Registration Dossier

Administrative data

Description of key information

Skin irritation/corrosion (OECD 439 and OECD 431): irritating

Eye irritation (OECD 405): irritating

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12 May - 16 Jun 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 28 July 2015
Deviations:
no
Qualifier:
according to
Guideline:
other: EU B.40 bis (In Vitro Skin Corrosion: Human Skin Model Test)
Version / remarks:
Commission Regulation (EC) No 440/2008 of 30 May 2008
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™ (EPI-200)
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: 23341
- Delivery date: 14 June 2016
- Date of initiation of testing: 14 June 2016

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature (3 ± 0.5 min exposure), 37 ± 1.5 °C (60 ± 5 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. The determined OD (540 - 570 nm) was 1.908 ± 0.185 (acceptance criteria: 1.0 - 3.0).
- 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 6.17 h (acceptance criteria: 4.77-8.72 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
Since the test substance did not directly reduce MTT, an additional test with freeze-killed or viable tissues was not performed.

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: single experiment

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%.
- The test substance is considered to be non-corrosive to skin if the viability after 3 minutes exposure is greater or equal than 50% and the viability after 1 hour exposure is greater or equal 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 ± 0.5 min and 60 ± 5 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 min exposure
Value:
114.8
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Remarks:
mean value of 2 tissues
Run / experiment:
60 min exposure
Value:
22
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
OTHER EFFECTS
- Direct-MTT reduction: Since the test substance did not directly reduce MTT, an additional test with freeze-killed or viable tissues was not performed.
- Colour interference with MTT: The test substance did not change colour, when mixed with deionised water and thus passed the colour interference pre-test.

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 thereby confirming the acceptable quality of the tissues.
- 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 (28.8%) and for the 60 min exposure period (7.4%) 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 < 30%, thus the validity of the test system and the specific batch of the tissue models is confirmed.

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

1.822

1.740

1.781

3.3

100.0

Positive control

0.412

0.615

0.513

27.8

28.8

Test substance

2.028

2.061

2.045

1.2

114.8

60 minutes treatment

Negative control

1.789

1.904

1.846

4.4

100.0

Positive control

0.141

0.132

0.136

4.8

7.4

Test substance

0.467

0.346

0.407

21.1

22.0

* Mean of three replicate wells after blank correction (blank after 3 and 60 min exposure was 0.039 and 0.036, respectively)

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

Interpretation of results:
other: non-corrosive according to Regulation (EC) No 1272/2008
Conclusions:
Under the conditions of the conducted test, the test substance did not possess corrosive properties towards reconstructed human epidermis tissue in the EpiDerm™ model.
Executive summary:

The skin corrosion potential of the test substance was assessed by an in vitro skin corrosion test using a human skin model according to OECD Guideline 431 and in compliance with GLP (2016). After treatment of the tissues with the test substance the mean relative absorbance value did not decrease (114.8%) after 3 min exposure compared to the negative control. After 1 h exposure the relative absorbance value was reduced to 22.0%. Both values did not exceed the threshold for corrosivity and therefore the test substance is not considered to be corrosive to the skin.

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
06 Jan - 03 Feb 2017
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)
Version / remarks:
adopted 28 Jul 2015
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Version / remarks:
(Commission Regulation (EC) No 440/2008 of 30 May 2008, 1st ATP 2009)
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™ (EPI-200)
Source strain:
not specified
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™ (EPI-200) (MatTek Corporation, Bratislava, Slovakia)
- Tissue batch number: 23390
- Delivery date: 31 Jan 2017
- Date of initiation of testing: 31 Jan 2017

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 ± 1.5 °C for 35 min in the incubator; thereafter at room temperature for 25 min in a sterile bench

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: Tissues were gently rinsed with DPBS at least 15 times in order to remove any residual test material. After the rinsing the inserts were submerged in DPBS at least 3 times. Afterwards the inserts were once again rinsed with DPBS from the inside and the outside.

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 v.4.7.1)
- Wavelength: 570 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: The quality of the EpiDerm tissue was assessed by undertaking an MTT cell viability test. The determined OD (540 - 570 nm) was 1.581 ± 0.226 (acceptance criteria: 1.0 - 3.0).
- 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 6.79 h (acceptance criteria: 4.77-8.72 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: 3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
Since the test substance did not directly reduce MTT, an additional test with freeze-killed tissues was not performed.

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: single experiment

PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be irritant to skin if the viability after 1 hour exposure is less or equal than 50%.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount applied: 30 µL

NEGATIVE CONTROL
- Amount applied: 30 µL

POSITIVE CONTROL
- Amount applied: 30 µL
- Concentration: 5% aqueous solution
Duration of treatment / exposure:
60 min
Duration of post-treatment incubation (if applicable):
approx. 42 h
Number of replicates:
triplicates for each treatment and control group
Irritation / corrosion parameter:
% tissue viability
Remarks:
mean value of 3 tissues
Run / experiment:
60 min exposure
Value:
2.7
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
OTHER EFFECTS:
- Direct-MTT reduction: The test substance was not considered to be a MTT reducer.
- Colour interference with MTT: The test substance did not dye water when mixed with it. Also its intrinsic colour was not intensive.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: After treatment with the negative control, the absorbance values (1.522, 1.439 and 1.509) were well within the required acceptability criterion of mean OD ≥ 0.8 and ≤ 2.8 for the 60 min treatment interval thus showing the quality of the tissues.
- Acceptance criteria met for positive control: Treatment with the positive control induced a decrease in the relative absorbance as compared to the negative control to 2.2% thus confirming the validity of the test system.
- Acceptance criteria met for variability between replicate measurements: The relative standard deviations between the % variability values of the test substance, the positive and negative controls in the main test were 3% at the highest (threshold of OECD 439: <18%), thus ensuring the validity of the study.

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

 

 

Mean absorbance at 570 nm *

 

 

Mean absorbance

of 3 tissues blank corrected

Rel. absorbance (%) **

Rel. SD (%)

Mean Rel. absorbance (% of negative control)***

Tissue 1

Tissue 2

Tissue

3

Tissue 1

Tissue 2

Tissue

3

Negative control

1.522

1.439

1.509

1.490

102.2

96.6

101.3

3.0

100.0

Positive control

0.030

0.037

0.033

0.033

2.0

2.5

2.2

0.2

2.2

Test substance

0.045

0.038

0.039

0.041

3.0

2.6

2.6

0.2

2.7

* Mean of 3 replicate wells after blank correction (mean blank value: 0.037)

** Relative absorbance per tissue (rounded values): 100 × (absorbance tissue) / (mean absorbance negative control)

*** Relative absorbance per treatment group (rounded values): 100 × (mean absorbance test item/positive control) / (mean absorbance negative control)

Interpretation of results:
other: irritating potential (Skin Irrit. 2 or Skin Corr. 1 according to Regulation (EC) No 1272/2008)
Conclusions:
Under the conditions of the conducted test, the test substance possessed irritating properties towards reconstructed human epidermis tissue in the EpiDerm™ model.
Executive summary:

The skin irritation potential of the test substance was determined by an in vitro skin irritation test using a reconstructed human skin model according to OECD Guideline 439 and in compliance with GLP (2017). After treatment with the test substance for 60 min the tissue viability decreased to 2.7% compared to the negative control (threshold for irritancy ≤ 50%).Therefore, the test substance is considered to possess an irritant potential towards human-derived epidermal keratinocytes.

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:
6 - 21 Nov 2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
Feb 1987
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
Jan 1997
Deviations:
no
GLP compliance:
yes
Remarks:
Ministry of Agriculture, Environmental Protection and Regional Planning, Potsdam, Germany
Species:
rabbit
Strain:
other: Chbb:HM(SPF) - Littlerussian
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Weight at study initiation: 2.0 - 2.1 kg
- Housing: individually in PPO cages (floor area: 2576 cm²) with perforated floor
- Diet: Altromin 2123 (Altromin, Lage, Germany), ad libitum
- Water: domestic quality drinking water acidified with hydrochloric acid to pH 2.5, ad libitum
- Acclimation period: at least one week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3
- Humidity (%): 55 ± 15
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 / 12

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount applied: 0.1 mL
Duration of treatment / exposure:
24 hours
Observation period (in vivo):
7 days
Reading time points: 1, 24, 48 and 72 hours and 7 days after treatment
Number of animals or in vitro replicates:
4 females
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing: 20 mL 0.9% sodium chloride solution was used
- Time after start of exposure: 24 hours

SCORING SYSTEM: Draize scoring system

TOOL USED TO ASSESS SCORE: fluorescein
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0.7
Max. score:
4
Reversibility:
fully reversible within: 72 h
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 48 h
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1.3
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
cornea opacity score
Basis:
animal #4
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
iris score
Basis:
animal: #1, #3 and #4
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 7 days
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.7
Max. score:
2
Reversibility:
fully reversible within: 72 h
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2.33
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Basis:
animal: #2 and #4
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2.7
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2.3
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
animal #4
Time point:
24/48/72 h
Score:
1.7
Max. score:
4
Reversibility:
fully reversible within: 7 days
Irritant / corrosive response data:
One hour after application of the test substance animal No. 1 showed easily discernible translucent areas with slightly obscured details of iris on more than one quarter, but less than half of the cornea, an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light, a diffuse beefy red conjunctiva, a swelling with lids more than half closed and a discharge with moistening of the lids and hairs, and considerable area around the eye. In animal No. 2 a scattered or diffuse area of opacity on more than one quarter, but less than half of the cornea, an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light, a diffuse, crimson red conjunctiva with individual vessels not easily discernible, an obvious swelling with partial eversion of lids and a discharge with moistening of the lids and hairs, and considerable area around the eye were observed. Animal No.3 showed easily discernible translucent areas with slightly obscured details of iris on more than one quarter, but less than half of the cornea, an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light, a diffuse beefy red conjunctiva, a swelling with lids about half closed and a discharge with moistening of the lids and hairs, and considerable area around the eye. In animal No. 4 a scattered or diffuse area of opacity on more than one quarter, but less than half, an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction lights, a diffuse beefy red conjunctiva, a swelling with lids about half closed and a discharge with moistening of the lids and hairs, and considerable area around the eye were observed.

24 hours after application of the test substance animal No. 1 showed a scattered or diffuse area of opacity on more than three quarters of the cornea, an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light, a diffuse beefy red conjunctiva, a swelling with lids about half closed and a discharge different from normal. In animal No. 2 a scattered or diffuse area of opacity on one quarter or less of the cornea, an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light, a diffuse, crimson red conjunctiva with individual vessels not easily discernible as well as a swelling above normal were observed. Animal No. 3 showed easily discernible translucent areas with slightly obscured details of iris on more than one quarter, but less than half of the cornea, an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light, a diffuse beefy red conjunctiva, a swelling with lids about half closed and a discharge with moistening of the lids and hairs just adjacent to lids. In animal No. 4 a scattered or diffuse area of opacity on more than one quarter, but less than half of the cornea, an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light, a diffuse beefy red conjunctiva, an obvious swelling with partial eversion of lids and a discharge different from normal were observed. After instillation of fluorescein animal No. 1 showed a scattered or diffuse area of opacity on more than three quarters of the cornea. A scattered or diffuse area of opacity was observed on one quarter or less of the cornea in animal No. 2 and on more than half, but less than three quarters of the cornea in animal No. 3. Animal No. 4 showed a scattered or diffuse area of opacity on more than one quarter, but less than half of the cornea.

48 hours alter application of the test substance in animal No. 1 showed a scattered or diffuse area of opacity on more than three quarters of the cornea, an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light, a diffuse, crimson red conjunctiva with individual vessels not easily discernible as well as an obvious swelling with partial eversion of lids. In animal No. 2 an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light, a diffuse, crimson red conjunctiva with individual vessels not easily discernible as well as a swelling above normal were observed. In animal No. 3 a scattered or diffuse area of opacity on more than one quarter, but less than half of the cornea, an iris with markedly deepened folds, congestion swelling, moderate circumcorneal injection and existence of reaction to light, a diffuse beefy red conjunctiva, an obvious swelling with partial eversion of lids and a discharge with moistening of the lids and hairs just adjacent to lids were observed. Animal No. 4 showed a scattered or diffuse area of opacity on more than one quarter, but less than half of the cornea, an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light, a diffuse, crimson red conjunctiva with individual vessels not easily discernible, an obvious swelling with partial eversion of lids and a discharge different from normal. After instillation of fluorescein in animal No. 1 a scattered or diffuse area of opacity on more than one quarter, but less than half of the cornea, was observed. Animals No. 3 and No. 4 showed a scattered or diffuse area of opacity on one quarter or less of the cornea.

At 72 hours an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light, a diffuse, crimson red conjunctiva with individual vessels not easily discernible as well as an obvious swelling with partial eversion of lids were observed in animal No. 1. Animal No. 2 showed a diffuse, crimson red conjunctiva with individual vessels not easily discernible as well as a swelling above normal. In animal No. 3 a scattered or diffuse area of opacity on more than one quarter, but less than half of the cornea, an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light, a diffuse, crimson red conjunctiva with individual vessels not easily discernible, a swelling above normal and a discharge different from normal were observed. Animal No. 4 showed a scattered or diffuse area of opacity on more than one quarter, but less than half of the cornea, an iris with markedly deepened folds, congestion, swelling, moderate circumcorneal injection and existence of reaction to light, some conjunctival vessels definitely injected and a swelling above normal. After instillation of fluorescein in animal No. 1 a scattered or diffuse area of opacity on one quarter or less of the cornea was observed.

7 days after application of the test substance all four animals were free of any signs of eye Irritation.

Table 1. Results of the eye irritation study

Animal

Time (h)

Conjunctivae

Cornea

Iris

Redness

Chemosis

1

1

3

4

2

1

24

3

3

1

1

48

2

2

1

1

72

2

2

0

1

Average 24+48+72

2.3

2.3

0.7

1.0

2

1

2

2

1

1

24

2

1

1

1

48

2

1

0

1

72

2

1

0

0

Average 24+48+72

2.0

1.0

0.3

0.7

3

1

3

3

2

1

24

3

3

2

1

48

3

2

1

1

72

2

1

1

1

Average 24+48+72

2.7

2.0

1.3

1.0

4

1

3

3

1

1

24

3

2

1

1

48

2

2

1

1

72

1

1

1

1

Average 24+48+72

2.0

1.7

1.0

1.0

Interpretation of results:
other: Eye Irrit. Cat. 2 according to Regulation (EC) 1272/2008
Conclusions:
Under the conditions of this eye irritation study in rabbits the test substance was irritating to the eye.
Executive summary:

The individual mean scores after 24, 48 and 72 h are for corneal opacity 0.7, 0.3, 1.3, 1.0, for iris lesion 1.0, 0.7, 1.0, 1.0, for conjunctival redness 2.3, 2.0, 2.7, 2.00 and for chemosis 2.3, 1.0, 2.0, 1.7. All ocular lesions were fully reversible within 7 days. Therefore, the test substance is considered to be irritating to the eye.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin

The skin corrosion potential of the test substance was assessed by an in vitro skin corrosion test using a human skin model according to OECD Guideline 431 and in compliance with GLP (2016). After treatment of the tissues with the test substance the mean relative absorbance value did not decrease (114.8%) after 3 min exposure compared to the negative control. After 1 h exposure the relative absorbance value was reduced to 22.0%. Both values did not exceed the threshold for corrosivity and therefore the test substance is not considered to be corrosive to the skin.

In a next step, the skin irritation potential of the test substance was determined by an in vitro skin irritation test using a reconstructed human skin model according to OECD Guideline 439 and in compliance with GLP (2017). After treatment with the test substance for 60 min the tissue viability decreased to 2.7% compared to the negative control (threshold for irritancy 50%).Therefore, the test substance is considered to possess an irritant potential towards human-derived epidermal keratinocytes.

In conclusion, based on the available data on skin irritation/corrosion, the test substance is considered to be irritating to skin and therefore classified as Skin Irrit. 2 (H315) according to CLP.

Eye

The eye irritation potential was tested in an acute eye irritation/corrosion test in four rabbits according to OECD Guideline 405 and in compliance with GLP (2000). After treatment with 0.1 mL of the test substance

The individual mean scores after 24, 48 and 72 h are for corneal opacity 0.7, 0.3, 1.3, 1.0, for iris lesion 1.0, 0.7, 1.0, 1.0, for conjunctival redness 2.3, 2.0, 2.7, 2.0 and for chemosis 2.3, 1.0, 2.0, 1.7. All ocular lesions were fully reversible within 7 days. Therefore, the test substance is considered to be irritating to the eye.

Justification for classification or non-classification

The available data on skin irritation of the test substance meet the criteria for classification as Skin Irrit. 2 (H315) according to Regulation (EC) 1272/2008.

The available data on eye irritation of the test substance meet the criteria for classification as Eye Irrit. 2 (H319) according to Regulation (EC) 1272/2008.