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

Eye irritation

Administrative data

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

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted 1987

Deviations:

no

Test guideline 2

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

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

431-770-1

EC Name:

-

Cas Number:

216698-07-6

Molecular formula:

C32 H44 O4

IUPAC Name:

2-[2-oxo-5-(2,4,4-trimethylpentan-2-yl)-2,3-dihydro-1-benzofuran-3-yl]-4-(2,4,4-trimethylpentan-2-yl)phenyl acetate

Details on test material:

- Description: White solid
- Purity: 98.7%
- Storage condition of test material: At room temperature in the dark

Test animals / tissue source

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

Test system

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.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: mean (24+48+72h)

Score:

0

Max. score:

4

Results 2

Irritation parameter:

iris score

Basis:

mean

Time point:

other: mean (24+48+72h)

Score:

0

Max. score:

2

Results 3

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

Results 4

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.

Any other information on results incl. tables

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

Applicant's summary and conclusion

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.