Registration Dossier

Administrative data

Description of key information

Skin corrosion (OECD 431): corrosive

Eye irritation/corrosion (WoE: OECD 431, 437 and 492): corrosive

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
26 Jan - 18 Feb 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Version / remarks:
adopted 27 July 2015
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Hess. Ministerium für Umwelt, Klimaschutz, Landwirtschaft und Verbraucherschutz, Wiesbaden, Germany
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: EpiDerm™
Source strain:
not specified
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™ (MatTek Corporation, Bratislava, Slovakia)
- Tissue batch number: 23314
- Delivery date: 16 February 2016
- Date of initiation of testing: 16 February 2016

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature (3 minutes exposure), 37 ± 1.5 °C (60 min exposure)

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: Tissues were gently rinsed using a wash bottle containing DPBS to remove any residual test material (20 times).

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3 h
- Spectrophotometer: microplate reader (Versamax, Molecular Devices, SoftMax Pro Enterprise v.4.7.1)
- Wavelength: 570 nm
- Filter: without reference filter

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: The quality of the EpiDerm™ tissue was assessed by undertaking an MTT cell viability test.
- Barrier function: The barrier function was assessed by determination of the exposure time required to reduce tissue viability by 50% (ET-50) upon application of 100 µL of 1% Triton X-100. The ET-50 value was determined to be 6.55 h.
- Contamination: The cells used to produce the EpiDerm™ tissue were screened for the presence of viruses, bacteria, yeast and other fungi.

NUMBER OF REPLICATE TISSUES: 2

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- killed tissues
- Procedure used to prepare the killed tissues: freezing
- N. of replicates : 2
- Method of calculation used: Since the MTT reducing test substance extract was classified as corrosive by the skin irritation test (tissue viability <50 %), the correction procedures were not necessary.

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: single experiment with 2 incubation periods

PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be corrosive to skin if the viability after 3 minutes exposure is less than 50%, or if the viability after 3 minutes exposure is greater than or equal to 50% and the viability after 1 hour exposure is less than 15%.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount applied: 50 µL

NEGATIVE CONTROL
- Amount applied: 50 µL

POSITIVE CONTROL
- Amount applied: 50 µL
- Concentration: 8 N
Duration of treatment / exposure:
3 and 60 min
Number of replicates:
in duplicates for each treatment and control group
Irritation / corrosion parameter:
% tissue viability
Remarks:
mean value of 2 tissues
Run / experiment:
3 minutes exposure
Value:
39.3
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Remarks:
mean value of 2 tissues
Run / experiment:
60 minutes exposure
Value:
10.9
Vehicle controls validity:
not examined
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
OTHER EFFECTS
- Direct-MTT reduction: Optical evaluation of the MTT-reducing capacity of the test substance after 1 h incubation with MTT-reagent showed blue colour. An additional test with freeze-killed tissues had to be performed, but correction of the viability values was not necessary, since the test substance proved to be corrosive even without correction.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean negative control OD, both for the 3 and 60 min exposure period, was in the range of ≥ 0.8 and ≤ 2.8 for every exposure time.
- Acceptance criteria met for positive control: Exposure to the positive control induced a decrease in the relative absorbance as compared to the negative control, both for the 3 min exposure period (23.0%) and for the 60 min exposure period (14.8%) thus confirming the validity of the test system and the specific batch of tissue models.
- Acceptance criteria met for variability between replicate measurements: The coefficient of variation in the range 20 – 100% viability between tissue replicates was < 14%.

Table 2. Results after treatment with the test substance and controls

Test group

Absorbance at 570 nm*

Mean absorbance of 2 tissues

Coefficient of variation (%)

Rel. absorbance (% of negative control)**

Tissue 1

Tissue 2

3 minutes treatment

Negative control

1.559

1.359

1.459

13.7

100.0

Positive control

0.359

0.311

0.335

3.3

23.0

Test substance

0.559

0.589

0.574

2.0

39.3***

60 minutes treatment

Negative control

1.337

1.452

1.394

7.9

100.0

Positive control

0.236

0.176

0.153

4.1

14.8

Test substance

0.157

0.148

0.153

0.6

10.9***

* Mean of three replicate wells after blank correction

** Relative absorbance (rounded values): 100 × (absorbance test substance/positive control) / (absorbance negative control)

*** Since the MTT reducing test substance extract was classified as irritant by the skin irritation test (tissue viability <50% after 3 minutes exposure), the correction procedures were not necessary.

Interpretation of results:
other: Skin Corr. Cat. 1B (H314)
Remarks:
according to Regulation (EC) 1272/2008
Conclusions:
Under the conditions of the reconstructed human epidermis test the test substance was corrosive to skin. According to the EpiSkinTM prediction model as described in OECD TG 431 a substance which causes cell viability >= 35% after 3 min exposure and < 35% after 60 min exposure, shall be regarded as a skin corrosive Category 1B / 1C. Thus, the test substance was classified as Skin Corr. Cat. 1B according to Regulation (EC) 1272/2008.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (corrosive)

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
6 - 28 Apr 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)
Version / remarks:
July 2015
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Hess. Ministerium für Umwelt, Klimaschutz, Landwirtschaft und Verbraucherschutz, Wiesbaden, Germany
Species:
human
Strain:
other: EpiOcular™
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount applied: 50 µL

NEGATIVE CONTROL
- Amount applied: 50 µL

POSITIVE CONTROL
- Amount applied: 50 µL
Duration of treatment / exposure:
30 min
Duration of post- treatment incubation (in vitro):
120 min
Number of animals or in vitro replicates:
in duplicates for each treatment and control group
Details on study design:
- RhCE tissue construct used, including batch number: EpiOcular™ tissue (MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia), batch number: 23707
- Viability: The quality of the final product was assessed by undertaking an MTT cell viability test.
- Barrier function: The barrier function was assessed by determination of the exposure time required to reduce tissue viability by 50% (ET-50) upon application of 100 µL of 0.3% Triton X-100. The ET-50 value was determined to be 15.12 min.
- Contamination: The cells used to produce the EpiOcular tissue were screened for the presence of viruses, bacteria, yeast and other fungi.
- Duration and temperature of exposure, post-exposure immersion and post-exposure incubation periods: 30 min exposure, 12 min post-exposure immersion and 120 min post-exposure incubation at 37 °C
- Indication of controls used for direct MTT-reducers and/or colouring test chemicals: The ability of the test substance to directly reduce MTT and to form a blue/purple reaction product was assessed in a pre-experiment. Since the test substance proved to reduce MTT a functional check on killed controls was performed in the definitive assay to show that the test substance was not binding to the tissue and leading to a false MTT reduction signal.
- Number of tissue replicates used per test chemical and controls: 2
- Wavelength: 570 nm
- Filter: without reference filter
- Description of the method used to quantify MTT formazan: The optical density was determined in a microplate reader (Versamax, Molecular Devices) without reference filter.
- Description of evaluation criteria used including the justification for the selection of the cut-off point for the prediction model: The test substance was considered to be not irritating to eye if the tissue viability after 30 min exposure, 12 min post-exposure immersion and 120 min post-exposure incubation is >60%.
- Acceptance criteria: The results are acceptable if the negative control OD is >0.8 and <2.5 and if the mean relative viability of the positive control is below 50% of the negative control viability.
- Reference to historical data of the RhCE tissue construct: Historical control data was used to assess the validity of the test.
- Acceptable variability between tissue replicates for the test chemical, positive and negative controls: The difference of viability between the two relating tissues of a single test substance is < 20% in the same run.
Irritation parameter:
other: % tissue viability
Remarks:
mean value of 2 tissues
Run / experiment:
30 min exposure
Value:
47.3
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The negative control OD (1.296 and 1.446) was in the range of > 1.0 and < 2.6.
- Acceptance criteria met for positive control: Treatment with the positive control induced a decrease in the mean relative absorbance compared with the negative control to 32.5%, thus the validity of the test system is ensured.

The difference of viability between the two relating tissues of a single substance is < 20% (values between 1.1% to 7.3%) in the same run (for positive and negative control tissues and tissues of single test substance).

- Other observations: The test substance proved to be an MTT reducer in the MTT pre-test. It was not coloured and it did not dye water or isopropanol in the colour interference pre-test. Therefore, an additional test with freeze-killed tissues to gain a correction factor for evaluation of the main experiment was performed, but an additional test with a viable tissue was not necessary. Results did not have to be considered for correction of the mean relative absorbance of the test substance in the main experiment, since the viability resulted from the test substance exposed tissues was lower than the cut-off value of 60%, because any MTT reduction would not change the classification for the test substance.

Table 1. Results after 30 min incubation time

Test group

Absorbance*

Mean absorbance of 2 tissues*

Rel. absorbance (%)**

Absolute value of the difference of the rel. absorbance (%) Tissue 1 and 2

Rel. absorbance (% of negative control)**

Tissue 1

Tissue 2

Tissue 1

Tissue 2

Blank

0.036

0.035

0.000

-

-

-

-

Negative control

1.289

1.388

1.338

96.3

103.7

7.3

100

Positive control

0.443

0.428

0.435

33.1

32.0

1.1

32.5

Test substance

0.619

0.646

0.633

46.3

48.2

2.0

47.3

Additional test with freeze killed tissues without MTT reduction***

Blank

0.036

0.035

0.000

-

-

-

-

Negative control

0.049

0.055

0.052

94.0

106.0

11.9

100.0

Test substance

0.717

0.706

0.712

1380.1

1359.3

20.8

1359.3

* Mean of two replicate wells after blank correction

** Relative absorbance (rounded values): 100 × (absorbance test substance/positive control) / (absorbance negative control)

*** Results did not have to be considered for correction of the mean relative absorbance of the test substance, since the viability resulted from the test substance exposed tissues was lower than the cut-off value of 50%, because any MTT reduction would not change the classification for the test substance.

Interpretation of results:
other: Eye Irrit. Cat. 2 according to Regulation (EC) 1272/2008
Conclusions:
Under the conditions of the conducted test, the test substance possessed irritating properties towards human-derived epidermal keratinocytes in the EpiOcular™ model.
Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
22 Jan 2016
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)
Version / remarks:
July 2013
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Hess. Ministerium für Umwelt, Klimaschutz, Landwirtschaft und Verbraucherschutz, Wiesbaden, Germany
Species:
other: cattle
Strain:
not specified
Details on test animals or tissues and environmental conditions:
TEST METHOD
The bovine corneal opacity and permeability (BCOP) test is an in-vitro test method used for identifying i) chemicals inducing serious eye damage and ii) chemicals not requiring classification for eye irritation or serious eye damage. The potential of a test substance to cause ocular corrosivity or severe irritancy is measured by its ability to induce opacity and increased permeability in an isolated bovine cornea. The opacity and permeability assessments of the cornea are combined to derive an in-vitro irritancy score (IVIS), which is used to classify the irritancy level of the test substance.

IDENTIFICATION OF THE SOURCE OF THE EYES, STORAGE AND TRANSPORT CONDITIONS
- Source: Schlachthof Aschaffenburg, Aschaffenburg, Germany
- Donor animals: at least 9 month old
- Time interval prior to initiating testing: The corneae were isolated after delivery of the eyes and directly used in the BCOP test on the same day.
- Transport medium and temperature conditions: The isolated eyes were transported in Hank's Buffered Salt Solution (HBSS) at ambient temperature.

PREPARATION OF THE EYES (BEFORE EXPOSURE)
- Eyes free of defects (scratches, neovascularisation, pigmentation, opacity): yes
- Dissection of the eyes and treatment: 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 cornea was mounted in a specially designed cornea holder.
- Description of the cornea holder: The cornea holder consists of anterior and posterior compartments, which interface with the epithelial and endothelial sides of the cornea, respectively.
- Test medium used in the cornea holder: The incubation medium consisted of MEM, supplemented with 1.1 g/500 mL sodium bicarbonate, 5 mL/500 mL L-glutamine, 5 mL penicillin/streptomycin (500 U penicillin, and 500 µg streptomycin per 5 mL). Immediately before starting the test, MEM was supplemented with 1% fetal calf serum.
- Equilibration time: 1 h at 32 ± 1 °C
- Quality check of the equilibrated corneas: At the end of the equilibration period, the basal opacity was determined (t0). Each cornea with a value of the basal opacity >7 was discarded.

DETERMINATION OF THE INITIAL OPACITY
- Method: Corneal opacity was determined by the amount of light transmission passing through the cornea via an opacitometer.
- Specification of the device: OP_KiT opacitometer (Electro Design, France)
Vehicle:
unchanged (no vehicle)
Controls:
other: number of eyes for the negative control: 3; number of eyes for the positive control: 3
Amount / concentration applied:
TEST MATERIAL
- Applied volume: 0.75 mL

POSITIVE SUBSTANCE
- Substance: 2-ethoxyethanol
- Applied volume: 0.75 mL
- Purity: 99%

NEGATIVE CONTROL
- 0.9% NaCl (w/v) solution in deionised water (saline)
- Applied volume: 0.75 mL
Duration of treatment / exposure:
10 min at 32 ± 1 °C
Observation period (in vivo):
not applicable
Number of animals or in vitro replicates:
number of eyes for the test item: 3
Details on study design:
TEST CONDITIONS
- Short description of the method used: The endothelial side of the cornea was positioned against the sealing ring (O-ring) of the posterior part of the holder. The cornea was gently flattened over the O-ring but stretching was avoided. The anterior part of the holder was positioned on the top of the cornea and fixed in place with screws. 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. The anterior compartment received the test item or the controls on the surface of the corneae. The corneae were incubated in a horizontal position at 32 ± 1 °C in the water-bath for 10 min.

POST-EXPOSURE TREATMENT
- Removal of the test substance: The test substance was rinsed off from the application side with saline.

DETERMINATION OF THE FINAL OPACITY
- Method: Corneal opacity was determined by the amount of light transmission through the cornea via an opacitometer.
- Time of determination: After the test item was rinsed off from the application side, the corneae were incubated for further 2 h in a vertical position, followed by a second opacity reading (t130).
- Specification of the device: OP_KiT opacitometer, Electro Design, France

DETERMINATION OF THE CORNEAL PERMEABILITY:
- Method: After the final opacity measurement was performed, the incubation medium was removed from the anterior compartment and replaced by sodium fluorescein solution. Corneae were incubated again in horizontal position for 90 min in a water-bath at 32 ± 1 °C. The amount of sodium fluorescein that crosses into the posterior chamber was quantitatively measured with a spectrophotometer as optical density at 490 nm (OD490).
- Amount and concentration of the dye: 1 mL sodium fluorescein solution (0.4% (w/v); dissolved in HBSS)
- Incubation time: 90 min at 32 ± 1 °C
- Treatment for measuring: Complete medium from the posterior compartment was removed, well mixed, transferred into a 96 well plate and OD490 was determined.
- Specification of the spectrophotometer: Versamax Molecular Devices
Irritation parameter:
in vitro irritation score
Remarks:
mean out of 3 eyes
Run / experiment:
10 min
Value:
46.42
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: Test substance
Irritation parameter:
in vitro irritation score
Remarks:
mean out of 3 eyes
Run / experiment:
10 min
Value:
65.57
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: Positive control
Irritation parameter:
in vitro irritation score
Run / experiment:
10 min
Value:
0.79
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: Negative control
Other effects / acceptance of results:
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: With the negative control (0.9% (w/v) NaCl solution in deionised water) neither an increase of opacity nor permeability of the corneae could be observed.
- Acceptance criteria met for positive control: The positive control (2-ethoxyethanol) showed clearly increased opacity and distinctive permeability of the corneae fulfilling the criteria for classification as severe irritating/corrosive.

Relative to the negative control, the test substance caused an increase of the corneal opacity and permeability. The calculated mean in vitro irritancy score (IVIS) was 46.42. In regard to the defined evaluation criteria, no prediction on the irritation potential of the test substance can be made.

Table 2. Results after 10 min incubation time.

Test group

Opacity value =
Difference (t130-t0) of Opacity

Permeability at 490 nm (OD490)

IVIS

Mean IVIS

 

Mean

 

Mean

 

 

Negative

control

0

0.00

0.048

0.053

0.72

0.79

0

0.062

0.93

0

0.048

0.72

Positive

control

53*

0.601*

62.02

65.57

61*

0.744*

72.17

55*

0.501*

62.52

Test substance

35*

0.843*

47.65

46.42

31*

0.910*

44.66

30*

1.130*

46.96

*: corrected values

Interpretation of results:
other: no prediction can be made
Conclusions:
The irritation potential of the test substance was assessed in the BCOP assay. Application of the test substance to bovine corneae resulted in a calculated mean IVIS of 46.42. According to OECD Guideline 437 no prediction on the irritation/corrosion potential can be made.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin

The skin corrosion potential of the test substance was determined by an in vitro skin corrosion test in human keratinocytes according to OECD Guideline 431 and in compliance with GLP (2016). The test substance did not lead to a colour change in water or isopropanol (pre-test for colour interference) but was proved to be an MTT reducer. Therefore, an additional test with freeze-killed tissues was performed. Human skin tissues (EpiDerm™) were treated in duplicates with 50 µL of the test substance, the negative control (deionized water) or the positive control (potassium hydroxide 8 N), respectively. The test substance and the positive and negative controls were washed off the skin tissues after 3 or 60 min treatment. For cell viability assessment tissues were treated with the MTT solution for 3 h following an extraction time of 19.5 h. The amount of extracted colorant was determined photometrically at 570 nm. 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 both treatment intervals, thus showing the quality of the tissues. Exposure to the positive control induced a decreased absorbance compared to the negative control, both for the 3 min exposure period (23.0%) and for the 1 h exposure period (14.8%) thus confirming the validity of the test system. After exposure to the test substance the relative absorbance value decreased to 39.3% (3 minutes exposure) and 10.9% (1 h exposure). Both values are below the threshold for corrosivity which is defined to be 50% (3 min exposure) and 15% (1 h exposure). Thus, corrections procedure by OD values of freeze killed tissues was not necessary. In conclusion, under the conditions of the reconstructed human epidermis test the test substance was corrosive to skin.

Eye

The eye irritation potential of the test substance was determined in a bovine corneal opacity and permeability (BCOP) test according to OECD Guideline 437 and in compliance with GLP (2016). After a first opacity measurement of the fresh bovine corneae, the neat test substance was applied directly to the epithelial surface of three cattle corneae in an incubation chamber in horizontal position for 10 min at 32 ± 1 °C. After the incubation phase the test substance was rinsed from the corneae. Further, the corneae were incubated for another 2 hours at 32 ± 1 °C in a vertical position, while the anterior chamber contained incubation medium as well. Afterwards, opacity was measured a second time. In addition, the 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. With the negative control, 0.9% (w/v) NaCl solution in deionised water, neither an increase of opacity nor permeability of the corneae could be observed. The mean in vitro irritation score (IVIS) of the negative control was 0.79. The positive control, 2-ethoxyethanol, showed clear opacity and distinctive permeability of the corneae fulfilling the criteria for classification as severe irritating/corrosive. The mean IVIS of the positive control was 65.57. All three values of the negative and positive controls, respectively, were within the range of historical data of the test facility. Relative to the negative control, the test substance caused an increase of the corneal opacity and permeability. The calculated mean IVIS was 46.42. According to OECD Guideline 437 no prediction on the irritation potential can be made.

In a second study, the eye irritation potential of the test substance was determined by an in vitro eye irritation test using a human cornea model according to OECD Guideline 492 and in compliance with GLP (2016). The test susbtance did not lead to a colour change in water or isopropanol (pre-test for colour interference) but was proved to be an MTT reducer. Therefore, an additional test with freeze-killed tissues was performed. EpiOcular ™ tissues were treated in duplicates with 50 mg of the test substance, negative (deionized water) or positive control (methyl acetate), respectively, for 30 min. After rinsing with Ca++Mg++-free DPBS, tissues were incubated for approx. 120 min in assay medium. For cell viability assessment tissues were treated with the MTT solution for 180 min following an extraction time of 2 - 3 h. The amount of extracted colorant was determined photometrically at 570 nm. After treatment with the negative control the absorbance values (1.296 and 1.446) were well within the required range of the acceptability criterion of mean OD ≥ 1.0 and ≤ 2.6, thus showing the quality of the tissues. Exposure to the positive control induced a decreased absorbance compared to the negative control (32.5%) thus confirming the validity of the test system. After incubation with the test substance the relative mean absorbance was 47.3% (threshold for irritancy: ≤ 60%) compared with the value of the negative control. Therefore, the test substance is considered to possess an eye irritating potential.

In conclusion, based on the available in vitro data on eye irritation/corrosion, the test substance showed equivocal results in the BCOP assay (OECD 437) and therefore corrosivity cannot be excluded. The EpiOcular Test (OECD 492) showed irritating properties to the eye. Due to the skin corrosive properties of the test substance (Skin corr. 1B, H314), the risk of severe damage to eyes is considered implicit (Serious Eye Damage Cat. 1) (Column 2 adaptation of Annexes VII and VIII of REACH Regulation). Following a worst case assumption the test substance is therefore classified as Eye Dam. 1.

Justification for classification or non-classification

The available data on skin irritation/corrosion meet the criteria for classification as Skin Corr. 1B (H314) according to Regulation (EC) 1272/2008.

The available data on eye irritation/corrosion meet the criteria for classification as Eye Dam. 1 (H318) according to Regulation (EC) 1272/2008.