Registration Dossier

Administrative data

Description of key information

Skin irritation / corrosion (OECD 404), rabbit: irritating

Eye irritation / corrosion (OECD 405), rabbit: severe damages to the eye

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:
11 Oct - 02 Nov 2000
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:
adopted in 1992
Deviations:
no
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
adopted in 2015
Deviations:
yes
Remarks:
directly 3 animals tested instead of 1 animal in the initial test and 2 in the confirmatory test, 21 days instead of 14 days observation period, first scoring 30 min instead of 60 min after patch removal
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
Version / remarks:
adopted in 1998
Qualifier:
according to guideline
Guideline:
other: JMAFF, 59 NohSan No. 4200
Version / remarks:
adopted in 1985
GLP compliance:
yes
Species:
rabbit
Strain:
other: Hra:(NZW)SPF
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Covance Research Products Inc., Kalamazoo, Michigan
- Age at study initiation: approximately 16 weeks
- Weight at study initiation: 2.382 - 2.479 g
- Housing: individually in stainless steel cages
- Diet: measured amount of high-fiber rabbit diet (#5326, Purina Mills, Inc.)
- Water: ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16 - 22 °C
- Humidity (%): 30 - 70%
- 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): 0.5 g moistened with approximately 0.4 mL distilled water
Duration of treatment / exposure:
4 h
Observation period:
21 days
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: 6.25 cm²
- % coverage: 100
- Type of wrap if used: 8-ply gauze patch secured with paper tape, loosely overwrapped with Saran Wrap®, and secured with Elastoplast® tape

REMOVAL OF TEST SUBSTANCE
- Washing: The test sites were washed using water and disposable paper towels. Any residual test substance was removed from the test sitesas thoroughly as possible.
- Time after start of exposure: 4 h

OBSERVATION TIME POINTS
4, 24, 48, 72, 96 hours and 7, 14, 21 days (The degree of erythema and edema was read approximately 30 min after removal of the test substance. This time point is recorded as the 4 h score.)

SCORING SYSTEM:
- Method of calculation: Draize scoring system
Irritation parameter:
erythema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
3
Max. score:
4
Reversibility:
fully reversible within: 21 days
Irritation parameter:
erythema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2.67
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
erythema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2.67
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
erythema score
Basis:
mean
Remarks:
of 3 animals
Time point:
24/48/72 h
Score:
2.78
Max. score:
4
Reversibility:
fully reversible within: 21 days
Irritation parameter:
edema score
Basis:
animal #1
Time point:
24/48/72 h
Score:
4
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
edema score
Basis:
animal #2
Time point:
24/48/72 h
Score:
4
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
edema score
Basis:
animal #3
Time point:
24/48/72 h
Score:
4
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritation parameter:
edema score
Basis:
mean
Remarks:
of 3 animals
Time point:
24/48/72 h
Score:
4
Max. score:
4
Reversibility:
fully reversible within: 14 days
Irritant / corrosive response data:
Beside erythema and edema reactions blanching, desquamation, necrotic appearing areas, eschar, scar-like tissue and exfoliation were observed. Comparing these findings between day 7, 14 and 21 a clear tendency to reversibility and recovery of the skin is apparent, with only scar-like tissue remaining at the end.

Table 1: Results of skin irritation study

Animal number

Hour

Day

4*

24

48

72

96

7

14

21

Erythema

1

2b

2b

3b

4n

4n

4x

1d,s

0s

2

2b

2b

2b

4n

4n

4x

0s

0s

3

2b

2b

2b

4n

4n

4x

0s

0s

Edema

1

4

4

4

4

4

3

0

0

2

4

4

4

4

4

2

0

0

3

4

4

4

4

4

4

0

0

* 30 min after removal of the test substance

b = blanching

d = desquamation

e = eschar

n = necrotic appearing area

s = scar-like tissue

x = exfoliation

Interpretation of results:
Category 2 (irritant) based on GHS criteria
Conclusions:
CLP: Skin Irrit. 2, H315
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
24 - 27 Jun 1983
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
adopted in 2017
Deviations:
yes
Remarks:
temperature: 16°C (10- 25°C acceptable range) instead of 20+/-3°C; artificial light: 10 h dark / 14 h light instead of 12 h dark / 12 h light; no anesthetics/analgesics; directly 3 animals tested instead of 1 animal in the initial test
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
adopted in 1981
Deviations:
yes
Remarks:
temperature: 16°C (10 - 25°C acceptable range) instead of 20+/-3°C; artificial light: 10 h dark / 14 h light instead of 12 h dark / 12 h light
GLP compliance:
no
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Morton Commercial Rabbits, Parsonage Farm, Stansted, England
- Age at study initiation: approximately 3 months
- Weight at study initiation: 2.90 - 3.02 kg
- Housing: individually in suspended galvanised steel cages (55 x 48 x 40 cm), mesh floors
- Diet: S.Q.C. Rabbit Standard, pelleted (Special Diet Services Limited), ad libitum
- Water: tap water, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 16 (10 - 25 acceptable limits)
- Humidity (%): 50
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 10 / 14
Vehicle:
unchanged (no vehicle)
Controls:
not required
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 100 mg
Duration of treatment / exposure:
72 h
Observation period (in vivo):
72 h
Reading time points: 1, 24, 48 and 72 h
Number of animals or in vitro replicates:
3
Details on study design:
SCORING SYSTEM: Draize; additional assessment of pain response

TOOL USED TO ASSESS SCORE: pencil beam torch / ophthalmoscope / fluorescein
Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2.67
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
3
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2.67
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
not reversible
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.67
Max. score:
2
Reversibility:
not reversible
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
not reversible
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2.67
Max. score:
3
Reversibility:
not reversible
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2.67
Max. score:
3
Reversibility:
not reversible
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2.67
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2
Max. score:
4
Reversibility:
not reversible
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1.33
Max. score:
4
Reversibility:
not reversible
Irritant / corrosive response data:
Irritation responses evident as diffuse opacity or translucency of the whole visible corneal surface, injection of the conjunctival blood vessels and eversion of the eyelids due to moderate chemosis became more marked at each subsequent examination and culminated in a state of severe ocular irritation 4 days after treatment. At this time there was an ocular discharge, the conjunctivae had a beefy-red appearance, the eyelids were everted by moderate chemosis, the whole surface of the cornea was opalescent and the colour of the iris indicated marked congestion of that tissue. In addition, areas of ulceration had exposed part of the corneal stroma of 2 rabbits. After consideration of the deteriorating condition of the treated
eye, particularly the cornea, the study was terminated 72 h after treatment.

Table 1: Grades of ocular irritation responses following instillation of Chlorophen

Region of the eye

Response

Grade of response at time after instillation

1 h

24 h

48 h

72 h

Animal #1

Cornea

Opacity

1

2

3

3

Area

4

4

4

4

Iris

Value

0

1

1

1

Conjunctiva

Redness

1

2

3

3

Chemosis

2

2

2

2

Discharge

0

2

2

2

Animal #2

Cornea

Opacity

2

3

3

3

Area

4

4

4

4

Iris

Value

0

0

1

1

Conjunctiva

Redness

1

2

3

3

Chemosis

2

2

2

2

Discharge

0

2

3

2

Animal #3

Cornea

Opacity

2

2

3

3

Area

4

4

4

4

Iris

Value

0

1

1

1

Conjunctiva

Redness

1

2

3

3

Chemosis

2

1

1

2

Discharge

0

1

1

2

Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Conclusions:
CLP: Eye Dam. 1, H318
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Reliable data regarding skin irritation/ corrosion and eye irritation/ corrosion are available for the test substance. Therefrom, the tests reported by Glaza 2000 (skin irritation/ corrosion) as well as by Cummins and Gardner 1983 (eye irritation) are considered the most suitable as key studies for both endpoints.

 

Skin:

With respect to the key study, the dermal irritancy of the test substance was tested in 3 New Zealand White rabbits under semi-occlusive conditions (Glaza, 2000). The procedure followed OECD Guideline 404 and was conducted in compliance with GLP. The test was conducted by applying a paste containing 0.5 g test substance to the skin of rabbits for 4 h. The animals were observed for 21 days. The Draize scale was used for scoring the skin reactions. Severe erythema and edema were caused by the test material. The test substance is irritating to skin. Thus, under the conditions of this study, the test substance is classified as Skin Irritating Category 2 according to Regulation (EC) No 1272/2008.

 

In a further study, the dermal irritancy of the test substance was assessed in 3 New Zealand White rabbits under semi-occlusive conditions (Cummins and Gardner, 1983). The procedure followed OECD Guideline 404. After application of 0.5 g of the test substance the skin was exposed for 4 h. The animals were observed for 13 days. Skin reactions were scored according to the Draize scale. Very slight or well-defined erythema was apparent at all dermal test sites. 2/3 animals also developed very slight edema. In the study report it was concluded that the test substance was irritant to the skin. However, evaluation of the results according to Regulation (EC) 1272/2008 (CLP Regulation) reveals that the criteria for classification as skin irritant are not fulfilled. Thus, the test substance is to be considered as non irritant to the skin under the conditions of this study.

 

Additionally, the irritative effect of the test substance was investigated after application of 0.5 g on the intact skin on the inner side of one ear of 2 New Zealand White rabbits (Pauluhn, 1983). Exposure duration was 24 h. The animals were observed for 7 days. Skin reactions were graded according to the Draize scale. The test substance caused strong irritation on the intact epidermis of the ear of rabbits. However, since the study was not conducted in compliance with a guideline and GLP the study is not used for classification. Exposure duration and site as well as reading time points significantly differ from the criteria laid down for classification.

 

Eye:

With respect to the key study, the eye irritation potential of the test substance was tested in 3 New Zealand White rabbits (Cummins and Gardner, 1983). The test substance (100 mg) was instilled into the conjunctival sac of the lower eyelid of the rabbits. Eyes were not rinsed. Ocular reactions were scored at 1, 24, 48 and 72 h after test substance administration. The Draize system was used for scoring eventual lesions. The method used in this study is in general accordance with OECD Guideline 405. The test substance produced ocular irritation characterised by diffuse opacity or translucency of the whole visible corneal surface, injection of the conjunctival blood vessels and eversion of the eyelids due to moderate chemosis. Positive reactions were observed in all animals. Irritation responses of the treated eyes and other effects of treatment became more marked at each subsequent examination and culminated in a state of severe ocular irritation four days after treatment. The study was terminated 72 h after treatment in light of the deteriorating condition of the treated eyes, especially the cornea. It was considered that significant resolution of the treatment effects was most improbable within the period of extended observation allowed by the OECD test method. The test substance is strongly irritating to eyes. Thus, under the conditions of this study, the test substance is classified as Eye Damaging Category 1 according to Regulation (EC) No 1272/2008.

 

In a further study, the effect of the test substance on the mucosa of rabbits was examined (Pauluhn, 1983). The test substance was applied on the conjunctiva of one eye of 2 New Zealand White rabbits (single application). The animals were observed for 7 days. Eye reactions were graded according to the Draize scale. The test material caused strong effects on conjunctiva, iris and cornea. However, since the study was not conducted in compliance with a guideline and GLP the study is not used for classification. Methods of examination significantly differ from the criteria laid down for classification.

Justification for classification or non-classification

The available data on skin irritation / corrosion and eye irritation meet the criteria for classification according to Regulation (EC) 1272/2008. The test substance is classified as Skin Irritating Category 2, H315: Causes skin irritation as well as Eye Damaging Category 1, H319: Causes serious eye irritation.

In fact, this is further in line with the conclusion of the ECHA Committee for Risk Assessment (RAC) Opinion for harmonised EU classification and labelling of Chlorophene based on the same reliable data, which was adopted in 2015 ( RAC CLH-O-0000001412-86-58/F, 12 March 2015). As a result, and referring to the Adaptation to Technical Progress (ATP) to CLP Regulation, Chlorophene was inserted in ATP10 with following classification and labelling for skin and eye irritation: Skin irrit. 2, H315 and Eye Dam. 1, H318.