Registration Dossier

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Administrative data

Description of key information

Skin irritation: not irritating (OECD 404, GLP, K, Rel.1) and (OECD 439, GLP, K, Rel.1)


Eye irritation: not irritating (OECD 405, GLP, K, Rel.1)

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:
20 July - 14 August 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
GLP study performed according to OECD Guideline 439.
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Version / remarks:
26 July 2013
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Version / remarks:
24 August 2009
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. QA statement)
Remarks:
08 April 2015.
Test system:
human skin model
Remarks:
Epi-200 Kit
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: reconstructed epidermis
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE:
Supplier: MatTek In Vitro Life Science Laboratories, Bratislava.
Date received: 14 August 2015
EpiSkinTM Tissues (0.38cm2) lot number: 21685
Maintenance Medium lot number: 1660120
Assay Medium lot number: 080615ZSD

EVALUATION OF DIRECT INTERACTION WITH MTT
- For correct interpretation of results it was necessary to assess the ability of the test item to directly reduce MTT. The direct interaction of MTT with the test item was checked by adding 30 µL of the test item to 1mL of the solution of MTT. No colour change could be observed in the present study. Therefore, there is no direct interaction between the test item and MTT

TREATMENT
- After approximately 18 hours incubation of the tissues, they were treated with the test item.
- 1 plate (3 tissues) was used as negative control; each tissue was treated with 30 μL DPBS buffer, a nylon mesh was added in order to ensure sufficient contact with the tissue sur-face.
- 1 plate was used as positive control; each tissue was treated with 30 μL SDS-solution, a nylon mesh was added in order to ensure sufficient contact with the tissue surface.
- 1 plate was used for treatment with the test item: 30 μL test item were applied, and a nylon mesh was added in order to ensure sufficient contact with the tissue surface.
Tissues were dosed in 1-min-intervals. After dosing the last tissue, all plates are trans-ferred into the incubator for 35 min at 37 ± 1°C and 5.0 ± 0.5% CO2. 60 min after the first application, the inserts were removed from the plates using sterile forceps and rinsed im-mediately in 1-min-intervals. After rinsing, each tissue was dried with a sterile cotton tip and then transferred into a new 6-well-plate with fresh assay medium (0.9 mL). Then, the tissues were set in the incubator for 24 h.

REMOVAL OF TEST MATERIAL AND CONTROLS
- For 3 incubated tissues, a new 6-well-plate with 0.9 mL assay medium in the upper row was prepared. The tissues were removed from the incubator and shaken for 10 min (500 rpm). Then the inserts were transferred into the new 6-well-plate and set into the in-cubator for 18 h for post-incubation.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE

- After a total incubation time of 42 h, a 24-well-plate was prepared with 300 μL freshly pre-pared MTT-reagent in each well. The tissues were blotted on the bottom and then trans-ferred into the 24-well-plate. Then the 24-well-plate was set into the incubator for 3 h at 37 ± 1°C and 5.0 ± 0.5% CO2.
- After 2 h, the inserts in which formazan had been produced were pierced with an injection needle, taking care that all colour was extracted. The inserts were then discarded and the content of each well was thoroughly mixed in order to achieve homogenisation.
- From each well, two replicates with 200 μL solution (each) were pipetted into a 96-well-plate which was read in a plate spectral photometer at 570 nm.

VIABILITY CALCULATION:

- Data from individual replicate tissues (OD values and calculated percent tissue viability data for the test item and controls), mean percent tissue viability and standard deviation for each individual test item and control were reported in Table 7.3.1/1. The results were expressed as a viability percentage compared with the negative control: viability % = (mean OD test item / mean OD negative control) * 100

ACCEPTABILITY OF THE ASSAY
- The absolute OD 570 nm of the negative control tissues in the MTT test is an indicator of tissue viability obtained after the shipping procedure and under specific conditions of the assay. Tissue viability is meeting the acceptance criterion if the mean OD of the two tissues is ≥ 0.8 and <=2.8.
The % Formazan production of positive control SDS is <=20% of negative control and the variation within replicates (RSD) is < 18%.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 µL
- Concentration (if solution): Undiluted
Duration of treatment / exposure:
60 minutes at 37 +/-1°C.
Duration of post-treatment incubation (if applicable):
18 hours post-incubation period at 37°+/-1°C, 5 +/-0.5% CO2
Number of replicates:
3
Irritation / corrosion parameter:
% tissue viability
Remarks:
mean
Run / experiment:
mean value
Value:
100.9
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
MTT VIABILITY ASSAY RESULTS
- The test item is considered as not skin irritant.
- After the treatment, the relative absorbance values were increased to 100.9%. This value is well above the threshold for skin irritation (50%).The optical density of the negative control was well within the required acceptability criteri-on of 0.8 ≤ mean OD ≤ 2.8, OD was 1.8. The positive control induced a decrease in the relative absorbance as compared to the negative control to 3.3 % (required: ≤ 20%) for thus ensuring the validity of the test system. Variation within replicates was within the ac-cepted range for negative control, positive control and test item.

Table 7.3.1/&: Absorbance Values negative control, test item and positive control (OD at 570 nm):




















































Designation



Measurement



Negative Con-trol



Test susbtance



Positive Control



Tissue 1



1



1.925



1.718



0.101



2



1.922



1.727



0.102



Tissue 2



1



1.830



1.900



0.098



2



1.857



1.924



0.097



Tissue 3



1



1.804



1.977



0.087



2



1.800



1.989



0.088



Table 7.3.1/2 Mean Absorbance Values










































Designation



Negative Control



Test susbtance



Positive Control



Mean – blank (tissue 1)



1.888



1.687



0.066



Mean – blank (tissue 2)



1.808



1.876



0.062



Mean – blank (tissue 3)



1.766



1.947



0.052



Mean of the three tissues



1.821



1.837



0.060



Relative standard deviation of the three tissues



3.4%



7.3%



12.0%



Table 7.3.1/3 % Formazan Production































Designation



Test susbtance



Positive Control



% Formazan production (tissue 1)



92.6%



3.6%



% Formazan production (tissue 2)



103.0%



3.4%



% Formazan production (tissue 3)



106.9%



2.9%



% Formazan production (mean)



100.9%



3.3%



 


Table 7.3.1/4 Validity criteria and results


























Criterion



Demanded



Found



OD of negative control



≥ 0.8 and ≤ 2.8



1.8



% Formazan production of positive control SDS



≤ 20% of negative control



3.3%



Variation within replicates (RSD)



< 18%



3.4% (negative control) 12.0% (positive control) 7.3% (test item)


Interpretation of results:
GHS criteria not met
Conclusions:
Under the test conditions, test material is not classified according to Regulation (EC) No. 1272/2008 (CLP).
Executive summary:

An in vitro skin irritation test using the Reconstructed Human Epidermis (Episkin Standard model) was performed according to the OECD Guideline 439 and in compliance with GLP to predict the acute skin irritation potential of the test item.


 


The test item was applied at the dose of 30 µL, to 3 tissues of the Human skin model EpiDerm during 60 minutes, followed by a rinse with 25 mL of PBS and a 18 hours post-incubation period at 37+/-1°C, 5 +/-0.5% CO2. Cell viability was then measured by enzymatic conversion of the vital dye MTT into a blue formazan salt that was quantitatively measured after extraction from tissues.


 


The mean corrected percent viability of the treated tissues was 100.9%, versus 3.3% in the positive control (5% Sodium Dodecyl Sulfate).


 


Under the test conditions and in accordance with Regulation EC No. 1272/2008, the test item was considered as non-irritant to skin.

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Between 21 July and 11 August, 2015.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study conducted in compliance with OECD Guideline No. 404 without any deviation.
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
24 April 2002
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Version / remarks:
30 May 2008
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. QA statement)
Remarks:
Inspected on 2015-04-23&24 / Signed on 2015-10-23
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: EUROLAP (F-35140 GOSNE)
- Weight at study initiation: 2.10 - 3.37 kg
- Housing: Each animal was kept in an individual box installed in conventional air conditioned animal husbanding.
- Diet: foodstuff (SAFE - 112), ad libitum
- Water: tap-water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 17-23 °C
- Humidity: 30-70 %
- Air changes: at least 10 changes per hour.
- Photoperiod: 12 hours light/12 hours dark.

IN-LIFE DATES: From 21 July to 11 August, 2015.
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
yes
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration (if solution): Test item was applied as supplied.
Duration of treatment / exposure:
4 hours
Observation period:
Skin reactions were appreciated 1, 24, 48 and 72 hours after removal of the patch and observations were continued from Days 4 to 14 (in case of persistent reactions were observed).
Number of animals:
3 females
Details on study design:
PRETEST
Initially, a single animal was treated. After consideration of the cutaneous responses produced in the first treated animal, two additional animals were treated.

TEST SITE
- Area of exposure: Undamaged skin area of the right flank of each animal
- Type of wrap if used: Patch was secured in position with a strip of surgical adhesive tape.

REMOVAL OF TEST SUBSTANCE
- Washing: After removal of the patch, the treated area was rinsed with distilled water.
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS
1, 24, 48 and 72 hours after removal of the patch and observations were continued from Days 4 to 14 (in case of persistent reactions were observed).

SCORING SYSTEM:
- Method of calculation: Skin irritant reaction was scored as per Draize scale according to OECD guideline 404
Irritation parameter:
erythema score
Basis:
animal #1
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
probability of mild irritation
Irritation parameter:
edema score
Basis:
animal #1
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
probability of mild irritation
Irritation parameter:
erythema score
Basis:
animal #2
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
not fully reversible within: 7 days.
Remarks on result:
probability of mild irritation
Irritation parameter:
edema score
Basis:
animal #2
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
probability of weak irritation
Irritation parameter:
erythema score
Basis:
animal #3
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
probability of weak irritation
Irritation parameter:
edema score
Basis:
animal #3
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
probability of weak irritation
Irritant / corrosive response data:
- Slight to well - defined erythema associated with a very slight to slight oedema was noted on the treated area of all animals, 1 hour after the patch removal. All reactions were totally reversible on day 7.
- On the cutaneous structure, loss of litheness was noted in one animal on day 3 and dryness of the skin in all animals on day 7. The skin recovered a normal aspect on day 14 in two animals but dryness of the skin was still noted on the day 14 in one animal.
Other effects:
None

Table 7.3.1/1: Individual and mean skin reactions/Erythema - Eschar formation following 4 hour exposure

Skin reaction

Observation time (following patch removal)

Individual Scores – Rabbit Number and Sex

A5225/Female

A5203/Female

A5264/Female

Erythema/Eschar formation

1 h

2

1

1

24 h

2

2

1

48 h

2

2

1

72 h

2

2

1

Day 7

0

0

0

Total (24, 48 and 72 hours)

6

6

3

Mean (24, 48 and 72 hours)

2

2

1

Table 7.3.1/2: Individual and mean skin reactions/Oedema formation following 4 hour exposure

Skin reaction

Observation time (following patch removal)

Individual Scores – Rabbit Number and Sex

A5225/Female

A5203/Female

A5264/Female

Oedema formation

1 h

2

2

1

24 h

2

1

1

48 h

2

1

1

72 h

2

1

1

Day 7

0

0

0

Total (24, 48 and 72 hours)

6

3

3

Mean (24, 48 and 72 hours)

2

1

1

Note:

A5225: dryness from D7 to D14

A5203: dryness at D7

A5264: dryness at D7. Loss of litheness at D3.

Interpretation of results:
Category 3 (mild irritant) based on GHS criteria
Conclusions:
Under the test conditions, test item must not be classified according to CLP Regulation (EC) No. 1272/2008 and classified as "Category 3 (mild irritant)" based on Globally Harmonized System (GHS).
Executive summary:

In a dermal irritation study performed according to the OECD Guideline No. 404, and in compliance with GLP, 0.5 mL of undiluted test item was applied on an undamaged skin area of the flank of 3 female New Zealand White rabbits. On the other flank an untreated area was served as the control. Test sites were covered with a semi-occlusive dressing for 4 h. Skin irritation was assessed and scored according to the Draize scale at 1, 24, 48 and 72 hours after removal of the patch and observations were continued from Days 4 to 14. Initially, a single animal was treated. After consideration of the cutaneous responses produced in the first treated animal, two additional animals were treated.

 

Slight to well - defined erythema associated with a very slight to slight oedema was noted on the treated area of all animals, 1 hour after the patch removal. All reactions were totally reversible on day  7. On the cutaneous structure, loss of litheness was noted in one animal on day 3 and dryness of the skin in all animals on day 7. The skin recovered a normal aspect on day 14 in two animals but dryness of the skin was still noted on the day 14 in one animal.

 

The mean scores for each animal within 3 scoring times (24, 48 and 72 h) were 2.0 / 2.0 / 1.0 for erythema and 2.0 / 1.0 / 1.0 for oedema. These effects are reversible on day 7.

Under the test conditions, test item must not be classified according to CLP Regulation (EC) No. 1272/2008 and classified as "Category 3 (mild irritant)" based on Globally Harmonized System (GHS).

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

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

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Between 27 July and 06 August, 2015.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
GLP study conducted in compliance with OECD Guideline No. 405 without any deviation.
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
02 October 2012.
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Deviations:
no
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. QA statement)
Remarks:
Inspected on 2015-04-23&24 / Signed on 2015-10-23
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: EUROLAP (F-35140 GOSNE)
- Weight at study initiation: no data.
- Housing: Each animal was kept in an individual box installed in conventional air conditioned animal husbanding.
- Diet: foodstuff (SAFE - 112), ad libitum
- Water: tap-water, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature: 17-23 °C
- Humidity: 30-70 %
- Air changes: at least ten changes per hour.
- Photoperiod: 12 hours light/12 hours dark.

IN-LIFE DATES: from 27 July to 06 August, 2015.
Vehicle:
unchanged (no vehicle)
Controls:
yes
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL

- Concentration (if solution): Test item was instilled, as supplied
Duration of treatment / exposure:
No washing was done
Observation period (in vivo):
Ocular examinations were performed on both right and left eyes 1, 24, 48 and 72 h & Days 7, 14 and 21 following treatment, according to a numerical evaluation.
Number of animals or in vitro replicates:
3 females
Details on study design:
TREATMENT
- 0.1 mL of the test item was instilled, as supplied, into the conjunctival sac of one eye. The other eye remained untreated serving as control.
- Initially, a single animal was treated. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated.

REMOVAL OF TEST SUBSTANCE
- Washing: No

SCORING SYSTEM: According to OECD guideline 405
Irritation parameter:
cornea opacity score
Basis:
animal #1
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
0.7
Max. score:
4
Reversibility:
fully reversible within: 2 days
Remarks on result:
probability of weak irritation
Irritation parameter:
iris score
Basis:
animal #1
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #1
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
1
Max. score:
3
Reversibility:
fully reversible within: 7 days
Remarks on result:
probability of weak irritation
Irritation parameter:
chemosis score
Basis:
animal #1
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 7 days
Remarks on result:
probability of weak irritation
Irritation parameter:
cornea opacity score
Basis:
animal #2
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 2 days.
Remarks on result:
probability of weak irritation
Irritation parameter:
iris score
Basis:
animal #2
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: not applicable.
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #2
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
0.7
Max. score:
3
Reversibility:
fully reversible within: 2 days
Remarks on result:
probability of weak irritation
Irritation parameter:
chemosis score
Basis:
animal #2
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
0.7
Max. score:
4
Reversibility:
fully reversible within: 2 days.
Remarks on result:
probability of weak irritation
Irritation parameter:
cornea opacity score
Basis:
animal #3
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 2 days
Remarks on result:
probability of weak irritation
Irritation parameter:
iris score
Basis:
animal #3
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: not applicable
Remarks on result:
no indication of irritation
Irritation parameter:
conjunctivae score
Basis:
animal #3
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
0.3
Max. score:
3
Reversibility:
fully reversible within: 2 days
Remarks on result:
probability of weak irritation
Irritation parameter:
chemosis score
Basis:
animal #3
Remarks:
mean individual score
Time point:
24/48/72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 2 days
Remarks on result:
probability of weak irritation
Irritant / corrosive response data:
The ocular reactions observed during the study have been slight to moderate and totally reversible:
- at the conjunctivae level: a slight to moderate redness noted 1 hour after the test item instillation and totally reversible between days 2 and 7, associated with a slight to moderate chemosis noted 1 or 24 hours after the test item instillation and totally reversible between days 2 and 7;
- at the iris level: a congestion, noted 1 hour after the test item instillation in one animal, and totally reversible on day 1.
- at the corneal level: a slight to moderate corneal opacity, noted 24 hours after the test item instillation and totally reversible on day 2.
Other effects:
None

Table 7.3.2/1: Individual and Mean Scores for Cornea, Iris and Conjunctivae

Rabbit Number and Sex

 

Time After Treatment

 

Corneal Opacity

 

Iris lesion

Conjunctival Redness

 

Conjunctival Chemosis

 

A5235 Female

 

1 h

0

1

1

1

24 h

2

0

1

1

48 h

0

0

1

1

72 h

0

0

1

1

Day 7

0

0

0

0

Total (24, 48 and 72 h)

2

0

3

3

Mean (24, 48 and 72 h)

0.7

0.0

1

1

Rabbit Number and Sex

 

Time After Treatment

 

Corneal Opacity

 

Iris lesion

Conjunctival Redness

 

Conjunctival Chemosis

 

A5266 Female

 

1 h

0

0

2

1

24 h

1

0

2

2

48 h

0

0

0

0

72 h

0

0

0

0

Day 7

0

0

0

0

Total (24, 48 and 72 h)

1

0

2

2

Mean (24, 48 and 72 h)

0.3

0.0

0.7

0.7

Rabbit Number and Sex

 

Time After Treatment

 

Corneal Opacity

 

Iris lesion

Conjunctival Redness

 

Conjunctival Chemosis

 

A5270 Female

 

1 h

0

0

1

1

24 h

1

0

1

1

48 h

0

0

0

0

72 h

0

0

0

0

Day 7

0

0

0

0

Total (24, 48 and 72 h)

1

0

1

1

Mean (24, 48 and 72 h)

0.3

0

0.3

0.3

Interpretation of results:
GHS criteria not met
Conclusions:
Under the test conditions, the test material is not classified as irritating to eyes according to the annex I of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.
Executive summary:

In an eye irritation study conducted according to OECD 405 guideline and in compliance with GLP, 3 New Zealand White female rabbits were exposed to 0.1 mL of test item in one eye while the contralateral eye remained untreated and served as control. The eyes were examined and the changes were observed at 1, 24, 48, 72 h and day 7 following treatment and graded according to the Draize method. Initially, a single animal was treated. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated.

A slight to moderate redness noted 1 hour after the test item instillation and totally reversible between days 2 and 7, associated with a slight to moderate chemosis noted 1 or 24 hours after the test item instillation and totally reversible between days 2 and 7. At the iris level, a congestion was noted 1 hour after the test item instillation in one animal, and totally reversible on day 1. A slight to moderate corneal opacity, noted 24 hours after the test item instillation and totally reversible on day 2.

 

 

Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals were 0.7, 0.3, 0.3 for cornea score; 0.0, 0.0, 0.0 for iris score; 1.0, 0.7, 0.3 for conjunctivae score and 1.0, 0.7, 0.3 for chemosis score. These effects are reversible between days 2 and 7.

 

Under the test conditions, the test material is not classified as irritating to eyes according to the annex I of the Regulation (EC) No. 1272/2008 (CLP) and to the GHS.

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

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:
- In vitro:
A key study was identified (Laus, 2015). This in vitro skin irritation test using the Reconstructed Human Epidermis (Standard model) was performed according to the OECD Guideline 439 and in compliance with GLP to predict the acute skin irritation potential of the test item.
The mean corrected percent viability of the treated tissues was 100.9%, versus 3.3% in the positive control (5% Sodium Dodecyl Sulfate).
Under the test conditions, the test item was considered as non-irritant to skin.


- In vivo:
A key study was identified (Phycher, 2015). In this dermal irritation study performed according to the OECD Guideline No. 404, and in compliance with GLP, 3 male New Zealand White rabbits were exposed for 4 hours to a semi-occluded application of 0.5 mL of undiluted test material to the intact skin. Initially, a single animal was treated. After consideration of the cutaneous responses produced in the first treated animal, two additional animals were treated.

Slight to well - defined erythema associated with a very slight to slight oedema was noted on the treated area of all animals, 1 hour after the patch removal. All reactions were totally reversible on day 7. On the cutaneous structure, loss of litheness was noted in one animal on day 3 and dryness of the skin in all animals on day 7. The skin recovered a normal aspect on day 14 in two animals but dryness of the skin was still noted on the day 14 in one animal.

The mean scores for each animal within 3 scoring times (24, 48 and 72 h) were 2.0 / 2.0 / 1.0 for erythema and 2.0 / 1.0 / 1.0 for oedema. These effects are reversible on day 7.

Eye irritation:
A key study was identified (Phycher, 2015). In this eye irritation study performed according to the OECD guideline 405, and in compliance with GLP, 0.1 mL of undiluted test material was instilled into one eye of 3 female New Zealand White rabbits. The other eye remained untreated and served as control. The eyes were examined and the changes were observed at 1, 24, 48 and 72 h and Day 7 following treatment and graded according to the Draize method. Initially, a single animal was treated. After consideration of the ocular responses produced in the first treated animal, two additional animals were treated.
A slight redness noted 1 hour after the test item instillation and totally reversible between the 3rd and the 5th day of the test, associated with a slight to moderate chemosis, noted 1 hour after the test item instillation and totally reversible between the 2nd and the 3rd day of the test. At the corneal level, a moderate opacity, registered 24 hours after the test item instillation, in only one animal, and totally reversible the 4th day of the test.

Mean individual scores at 24, 48 and 72 h after exposure for the 3 animals were 0.7, 0.3, 0.3 for cornea score; 0.0, 0.0, 0.0 for iris score; 1.0, 0.7, 0.3 for conjunctivae score and 1.0, 0.7, 0.3 for chemosis score. These effects are reversible between days 2 and 7.


 

Justification for classification or non-classification

Harmonized classification:


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


 


Self classification:


Skin irritation:


Based on the available data:


- no additional self-classification is proposed regarding skin irritation according to the CLP.


- the substance is classified in "Category 3 (mild irritant)" according to the GHS.


 


Eye irritation:


Based on the available data, no additional self-classification is proposed regarding eye irritation according to the CLP and to the GHS.


 


Respiratory irritation:


No data was available regarding respiratory irritation.