Registration Dossier

Administrative data

Description of key information

Skin irritant

Eye irritant

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
28 August 2012 to 3 September 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
other: in vitro Reconstructed Human Epidermis (RHE) Model
Details on test animals and environmental conditions:
The EPISKIN model is a three-dimensional reconstructed human epidermis model consisting of adult human-derived epidermal keratinocytes seeded on a dermal substitute consisting of a collagen type I matrix coated with type IV collagen. A highly differentiated and stratified epidermis model is obtained after a 13-Day culture period comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum.

EPISKIN Model Kit
Supplier: SkinEthic Laboratories, Lyon, France
Type of coverage:
other: not applicable
Preparation of test site:
other: not applicable
Vehicle:
unchanged (no vehicle)
Controls:
other: in vitro controls were used
Amount / concentration applied:
10 µL
Duration of treatment / exposure:
A treatment period of 15 minutes was followed by a post-exposure incubation period of 42 hours.
Observation period:
Not applicable
Number of animals:
Not applicable
Details on study design:
MATERIAL PREPARATION
-Dulbecco’s Phosphate Buffered Saline (DPBS) with Ca++ and Mg++ was used as the negative control.
-Sodium Dodecyl Sulphate (SDS) 5 % w/v aqueous dilution was used as the positive control.
-A 3 mg/mL MTT stock solution was prepared in DPBS. The stock solution was diluted to 0.3 mg/mL with assay medium when required.
-A 0.04 N concentration of hydrochloric acid in isopropanol was prepared when required.

PRE-TEST
Assessment of Direct Test Material Reduction of MTT

MTT dye metabolism, cell viability assay
The MTT assay, a colourimetric method of determining cell viability, is based on reduction of the yellow tetrazolium salt (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) to a blue formazan dye by mitochondrial succinate dehydrogenase in viable cells.
One limitation of the assay is possible interference of the test material with MTT. A test material may directly reduce MTT, thus mimicking dehydrogenase activity of the cellular mitochondria. This property of the test material is only a problem, if at the time of the MTT test (after rinsing) there are still sufficient amounts of the test material present on or in the tissues. In this case, the true metabolic MTT reduction and the false direct MTT reduction can be differentiated and quantified.

Test for Direct MTT Reduction
The test material was checked for the ability to directly reduce MTT according to the following procedure:
10 µL of the test material was added to 2 mL of a 0.3 mg/mL MTT solution freshly prepared in assay medium. The solution was incubated in the dark at 37 °C, 5 % CO₂ in air for 3 hours. Untreated MTT solution was used as a control.
If the MTT solution containing the test material turns blue, it is presumed to have reduced the MTT.

The test material was shown to directly reduce MTT. There was a possibility that if the test material could not be totally rinsed off the tissues, any residual test material present on or in the tissue may have given rise to a false negative result. Therefore, the determination of skin irritation potential was performed in parallel on viable and water-killed tissues. This step was a functional check which employs water-killed tissues that possess no metabolic activity but absorb and bind the test material like viable tissues.

Water-killed tissues were prepared by placing untreated EPISKIN tissues in a 12-well plate containing 2.2 mL of sterile distilled water in each well. The tissues were incubated at 37 °C, 5 % CO₂ in air for 48 ± 1 hours. At the end of the incubation the water was discarded. Once killed, the tissues were stored in a freezer (-14 to -30 °C) for up to 6 months. Before use, each tissue was thawed by placing in 2.2 mL of maintenance medium for approximately 1 hour at room temperature.
In addition to the normal test procedure, the test material was applied to a water-killed tissue. In addition, one water-killed tissue remained untreated. The untreated water-killed control showed a small amount of MTT reduction due to residual reducing enzymes within the killed tissue.

PRE-INCUBATION (Day 0: Tissue Arrival)
2 mL of maintenance medium, warmed to approximately 37 °C, was pipetted into the first column of 3 wells of a pre-labelled 12-well plate. Each epidermis unit was transferred into the maintenance medium filled wells (3 units per plate). A different 12-well plate was used for the test material and each control material. The tissues were incubated at 37 °C, 5 % CO₂ in air overnight.

MAIN TEST
APPLICATION OF TEST MATERIAL AND RINSING (Day 1)
2 mL of maintenance medium, warmed to approximately 37 °C, was pipetted into the second column of 3 wells of the 12-well plate.
Triplicate tissues were treated with the test material for an exposure period of 15 minutes. It was applied topically to the corresponding tissues, ensuring uniform covering. 10 µL of the test material was applied to the epidermis surface. Triplicate tissues treated with 10 µL of DPBS served as the negative controls and triplicate tissues treated with 10 µL of SDS 5 % w/v served as the positive controls. To ensure satisfactory contact with the positive control material, the SDS solution was spread over the entire surface of the epidermis using a pipette tip (taking particular care to cover the centre). After 7 minutes of contact time, the SDS solution was re-spread with a pipette tip to maintain the distribution for the remainder of the contact period. The plate(s) were kept in a biological safety cabinet at room temperature for 15 minutes.

At the end of the exposure period, each tissue was removed from the well using forceps and rinsed using a wash bottle containing DPBS with Ca++ and Mg++. Rinsing was achieved by filling and emptying each tissue insert for approximately 40 seconds using a constant soft stream of DPBS to gently remove any residual test material. The rinsed tissues were transferred to the second column of 3 wells containing 2 mL of maintenance medium in each well. The rinsed tissues were incubated at 37 °C, 5 % CO₂ in air for 42 hours.

MTT LOADING/FORMAZAN EXTRACTION (Day 3)
Following the 42 hour post-exposure incubation period, each 12-well plate was placed onto a plate shaker for 15 minutes to homogenise the released mediators in the maintenance medium. 1.6 mL of the maintenance medium from beneath each tissue was transferred to pre-labelled micro tubes and stored in a freezer at -14 to -30 °C for possible inflammatory mediator determination.

2 mL of a 0.3 mg/mL MTT solution, freshly prepared in assay medium, was pipetted into the third column of 3 wells of the 12 well plate(s). The tissues were transferred to the MTT filled wells, being careful to remove any excess maintenance medium from the bottom of the tissue insert by blotting on absorbent paper. The tissues were incubated for 3 hours at 37 °C, 5 % CO₂ in air. At the end of the 3 hour incubation period, each tissue was placed onto absorbent paper to dry. A total biopsy of the epidermis was made using the EPISKIN biopsy punch. The epidermis was carefully separated from the collagen matrix using forceps and both parts (epidermis and collagen matrix) placed into labelled 1.5 mL micro tubes containing 500 µL of acidified isopropanol, ensuring that both the epidermis and collagen matrix were fully immersed. Each tube was plugged to prevent evaporation and mixed thoroughly on a vortex mixer. The tubes were refrigerated at 1 to 10 °C until Day 6 of the experiment, allowing the extraction of formazan crystals out of the MTT-loaded tissues.

ABSORBANCE/OPTICAL DENSITY MEASUREMENTS (Day 6)
At the end of the formazan extraction period, each tube was mixed thoroughly on a vortex mixer to produce an homogenous coloured solution.
For each tissue, duplicate 200 µL samples were transferred to the appropriate wells of a pre-labelled 96-well plate. 200 µL of acidified isopropanol alone was added to the two wells designated as ‘blanks’. The optical density was measured (quantitative viability analysis) at 540 nm (without a reference filter) using the Anthos 2001 microplate reader.

INTERPRETATION OF RESULTS
-Quantitative MTT Assessment (percentage tissue viability)
For the test material, the relative mean tissue viabilities obtained after the 15 minute exposure period followed by the 42 hour post-exposure incubation period were compared to the mean of the negative control treated tissues (n=3). The relative mean viabilities were calculated in the following way:
Relative mean viability (%) = (mean OD540 of test material / mean OD540 of negative control) x 100

Classification of irritation potential is based upon relative mean tissue viability according to the following:

Criteria for in vitro interpretation EU DSD Classification EU CLP Classification
Relative Mean Tissue Viability is ≤50 % Irritant (I) R38 Category 2
Relative Mean Tissue Viability is >50 % Non-Irritant (NI) No Category

QUALITY CRITERIA
The results of the assay are considered acceptable if the following assay acceptance criteria are achieved:

-Positive Control
The assay establishes the acceptance criterion for an acceptable test if the relative mean tissue viability for the positive control treated tissues was ≤40 % relative to the negative control treated tissues, and the standard deviation value of the percentage viability is ≤18 %.
-Negative Control
The assay establishes the acceptance criterion for an acceptable test if the mean OD540 for the negative control treated tissues was ≥0.6, and the standard deviation value of the percentage viability is ≤18 %.
-Test Material
The assay establishes the acceptance criterion for an acceptable test if the standard deviation calculated from individual percentage tissue viabilities of the three identically treated tissues is ≤18 %.
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
OD540 0.061
Value:
7.9
Remarks on result:
positive indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
OD540 0.083
Value:
10.7
Remarks on result:
positive indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
OD540 0.092
Value:
11.9
Remarks on result:
positive indication of irritation

Direct MTT Reduction

An assessment found that the test material was able to directly reduce MTT. Therefore, an additional procedure using water-killed tissues was performed during the determination of skin irritation potential. However, the results obtained showed that a negligible degree of interference due to direct reduction of MTT occurred. It was therefore considered unnecessary to use the results of the water-killed tissues for quantitative correction of results or for reporting purposes.

 

Quality Criteria

-The relative mean tissue viability for the positive control treated tissues was 8.9 % relative to the negative control treated tissues and the standard deviation value of the percentage viability was 1.9 %. The positive control acceptance criterion was therefore satisfied.

 -The mean OD540 for the negative control treated tissues was 0.774 and the standard deviation value of the percentage viability was 15.0 %. The negative control acceptance criterion was therefore satisfied.

-The standard deviation calculated from individual percentage tissue viabilities of the three identically treated tissues was 2.1 %. The test material acceptance criterion was therefore satisfied.

 

Table 1 Mean OD540 Values and Percentage Viabilities for the Negative and Positive Controls and the Test Material

Material

OD540 of Tissues

Mean OD540 of Triplicate Tissues

± SD of OD540

Relative Individual Tissue Viability (%)

Relative Mean Viability (%)

± SD of Relative Mean Viability (%)

Negative Control

0.674

 

0.774

 

0.116

87.1

 

100*

 

15.0

0.747

96.5

0.901

116.4

Positive Control

0.052

 

0.069

 

0.015

6.7

 

8.9

 

1.9

0.079

10.2

0.077

9.9

Test

Material

0.061

 

0.079

 

0.016

7.9

 

10.2

 

2.1

0.083

10.7

0.092

11.9

*The mean viability of the control group is set at 100 %

SD = Standard deviation

Interpretation of results:
other: Skin irritant Cat. 2
Remarks:
according to the criteria set up on the OECD guideline 439
Conclusions:
Skin irritant
Executive summary:

The skin irritation potential of the test material was evaluated in vitro using the EPISKIN reconstructed human epidermis model in accordance with the standardised guidelines OECD 439 and EU Method B.46 under GLP conditions.

Triplicate tissues were treated with the test material for an exposure period of 15 minutes. At the end of the exposure period, each tissue was rinsed before being incubated for 42 hours, after which each tissue was taken for MTT loading. After MTT loading, a total biopsy of each epidermis was made and formazan crystals were extracted out of the MTT-loaded tissues. Duplicate tissues treated with Dulbecco’s Phosphate Buffered Saline with Ca++ and Mg++ served as the negative control and duplicate tissues treated with 5 % w/v aqueous Sodium Dodecyl Sulphate served as the positive control. The optical density of all treated tissues was measured at 540 nm.

The relative mean viability of the test material treated tissues was 10.2 % after the 15 minute exposure period.

Under the conditions of this study, the test material was determined to be a Category 2 irritant in accordance with the criteria set out in Annex I of Regulation (EC) No 1272/2008 (CLP). It also classified as irritant according to EU labelling regulations Commission Directive 2001/59/EC.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (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:
17 September 2012 to 24 October 2012
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline 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
GLP compliance:
yes (incl. certificate)
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Strain: Hsdlf:NZW
- Age at study initiation: 12 to 20 weeks
- Weight at study initiation: 2.25 or 2.95 kg
- Housing: The animals were individually housed in suspended cages.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): Free access to mains drinking water.
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23 °C
- Humidity (%): 30 to 70 %
- Air changes (per hr): At least 15 per hour.
- Photoperiod (hrs dark / hrs light): The lighting was controlled by a time switch to give twelve hours continuous light (0600 to 1800) and twelve hours darkness.

IN-LIFE DATES: From: 17 September 2012 To: 24 October 2012
Vehicle:
unchanged (no vehicle)
Controls:
other: The left eye remained untreated and was used for control purposes.
Amount / concentration applied:
0.1 mL
Duration of treatment / exposure:
The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test material, and then released.
Observation period (in vivo):
21 days
Number of animals or in vitro replicates:
2
Details on study design:
SCORING SYSTEM: Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the Draize numerical evaluation.
Additional observations were made on Days 7, 14 and 21 to assess the reversibility of the ocular effects. Individual bodyweights were recorded on Day 0 (the day of dosing) and at the end of the observation period. Any clinical signs of toxicity, if present, were also recorded.

Draize Scale for Scoring Ocular Irritation

1. CONJUNCTIVAE

Redness (refers to palpebral and bulbar conjunctivae excluding cornea and iris)
Vessels normal............................................................................................................................................0
Vessels definitely injected above normal................................................................................................1
More diffuse, deeper crimson red, individual vessels not easily discernible....................................2
Diffuse beefy red.........................................................................................................................................3

Chemosis
No swelling..................................................................................................................................................0
Any swelling above normal (includes nictitating membrane).............................................................1
Obvious swelling with partial eversion of lids.......................................................................................2
Swelling with lids about half closed.........................................................................................................3
Swelling with lids half closed to completely closed...............................................................................4

Discharge
No discharge...............................................................................................................................................0
Any amount different from normal (does not include small amounts observed in inner
canthus of normal animals)......................................................................................................................1
Discharge with moistening of the lids and hairs just adjacent to lids...............................................2
Discharge with moistening of the lids and hairs a considerable area around the eye...................3

2. IRIS

Values
Normal.........................................................................................................................................................0
Folds above normal, congestion, swelling, circumcorneal injection (any or all
of these or combination of any thereof) iris still reacting to light
(sluggish reaction is positive).................................................................................................................1
No reaction to light, haemorrhage, gross destruction (any or all of these)....................................2

3. CORNEA

Degree of Opacity (most dense area used)
No opacity...................................................................................................................................................0
Scattered or diffuse areas, details of iris clearly visible......................................................................1
Easily discernible translucent areas, details of iris slightly obscured..............................................2
Opalescent areas, no details of iris visible, size of pupil barely discernible....................................3
Opaque, iris not discernible through the opacity................................................................................4

Area of Cornea Involved
One quarter (or less) but not zero..........................................................................................................1
Greater than one quarter but less than half...........................................................................................2
Greater than half but less than three quarters......................................................................................3
Greater than three quarters, up to whole area.......................................................................................4

TOOL USED TO ASSESS SCORE: Any additional ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
of 2 animals
Time point:
24/48/72 h
Score:
1
Max. score:
1
Reversibility:
fully reversible within: 14 days
Irritation parameter:
iris score
Basis:
mean
Remarks:
of 2 animals
Time point:
24/48/72 h
Score:
0.83
Max. score:
1
Reversibility:
fully reversible within: 14 days
Irritation parameter:
conjunctivae score
Remarks:
(redness)
Basis:
mean
Remarks:
of 2 animals
Time point:
24/48/72 h
Score:
2
Max. score:
2
Reversibility:
fully reversible within: 21 days
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
of 2 animals
Time point:
24/48/72 h
Score:
2
Max. score:
2
Reversibility:
fully reversible within: 21 days
Irritation parameter:
other: discharge score
Basis:
mean
Remarks:
of 2 animals
Time point:
24/48/72 h
Score:
1.67
Max. score:
3
Reversibility:
fully reversible within: 14 days
Irritant / corrosive response data:
Individual scores for ocular irritation are given in Table 1.
Scattered or diffuse corneal opacity was noted in both treated eyes one hour after treatment and at the 24, 48, 72 hour and 7 day observations.
Iridial inflammation was noted in all treated eyes one hour after treatment and at the 24 and 48 hour observations. Iridial inflammation was noted in one treated eye at the 72 hour observation and in both treated eyes at the 7 day observation.
Moderate conjunctival irritation was noted in both treated eyes one hour after treatment and at the 24, 48, 72 hour observations. Moderate conjunctival irritation was noted in one treated eye and minimal conjunctival irritation was noted in the other treated eye at the 7 day observation. Minimal conjunctival irritation was noted in one treated eye at the 14 day observation.

One treated eye appeared normal at the 14 day observation and the other treated eye appeared normal at the 21 day observation.
Other effects:
BODYWEIGHT
Both animals showed the expected gain in bodyweight during the study.

Table 1 Individual Scores for Ocular Irritation

Rabbit Number and Sex

72425 Male

72537 Male

IPR = 2

IPR = 2

Time After Treatment

1

hour

24 hours

48 hours

72 hours

7

days

14 days

1

hour

24 hours

48 hours

72 hours

7

days

14 days

21 days

Cornea

Degree

Area

 

1

1

 

1

1

 

1

2

 

1

2

 

1

1

 

0

0

 

1

2

 

1

2

 

1

2

 

1

2

 

1

2

 

0

0

 

0

0

Iris

1

1

1

0

1

0

1

1

1

1

1

0

0

Redness

2

2

2

2

1

0

2

2

2

2

2

2

0

Chemosis

2

2

2

2

1

0

2

2

2

2

2

1

0

Discharge

2

2

1

1

0

0

3

3

2

1

1

0

0

IPR = Initial pain reaction; a score of 2 indicates slight initial pain. The rabbit blinks and tries to open the eye but reflex closes it.

Interpretation of results:
other: Category 2 (irritating to eyes) based on CLP criteria
Conclusions:
Eye irritant
Executive summary:

The irritancy potential of the test material was assessed in vivo in accordance with the standardised guidelines OECD 405 and EU Method B.5 under GLP conditions.

A single application of the test material was sequentially administered to the right eye of two New Zealand White rabbits. The treated eyes were not irrigated; the left eye remained untreated and served as the control. The animals were observed for 21 days.

Administration of the test material produced scattered or diffuse corneal opacity, iridial inflammation and moderate conjunctival irritation. One treated eye appeared normal at the 14 day observation; the other treated eye appeared normal at the 21 day observation.

Under the conditions of this study, the test material causes irritation to the eye and requires classification as Category 2 in accordance with EU criteria.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin Irritation

In the first key study, the corrosivity potential of the test material was evaluated in vitro using the EPISKIN in vitro Reconstructed Human Epidermis (RHE) Model in accordance with the standardised guidelines OECD 431 and EU Method B.40 under GLP conditions.

Duplicate tissues were treated with the test material for exposure periods of 3, 60 and 240 minutes. At the end of the exposure period, the test material was rinsed from each tissue before each tissue was taken for MTT-loading. Following this, a total biopsy of each epidermis was made and formazan crystals extracted from the MTT-loaded tissues. Duplicate tissues treated with 0.9 % w/v sodium chloride solution served as negative controls and duplicate tissues treated with glacial acetic acid served as positive controls. The optical density of all treated tissues was measured at 540 nm.

The relative mean viability of the test material treated tissues, relative to the negative control, was as follows:

240 minutes exposure: 114.2 %

60 minutes exposure: 107.1 %

3 minutes exposure: 100.8 %

Under the conditions of this study, the test material was considered to be non-corrosive and required no classification in accordance with EU criteria.

 

Following on from the negative result for corrosivity, the skin irritation potential of the test material was evaluated in a second key study in vitro using the EPISKIN reconstructed human epidermis model in accordance with the standardised guidelines OECD 439 and EU Method B.46 under GLP conditions.

Triplicate tissues were treated with the test material for an exposure period of 15 minutes. At the end of the exposure period, each tissue was rinsed before being incubated for 42 hours, after which each tissue was taken for MTT loading. After MTT loading, a total biopsy of each epidermis was made and formazan crystals were extracted out of the MTT-loaded tissues. Duplicate tissues treated with Dulbecco’s Phosphate Buffered Saline with Ca++ and Mg++ served as the negative control and duplicate tissues treated with 5 % w/v aqueous Sodium Dodecyl Sulphate served as the positive control. The optical density of all treated tissues was measured at 540 nm.

The relative mean viability of the test material treated tissues was 10.2 % after the 15 minute exposure period.

Under the conditions of this study, the test material was determined to be a Category 2 irritant in accordance with EU criteria. It also classified as irritant according to EU labelling regulations Commission Directive 2001/59/EC.

 

Eye Irritation

In the first key study, the corrosion and irritancy potential of the test material was investigated in vitro using the Bovine Corneal Opacity and Permeability Assay (BCOP) in accordance with the standardised guideline OECD 437 under GLP conditions.

The undiluted test material was applied for 10 minutes, followed by an incubation period of 120 minutes. Negative (0.9 % w/v sodium chloride solution) and positive controls (ethanol) were tested concurrently. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability) were combined in an empirically derived formula to generate an In Vitro Irritancy Score (IVIS).

The IVIS for the test material was found to be 47.6.

Under the conditions of this study, no prediction can be made about the toxicity to the eye of the test substance.

 

Following this results, the irritancy potential of the test material was assessed in a second key study in vivo in accordance with the standardised guidelines OECD 405 and EU Method B.5 under GLP conditions.

A single application of the test material was sequentially administered to the right eye of two New Zealand White rabbits. The treated eyes were not irrigated; the left eye remained untreated and served as the control. The animals were observed for 21 days.

Administration of the test material produced scattered or diffuse corneal opacity, iridial inflammation and moderate conjunctival irritation. One treated eye appeared normal at the 14 day observation; the other treated eye appeared normal at the 21 day observation.

Under the conditions of this study, the test material causes irritation to the eye and requires classification as Category 2 in accordance with EU criteria.

In accordance with the principles set out in Section 1.4 of Annex XI of Regulation (EC) 1907/2006 (REACH), it is considered scientifically justified to omit any in vivo testing to confirm the substance's skin irritation or corrosive potential as these endpoints are adequately addressed by the in vitro testing summarised in this dossier.


Justification for selection of skin irritation / corrosion endpoint:
Two studies are provided to address this endpoint: an in vitro study to determine corrosivity conducted in accordance with the standardised guidelines OECD 431 and EU Method B.40 and an in vitro irritation study conducted in accordance with the standardised guidelines OECD 439 and EU Method B.46.
Both were carried out under GLP conditions and were assigned a reliability score of 1 in accordance with the criteria detailed by Klimisch (1997). The irritation study is selected as key on the basis that this result will be the basis for classification of the registered substance.

Justification for selection of eye irritation endpoint:
Two studies are provided to address this endpoint: an in vitro study to determine corrosivity and severe irritation conducted in accordance with the standardised guideline OECD 437 and an in vivo irritation study conducted in accordance with the standardised guidelines OECD 405 and EU Method B.5.
Both were carried out under GLP conditions and were assigned a reliability score of 1 in accordance with the criteria detailed by Klimisch (1997). The irritation study is selected as key on the basis that this result will be the basis for classification of the registered substance.

Effects on skin irritation/corrosion: irritating

Effects on eye irritation: irritating

Justification for classification or non-classification

In accordance with the criteria for classification as defined in Annex I, Regulation 1272/2008, the test material requires classification for both skin and eye irritation.

Skin Irritation

The test material was determined to be a Category 2 irritant in accordance with EU criteria. It also classified as irritant according to EU labelling regulations Commission Directive 2001/59/EC.

Eye Irritation

The test material causes irritation to the eye and requires classification as Category 2 in accordance with EU criteria.