Registration Dossier

Administrative data

Description of key information

- Skin irritation/corrosion: not irritating (OECD 404, GLP, K, rel. 1)
- Eye irritation: not irritating (OECD 437 & 405, GLP, K, rel. 1)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 1994-12-12 to 1994-12-15
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study performed according to OECD test guideline No. 404 and in compliance with GLP.
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Deviations:
no
Principles of method if other than guideline:
The study was extended to investigate 5 different concentrations of the test article and a vehicle control on each animal
GLP compliance:
yes (incl. certificate)
Remarks:
OECD GLP (inspection date: 6-7 April, 1994)
Species:
rabbit
Strain:
other: SPF albino
Details on test animals and environmental conditions:
TEST ANIMALS:
- Source: stock Mol:Russian from Mollegaard Breeding and Research Centre A/S, Ebjy, DK-4623 Lille Skensved
- Age at study initiation: no data
- Weight at study initiation: 2.0 - 2.1 kg bw
- Housing: individually, in 62 x 52 cm PPO cages with perforated floor
- Diet (e.g. ad libitum): pelleted complete diet ad libitum ("Altromin 2123" from Chr. Petersen, DK-4100 Ringsted)
- Water (e.g. ad libitum): domestic quality drinking water acidified with hydrochloric acid to pH 2.5 in order to prevent microbial growth
- Acclimation period: at least 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 +/- 3 °C
- Humidity (%): 55 +/- 5 %
- Air changes (per hr): 10 per hr
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Preparation of test site:
shaved
Vehicle:
other: ethanol:diethyl phtalate (1:1 w/w)
Controls:
other: a site of each animal served as vehicle control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
- Concentration (if solution): 100, 20, 10, 5, 1 and 0 % on each animal
Duration of treatment / exposure:
4 hours
Observation period:
1, 24, 48 and 72 h after termination of the exposure
Number of animals:
4 females
Details on study design:
TEST SITE
- Area of exposure: an area of 10 x 10 cm on the back was clipped and divided into 6 test sites: 2 anterior, 2 centrally and 2 posterior. Each site had an area of 2.5 x 2.5 cm
- Type of wrap if used: gauze patches were secured with a semi-occlusive dressing by means of a 1 cm wide adhesive tape and fixed with Scanpor tape, 4.5 cm width, loosely wound round the trunk

REMOVAL OF TEST SUBSTANCE
- Washing (if done): soap and lukewarm water
- Time after start of exposure: 4 h

SCORING SYSTEM: Draize scale according to the OECD 404
Irritation parameter:
erythema score
Basis:
mean
Remarks:
4 animals
Time point:
other: mean 24-48-72h
Score:
0
Max. score:
4
Remarks on result:
other: for each test concentration
Irritation parameter:
edema score
Basis:
mean
Remarks:
4 animals
Time point:
other: mean 24-48-72h
Score:
0
Max. score:
4
Remarks on result:
other: for each test concentration
Irritant / corrosive response data:
Slight dose related (100 %, 20 % and 10 % test concentrations) skin erythema was observed 1 hour after termination of exposure. Thereafter no abnormalities were observed.
Other effects:
none

Table 7.3.1/1: Irritant/corrosive response data for each animal at each observation time up to removal of animals from the test – 100 %

 

Score at time point / Reversibility

Erythema

Max. score 4

Oedema

Max. score 4

1 h

1 / 1 / 1 / 1

0 / 0 / 0 / 0

24 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

48 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

72 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

Average 24h, 48h, 72h

0.0 / 0.0 / 0.0

0.0 / 0.0 / 0.0

Reversibility*)

-

-

Average time (day) for reversion**

-

-

 *) Reversibility: c. = completely reversible; n.c. = not completely reversible; n. = not reversible

**): correspond to the last day for which skin irritation signs in the last animal were observed

Table 7.3.1/2: Irritant/corrosive response data for each animal at each observation time up to removal of animals from the test – 20 %

 

Score at time point / Reversibility

Erythema

Max. score 4

Oedema

Max. score 4

1 h

1 / 1 / 1 / 0

0 / 0 / 0 / 0

24 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

48 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

72 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

Average 24h, 48h, 72h

0.0 / 0.0 / 0.0

0.0 / 0.0 / 0.0

Reversibility*)

-

-

Average time (day) for reversion**

-

-

 *) Reversibility: c. = completely reversible; n.c. = not completely reversible; n. = not reversible

**): correspond to the last day for which skin irritation signs in the last animal were observed

Table 7.3.1/3: Irritant/corrosive response data for each animal at each observation time up to removal of animals from the test – 10 %

 

Score at time point / Reversibility

Erythema

Max. score 4

Oedema

Max. score 4

1 h

1 / 1 / 1 / 0

0 / 0 / 0 / 0

24 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

48 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

72 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

Average 24h, 48h, 72h

0.0 / 0.0 / 0.0

0.0 / 0.0 / 0.0

Reversibility*)

-

-

Average time (day) for reversion**

-

-

 *) Reversibility: c. = completely reversible; n.c. = not completely reversible; n. = not reversible

**): correspond to the last day for which skin irritation signs in the last animal were observed

Table 7.3.1/4: Irritant/corrosive response data for each animal at each observation time up to removal of animals from the test – 5 %

 

Score at time point / Reversibility

Erythema

Max. score 4

Oedema

Max. score 4

1 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

24 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

48 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

72 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

Average 24h, 48h, 72h

0.0 / 0.0 / 0.0

0.0 / 0.0 / 0.0

Reversibility*)

-

-

Average time (day) for reversion**

-

-

 *) Reversibility: c. = completely reversible; n.c. = not completely reversible; n. = not reversible

**): correspond to the last day for which skin irritation signs in the last animal were observed

Table 7.3.1/1: Irritant/corrosive response data for each animal at each observation time up to removal of animals from the test – 1 %

 

Score at time point / Reversibility

Erythema

Max. score 4

Oedema

Max. score 4

1 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

24 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

48 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

72 h

0 / 0 / 0 / 0

0 / 0 / 0 / 0

Average 24h, 48h, 72h

0.0 / 0.0 / 0.0

0.0 / 0.0 / 0.0

Reversibility*)

-

-

Average time (day) for reversion**

-

-

 *) Reversibility: c. = completely reversible; n.c. = not completely reversible; n. = not reversible

**): correspond to the last day for which skin irritation signs in the last animal were observed

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the condition of the study the test material does not require a classification as irritant to the skin according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and the Directive 67/548/EC.
Executive summary:

A dermal irritation study was performed according to the OECD guideline No. 404 and in compliance with GLP. The study was extended to investigate 5 different concentrations of the test material (100, 20, 10; 5, 1 %) and a vehicle control on each animal. 4 SPF rabbits were exposed to different concentrations of the test article (as provided or diluted in ethanol:DEP) at 6 skin sites on the back.

Test sites were covered with a semi-occlusive dressing for 4 hours. Skin irritation was assessed and scored according to the Draize scale at 1, 24, 48 and 72 h after the removal of the patch.

The mean scores calculated over the 4 animals tested within 3 scoring times (24, 48 and 72 h) were 0.00 for erythema and 0.00 for oedema.

 

Slight dose related (100 %, 20 % and 10 % test concentrations) skin erythema was observed 1 hour after termination of exposure. Thereafter no abnormalities were observed.

 

Therefore the test material does not require a classification as irritant to the skin according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and the Directive 67/548/EC.

This study is considered as acceptable and satisfies the requirement for skin irritation endpoint.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Between 2012-10-09 and 2012-10-18
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study performed according to OECD test guideline No. 405 and in compliance with GLP.
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)
Remarks:
UK GLP Compliance Monitoring Programme (inspected on 2012-07-10)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS: New Zealand White (Hsdlf:NZW) strain male rabbits.
- Source: Harlan Laboratories UK Ltd., Leicestershire, UK.
- Age at study initiation: twelve to twenty weeks old.
- Weight at study initiation: 2.56 to 3.35 kg bw.
- Housing: individually housed in suspended cages.
- Diet (e.g. ad libitum): free access to food (2930C Teklad Global Certified Rabbit diet supplied by Harlan Laboratories UK Ltd., Oxon, UK)
- Water (e.g. ad libitum): free access to drinking water.
The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.
- Acclimation period: at least five days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23
- Humidity (%): 30-70
- Air changes (per hr): at least 15
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
other: the left eye of each rabbit served as control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
Duration of treatment / exposure:
The eyes were not rinsed after administration of the test item.
Observation period (in vivo):
approximately 1 hour and 24, 48 and 72 hours following treatment
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): The eyes were not rinsed after administration of the test item.

SCORING SYSTEM: Draize J H (1977) "Dermal and Eye Toxicity Tests" In: Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, Washington DC p.48 to 49.

TOOL USED TO ASSESS SCORE: light source from a standard ophthalmoscope.

OTHER
Any clinical signs of toxicity, if present, were also recorded.
Individual bodyweights were recorded on Day 0 (the day of dosing) and at the end of the observation period.
Irritation parameter:
cornea opacity score
Basis:
animal: #1, #2 and #3
Time point:
other: mean 24-48-72h
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
animal: #1, #2 and #3
Time point:
other: mean 24-48-72h
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
animal: #1 and #2
Time point:
other: mean 24-48-72h
Score:
0.3
Max. score:
3
Reversibility:
fully reversible within: 48h
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
other: mean 24-48-72h
Score:
0.7
Max. score:
3
Reversibility:
fully reversible within: 72h
Irritation parameter:
chemosis score
Basis:
animal: #1, #2 and #3
Time point:
other: mean 24-48-72h
Score:
0
Max. score:
4
Irritant / corrosive response data:
No corneal or iridial effects were noted during the study.
Moderate conjunctival irritation (grade 2) was noted in one treated eye with minimal conjunctival irritation (grade 1) noted in two treated eyes one hour after treatment. Minimal conjunctival irritation (grade 1) was noted in all treated eyes at the 24-Hour observation and persisted in one treated eye at the 48-Hour observation.
Two treated eyes appeared normal at the 48-Hour observation and one treated eye appeared normal at the 72-Hour observation, therefore the study was terminated on Day 3 post instillation.
Other effects:
All animals showed expected gain in bodyweight during the study.

Table 7.3.2/1: Irritant/corrosive response data each animals at each observation time up to removal from the test

 

Score at time point / Reversibility

Cornea

Iris

(/2)

Conjunctivae

Opacity

(/4)

Area

(/4)

Redness

(/3)

Chemosis

(/4)

Discharge

(/3)

1 h

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

2 / 1 / 1

1 / 1 / 1

1 / 0 / 0

24 h

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

1 / 1 / 1

0 / 0 / 0

0 / 0 / 0

48 h

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

0 / 0 / 1

0 / 0 / 0

0 / 0 / 0

72 h

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

Average 24h, 48h, 72h

0 / 0 / 0

0 / 0 / 0

0 / 0 / 0

0.3 / 0.3 / 0.7

0 / 0 / 0

0 / 0 / 0

Reversibility*)

-

-

-

c.

c.

c.

Average time (unit) for reversion

-

-

-

72 h

24 h

24 h

*) Reversibility: c. = completely reversible; n.c. = not completely reversible; n. = not reversible

 

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Mean scores calculated for each animal over 24, 48 and 72 hours were 0.0/0.0/0.0 for cornea opacity, 0.0/0.0/0.0 for iris lesions, 0.3/0.3/0.7 for redness of the conjunctivae and 0.0/0.0/0.0 for chemosis.
Executive summary:

In an eye irritation study performed according to the OECD guideline No. 405, and in compliance with GLP, 0.1 mL of undiluted Ethylene Brassylate was instilled initially into the right eye of a single New Zealand White rabbit. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated. The eyes were not rinsed after administration of Ethylene Brassylate. The left eye of each rabbit served as control. Animals were observed 1, 24, 48 and 72 hours after dosing under a light source from a standard ophthalmoscope. The reactions in the conjunctiva (redness, chemosis and discharge), the iris and the cornea (opacity and area involved) were scored according to the Draize scale. 

No ocular irritation was noted in any animal at the 72-Hour observation, therefore no further observations were required and the study was terminated.

No corneal or iridial effects were noted during the study. Moderate conjunctival irritation (grade 2) was noted in one treated eye with minimal conjunctival irritation (grade 1) noted in two treated eyes one hour after treatment. Minimal conjunctival irritation (grade 1) was noted in all treated eyes at the 24-hour observation and persisted in one treated eye at the 48-hour observation. Two treated eyes appeared normal at the 48-hour observation and one treated eye appeared normal at the 72-hour observation, therefore the study was terminated on Day 3 post instillation.

Mean scores calculated for each animal over 24, 48 and 72 hours were 0.0/0.0/0.0 for cornea opacity, 0.0/0.0/0.0 for iris lesions, 0.3/0.3/0.7 for redness of the conjunctivae and 0.0/0.0/0.0 for chemosis.

Under the test conditions, Ethylene Brassylate is not classified as irritating to eyes according to the criteria of the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and of the Directive 67/548/EEC.

This study is considered as acceptable and satisfies the requirement for eye irritation endpoint.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation/corrosion:

A key study was identified (Aarup, 1994, rel. 1). In this dermal irritation study performed according to the OECD guideline No. 404 and in compliance with GLP, the substance was applied on the skin of 4 rabbits. The study was extended to investigate 5 different concentrations of the test material (undiluted and diluted in ethanol:DEB at 20%, 10%, 5% and 1 %). The mean scores calculated over the 4 animals tested within 3 scoring times (24, 48 and 72 h) for the 5 concentrations investigated were 0.00 for erythema and 0.00 for oedema.  Slight dose related (100 %, 20 % and 10 % test concentrations) skin erythema was observed 1 hour after termination of exposure. Thereafter no abnormalities were observed.   Therefore, the substance does not require a classification as a skin irritant.

Eye irritation:

The Integrated Testing Strategy as described in ECHA guidance chapter R.7A was followed to assess the eye irritation potential of the substance. In the absence of any alert based on physico-chemical properties, existing human data, existing animal studies and structurally-related substances a new in vitro test was performed (Warren, 2013, Rel.1). This BCOP assay was conducted according to the OECD guideline No. 437 and in compliance with GLP. Under the conditions of the test, with an In Vitro Irritancy Score < 55.1, the test item was considered not to be an ocular corrosive or severe irritant. Therefore an in vivo assay was performed (Sanders, 2013). In this in vivo eye irritation study performed according to the OECD guideline No. 405, and in compliance with GLP, undiluted Ethylene Brassylate was instilled initially into the right eye of a single New Zealand White rabbit. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated. No corneal or iridial effects were noted during the study. Moderate conjunctival irritation (grade 2) was noted in one treated eye with minimal conjunctival irritation (grade 1) noted in two treated eyes one hour after treatment. Minimal conjunctival irritation (grade 1) was noted in all treated eyes at the 24-hour observation and persisted in one treated eye at the 48-hour observation. Two treated eyes appeared normal at the 48-hour observation and one treated eye appeared normal at the 72-hour observation, therefore the study was terminated on Day 3 post instillation. Mean scores calculated for each animal over 24, 48 and 72 hours were 0.0/0.0/0.0 for cornea opacity, 0.0/0.0/0.0 for iris lesions, 0.3/0.3/0.7 for redness of the conjunctivae and 0.0/0.0/0.0 for chemosis.

Following the ITS strategy, Ethylene Brassylate does not require classification as eye irritant.


Justification for selection of skin irritation / corrosion endpoint:
Only one study available, GLP-compliant and of high quality (Klimisch score = 1).

Justification for selection of eye irritation endpoint:
The in vivo study was selected as the BCOP does not allow predicting the absence of classification. The in vivo study is GLP-compliant and of high quality (Klimisch score = 1).

Justification for classification or non-classification

Harmonized classification:

The substance has no harmonized classification according to the Regulation (EC) No. 1272/2008 including ATP3.

Self-classification:

Based on the available information no additional self-classification is proposed regarding both skin and eye irritation according to the Annex VI of the Regulation (EC) No. 1272/2008 (CLP) and of the Directive 67/548/EC. No data is available regarding respiratory irritation.