noted in both males and females in any groups.

Based on the above results, the LD₅₀ of HBM-AV was estimated to be 2026 mg/kg for males and 1790 mg/kg for females under the conditions of the study.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

Based on the results as detailed in the discussion above, the LD50 of HBM-AV was estimated to be 2026mg/kg for males and 1790 mg/kg for females under the conditions of the study.

Executive summary:

HBM-AV was administered by gavage to SD [Crj:CD(SD)] rats (five rats of each sex/group) at doses of 1024, 1280, 1600, 2000 and 2500 mg/kg. After administration, observations were performed for 15 days (including the day of treatment) and the potential acute toxicity and the LD₅₀ of HBM-AV were studied.

All of the deaths were observed on day 2 after administration. Deaths occurred in 1 male and 2 females at 1600 mg/kg, 1 male and 3 females at 2000 mg/kg. Meanwhile, no death occurred in the 1024 nor the 1280 mg/kg group.

Based on the results of the test VEEA is classified as harmful according to the EU classification and labelling system..

At clinical signs, in 1280 mg/kg or higher groups decreased locomotor activity and abnormal gait were observed in both males and females at 3 - 6 hours after administration. In 2000 mg/kg or higher groups, prone position, panting sedation and hypothermia were observed in either males or females. Meanwhile, in the 1024 mg/kg group, no adnormal findings were noted through the obsevration period.

The body weights of all survivng animals in each group increased satisfactorily throughout the observation period.

At necropsy, in all animals that died, stomach distention and dark redness of glandular stomach were observed. But in the animals that survived, no special changes were noted in both males and females in any groups.

Based on the above results, the LD50 of HBM-AV was estimated to be 2026mg/kg for males and 1790 mg/kg for females under the conditions of the study.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

300 mg/kg bw

Quality of whole database:

Key study follows OECD guideline and is to GLP.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

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.

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Deviations:

no

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

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

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.

Preliminary study:

Not applicable

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.

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

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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

5 820 mg/m³ air

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

between 22nd February 2006 and 11th April 2006

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.

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Date of inspection: April to May 2005 Date of signature: November 2005

Test type:

fixed dose procedure

Limit test:

yes

Species:

rat

Strain:

other: HanRcc:WIST (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: RCC Ltd, Laboratory Animal Services, CH-4414 Füllinsdorf, Switzerland
- Age at study initiation:
Males: 9 - 10 weeks
Females: 12 - 13 weeks
- Weight at study initiation: The body weight of the animals was within the range commonly recorded for this strain and
age.

- Housing: Individually in Makrolon type-3 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz)
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3433 rat/mouse maintenance diet, batch no. 63/05
- Water (e.g. ad libitum): Community tap water from Füllinsdorf ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25 deg C
- Humidity (%): between 30 and 70%
- Air changes (per hr): 10 - 15 air changes per hour
- Photoperiod (hrs dark / hrs light): light cycle of 12 hours light and 12 hours dark, music during the daytime light period.


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

Type of coverage:

semiocclusive

Vehicle:

polyethylene glycol

Details on dermal exposure:

TEST SITE
- Area of exposure: not stated
- % coverage: approximately 10 %
- Type of wrap if used: covered with a semi-occlusive dressing. he dressing was wrapped around the abdomen and fixed with an elastic adhesive bandage.


REMOVAL OF TEST SUBSTANCE
- Washing (if done): Twenty-four hours after the application the dressing was removed and the skin was flushed with lukewarm tap water and dried with disposable paper towels.
- Time after start of exposure: 24 hours


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 4 mL
- Concentration (if solution): 2000 mg/kg


VEHICLE:

The following information was provided by RCC Ltd:

Identification: Polyethylene glycol 300 (PEG 300)
Description: Colorless viscous liquid
Lot number: 11577905 1105271
Source: FLUKA Chemie GmbH, CH-9471 Buchs
Stability of vehicle: Stable under storage conditions;
expiration date: 19-JUL-2006
Storage conditions: At room temperature (range of 20 ± 5 °C), light protected.
Safety precautions: Routine hygienic procedures were used to ensure the health and safety of the personnel.

The vehicle was chosen after a non-GLP solubility trial which was performed before the study initiation date. This formulation trial is excluded from the GLP statement of compliance.

The test item was diluted in vehicle (PEG 300) at a concentration of 0.5 g/mL and administered at a volume dosage of 4 mL/kg. The application period was 24 hours.

Duration of exposure:

24 hours

Doses:

On test day 1, the test item was applied at a dose of 2000 mg/kg body weight evenly on the intact skin with a syringe and covered with a semi-occlusive dressing.

No. of animals per sex per dose:

5 Males and 5 Females were treated with 2-(2-Vinyloxyethoxy)ethyl acrylate at 2000 mg/kg by dermal application.

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days (or other?)
- Frequency of observations and weighing:
Mortality / Viability: Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on test day 1 (along with the clinical signs) and twice daily during days 2-15.

Body weights: On test days 1 (prior to administration), 8 and 15.

Clinical signs: Daily during the acclimatization period, during the first 30 minutes and at approximately 1, 2, 3 and 5 hours after administration on te
st day 1. Once daily during days 2-15. All abnormalities were recorded.

Local signs: Once daily during days 2-15. All abnormalities were recorded.

- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs and body weight.

Statistics:

No statistical analysis was used.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

No deaths occurred during the study.

Clinical signs:

other: No systemic signs of toxicity were observed during the study period. Vocaliation was noted in all animals during removal of the dressing on test day 2. Four males animals also vocalized during the examination of the skin on test day 2 and 3. The same was

Gross pathology:

All animals were killed at the end of the observation period by Carbon dioxide asphyxiation and discarded after macroscopic examinations were performed. No organs or tissues were retained. No macroscopic findings were observed at necropsy

Other findings:

- Organ weights: No organs or tissues were retained.
- Other observations: None

Please see attachment 1 for the individual findings and the bodyweight tables.

Interpretation of results:

GHS criteria not met

Conclusions:

The median lethal dose of 2-(2-Vinyloxyethoxy)ethyl acrylate after single dermal administration to rats of both sexes, observed over a period of 14 days is:
LD50 (rat): greater than 2000 mg/kg body weight

Executive summary:

Method:

The following guidelines were followed during this study:

OECD Guidelines for Testing of Chemicals, Section 4, Number 402 "Acute Dermal Toxicity", adopted February 24, 1987. Directive 92/69/EEC, B.3. "Acute Toxicity-Dermal", July 31, 1992.

Procedure:

Five male and five female HanRcc:WIST (SPF) rats were treated with 2-(2- Vinyloxyethoxy)ethyl acrylate at 2000 mg/kg by dermal application. The test item was diluted in vehicle (PEG 300) at a concentration of 0.5 g/mL and administered at a volume dosage of 4 mL/kg. The application period was 24 hours.

Results:

No deaths occurred during the study. The body weight of the animals was within the range commonly recorded for this strain and age and no macroscopic findings were observed at necropsy.

Conclusion:

The median lethal dose of 2-(2-Vinyloxyethoxy)ethyl acrylate after single dermal administration to rats of both sexes, observed over a period of 14 days is: LD50 (rat): greater than 2000 mg/kg body weight

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Additional information

Acute toxicity Oral (Key Study)

Three groups, each of three female HanBrl: WIST (SPF) rats were treated with 2-(2'Vinyloxy ethoxy) ethyl acrylate by oral gavage administration at a dosage of 2000 mg/kg or 300 mg/kg body weight. The test item was diluted in vehicle (PEG 300) at concentrations of 0.2 g/mL and 0.03 g/ml, respectively and administered at a volume dosage of 10 mL/kg. The animals were examined daily during the acclimatization period and mortality, viability and clinical signs were recorded. All animals were examined for clinical signs at approximately 1, 2, 3 and 5 hours after treatment on day 1 and once daily during test days 2-15. Mortality/viability was recorded twice daily during test days 1-15. Body weights were recorded on day 1 (prior to administration) and on days 8 and 15. All animals were necropsied and examined macroscopically. The following females were treated and percentage of mortality was observed:

2000 mg/kg -       100%

300 mg/kg - 0%

All 2000 mg/kg treated animals were found dead 5 hours after treatment. Slightly ruffled fur with hunched posture was noted in all 2000 mg/kg treated animals 3 hours after administration. No clinical signs were observed in the 300 mg/kg treated animals during the course of the study. The body weight of the animals was within the range commonly recorded for this strain and age. No macroscopic findings were recorded at necropsy.

Acute inhalation:

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 patternSpleen - paleStomach - gaseous distension.

Conclusion: 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 the test material, in the HsdRccHan : WIST strain rat, was greater than 5.82 mg/L.

Acute Dermal:

Method:

The following guidelines were followed during this study:

OECD Guidelines for Testing of Chemicals, Section 4, Number 402 "Acute Dermal Toxicity", adopted February 24, 1987. Directive 92/69/EEC, B.3. "Acute Toxicity-Dermal", July 31, 1992.

Procedure:

Five male and five female HanRcc:WIST (SPF) rats were treated with 2-(2- Vinyloxyethoxy)ethyl acrylate at 2000 mg/kg by dermal application. The test item was diluted in vehicle (PEG 300) at a concentration of 0.5 g/mL and administered at a volume dosage of 4 mL/kg. The application period was 24 hours.

Results:

No deaths occurred during the study. The body weight of the animals was within the range commonly recorded for this strain and age and no macroscopic findings were observed at necropsy.

Conclusion:

The median lethal dose of 2-(2-Vinyloxyethoxy)ethyl acrylate after single dermal administration to rats of both sexes, observed over a period of 14 days is: LD50 (rat): greater than 2000 mg/kg body weight.

Justification for classification or non-classification

The median lethal dose of 2-(2'-Vinyloxy ethoxy) ethyl acrylate after single oral administration to female rats, observed over a period of 14 days is: 300 mg/kg body weight < LD50 (female rat) < 2000 mg/kg body weight.

The substance is therefore classified under CLP for acute toxicity (oral) as Category 4 (H302: Harmful if swallowed).

This classification is required for substances with oral LD50 in the range 300 to <= 2000 mg/kg bw.

The substance VEEA is not classified based on the results of the acute toxicity inhalation and acute toxicity dermal studies.