Registration Dossier

Administrative data

Description of key information

The substance was investigated in the OECD Guideline No. 439 (EU Commission Directive 440/2008 B.46) and OECD Guideline 431 (EU Method B.40). Under the experimental conditions the test item is considered to be irritant to skin but was not skin corrosive.

The local effect of the test substance in the in vitro assay on the bovine cornea according to OECD Guideline 437 proved negative. No classification as eye irritant is required.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017-04-28 to 2017-08-18
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
Qualifier:
according to
Guideline:
other: UN GHS
Version / remarks:
published 2003, 7h revision, 2017
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: human-derived epidermal keratinocytes cultured from a multilayered, hughly differentiated model of the human epidermis
Source strain:
not specified
Vehicle:
unchanged (no vehicle)
Details on test system:
TEST SYSTEM
- Source: MatTek Corporation (Bratislava, Slovakia)
The EpiDerm™ tissue consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo.

- Procedure used: The plastic bag containing the 24-well plate with epidermal tissues was opened under sterile conditions and the gauze was removed and the inserts were taken out under an airflow using forceps. Any remaining agarose was removed by gentle blotting on the sterile filter paper or gauze, and the tissues were placed in the empty, sterile 6-well plate. Prior to the exposure of the test item and of the controls the EpiDerm™ tissues were inspected for quality.
- Quality control for skin discs: The tissues were checked for air bubbles between agarose and insert (should be less than 30% of the total surface), liquid on top of the insert was removed with sterile cotton tips, if again moisture was observed on top of the inserts after the pre-incubation or in case of visible defects the respective skin models were discarded.

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 ± 1.5 °C,
- Temperature of post-treatment incubation (if applicable): 37 ± 1.5 °C,

REMOVAL OF TEST MATERIAL AND CONTROLS
- Number of washing steps: The inserts were removed immediately from the 6-well plate and tissues were gently rinsed with DPBS at least 15 times in order to remove any residual test material and then submerged in DPBS at least three times. Afterwards the inserts were once again rinsed with sterile DPBS from the inside and the outside.
- Observable damage in the tissue due to washing: none reported
- Modifications to validated SOP: none reported

DYE BINDING METHOD
- Dye used in the dye-binding assay: MTT
- Spectrophotometer: microplate reader (Versamax® Molecular Devices, Softmax Pro, version 4.7.1)
- Filter: 570 nm

NUMBER OF INDEPENDENT TESTING RUNS AND DECISION CRITERIA
- The test chemical is considered to be irritant to skin in accordance with UN GHS and EU CLP Category 2 if the tissue viability of three individual tissues after exposure and post-treatment incubation is less than or equal (≤) to 50%.
- The test substance is considered to be non-irritant to skin if the mean relative tissue viability of three individual tissues is reduced > 50% of the negative control.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
yes, concurrent MTT non-specific colour control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 μL (47 μL/cm2)
- Concentration (if solution): test item was applied undiluted

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 μL of DPBS was applied to triplicate tissue.

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 μL of SLS was applied to triplicate tissue.
- Concentration (if solution): 5% SLS solution in deionised water
Duration of treatment / exposure:
60 minutes
Duration of post-treatment incubation (if applicable):
41 hours
Number of replicates:
Three replicates per test item, negative and positive controls were used.
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1
Value:
26.5
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
2
Value:
30.3
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3
Value:
30.4
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
mean
Value:
29.1
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Other effects / acceptance of results:
The optical pre-experiment (colour interference pre-experiment) to investigate the test item’s colour change potential in water did not led to a change in colour of water. Optical evaluation of the MTT-reducing capacity of the test item after 1 hour incubation with MTT-reagent did not show blue colour. Therefore, the test item passed the MTT- and the colour interference pre-tests.

The is considered valid as all the acceptability criteria were met:

In the negative control the absorbance values were well within the required acceptability criterion of mean OD ≥ 0.8 and ≤ 2.8 for the 60 minutes treatment interval thus showing the quality of the tissues.
Treatment with the positive control induced a decrease in the relative absorbance compared with the negative control to 3.3% thus ensuring the validity of the test system.
The relative standard deviations between the % variability values of the test item, the positive and negative controls in the main test were below 8% (threshold of the "OECD Guideline for the Testing of Chemicals 439: In vitro Skin Irritation: Reconstructed Human Epidermis Test Method”: < 18%), thus ensuring the validity of the study.

Table 1. Results

Dose group

Tissue No.

Absorbance 570 nm

Mean absorbance of 3 wells

Mean absorbance of 3 wells blank corrected

Mean absorbance of 3 tissues

Rel. absorbance [%] tissue 1, 2+3

Rel. standard deviation

Mean Rel. absorbance [%]

Well 1

Well 2

Well 3

Blank

 

0.038

0.038

0.037

0.037

0.000

 

 

 

 

Negative control

1

1.708

1.718

1.740

1.722

1.684

1.670

100.8

4.6

100.0

2

1.804

1.757

1.767

1.776

1.738

104.1

3

1.659

1.606

1.611

1.625

1.588

95.1

Positive control

1

0.090

0.095

0.094

0.093

0.055

0.055

3.3

0.6

3.3

2

0.093

0.093

0.091

0.092

0.055

3.3

3

0.092

0.093

0.093

0.093

0.055

3.3

Blank

 

0.037

0.037

0.036

0.037

0.000

 

 

 

 

Test item

1

0.489

0.473

0.477

0.480

0.443

0.485

26.5

7.5

29.1

2

0.557

0.537

0.535

0.546

0.506

30.3

3

0.555

0.540

0.536

0.544

0.507

30.4

Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
The test item is considered as irritating to skin under test conditions as the mean relative absorbance value was reduced to 29.1% after exposure of the skin tissues compared to the relative absorbance value of the negative control.
Executive summary:

An in vitro study was performed to assess the irritation potential of test item by means of the Human Skin Model Test. The study was conducted according to OECD 439 and EU B.46 guidelines and is GLP-compliant.

Each three tissues of the human skin model EpiDerm were treated with the test item, the negative control (DPBS) or the positive control (5% SLS) for 60 minutes.

After treatment with the negative control the absorbance values were well within the required range of the acceptability criterion of mean OD ≥ 0.8 and ≤ 2.8 for the 60 minutes treatment interval, thus assuring the quality of the tissues. Treatment with the positive control induced a sufficient decrease in the relative absorbance as compared to the negative control for the 60 minutes treatment interval, and thus assuring the validity of the test system. For the test item, the mean relative absorbance value decreased to 29.1% compared to the relative absorbance value of the negative control. This value is below the threshold for irritancy of ≤ 50%. Therefore, under the experimental conditions the test item is considered to be irritant to skin.

The study is reliable without restrictions (Klimisch 1) as it was conducted according to the standard methods and to GLP. It is therefore considered as relevant for key study for this endpoint.

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017-08-25 to 2018-01-10
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
EU Method B.40 (In Vitro Skin Corrosion: Transcutaneous Electrical Resistance Test (TER))
Deviations:
no
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test system:
human skin model
Source species:
human
Cell type:
other: keratinocytes
Vehicle:
unchanged (no vehicle)
Details on test system:
TEST SYSTEM
- Source: MatTek Corporation
- Cell culture: The EpiDerm™ tissues (surface 0.63 cm²) are cultured on specially prepared cell culture inserts (MILLICELLs, 10 mm Ø).
- Pre-incubation period: pre-incubation phase of the EpiDerm™ tissues started on the day of receipt.
- Treatment: EpiDerm tissues were treated with the test substance
- Amount/test concentration: 50 µL
- Duration of treatment: 3 minutes, 60 minutes
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure : room tempeature
- Temperature of post-treatment incubation (if applicable): 37+-1.5°C
CONTROL
- Negative Control: 50 µL deionised water was used as negative control per tissue;
- Positive Control: 50 µL 8.0 N potassium hydoxide (Sigma) was used a positive control per tissue
REMOVAL OF TEST MATERIAL AND CONTROLS
- Washing (if done): Tissues washed with DPBS at least 20 times in order to remove any residual test material. Excess DPBS was removed by gently shaking the tissue inserts and blotting the lower surface with blotting paper.
SCORING SYSTEM:
Viability measured using MTT assay
mean tissue viability < 50% after 3 minutes exposure; corrosive: optional sub-category 1A
mean tissue viability = 50% after 3 minutes exposure AND < 15% after 60 minutes exposure; Corrosive: Optional Sub-category 1B and 1C
mean tissue viability = 50% after 3 minutes exposure AND = 15% after 60 minutes exposuref; Non-corrosive
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
undiluted
Duration of treatment / exposure:
3 minutes; 60 minutes
Duration of post-treatment incubation (if applicable):
3 hours
Number of replicates:
Duplicates
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
3 minutes
Value:
118.4
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
60 minutes
Value:
121.6
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
- OTHER EFFECTS:
- Direct-MTT reduction: test item did not reduce MTT
- Colour interference with MTT:it did not change colour

Dose Group

Exposure Interval

Absorbance Well 1 (Tissue ½)

Absorbance Well 2 (Tissue ½)

Absorbance Well 3 (Tissue ½)

Mean Absorbance (Tissue ½)

Absorbance 570 nm Tissue 1*

Absorbance 570 nm Tissue 2*

Mean Absorbance of 2 Tissues

CV [%]

Rel Absorbance [% of Negative Control]**

Negative control

3 min

1.237

1.341

1.217

1.343

1.221

1.335

1.225

1.340

1.190

1.305

1.247

6.5

100.0

Positive control

0.281

0.231

0.274

0.227

0.275

0.227

0.277

0.228

0.242

0.193

0.218

15.8

17.4

Test item

1.547

1.440

1.554

1.444

1.561

1.527

1.554

1.470

1.519

1.435

1.477

4.0

118.4

Negative control

60 min

1.288

1.220

1.304

1.220

1.310

1.224

1.301

1.222

1.266

1.187

1.226

4.6

100.0

Positive control

0.073

0.077

0.073

0.077

0.073

0.077

0.073

0.077

0.038

0.042

0.040

7.6

3.3

Test item

1.578

1.523

1.563

1.498

1.503

1.493

1.548

1.505

1.513

1.470

1.491

2.1

121.6

* Mean of three replicates wells after blank correction

** relative absorbance [rounded values]: 100 * (absorbance test item / positive control) / (absorbance negative control)

Interpretation of results:
GHS criteria not met
Conclusions:
In the test study the skin corrosion potential of the test item was determined by means of the Human Skin Model Test (OECD 431). The relative absorbance value of the negative control the mean relative absorbance value was 118.4% after 3 minutes exposure and 121.6% after 60 minutes exposure of the skin tissues to the test item. This value is above the threshold for corrosion of 50%. Therefore, the test item is not considered to possess corrosion potential.
Under the experimental conditions reported, the test substance is not corrosive to skin.
Executive summary:

This in vitro study was performed to assess the corrosive potential of the test substance by means of the Human Skin Model Test with EpiDerm™ tissues models. The test item did not reduce MTT (test for direct MTT reduction), and it did not change colour when mixed with deionised water (test for colour interference). Also its intrinsic colour was not intensive. Consequently, additional tests with freeze-killed or viable tissues were not necessary.

 

Independent duplicate tissues of EpiDerm™ were exposed to either the test item, the negative control (deionised water) or the positive control (8.0 N KOH) for 3 minutes and 1 hour, respectively. The test item (50 µL) was dispensed directly onto duplicate EpiDermTM tissue surface, and spread to match the surface of the tissue. After exposure to the negative control the absorbance values met the required acceptability criterion of a mean OD570 =0.8 and =2.8 for both treatment intervals thereby confirming the acceptable quality of the tissues. Exposure to the positive control induced a decrease in the relative absorbance as compared to the negative control, both for the 3 minutes exposure period and for the 1 hour exposure period. The 1 hour exposure caused a decrease of the cell viability <15% of the negative control. The CV in the range 20 – 100% viability between the tissue replicates is =30%, thus the validity of the test system and the specific batch of the tissue models is confirmed. After exposure of the tissues to the test item the relative absorbance values were not reduced compared with the negative control value, neither after 3 minutes exposure, nor after 1 hour exposure (118.4% and 121.6%, respectively). Both values did not touch the threshold for corrosivity, which is defined to be <50% after the 3 minutes exposure and <15% after the 1 hour exposure. Therefore, the test item was not considered to be corrosive.

 

In conclusion, it can be stated that in this study and under the reported experimental conditions, the test item was non corrosive to skin according to EU CLP and UN GHS.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
Deviations:
no
Qualifier:
according to
Guideline:
EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Species:
cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
- Justification of the test method and considerations regarding applicability: The purpose of this study was to assess the possible corneal damage potential when fresh bovine corneae are exposed to the test item. This study should provide a rational basis for risk assessment in humans.
SOURCE OF COLLECTED EYES
- Source: AB Schlachthof GmbH & Co. KG, 63739 Aschaffenburg, Germany
- Number of animals: not reported, but sets of three corneae were used for treatment with the test item and the negative and positive controls.
- Characteristics of donor animals (e.g. age, sex, weight): at least 9 month old donor cattle
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): The isolated eyes were stored in HBSS containing 1% (v/v) Penicillin/Streptomycin (100 units/mL penicillin and 100 μg/mL streptomycin) in the cooled slaughter-house until transportation on the same morning to the laboratory using a Styrofoam box.
- Time interval prior to initiating testing: The corneae were isolated on the same day after delivery of the eyes.
- indication of any existing defects or lesions in ocular tissue samples: All eyes were carefully examined macroscopically for defects. Those presenting defects such as vascularization, pigmentation, opacity and scratches were discarded.
- Indication of any antibiotics used: 1 % (v/v) Penicillin/Streptomycin (100 units/mL penicillin and 100 μg/mL streptomycin) was used during storing freshly isolated eyes.
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL
Duration of treatment / exposure:
10 minutes
Details on study design:
SELECTION AND PREPARATION OF CORNEAS: Freshly isolated bovine eyes were carefully examined macroscopically for defects and eyes showing defects (such as vascularization, pigmentation, opacity and scratches) were discarded. The cornea was carefully removed from the eye using scalpel and rounded scissors. A rim of about 2 mm of tissue (sclera) was left for stability and handling of the isolated cornea. Each isolated cornea was mounted in a specially designed cornea holder as described in the guideline.

QUALITY CHECK OF THE ISOLATED CORNEAS: After mounting the corneas in the holder, both compartments of the holder were filled with incubation medium. The posterior compartment was filled first to return the cornea to its natural convex position. Care was taken to assure no air bubbles were present within the compartments. For equilibration, the corneae in the holder were incubated in a vertical position for about one hour at 32 ± 1 °C in a water-bath.
At the end of the incubation period, the basal opacity was determined (t0). The basal opacity of all corneae was recorded. Each cornea with a value of the basal opacity > 7 was discarded.

NUMBER OF REPLICATES: 3

NEGATIVE CONTROL USED: Saline (0.9% NaCl in deionised water, using ultrasonic technique)

POSITIVE CONTROL USED: 2-Ethoxyethanol (purity: 99%)

APPLICATION DOSE AND EXPOSURE TIME: The test item was applied in a volume of 0.75 mL on the surface of the corneae. The corneae were incubated in a horizontal position at 32 ± 1 °C in the water-bath for ten minutes.

TREATMENT METHOD: the test item or negative or positive control was applied into the anterior compartment on the surface of the corneae.

POST-INCUBATION PERIOD: no.

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: 1
- POST-EXPOSURE INCUBATION: After the test item or control items, respectively, were rinsed off from the application side with saline, fresh cMEM was added into the anterior compartment. Then the corneae were incubated at 32 ± 1 °C for further two hours in a vertical position.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Opacitometer OP_KiT opacitometer (Electro Design, 63-Riom France) calibrated as described in the manual and the opacity of each of the corneae was determined by reading each holder placed in the photoreceptor compartment for treated cornea.
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of spectrophotometry (OD490)

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA:
The test item was classified into the following category according to OECD guideline 437:
≤ 3: No category
>3 - ≤ 55 No prediction can be made
> 55 Category 1
Irritation parameter:
in vitro irritation score
Run / experiment:
1
Value:
1.81
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
in vitro irritation score
Run / experiment:
2
Value:
3.93
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
in vitro irritation score
Run / experiment:
3
Value:
1.89
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
in vitro irritation score
Run / experiment:
mean
Value:
2.54
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes, neither an increase of opacity nor permeability of the corneae could be observed (mean IVIS =1.09).
- Acceptance criteria met for positive control: yes, clear opacity and distinctive permeability of the corneae observed (mean IVIS = 83.62) corresponding to a classification as serious eye damaging (CLP/EPA/GHS (Cat 1)).
Interpretation of results:
GHS criteria not met
Conclusions:
The calculated mean in vitro irritancy score for the test item was 2.54. The test item is not classified as eye irritant according to GHS.
Executive summary:

An in vitro study was performed according to the OECD 437 guideline to assess the corneal damage potential of test item by means of the BCOP assay using fresh bovine corneae.

The undiluted test item, the positive, and the negative controls were applied to corneae and incubated for 10 minutes at 32 ± 1 °C. After the incubation phase, treatment and controls were each rinsed and the corneae were incubated for

another 120 minutes at 32 ± 1 °C in incubation medium, and opacity was measured. After the opacity measurements, permeability of the corneae was determined by measuring spectrophotometrically the transfer of sodium fluorescein after incubation in a horizontal position for 90 minutes at 32 ± 1 °C.

Relative to the negative control, the test item did not cause a relevant increase of the corneal opacity or permeability. The calculated mean in vitro irritancy score was 2.54. The test item is not classified as eye irritant according to GHS.

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

The skin irritant effect of the test substance was investigated under REACH Annex VII requirements according to the in vitro method recommended in the OECD Guideline No. 439 (EU Commission Directive 440/2008 B.46) and OECD Guideline 431 (EU Method B.40).

In the OECD 439, a positive result was obtained. For the test item, the mean relative absorbance value decreased to 29.1% compared to the relative absorbance value of the negative control. This value is below the threshold for irritancy of ≤ 50%. Therefore, under the experimental conditions the test item is considered to be irritant to skin.

To exclude corrosive properties, OECD 431 was perforemd.After exposure of the tissues to the test item the relative absorbance values were not reduced compared with the negative control value, neither after 3 minutes exposure, nor after 1 hour exposure (118.4% and 121.6%, respectively). Both values did not touch the threshold for corrosivity, which is defined to be <50% after the 3 minutes exposure and <15% after the 1 hour exposure. Therefore, the test item was not considered to be corrosive.A supporting skin irritation study conducted in humans produced a negative result for skin irritation with a 5% formulation and a phototoxicity test produced neagtive results at 5% in vaseline.


The local effect of the test substance in the in vitro assay on the bovine cornea according to OECD Guideline 437 proved negative. Here, the undiluted test item, the positive, and the negative controls were applied to corneae and incubated for 10 minutes at 32 ± 1 °C. After the incubation phase, treatment and controls were each rinsed and the corneae were incubated for another 120 minutes at 32 ± 1 °C in incubation medium. Relative to the negative control, the test item did not cause a relevant increase of the corneal opacity or permeability. The calculated meanin vitroirritancy score was 2.54 (≤ 3: No category, >3 - ≤55 No prediction can be made, > 55 Category 1). No classification as eye irritant is required. This is supported by an in vivo test performed similar to OECD Guideline 405. Here, a 5% formulation of the test substance in vaseline when instilled at dose of 0.1g into the conjuctival sac of six New Zealand White rabbits produced only very mild effects upto 48 hours after treatment. The mean scores after 24, 48 and 72 hours  were 0.3, 0.3, 0.3 and 0.3 respectively.

Justification for classification or non-classification

Skin : According to the available data, the substance is considered skin irritant Cat 2 (H315) according to Regulation (EC) No 1272/2008 (2017).

Eye : Based on the available data, no classification is required according to Regulation (EC) No 1272/2008 (2017).