2-Hexyl-2 ‘-hydroxy-5 ‘methyldecananilide

Endpoint summary

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.