2-(2-oxo-5-(1,1,3,3-tetramethylbutyl)-2,3-dihydro-1-benzofuran-3-yl)-4-(1,1,3,3-tetramethylbutyl)phenyl acetate

Endpoint summary

Description of key information

Skin Irritation: not irritating; Notox 1999, OECD Guideline Study
Eye Irritation: not irritating; Notox 1999, OECD Guideline Study

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:

21.06. - 04.09.1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline study, GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

adopted 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

GLP compliance:

yes

Remarks:

NOTOX B.V., Hambakenwetering 7, 5231 DD ‘s-Hertogenbosch, The Netherlands

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Nederland, The Netherlands.
- Age at study initiation: at least 6 weeks old.
- Weight at study initiation: less than 3.5 kg
- Housing: Individually housed in labelled cages with perforated floors (Scanbur, Denmark) and equipped with an automatic drinking system (ITL, Bergen, The Netherlands)
- Diet: Standard laboratory rabbit diet, ad libitum (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Germany)
- Water: Free access to tap-water
- Acclimation period: at least 5 days before start of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 50
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12

Type of coverage:

semiocclusive

Preparation of test site:

other: clipped

Vehicle:

water

Controls:

other: adjacent untreated areas served as control

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 0.5 g

Duration of treatment / exposure:

4 hrs

Observation period:

3 days. skin reactions were assessed 1, 24, 48 and 72 hrs after the removal of the dressings

Number of animals:

3

Details on study design:

TEST SITE
- Approximately 24 hours before treatment, the dorsal fur was clipped with electric clippers, exposing an area of approximately 150 square centimeters (10x15 cm2).Each animal was treated by dermal application of 0.5 gram of the moistened test substance to the skin of one flank, using a Metalline patch of 2x3 cm. The patch was mounted on Micropore tape, which was wrapped around the abdomen and secured with Coban elastic bandage.

REMOVAL OF TEST SUBSTANCE
- Washing: The skin was cleaned of residual test substance using water
- Time after start of exposure: 4 hrs

SCORING SYSTEM:
The exposed skin areas were examined for signs of erythema and edema and the responses were scored (according to the grading system given in the OECD guideline 404) approximately 1 hr, and 24, 48 and 72 hours after removal of the patches.

Irritation parameter:

erythema score

Basis:

mean

Time point:

other: mean (24+48+72h)

Score:

0

Max. score:

4

Irritation parameter:

edema score

Basis:

mean

Time point:

other: mean (24+48+72h)

Score:

0

Max. score:

4

Irritant / corrosive response data:

No skin irritation and no corrosion was caused by 4 hours of exposure to the test article. All scores were zero at any time point.

Other effects:

No staining of the treated skin by the test substance was observed. No symptoms of systemic toxicity were observed in the animals during the test
period and no mortality occurred.

Interpretation of results:

not irritating

Remarks:

Migrated information

Conclusions:

The test article did not cause any signs of irritation when applied semiocclusively for 4 hours to the rabbit skin.

Executive summary:

In a primary skin irritation/corrosion study following OECD guideline 404 and in compliance with GLP, three rabbits were exposed to 0.5 gram of the test substance for four 4 hours. The test substance was moistened with water and then applied onto clipped skin using a semi-occlusive dressing. After 4 hours the dressing was removed and the skin was cleaned of residual test substance using water. Observations were made 1, 24, 48 and 72 hours after exposure. No skin irritation was caused by 4 hours of exposure to the test article. All scores for edema and erythema were zero at any time point. Based on the results and according to the EC criteria for classification and labeling requirements for dangerous substances and preparations (Guidelines in Commission Directive 93/21/EEC), the test article is regarded as non-irritating to the rabbit skin and does not have to be 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:

12.07. - 04.09.1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline study in compliance with GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted 1987

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

GLP compliance:

yes

Remarks:

NOTOX B.V., Hambakenwetering 7, 5231 DD ‘s-Hertogenbosch, The Netherlands

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Charles River Nederland, The Netherlands.
- Age at study initiation: at least 6 weeks old.
- Weight at study initiation: less than 3.5 kg
- Housing: Individually housed in labelled cages with perforated floors (Scanbur, Denmark) and equipped with an automatic drinking system (ITL, Bergen, The Netherlands).
- Diet: Standard laboratory rabbit diet, ad libitum (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Germany) approx. 100 g. per day
- Water: Free access to tap-water
- Acclimation period: at least 5 days before start of treatment

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 50
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12

Controls:

other: the untreated eye served as control

Amount / concentration applied:

TEST MATERIAL
- Amount applied: 44.7 +/- 0.8 mg (approximately 0.1 ml)

Duration of treatment / exposure:

single instillation

Observation period (in vivo):

Three days. The eyes were examined after 1 hour and 1, 2, 3 days after instillation of the test substance.

Number of animals or in vitro replicates:

3 males

Details on study design:

SCORING SYSTEM:
According to the grading system given in the OECD guideline 405

TOOL USED TO ASSESS SCORE: Fluorescein
Immediately after the 24 hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. When considered necessary, this procedure was repeated to assess recovery. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: mean (24+48+72h)

Score:

0

Max. score:

4

Irritation parameter:

iris score

Basis:

mean

Time point:

other: mean (24+48+72h)

Score:

0

Max. score:

2

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

other: mean (24+48+72h)

Score:

0.67

Max. score:

3

Reversibility:

fully reversible within: 72 hrs

Irritation parameter:

chemosis score

Basis:

mean

Time point:

other: mean (24+48+72h)

Score:

0.1

Max. score:

4

Reversibility:

fully reversible within: 48 hrs

Irritant / corrosive response data:

Instillation of approximately 45 mg (a volume of approximately 0.1 ml) of the test substance into one eye of each of three rabbits resulted in effects on the cornea, iris and conjunctivae. Slight dulling of the normal lustre (opacity grade 0) and epithelial damage (10% of the total corneal area) were observed in one animal, 24 hours after instillation. The corneal injury had resolved within 72 hours after instillation. No corneal injury was observed in the other two animals. Iridial irritation (grade 1), seen in all three animals, had resolved within 24 hours after instillation. Irritation of the conjunctivae was seen as slight redness, chemosis and discharge, which had completely resolved within 48 hours in two animals and within 72 hours in the third animal.

Other effects:

Remnants of the test substance were present in the eyes of all animals on day 1. No staining of peri-ocular tissues by the test substance was observed. No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred.

Table 1: Eye irritation scores of the test substance

	time	animal 1	animal 2	animal 3
corneal opacity	24 h	0	0	0
	48 h	0	0	0
	72 h	0	0	0
iris	24 h	0	0	0
	48 h	0	0	0
	72 h	0	0	0
conjunctivae	24 h	2	1	2
	48 h	0	0	1
	72 h	0	0	0
chemosis	24 h	0	0	1
	48 h	0	0	0
	72 h	0	0	0

Interpretation of results:

not irritating

Remarks:

Migrated information

Conclusions:

The test article caused minimal effects to iris, cornea and conjunctivae below the level of regulatory significance and is therefore regarded as not irritating to the rabbit eye.

Executive summary:

In an acute eye irritation/corrosion study according to OECD guideline 405 and in compliance with GLP, single samples of approximately 45 mg of the undiluted test article (a volume of approximately 0.1 ml) were instilled into one eye of each of three rabbits. Observations were made 1, 24, 48 and 72 hours after instillation. Instillation of the test substance resulted in effects on the cornea, iris and conjunctivae. Slight dulling of the normal lustre (opacity grade 0) and epithelial damage (10% of the total corneal area) were observed in one animal, 24 hours after instillation. The corneal injury had resolved within 72 hours after instillation. No corneal injury was observed in the other two animals. Iridial irritation (grade 1), seen in all three animals, had resolved within 24 hours after instillation. Irritation of the conjunctivae was seen as redness, chemosis and discharge, which had completely resolved within 48 hours in two animals and within 72 hours in the third animal. Remnants of the test substance were present in the eyes of all animals on day 1. The average scores (24 -72 hours) for all animals were 0.67 and 0.1 for erythema and edema, respectively. Based on these results and according to the EC criteria for classification and labeling requirements for dangerous substances and preparations (Guidelines in Commission Directive 93/21/EEC), the test substance does not need to be classified.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

There are valid in vivo data available for the assessment of the skin and eye irritation potential of the test item.

 

Skin

A GLP-compliant skin irritation test was performed with three New Zealand White rabbits according to OECD 404 (Notox, 1999). A 4-hour semiocclusive exposure to approximately 0.5 g of the test material moistened with water produced no effects during the observation period of 3 days on the treated sites of the animals. The mean scores for the 24, 48 and 72 h reading for erythema and edema were zero in all animals.

 

Eye

A GLP-compliant eye irritation test was performed with three New Zealand White rabbits according to OECD 405 (Notox, 1999). Single samples of approximately 45 mg of the test article (a volume of approximately 0.1 ml) were instilled into one eye of each of the three rabbits. Observations were made 1, 24, 48 and 72 hours after instillation. Instillation of the test substance resulted in effects on the cornea, iris and conjunctivae. Slight dulling of the normal lustre (opacity grade 0) and epithelial damage (10% of the total corneal area) were observed in one animal, 24 hours after instillation. The corneal injury had resolved within 72 hours after instillation. No corneal injury was observed in the other two animals. Iridial irritation (grade 1), seen in all three animals, had resolved within 24 hours after instillation. Irritation of the conjunctivae was seen as redness, chemosis and discharge, which had completely resolved within 48 hours in two animals and within 72 hours in the third animal. Remnants of the test substance were present in the eyes of all animals on day 1. The scores recorded were well below the threshold of significance.

In conclusion, based on the data presented, the test substance is neither irritating to skin nor to the eye of New Zealand White rabbits.

Justification for selection of skin irritation / corrosion endpoint:
GLP compliant guideline study

Justification for selection of eye irritation endpoint:
GLP compliant guideline study

Justification for classification or non-classification

Dangerous Substance Directive (67/548/EEC)

The available experimental test data is reliable and suitable for the purpose of classification under Directive 67/548/EEC. Based on the data, classification for skin/eye irritation is not warranted under Directive 67/548/EEC.

 

Classification, Labeling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for the purpose of classification under Regulation (EC) No.1272/2008. Based on the data, classification for skin/eye irritation is not warranted under Regulation (EC) No.1272/2008.