Registration Dossier

Administrative data

Description of key information

Skin irritation/corrosion (OECD 404): not irritating

Eye irritation: (OECD 405): not irritating

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
occlusive instead of semi-occlusive dressing; test substance is used as 3% and 20% aqueous solution instead of moistened solid
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
occlusive instead of semi-occlusive dressing; test substance is used as 3% and 20% aqueous solution instead of moistened solid
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Broekman Instituut B.V.
- Age at study initiation: young adult
- Weight at study initiation: 1500-1900 g
- Acclimation period: 4 days
Type of coverage:
occlusive
Preparation of test site:
clipped
Vehicle:
water
Controls:
no
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 mL
- Concentration (if solution): 3% and 20%

Duration of treatment / exposure:
4 hrs
Observation period:
72 hrs
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: 2.5 x 2.5 cm
- Type of wrap if used: Patches fixed with adhesive tape, and covered with occlusive wrap.

REMOVAL OF TEST SUBSTANCE
- Washing: Yes, with water.
- Time after start of exposure: 4 hrs

OBSERVATION TIME POINTS
1, 24, 48, and 72 hrs after end of exposure.

SCORING SYSTEM:
- Method of calculation: Draize.
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritant / corrosive response data:
No signs of redness or swelling was seen at any of the observations at either the 3% or 20% concentration.
Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008
Conclusions:
The test substance is considered to be not irritating to skin.
Executive summary:

Three rabbits were exposed to 3% and 20% test substance in water for 4 hrs under occlusive conditions. The exposed skin was then examined at 1, 24, 48, and 72 hrs after the end of the exposure. No signs of irritation were seen at any of the observations. Thus, under the test conditions used, the test substance was not irritant to the skin of rabbit.

Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
16 - 23 Jan 1995
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
documentation insufficient for assessment
Remarks:
limited documentation; no analytical details on test substance given
Qualifier:
according to
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Deviations:
yes
Remarks:
limited documentation; no analytical details on test substance given; testing was done on scarified and non-scarified skin
GLP compliance:
not specified
Species:
rabbit
Strain:
other: New Zealand
Type of coverage:
not specified
Preparation of test site:
not specified
Vehicle:
not specified
Controls:
not specified
Observation period:
7 days
Number of animals:
3
Details on study design:
OBSERVATION TIME POINTS
1, 24, 48 and 72 h and 7 days

SCORING SYSTEM:
- Method of calculation: Draize scoring system
Irritation parameter:
erythema score
Basis:
mean
Remarks:
of 3 animals
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
other: results for scarified and non-scarified side
Irritation parameter:
edema score
Basis:
mean
Remarks:
of 3 animals
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Remarks on result:
other: results for scarified and non-scarified
Other effects:
No desquamative reaction and no loss of cutaneous extensive range index.

Table 1: Results of skin irritation test

Observation time

Rabbit no.

1

2

3

Erythema

Edema

Erythema

Edema

Erythema

Edema

1 h

1

0

1

0

0

0

24 h

0

0

0

0

0

0

48 h

0

0

0

0

0

0

72 h

0

0

0

0

0

0

7 days

0

0

0

0

0

0

Mean value
24 + 48 + 72 h

0.0

0.0

0.0

0.0

0.0

0.0

Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008
Conclusions:
Under the conditions of the Acute Dermal Irritation/Corrosion Test in rabbits the test substance does not possess any skin irritating potential.
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:
17 - 28 Feb 1997
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:
adopted Feb 24, 1987
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
EEC Directive 92/69/EEC, Dez 1992
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Remarks:
(SPF)
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Broekman Instituute B.V., Someren, Netherlands
- Age at study initiation: 8, 17 or 18 weeks
- Housing: individually in cages with perforated floors
- Diet: Standard laboratory rabbit diet (LKK-20, pellet diameter 4 mm, Hope Farms, Woerden, Netherlands), approx. 100 g per day; hay (BMI, Helmond, Netherlands) once a week
- Water: tap water diluted with decalcified water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 50
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent no treatment
Amount / concentration applied:
TEST MATERIAL
- Amount applied: 63.9 ± 2.6 mg (approx. 0.1 mL)
Duration of treatment / exposure:
single treatment without washing
Observation period (in vivo):
7 days
Reading time points: 1, 24, 48 and 72 h and 7 days
Number of animals or in vitro replicates:
3 males
Details on study design:
SCORING SYSTEM: Draize scoring system

TOOL USED TO ASSESS SCORE: fluorescein

OBSERVATIONS: Animlas were observed for mortality/viability twice daily and for signs of toxicity at least once a day. Body weight was determined on the day of treatment prior to instillation.
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
out of all 3 animals
Time point:
24/48/72 h
Score:
0
Max. score:
4
Reversibility:
other: not applicable
Irritation parameter:
iris score
Basis:
mean
Remarks:
out of all 3 animals
Time point:
24/48/72 h
Score:
0
Max. score:
2
Reversibility:
other: not applicable
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1.3
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.7
Max. score:
3
Reversibility:
fully reversible within: 72 h
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1.7
Max. score:
3
Reversibility:
fully reversible within: 7 days
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
out of all 3 animals
Time point:
24/48/72 h
Score:
0.3
Max. score:
4
Reversibility:
fully reversible within: 48
Irritant / corrosive response data:
Iridic irritation grade 1 was observed in one animal 1 hour after instillation. The irritation of the conjunctivae consisted of redness, chemosis and discharge and had completely resolved within 72 hours in one animal and within 7 days in the other two animals. No corneal opacity or corneal epithelial damage was observed in any of the animals.
Other effects:
- Other observations: No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Table 1. Results of eye irritation test

Alteration

Animal No.

Time (h)

 

24

 

48

 

72

mean 24/48/72 h

reversible within

Cornea

1

0

0

0

0.0

n.a.

2

0

0

0

0.0

n.a.

3

0

0

0

0.0

n.a.

Iris

1

0

0

0

0.0

n.a.

2

0

0

0

0.0

n.a.

3

0

0

0

0.0

n.a.

Conjunctivae Redness

1

2

1

1

1.3

7 d

2

1

1

0

0.7

72 h

3

2

2

1

1.7

7 d

Conjunctivae Chemosis

1

1

0

0

0.3

48 h

2

1

0

0

0.3

48 h

3

1

0

0

0.3

48 h

n.a.: not applicable

Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008
Conclusions:
Under the conditions of this eye irritation study in rabbits the test substance was not irritating to the eye.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin

In vivo

The skin irritation potential of the test substance was determined by an in vivo skin irritation test (Prinsen, 1997) in albino rabbits according to OECD Guideline 404 and in compliance with GLP. 0.5 mL of 3 and 20 % solutions of the test substance in water were applied to the skin of three rabbits under occlusive conditions for 4 hours. After the exposure period the patch was removed and residual test material was removed by water. Scores were taken 1, 24, 48 and 72 hours after patch removal using the Draize scoring system. No signs of redness or swelling were seen at any of the observations at either the 3% or 20% concentration. Based on the results, the undiluted test substance was not irritating to the skin under the conditions of the test.

In a second study, the skin irritation potential of the test substance was determined by an in vivo skin irritation test (Bouffechoux, 1995) in albino rabbits according to OECD Guideline 404. The test substance was applied to the scarified and non-scarified skin of three rabbits. No further details on test method were described. Scores were taken 1, 24, 48 and 72 hours and 7 days after patch removal using the Draize scoring system. No signs of redness or swelling were seen at any of the observation time points at any animal. Based on the results, the test substance was not irritating to the skin under the conditions of the test.

Based on a weight of evidence approach and considering two negative in vivo tests in rabbits, the test substance is not considered to be skin irritating.

 

Eye

In vivo

The eye irritation potential of the test substance was investigated in three albino rabbits according to OECD Guideline 405 (Busschers, 1997) and in compliance with GLP. 64 mg of the test substance was placed in one eye of each animal. The other eye remained untreated and served as control. The eyes were examined and scored according to the Draize scoring system 1, 24, 48, 72 hours and 7 days after instillation. Slight redness of the iris (score 1) was observed in one animal 1 hour after instillation reversible within 1 day. Redness of the conjuctivae (score 1-2) was observed in all animals 1 h after instillation of the test substance reversible within 72 h in one animal and within 7 days for two animals. Conjunctivae chemosis (score 2) was observed in all three animals 1 h after instillation reversible within 48 h. Cornea was not affected by the test substance. The mean scores for all animals at 24/48/72 h were 0.0, 0.0 and 0.3 for cornea, iris and chemosis, respectively. Scores for conjunctivae redness were 1.3, 0.7 and 1.7 for each animal respectively. Based on the results of this study, the test substance was not irritating to the eyes under the conditions of the test.

The eye irritation potential of the test substance was investigated in three albino rabbits according to OECD Guideline 405 (Prinsen, 1997) and in compliance with GLP. 0.1 mL of the test substance was placed in one eye of each animal. The other eye remained untreated and served as control. The eyes were examined and scored according to a 4-point scoring system 1, 24, 48 and 72 hours after instillation. Slight redness (score 1) and slight to moderate swelling (score 1-2) was noted in each animal at the 1 hour observation. At the 24 hour observation, all 3 animals showed slight redness (score 1), and 2 showed slight swelling (score 1) as well. 1 animal showed slight discharge. At the 48 hour observation, only 1 animal showed treatment related effects, slight redness. By the 72 hour observation, no animals showed any treatment related effects. The mean scores at 24/48/72 hour for all animals were 0.0 for cornea and iris, respectively. Mean scores at 24/48/72 hours for conjunctivae redness were 0.7, 0.3 and 0.3 and for chemosis 0.3 and 0.3, respectively for each animal. Based on the results of this study, the test substance was not irritating to the eyes under the conditions of the test.

 

In a further study the eye irritation potential of the test substance was investigated in three albino rabbits according to OECD Guideline 405 (Bouffechoux, 1995). An unknown amount of the test substance was placed in one eye of each animal. The eyes were examined and scored according to the Draize scoring system 1, 24, 48 and 72 hours and 7 days after instillation. Slight redness of conjunctivae (score 1) was noted in all animals 1 hour after instillation. The effects were reversible within 24 h. The mean scores for all animals at 24/48/72 hour were 0.0, 0.0 and 0.33 for cornea, iris and conjunctivae redness, respectively. Based on the results of this study, the test substance was not irritating to the eyes under the conditions of the test.

 

In vitro

The eye irritation potential of the test substance was determined in a bovine corneal opacity and permeability test (BCOP test) according to OECD Guideline 437 and in compliance with GLP (Sakakibara, 2016). After a first opacity measurement of the fresh bovine corneae, 750 µl of a 20% (w/v) solution of the test substance in saline was applied directly to the epithelial surface of three cattle corneae in an incubation chamber in horizontal position for 4 hours at 32 ± 1 °C. After the incubation phase the test substance was rinsed from the corneae. Afterwards, opacity was measured a second time. In addition, the permeability of the corneae was determined by measuring spectrophotometrically the transfer of sodium fluorescein after incubation in a horizontal position for 90 min at 32 ± 1 °C. The results of the opacity and permeability measurement of the test substance at 20% solution were used to calculate a mean in vitro irritation score (IVIS) of 0.3. With the negative control saline neither an increase of opacity nor permeability of the corneae could be observed. The mean IVIS of the negative control was -1.3. The mean IVIS of the positive control (imidazole 20% (w/v)) was 95.2, thus confirming the validity of the test system. Based on the results, a 20% solution of the test substance was not irritating to the eye under the conditions of the test.

Justification for classification or non-classification

The available data on skin and eye irritation do not meet the criteria for classification according to Regulation (EC) 1272/2008, and are therefore conclusive but not sufficient for classification.