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Administrative data

Description of key information

Skin (OECD 404): not irritating (0.5 g, rabbit, semi-occlusive, 4 h)
Eye (OECD 405): not irritating (0.1 mL/15.8 – 17.7 mg, rabbit, single application without washing)

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:
12 Aug - 22 Aug 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP - Guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
Version / remarks:
adopted 2002
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)
Version / remarks:
adopted 1992
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2500 (Acute Dermal Irritation)
Version / remarks:
adopted August 1998
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), November 2000, including the most recent partial revisions
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Kisslegg, Germany
- Age at study initiation: at least 6 weeks
- Weight at study initiation: 1.465 - 1.515 kg
- Housing: individually in labelled cages with perforated floors (Scanbur, Denmark)
- Diet: standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Germany), approximately 100 g per day and pressed hay twice a week
- Water: tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.4 - 23.9
- Humidity (%): 45 - 89
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12
Type of coverage:
semiocclusive
Preparation of test site:
clipped
Vehicle:
other: watery ethanol (50% v/v)
Controls:
other: not required, untreated site of the same animal served as control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.5 mL
(0.5 g test substance was moistened with 0.5 mL vehicle before application to ensure close contact to the skin)

Duration of treatment / exposure:
4 hours
Observation period:
72 hours
reading time points: 1, 24, 48 and 72 hours
Number of animals:
3 males
Details on study design:
TEST SITE
- Area of exposure: dorsal area, 6 cm²
- Type of wrap if used: metalline patch mounted on Micropore tape which is wrapped around the abdomen and secured with Coban elastic bandage

REMOVAL OF TEST SUBSTANCE
- Washing (if done): residual test substance was removed using water and ethanol
- Time after start of exposure: 4 hours

SCORING SYSTEM: Draize scoring system
Irritation parameter:
erythema score
Basis:
mean
Remarks:
out of all three animals
Time point:
other: mean over 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: reversibility not applicable
Irritation parameter:
edema score
Basis:
mean
Remarks:
out of all three animals
Time point:
other: mean over 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: reversibility: not applicable
Irritant / corrosive response data:
The test substance resulted in very slight erythema in one animal (score of 1 at 1 hour reading after removal of the test substance) which was fully reversible within 24 hours.
Other effects:
No symptoms of toxicity and no staining of the skin by the test substance were observed.
Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
DSD: not classified
CLP: not classified
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:
18 Aug - 28 Aug 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP - Guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
adopted in 2012
Deviations:
yes
Remarks:
no anesthetics and systemic analgesics used
Qualifier:
according to guideline
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Version / remarks:
adopted 2002
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)
Version / remarks:
adopted 1992
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2400 (Acute Eye Irritation)
Version / remarks:
adopted August 1998
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), November 2000
Deviations:
no
GLP compliance:
yes
Species:
rabbit
Strain:
New Zealand White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland, Kisslegg, Germany
- Age at study initiation: 8-10 weeks
- Weight at study initiation: 1.651 - 1.785 kg
- Housing: individually in labelled cages with perforated floors (Scanbur, Denmark)
- Diet: standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Germany), approximately 100 g per day and pressed hay twice a week
- Water: tap water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.4 - 22.2
- Humidity (%): 43 - 89
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
unchanged (no vehicle)
Controls:
other: not required: the untreated eye of the same animal served as control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 16.8 mg (15.8 - 17.7 mg) (corresponding to a volume of approximately 0.1 mL)
Duration of treatment / exposure:
single application without washing
Observation period (in vivo):
72 hours
reading time points: 1, 24, 48 and 72 hours
Number of animals or in vitro replicates:
3 males
Details on study design:
SCORING SYSTEM: Draize scoring system

TOOL USED TO ASSESS SCORE: 2% fluorescein in water (pH 7.0)
Irritation parameter:
cornea opacity score
Basis:
mean
Remarks:
out of all 3 animals
Time point:
other: mean over 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
other: reversibility: not applicable
Irritation parameter:
iris score
Basis:
mean
Remarks:
out of all 3 animals
Time point:
other: mean over 24, 48 and 72 hours
Score:
0
Max. score:
2
Reversibility:
other: reversibility: not applicable
Irritation parameter:
conjunctivae score
Basis:
mean
Remarks:
out of all 3 animals
Time point:
other: mean over 24, 48 and 72 hours
Score:
0.33
Max. score:
3
Reversibility:
fully reversible within: 48 hours
Irritation parameter:
chemosis score
Basis:
mean
Remarks:
out of all 3 animals
Time point:
other: mean over 24, 48 and 72 hours
Score:
0
Max. score:
4
Reversibility:
fully reversible within: 24 hours
Irritant / corrosive response data:
Irritation of the conjunctivae was seen as redness, chemosis and discharge for all animals 1 hour after instillation. Redness was still visible 24 hours after instillation but was completely reversed thereafter. No iridial irritation or cornea opacity were observed. Treatment of the eyes with 2%
fluorescein 24 hours after test substance instillation revealed no corneal epithelial damage.
Other effects:
No staining of ocular tissues by the test substance was observed.
Remnants of the test substance were present in the eyes of two animals and on the outside of the eyelid in one animal on day 1.
No mortality and no symptoms of systemic toxicity were observed.

Table 1. Results of eye irritation study.

 

Rabbit #

 

Time [h]

 

conjunctivae

 

iris

 

cornea

 

 

conjunctivae

 

iris

 

cornea

 

redness

swelling

redness

swelling

1

 

 

 

 

1

2

1

0

0

 

24

1

0

0

0

48

0

0

0

0

72

0

0

0

0

average

0.3

0.0

0.0

0.0

Time to reversion

48.0

24.0

0.0

0.0

2

 

 

 

 

1

1

1

0

0

 

 

24

1

0

0

0

48

0

0

0

0

72

0

0

0

0

average

0.3

0.0

0.0

0.0

Time to reversion

48.0

24.0

0.0

0.0

3

 

 

 

 

1

2

1

0

0

 

 

24

1

0

0

0

48

0

0

0

0

72

0

0

0

0

average

0.3

0.0

0.0

0.0

Time to reversion

48.0

24.0

0.0

0.0

 

 

 

Time [h]

conjunctivae

 

iris

cornea

 

redness

swelling

 

 

avg. time to reversion

48.0

24.0

0.0

0.0

average

score

1

1.67

1.00

0.00

0.00

 

24

1.00

0.00

0.00

0.00

48

0.00

0.00

0.00

0.00

72

0.00

0.00

0.00

0.00

24+48+72

0.33

0.00

0.00

0.00

 

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
DSD: not classified
CLP: not classified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

A study assessing the skin irritation potential of 2-Hexyl-2 ‘-hydroxy-5 ‘methyldecananilide (AF-654) was performed according to OECD Guideline 404 (Hooiveld, 2003b). 0.5 g test substance was, due to its insolubility in water, moistened with watery ethanol (50% v/v) to ensure close contact with the skin, and applied sequentially to the clipped skin of New Zealand White rabbits under semi-occlusive conditions for 4 hours. The scoring of skin reactions (erythema and edema) was performed 1, 24, 48 and 72 hours after removal of the dressing. Exposure to the test substance resulted in very slight erythema (grade 1) in one animal at the 1-hour reading time point. The erythema had resolved within 24 hours. No staining of the treated skin by the test substance was noted.

In the acute dermal toxicity study, AF-654 was administered to rats in a limit test according to OECD Guideline 402 (van Otterdijk, 2004a). A single dose of 2000 mg/kg bw of the test substance (in propylene glycol) was applied to the clipped skin of rats (5 per sex) under occlusive conditions for 24 hours. White staining of the treated skin area was noted among 9/10 animals between Days 2 and 9. In addition, slight erythema (Days 2 and/or 3) or scales (between Days 3 and 12) classified as grade 1 were noted on the treated skin area of 2/5 and 1/5 females, respectively.

The skin irritation study, performed according to the standard test (OECD Guideline 404 with 4-hour exposure period under semi-occlusive conditions) did not result in skin irritation. The longer exposure period (24 hour) employed in the acute dermal toxicity study shows that even under more rigorous conditions the test substance does not cause skin irritation that leads to a classification. Therefore, 2-Hexyl-2 ‘-hydroxy-5 ‘methyldecananilide (AF-654) is considered not to be a skin irritant.

To evaluate the eye irritation potential of AF-654, a study was performed according to OECD Guideline 405 (Hooiveld, 2003c). 15.8 – 17.7 mg (weight corresponding to 0.1 mL) was instilled into one eye each of 3 New Zealand White rabbits, without washing. The other eye remained untreated and served as control. Remnants of the test substance were still observed in the eyes of 2/3 animals and outside the eye of 1/3 animals on Day 1 after instillation. The eyes were examined and scored 1, 24, 48 and 72 hours after the treatment. Instillation of the test substance resulted in irritation of the conjunctivae seen as redness, chemosis and discharge for 3/3 animals 1 hour after instillation. Redness (grade 1) was still visible 24 hours after instillation but had resolved completely within the 48-hour reading time point. No iridial irritation or corneal opacity was observed, and treatment of the eyes with 2% fluorescein, 24 hours after test substance instillation, revealed no corneal epithelial damage in any of the animals. The mean score for conjunctivae after 24, 48, and 72 hours was 0.33 for all 3 animals. The effects on cornea, iris and chemosis after 24, 48, and 72 hours were scored 0 for 3/3 animals. Based on the results of this study, 2-Hexyl-2 ‘-hydroxy-5 ‘methyldecananilide (AF-654) is considered not to be an eye irritant.


Justification for selection of skin irritation / corrosion endpoint:
There is only one study available.

Justification for selection of eye irritation endpoint:
There is only one study available.

Justification for classification or non-classification

The available data on skin and eye irritation of the test substance do not meet the criteria for classification according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.