Registration Dossier

Administrative data

Description of key information

Based on the results of a study according to OECD Test Guideline 404 under GLP, the test substance is considered to be skin irritating.
Based on the results of a study according to OECD Test Guideline 431 under GLP, the test substance is considered to be not skin corrosive.
Based on the results of a study according to OECD Test Guideline 405 under GLP, the test substance is considered to be eye irritating.

Based on the results of a study according to OECD Test Guideline 437 under GLP, the test substance did not induce any effects warranting a classification as Category 1, however, a classification as Category 2 ccould not be excluded.

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:
02.2009 - 04.2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
Version / remarks:
April 2004
GLP compliance:
yes
Test system:
human skin model
Source species:
human
Cell type:
other: EpiDerm tissue
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm TM tissues
- Tissue batch number(s):
9640 Kit N
- Delivery date:
02/17/09
- Date of initiation of testing:
02/18/09

REMOVAL OF TEST MATERIAL AND CONTROLS
- tissue was rinsed with phosphate buffered saline

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration:
1mg/ml MTT diluted in Dulbecco´s Modified Eagle Medium (DMEM)
- Incubation time: 3h
- Spectrophotometer:
microplate reader
- Wavelength:
540 nm

NUMBER OF REPLICATE TISSUES:
duplicate

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Frozen tissues
- Method of calculation used:
% viability = 100 X (OD sample / OD negative control)

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

NEGATIVE CONTROL
- Amount applied: 50µl
- Concentration: not applicable

POSITIVE CONTROL
- Amount applied: 50µl
- Concentration: 8.0 N
Duration of treatment / exposure:
3min, 60min
Number of replicates:
2
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Basis: mean in %. Time point: 3 minutes
Value:
105.5
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Basis: mean in %. Time point: 60 minutes
Value:
102.5
Interpretation of results:
GHS criteria not met
Conclusions:
The test substance with a purity of 54.5% and in the presence of other chemicals originating from the production process (petroleum and nonylphenol) was not corrosive in the in vitro test according to OECD 431.
Executive summary:

The skin corrosion potential of the test substance with a purity of 54.5% and in the presence of other chemicals originating from the production process was evaluated in a study according to OECD Test Guideline 431 under GLP. The substance was applied unchanged for 60 minutes to in vitro skin tissue. After 60 minutes cell viability was not reduced. Interpreting these results by applying the evaluation system of the EU regulation 1272/2008, the test substance is considered as not corrosive to the skin.

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
1992
GLP compliance:
yes
Species:
rabbit
Strain:
other: Mol:Russian
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Mollegaard Breeding and Research Centre
- Weight at study initiation: 2.1-2.4 kg
- Housing: individually
- Diet: pelleted complete rabbiet diet "Altromin 2123"; ad libitum
- Water: domestic quality drinking water acidified with hydrochloric acid to pH 2.5; ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +- 3
- Humidity (%): 55 +- 15
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5ml
Duration of treatment / exposure:
4 h
Observation period:
21d
Number of animals:
3
Details on study design:
The day before the experiment was started the rabbits were weighed and an area of 10 x 10 cm on the back was clipped as closely as possible with an electric clipper. On the experimental day the rabbits were physically restrained on a test table, and the clipped area was divided into four fields. The two anterior fields were used for testing the experimental preparation. To each of two 16-layer gauze patches (2.5 x 2.5 cm) 0.5 ml of the
test article was applied, and the patches were placed on the appropriate test site on the back of each rabbit. The gauze patches were secured with 2.5 cm wide adhesive tape and fixed with Micropore tape, 5 cm width, loosely wound round the trunk.
After an exposure time of 4 hours the tape and patches were removed, the test fields were marked and the treated skin was cleaned with lukewarm water. The skin reactions were read 1 hour later.
Reading was also made 24, 48 and 72 hours as weIl as 7, 14 and 21 days after termination of exposure.
The respective scores for erythema and oedema formation for the 3 readings 24, 48 and 72 hours of each rabbit were summed up separately and divided by 6. The results are the mean scores for erythema and oedema formation of the individual rabbit.
The mean scores for erythema and oedema formation in 3 rabbits used were subsequently calculated.
Irritation parameter:
erythema score
Basis:
mean
Time point:
24/48/72 h
Score:
2.9
Max. score:
3
Reversibility:
not fully reversible within: 21 d
Irritation parameter:
edema score
Basis:
mean
Time point:
24/48/72 h
Score:
2.9
Max. score:
4
Reversibility:
fully reversible within: 21 d
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
other: 1 h
Score:
0
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24 h
Score:
2
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
48 h
Score:
3
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
72 h
Score:
3
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #1
Time point:
other: 1 h
Score:
1
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24 h
Score:
2
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #1
Time point:
48 h
Score:
3
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #1
Time point:
72 h
Score:
3
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
other: 1 h
Score:
1
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24 h
Score:
3
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
48 h
Score:
3
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
72 h
Score:
3
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #2
Time point:
other: 1 h
Score:
1
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24 h
Score:
3
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #2
Time point:
48 h
Score:
3
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #2
Time point:
72 h
Score:
3
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
other: 1 h
Score:
1
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24 h
Score:
3
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
48 h
Score:
3
Max. score:
4
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
72 h
Score:
3
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #3
Time point:
other: 1 h
Score:
1
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24 h
Score:
3
Max. score:
4
Remarks on result:
other: left test field
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24 h
Score:
4
Max. score:
4
Remarks on result:
other: right test field
Irritation parameter:
edema score
Basis:
animal #3
Time point:
48 h
Score:
3
Max. score:
4
Irritation parameter:
edema score
Basis:
animal #3
Time point:
72 h
Score:
3
Max. score:
4
Irritant / corrosive response data:
One hour after the end of application no erythema was seen in animal No. 833. In animal Nos. 671 and 678 a slight erythema was observed. In all rabbits a very slight oedema was seen at this time.
24 hours after removal of patches animal No. 833 showed a well-defined erythema and a slight oedema on both test sites. In animal No. 671 a moderate erythema and a moderate oedema was observed on both test sites. Animal No. 678 showed a moderate to severe erythema on
both test sites and a moderate oedema on left test area resp. a severe oedema on right test area.
The test areas of all three rabbits showed a moderate to severe erythema and a moderate oedema as weIl after 24 resp. 48 hours.
All test sites were completely covered with scales on day 7.
The test areas were easily discernible and the scales were almost completely fall off.
After 21 days the scales seemed to be removed from the test areas, in animal Nos. 671 and 833 a slight redness and an increased hair growth on the test fields were observed.
The mean individual erythema scores were 2.67, 3.00, 3.00, respectively.
The mean individual oedema scores were 2.67, 3.00, 3.17, respectively.
The mean score for the test article was 2.9 for erythema and 2.9 for oedema.
Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
Based on the results of a study according to OECD Test Guideline 404 under GLP, the test substance is considered as skin irritating (applying the evaluation system of the EU regulation 1272/2008).
Executive summary:

The skin irritation potential of the test substance was evaluated in a study according to OECD Test Guideline 404 under GLP.The amount of 0.5 ml of the substance was applied unchanged under semi-occlusive conditions for 4 hours to the shaved skin of three Mol:Russian rabbits, which were observed for 21 days post application. Over the first three days, clear erythema and oedema were observed in all rabbits, which almost fully reversed within 21 days. Interpreting these results by applying the evaluation system of the EU regulation 1272/2008, the test substance is considered irritating to the skin.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Referenceopen allclose all

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
adopted: 24th April 2002
GLP compliance:
yes (incl. QA statement)
Species:
rabbit
Strain:
other: Crl:CHBB (HM) Russian rabbit
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland GmbH
- Age at study initiation: 9-12 months
- Weight at study initiation: 2.3-3.3 kg
- Housing: individually
- Diet: ad libitum
- Water: ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +- 3
- Humidity (%): 30-70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
other: untreated eye of the same animals
Amount / concentration applied:
0.1 ml
Duration of treatment / exposure:
single application
Observation period (in vivo):
21 days
Number of animals or in vitro replicates:
3
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): no

SCORING SYSTEM: according to OECD 405 guideline

TOOL USED TO ASSESS SCORE: light and fluorescein
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
other: mean (24/48/72 hours)
Score:
1.33
Reversibility:
fully reversible within: 7d
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
other: mean (24/48/72 hours)
Score:
1
Reversibility:
fully reversible within: 7d
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
other: mean (24/48/72 hours)
Score:
0.67
Reversibility:
fully reversible within: 7d
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: mean (24/48/72 hours)
Score:
0.67
Reversibility:
fully reversible within: 7d
Irritation parameter:
iris score
Basis:
animal #2
Time point:
other: mean (24/48/72 hours)
Score:
0.67
Reversibility:
fully reversible within: 7d
Irritation parameter:
iris score
Basis:
animal #3
Time point:
other: mean (24/48/72 hours)
Score:
0.33
Reversibility:
fully reversible within: 7d
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
other: mean (24/48/72 hours)
Score:
2.33
Reversibility:
fully reversible within: 7d
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
other: mean (24/48/72 hours)
Score:
1.67
Reversibility:
fully reversible within: 7d
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
other: mean (24/48/72 hours)
Score:
1.67
Reversibility:
fully reversible within: 7d
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: mean (24/48/72 hours)
Score:
2
Reversibility:
fully reversible within: 7d
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: mean (24/48/72 hours)
Score:
1.33
Reversibility:
fully reversible within: 7d
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: mean (24/48/72 hours)
Score:
1.67
Reversibility:
fully reversible within: 7d
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
other: 1 h
Score:
1
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24 h
Score:
1
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24 h
Score:
2
Max. score:
4
Remarks on result:
other: reading after installation of Fluorescein
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
48 h
Score:
1
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
48 h
Score:
1
Max. score:
4
Remarks on result:
other: reading after installation of Fluorescein
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
72 h
Score:
1
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
72 h
Score:
1
Max. score:
4
Remarks on result:
other: reading after installation of Fluorescein
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
7 d
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
7 d
Score:
0
Max. score:
4
Remarks on result:
other: reading after installation of Fluorescein
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
14 d
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
21 d
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
animal #1
Time point:
other: 1 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24 h
Score:
1
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #1
Time point:
48 h
Score:
1
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #1
Time point:
72 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #1
Time point:
7 d
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #1
Time point:
14 d
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #1
Time point:
21 d
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
other: 1 h
Score:
1
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24 h
Score:
2
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
48 h
Score:
3
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
72 h
Score:
2
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
7 d
Score:
0
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
14 d
Score:
0
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
21 d
Score:
0
Max. score:
3
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
other: 1 h
Score:
1
Max. score:
4
Remarks on result:
other: moderate flow of tears
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24 h
Score:
3
Max. score:
4
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
48 h
Score:
2
Max. score:
4
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
72 h
Score:
1
Max. score:
4
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
7 d
Score:
0
Max. score:
4
Remarks on result:
other: tight scab on the upper eyelid
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
14 d
Score:
0
Max. score:
4
Remarks on result:
other: without scab, but with a hairless fringe of the eyelid
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
21 d
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
other: 1 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24 h
Score:
1
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24 h
Score:
2
Max. score:
4
Remarks on result:
other: reading after installation of Fluorescein
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
48 h
Score:
1
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
48 h
Score:
1
Max. score:
4
Remarks on result:
other: reading after installation of Fluorescein
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
72 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
72 h
Score:
0
Max. score:
4
Remarks on result:
other: reading after installation of Fluorescein
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
7 d
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
14 d
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
21 d
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
animal #2
Time point:
other: 1 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24 h
Score:
1
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #2
Time point:
48 h
Score:
1
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #2
Time point:
72 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #2
Time point:
7 d
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #2
Time point:
14 d
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #2
Time point:
21 d
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
other: 1 h
Score:
1
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24 h
Score:
2
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
48 h
Score:
2
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
72 h
Score:
1
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
7 d
Score:
0
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
14 d
Score:
0
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
21 d
Score:
0
Max. score:
3
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
other: 1 h
Score:
1
Max. score:
4
Remarks on result:
other: mild flow of tears
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24 h
Score:
2
Max. score:
4
Remarks on result:
other: moderate flow of tears
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
48 h
Score:
2
Max. score:
4
Remarks on result:
other: mild flow of tears
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
72 h
Score:
0
Max. score:
4
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
7 d
Score:
0
Max. score:
4
Remarks on result:
other: moderate scab on the upper eyelid and moderate loss of hair around the eye
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
14 d
Score:
0
Max. score:
4
Remarks on result:
other: without scab, but with a moderate loss of hair on the upper eyelid
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
21 d
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
other: 1 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24 h
Score:
1
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24 h
Score:
1
Max. score:
4
Remarks on result:
other: reading after installation of Fluorescein
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
48 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
48 h
Score:
1
Max. score:
4
Remarks on result:
other: reading after installation of Fluorescein
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
72 h
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
7 d
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
14 d
Score:
0
Max. score:
4
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
21 d
Score:
0
Max. score:
4
Irritation parameter:
iris score
Basis:
animal #3
Time point:
other: 1 h
Score:
1
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24 h
Score:
1
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #3
Time point:
48 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #3
Time point:
72 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #3
Time point:
7 d
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #3
Time point:
14 d
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #3
Time point:
21 d
Score:
0
Max. score:
2
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
other: 1 h
Score:
2
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24 h
Score:
2
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
48 h
Score:
2
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
72 h
Score:
1
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
7 d
Score:
0
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
14 d
Score:
0
Max. score:
3
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
21 d
Score:
0
Max. score:
3
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
other: 1 h
Score:
2
Max. score:
4
Remarks on result:
other: moderate flow of tears
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24 h
Score:
3
Max. score:
4
Remarks on result:
other: moderate flow of tears
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
48 h
Score:
2
Max. score:
4
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
72 h
Score:
0
Max. score:
4
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
7 d
Score:
0
Max. score:
4
Remarks on result:
other: mild scab on the upper eyelid and moderate loss of hair around the eye
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
14 d
Score:
0
Max. score:
4
Remarks on result:
other: without scab, but with mild loss of hair on the upper eyelid
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
21 d
Score:
0
Max. score:
4

One hour after the application of the test item, animal No. 4409 showed scattered or diffuse areas of opacity with details of iris clearly visible on about one quarter of the cornea area, some hyperaemic conjunctival blood vessels, a swelling above normal and a moderate flow of tears. Some hyperaemic conjunctival blood vessels, as welling above normal and a mild flow of tears were observed in animal No. 4410.

Animal No. 4414 showed an iris with markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia and an existing reaction to light, a conjunctiva with diffuse crimson colour and individual vessels not easily discernible, an obvious swelling with partial eversion of Iids as weil as a moderate flow of tears. 24 hours after the application of the test item, scattered or diffuse areas of opacity

with details of iris clearly visible on more than three quarters of the cornea area, an iris with markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia and an existing reaction to light, a conjunctiva with diffuse crimson colour and individual vessels not easily discernible, a swelling with Iids about half closed and a moderate flow of tears were observed in animal No. 4409. Animal No. 441 0 showed scattered or diffuse areas of opacity with details of iris clearly visible on about one quarter of the cornea area, an iris with markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia and an existing reaction to light, a conjunctiva with diffuse crimson colour and individual vessels not easily discernible, an obvious swelling with partial eversion of Iids and a moderate flow of tears. Scattered or diffuse areas of opacity with details of iris clearly visible on about one quarter of the cornea area, an iris with markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia and an existing reaction to light, a conjunctiva with diffuse crimson colour and individual vessels not easily discernible, a swelling with Iids about half closed and a moderate flow of tears were observed in animal No. 4414.

After the instillation of Fluorescein, animal No. 4409 showed an easily discernible translucent area with details of the iris slightly obscured on more than half, but less than three quarters of the cornea area. An easily discernible translucent area with details of the iris slightly obscured was observed on more than one quarter, but less than half of the cornea area of animal No. 4410, whereas animal No. 4414 showed

scattered or diffuse areas of opacity with details of iris clearly visible on more than one quarter, but less than half of the cornea area. 48 hours after the application of the test item, scattered or diffuse areas of opacity with details of iris clearly visible on more than three quarters of the cornea area, an iris with markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia and an existing reaction to light, a conjunctiva with diffuse beefy red colour and an obvious swelling with partial eversion of Iids were observed in animal No. 4409. Anima I No. 4410 showed scattered or diffuse areas of opacity with details of iris clearly visible on about one quarter of the cornea area, an iris with markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia and an existing reaction to light, a conjunctiva with diffuse crimson colour and individual vessels not easily discernible, an obvious swelling with partial eversion of Iids and a mild flow of tears. A conjunctiva with diffuse crimson colour and individual vessels not easily discernible as weil as an obvious swelling with partial eversion of Iids were observed in animal No. 4414.

After the instillation of Fluorescein, animal No. 4409 showed scattered or diffuse areas of opacity with details of iris clearly visible on more than one quarter, but less than half of the cornea area. Scattered or diffuse areas of opacity with details of iris clearly visible were observed on about one quarter of the cornea area of animal No. 4410 and No. 4414.

72 hrs after the application of the test item, animal No. 4409 showed scattered or diffuse areas of opacity with details of iris clearly visible on more than one quarter, but less than half of the cornea area, a conjunctiva with diffuse crimson colour and individual vessels not easily discernible as weil as a swelling above normal. Some hyperaemic conjunctival blood vessels were observed in animal No. 4410 and

No. 4414.

After the instillation of Fluorescein, animal No. 4409 showed scattered or diffuse areas of opacity with details of iris clearly visible on about one quarter of the cornea area.

Seven days after the application of the test item, no ocular reactions were observed in the eye of animal No. 4409, but tight scab was revealed on the upper eyelid.

Anima I No. 4410 showed moderate formation of scab on the upper eyelid and loss of hair around the eye. Mild formation o scab and loss of hair around the eye was observed in animal No. 4414.

Fourteen days after the application of the test item, animal No. 4409 still showed a hairless fringe of the eye but scab not any longer. The upper eyelid of animal No. 4410 was very hairless but free of scab. The loss of hair on the upper eyelid of animal No. 4414 was mild and scab was not present any longer. Twenty-one days after the application of the test item, animal No. 4409, No. 4410

and No. 4414 was free of signs of irritation. The hair was grown again in the adjacency of the eye.

Interpretation of results:
Category 2 (irritating to eyes) based on GHS criteria
Conclusions:
The test substance is classified as irritating to eyes according to category 2 of the regulation (EC) No 1272/2008 (2008)7, because of the data concerning the opacity of the cornea obtained after the use of fluorescein.
Executive summary:

The local effect of Ethanone, 1-(2-hydroxy-5-nonylphenyl)-, oxime, branched was investigated according to the method recommended in the OECD Guideline 405 (2002) and the council regulation (EC) No 440/2008 part B.5. (2008).

During an initial test, an amount of 0.1 ml of the undiluted test item was applied in one eye of a female albino rabbit. The eye was examined and the changes were graded according to a numerical scale after 1 hr, 24 hrs, 48 hrs and 72 hrs as well as on day 7,14 and 21. Well defined changes were observed in the treated eye. Since the animal used during the initial test did not show corneal defects on day 7, two more female albino rabbits were treated with 0.1 ml of the undiluted test item each. Likewise ocular defects were graded after 1 hr, 24 hrs, 48 hrs and 72 hrs as well as on day 7, 14 and 21. The two test animals showed also well defined changes concerning eyes. All observed effects were fully reversible within 7 days.

Based on the results of the study described in this report Ethanone, 1-(2-hydroxy-5-nonylphenyl)-, oxime, branched

• shall not be classified as irritating to eyes according to annex VI of the directive 67/548/EEC (2009)3 as well as the directive 2006/12 11EC (2006, 2007)4.S.

• shall be classified as irritating to eyes according to category 2B of the GHS (2007)6, because of the data concerning the opacity of the cornea obtained after the use of Fluorescein.

• shall be classified as irritating to eyes according to category 2 of the regulation (EC) No 1272/2008 (2008)7, because of the data concerning the opacity of the cornea obtained after the use of Fluorescein.

Endpoint:
eye irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
10.2009 - 01.2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)
Version / remarks:
September 2009
GLP compliance:
yes (incl. QA statement)
Species:
other: bovine
Details on test animals or tissues and environmental conditions:
SOURCE OF COLLECTED EYES
- Source: Odenwaldschlaohthof Brensbach, Germany
- Number of Corneae: 9
- Characteristics of donor animals (e.g. age, sex, weight): no data
- Storage, temperature and transport conditions of ocular tissue: Excess tissue was removed from the excised eyes and they were contained and transported in Hank's BSS supplemented with streptomycin / penicillin at room temperature. The corneae were isolated immediatey after delivery of the eyes to the laboratory.
- Time interval prior to initiating testing: The bovine eyes were fetched from the slaughterhouse and stored in the refrigerator at about 2 -8 'C until preparaton of the comeae on the same day.
- indication of any existing defects or lesions in ocular tissue samples: eyes presenting defects such as vascularization, pigmentation, opacity and scratches were discarded
- Indication of any antibiotics used: yes, streptomycin I penicillin
Controls:
yes, concurrent positive control
yes, concurrent negative control
Duration of treatment / exposure:
The incubation time lasted ten minutes (± 30 seconds).
Duration of post- treatment incubation (in vitro):
After the test item or control items, respectively, were rinsed off from the application side by changing cMEM several times, in minimum three times, fresh cMEM was added and opacity was measured (t10). The comeae were then incubated at 32 ± 2 *C for further two hours in a vertical position, followed by a third opacity reading (t130).
Number of animals or in vitro replicates:
3
Irritation parameter:
other: opacity and permeability
Run / experiment:
test substance
Value:
1.52
Irritation parameter:
other: opacity and permeability
Run / experiment:
negative control
Value:
0.91
Irritation parameter:
other: opacity and permeability
Run / experiment:
positive control
Value:
76.33
Interpretation of results:
GHS criteria not met
Conclusions:
Tested in the BCOP, the test substance did not induce any effects warranting a classification as Category 1.
However, this results does not allow to conclude that the test substance has not to be classified as Category 2.
Executive summary:

When tested using the BCOP test protocol, the test substance did not induce any effects warranting a classification as Category 1.However, this results does not allow to conclude that the test substance has not to be classified as Category 2. Consequently, a confirmation using an in vivo protocol is required.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation/corrosion:

An in vitro study according to OECD test guideline 431 and GLP showed that the test substance is not skin corrosive. This complements the results of an in vivo study according to OECD Test Guideline 404 and GLP. Three female albino rabbits were exposed to the test article at two skin sites on the back. After 4 hours of exposure the test article was removed and the skin was examined 1, 24, 48 and 72 hours as well as 7, 14 and 21 days after termination of exposure.

The test article caused a moderate to severe erythema and a moderate oedema in all three rabbits.

Under the experimental conditions described in this report, the mean score for erythema was 2.9 and for oedema was 2.9.

The test substance is thus irritating to the skin.

Eye irritation:

The local effect of the registered substance was investigated according to the method recommended in the OECD Guideline 405 (2002) and the council regulation (EC) No 440/2008 part B.5. (2008). During an initial test, an amount of 0.1 ml of the undiluted test item was applied in one eye of a female albino rabbit. The eye was examined and the changes were graded according to a numerical scale after 1 hr, 24 hrs, 48 hrs and 72 hrs as well as on day 7, 14 and 21. Well defined changes were observed in the treated eye. Since the animal used during the initial test did not show corneal defects on day 7, two more female albino rabbits were treated with 0.1 ml of the undiluted test item each. Likewise ocular defects were graded after 1 hr, 24 hrs, 48 hrs and 72 hrs as well as on day 7, 14 and 21. The two test animals showed also well defined changes concerning the eyes.

The mean scores (24/48/72 hours) in the individual animals were:

- corneal opacity (without fluorescein): 1.00/0.67/0.33

- corneal opacity (with fluorescein): 1.33/1.00/0.67

- iris lesions: 0.67/0.67/0.33

- redness of the conjunctiva: 2.33/1.67/1.67

- Oedema of the conjunctiva: 2.00/1.33/1.67

All observed effects were fully reversible within 7 days.

Additionally, an in vitro study was performed to assess the corneal irritation and damage potential of the test substance by means of the BCOP assay using fresh bovine corneae. After a first opacity measurement of the fresh bovine corneae (k), the neat test item, the positive, and the negative controls were applied to corneae and incubated for 10 minutes at 32 ± 2 °C in complete medium. After the incubation phase the test item, the positive, and the negative controls were each rinsed from the corneae and opacity was measured again (t10). Further, the corneae were incubated for another 120 minutes at 32 ± 2 °C in complete medium, and opacity was measured a third time (t130). After the opacity measurements permeability of the corneae was determined while application of 1 mL of a fluorescein solution for 90 minutes at 32 ± 2 °C in a horizontal position. The coming out liquid was measured spectrophotometricaly.

With the negative control (0.9% NaCI solution) neither an increase of opacity nor permeability of the corneae could be observed. The mean in vitro score was calculated as 0.91.

The positive control (2-Ethoxyethanol) showed clear opacity and distinctive permeability of the comeae and therefore, is classified as severe eye irritant. The mean in vitro score was calculated as 76.33.

The test item did not cause any opacity or permeability of the corneae compared with the results of the negative control. The calculated mean in vitro score was 1.52 and therefore, the test item was classified as not corrosive to the eye.


Justification for classification or non-classification

Skin irritation/corrosion:

Interpreting the results of both the in vitro and in vivo test by applying the evaluation system of the EU regulation 1272/2008 and EU directive 67/548/EEC, the test substance is considered irritating to the skin.

Eye irritation:

Based on the results of an in vivo study the registered substance • shall not be classified as irritating to eyes according to annex VI of the directive 67/548/EEC (2009) as well as the directive 2006/12 11EC (2006, 2007) • shall be classified as irritating to eyes according to category 2B of the UN-GHS (Rev. 8, 2019), because of the data concerning the opacity of the cornea obtained after the use of Fluorescein. • shall be classified as irritating to eyes according to category 2 of the regulation (EC) No 1272/2008 (2008)7, because of the data concerning the opacity of the cornea obtained after the use of Fluorescein.