Registration Dossier

Administrative data

Description of key information

Skin irritation: irritating to the skin, EU Method B.4, Toxicol Laboratories Limited 1992
Eye irritation: not severe irritant or corrosive to the eye, OECD 473, Harlan Laboratories Ltd. 2013
Eye irritation: not an eye irritant, EpiOcular EIT-100, Harlan CCR 2014

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1992
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Method followed similar to a recognised guideline; performed under GLP.
Qualifier:
equivalent or similar to
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: reognised animal source
- Age at study initiation: not reported
- Weight at study initiation: 2.3 - 2.5 kg
- Housing: individually housed in grid-bottomed metal cages.
- Diet (e.g. ad libitum): certified pelleted diet ad libitum
- Water (e.g. ad libitum): mains water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 - 21
- Humidity (%): 39 - 65
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12 hours dark / 12 hours light
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
no
Amount / concentration applied:
0.5ml test material applied to clipped backs of test animals.
Duration of treatment / exposure:
4 hours
Observation period:
skin reaction to the test article was assessed after one, 24, 48 and 72 hours. Additional assessments were carried out 7 and 14 days after dosing.
Number of animals:
4 females
Details on study design:
TEST SITE
- Area of exposure: doral surface of the trunk
- % coverage: not reported
- Type of wrap if used: Elastoplast elastic adhesive bandage 7.5cm wide.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes - test substance application site cleansed using cotton wool soaked in warm water
- Time after start of exposure: 4 hours

Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 1 hour
Score:
0.75
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 24 hour
Score:
2
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 48 hour
Score:
2.75
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 72 hour
Score:
3
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 7 days
Score:
2.75
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
erythema score
Basis:
mean
Time point:
other: 14 days
Score:
1.25
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 1 hour
Score:
2
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 24 hour
Score:
2.5
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 48 hour
Score:
3
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 72 hour
Score:
3
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 7 days
Score:
2.75
Max. score:
4
Reversibility:
fully reversible
Irritation parameter:
edema score
Basis:
mean
Time point:
other: 14 days
Score:
0
Max. score:
4
Reversibility:
fully reversible
Irritant / corrosive response data:
One hour after dosing all animals exhibited slight oedema (grade 2) and most exhibited very slight erythema (grade 1). By 24 hours all animals exhibited well-defined erythema (grade 2) and slight to moderate oedema (grades 2-3). Responses increased until at the 72 hour examination all animals exhibited moderate to severe erythema and moderate oedema (grade 3). Seven days after dosing most animals were noted to have the same degree of irritation, except one animal, which exhibited grade 2 erythema and oedema. At this examination all animals were also noted to have desquamation. Fourteen days after dosing all the animals still exhibited very slight to well-defined erythema (grades 1-2) and one animal had desquamation and skin thickening but no oedema was observed on any animal.
Interpretation of results:
irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of the study the test substance is considered to be irritating to skin.
Executive summary:

The study was performed to assess the irritancy potential of the test substance to the skin of the New Zealand White rabbit. The study was performed to GLP and followed a method similar to the EU Method B.4. The test substance was applied over an area of approximately 2.5cm square to the clipped backs of 4 albino rabbits. The test substance was held in contact with the skin under a semi-occlusive patch for a 4 hour period. After this time the patches were removed and the skin reaction to the test substance was assessed after one, 24, 48 and 72 hours. Additional assessments were carried out 7 and 14 days after dosing to determine the degree of reversibility of the skin reaction. One hour after dosing all animals exhibited slight oedema (grade 2) and most exhibited very slight erythema (grade 1). By 24 hours all animals exhibited well-defined erythema (grade 2) and slight to moderate oedema (grades 2-3). Responses increased until at the 72 hour examination all animals exhibited moderate to severe erythema and moderate oedema (grade 3). Seven days after dosing most animals were noted to have the same degree of irritation, except one animal, which exhibited grade 2 erythema and oedema. At this examination all animals were also noted to have desquamation. Fourteen days after dosing all the animals still exhibited very slight to well-defined erythema (grades 1-2) and one animal had desquamation and skin thickening but no oedema was observed on any animal. The calculated mean score for erythema or eschar formation was 2.6. The calculated mean score for oedema was 2.8. It was concluded that the test material is irritating to the skin of the albino rabbit.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Eye irritation

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

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation:

The study was performed to assess the irritancy potential of the test material to the skin of the New Zealand White rabbit. The study was performed to GLP and followed a method similar to the EU Method B.4. The test substance was evaluated in 4 female New Zealand white rabbits. A dose of 0.5 ml test substance (undiluted), was applied to the intact clipped dorsal skin site under a semi-occlusive dressing for 4 hours. The patch was then removed and the application site was washed to remove residual test material. Skin observations were made 1, 24, 48, 72 hours and 7, 14 days after patch removal. A single 4-hour, semi-occluded application of the test material to the intact skin produced irritation effects. One hour after dosing all animals exhibited slight oedema and most exhibited very slight erythema. Fourteen days after dosing all the animals still exhibited very slight to well-defined erythema and one animal had desquamation and skin thickening but no oedema was observed on any animal. Under the conditions of this study the test material is considered to be irritating to skin of New Zealand white rabbits.

 

Eye irritation:

OECD 473, 2013 - The study was performed to assess the eye irritancy potential of the test material in isolated bovine corneas. The study was performed under GLP and followed a method equivalent to OECD guideline 437. The ocular irritancy of the test substance was tested through topical application for 10 ± 1 minutes. The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (ethanol), was 36.2 and was within the historical positive control data range (22.5 to 60.9). The test substance did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of 23.5 after 10 minutes of treatment. Under the conditions of this study the test material is not considered to be a severe irritant or corrosive in the Bovine Corneal Opacity and Permeability test.

 

In vitro EpiOcular EIT-100, 2014 - The study was performed to assess the irritancy potential of the test material to the eye means of the Human Cornea Model Test using the EpiOcular™ model . The study was performed to GLP. The method was according to manufacturer guideline. Tissues of the human cornea model EpiOcular™ were pre-treated for 30 minutes with PBS. The tissues were exposed to each 50 uL of the test item and of the positive (1% (w/v) Benzalkonium chloride in deionised water) and the negative control (deionised water) for 30 minutes in triplicate. Afterwards the cell viability was determined using the MTT assay. After treatment with the negative control, the mean OD of the three tissues was >= 1.0. Compared with the value of the negative control, treatment with the positive control induced a sufficient decrease in the relative absorbance, and the mean tissue viability was <= 60% of the negative control. The relative standard deviation between each the three identically treated equivalents of each dose group was <= 20%. Consequently, the test met the acceptance criteria. No irritating effects were observed following incubation with test substance. The relative absorbance values of the test item, corresponding to the cell viability, did not decrease below 60% compared with the result of the negative control.


Justification for selection of skin irritation / corrosion endpoint:
Study selected is an in vivo study (Klimisch 2)

Justification for selection of eye irritation endpoint:
Two in vitro GLP compliant studies. No study is selected since a Weight of Evidence determination is made based on both in vitro studies.

Effects on skin irritation/corrosion: irritating

Justification for classification or non-classification

The substance meets classification criteria under EU Directive 67/548/EEC for dermal irritation: R38.

The substance meets classification criteria under Regulation (EC) No 1272/2008 for dermal irritation category 2.

The substance classification does not meet the classification criteria under EU Directive 67/548/EEC for eye irritation.

The substance classification does not meet the classification criteria under Regulation (EC) No 1272/2008 for eye irritation.

For eye irritation, the weight of evidence according to REACH Regulation (EC) 1907/2006: Annex XI section 1.2 and with regard to Article 13 (3): indicates the substance is predicted to be not irritating to the eye. The substance in a two tier testing strategy is confirmed to not be a severe irritant in a GLP guideline in vitro BCOP study according to OECD 473. The substance is confirmed to not be an irritant in an in vitro EpiOcular EIT-100 study under GLP using 60% viability cut-off. Additionally there is no evidence or history of worker incidents that the substance meets the classification criteria as eye irritant. Furthermore based on expert judgement and with regard to literature published by Scott et al. and Kolle et al. it is considered that the substance does not meet the classification criteria for eye. The information is deemed to meet the tonnage driven information requirements of the Regulation (EC) 1907/2006.

 

References:

ECHA Guidance on Information Requirements R7.a: section 7.2; 166-200, version 2.4, 2014

L. Scott et al., Toxicology in Vitro 24, 1–9, 2010

S. Kolle et al., ATLA 39, 365–387, 2011