Z-85

Eye irritation

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Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 October 2006 to 8 December 2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Reference

Name:

Unnamed

Type:

Constituent

Test material form:

liquid: viscous

Details on test material:

Description: tan, slightly viscous liquid
Storage: room temperature in dark

See justification for read-across in Section 13

Test animals / tissue source

Species:

rabbit

Strain:

not specified

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml

Duration of treatment / exposure:

Preliminarty Test: Rabbit Enucleated Eye Test (REET)
The test material was applied to the enucleated eye for 30 seconds and then washed away using saline.

Observation period (in vivo):

Corneal cloudiness: pre-enucleation, post equilibration, 60, 120, and 240 minutes following treatment.
Corneal thickness: pre-enucleation, post equilibration, 60, 120, and 240 minutes following treatment.
Corneal epithelium: 60, 120, and 240 minutes following treatment.
Fluorescein uptake: pre-enucleation, post equilibration, and 240 minutes following treatment.

Number of animals or in vitro replicates:

Test material: 3 enucleated eyes
Control: 2 enucleated eyes treated with saline

Details on study design:

Preliminarty Test: Rabbit Enucleated Eye Test (REET)

This test was completed prior to an in vivo testing in accordance with OECD Guideline 405 as a step-wise procedure in the interest of animal welfare. This study was completed to ascertain any ocular irritancy potential of the test material in the rabbit following application onto the cornea of the enucleated eye. The REET was completed to confirm information that the test material may cause irritation in a rabbit eye.

For the REET, 0.1 mL of test material was applied onto the cornea of three separate enucleated eyes. The enucleated eyes have been stored at 32 +/- 1.5 degrees C within a superfusion chamber. The superfusion chamber, a water heating circulator, gave a stable temperature. Also, a persistaltic pump was used to supply saline solution at a flow rate of 0.15 - 0.4 ml/minute into the reat of each chamber of the apparatus in order to irrigate the surface of the cornea.

The removed eyes were allowed acclimate for 30 minutes. Three eyes were treated with test material and two eyes were untreated and were controls. For test eyes, 0.1 mL of undiluted test substance was applied to the cornea and allowed to remain for 30 seconds. Then the test material was washed away using 20 mL of saline solution. Immediately following the saline wash, the treat eye was returned to the chamber and continued to be flushed wth saline.

Endpoints included corneal opacity, condition of corneal epithelium, fluorescein uptake, and percentage change of corneal thickness.

After analysis of REET results, it was determined that the test material has the potential to cause severe ocular irritancy in vivo. Therefore, the in vivo eye irritation study is not required.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

cornea opacity score

Run / experiment:

1

Value:

1

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Results 2

Irritation parameter:

cornea opacity score

Run / experiment:

2

Value:

1

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Results 3

Irritation parameter:

cornea opacity score

Run / experiment:

3

Value:

1

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Results 4

Irritation parameter:

percent corneal swelling

Run / experiment:

1

Value:

27

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Results 5

Irritation parameter:

percent corneal swelling

Run / experiment:

2

Value:

113.9

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Results 6

Irritation parameter:

percent corneal swelling

Run / experiment:

3

Value:

681.6

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Results 7

Irritation parameter:

other: Flourescein Uptake (Intensity)

Run / experiment:

1

Value:

1

Negative controls validity:

valid

Positive controls validity:

valid

Results 8

Irritation parameter:

other: Flourescein Uptake (Intensity)

Run / experiment:

2

Value:

1

Negative controls validity:

valid

Positive controls validity:

valid

Results 9

Irritation parameter:

other: Flourescein Uptake (Intensity)

Run / experiment:

3

Value:

1

Negative controls validity:

valid

Positive controls validity:

valid

Results 10

Irritation parameter:

other: Flourescein Uptake (Area)

Run / experiment:

1

Value:

2

Negative controls validity:

valid

Positive controls validity:

valid

Results 11

Irritation parameter:

other: Flourescein Uptake (Area)

Run / experiment:

2

Value:

2

Negative controls validity:

valid

Positive controls validity:

valid

Results 12

Irritation parameter:

other: Flourescein Uptake (Area)

Run / experiment:

3

Value:

2

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

For REET cut-off:
1. Max corneal opacity > or = 4
2. Max fluorescein uptake > or = 4
3. Mean Corneal swelling > or = 25%
4. Corneal epithelium observations: pitting, mottling, sloughing

-Corneal sloughing was seen in all test eyes at 2, 3, and 4 hr post treatment. The corneal epithelium did not demonstrate any symptons and was categorized as normal at all time points.

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

The test material has the potential to cause severe ocular irritancy ex vivo. In accordance with EU CLP Regulation No. 1272/2008 classification of this substance as Eye Dam. Category 1 is required for eye irritation.

Executive summary:

Test Guidance

The Rabbit Enucleation Eye Test (REET) was completed prior to an in vivo testing in accordance with OECD Guideline 405 as a step-wise procedure in the interest of animal welfare.

Method and materials

This study was completed to ascertain any ocular irritancy potential of the test material in the rabbit following application onto the cornea of the enucleated eye. The REET was completed to confirm information that the test material may cause irritation in a rabbit eye.

For the REET, 0.1 mL of test material was applied onto the cornea of three separate enucleated eyes. The enucleated eyes have been stored at 32 +/- 1.5 degrees C within a superfusion chamber. The superfusion chamber, a water heating circulator, gave a stable temperature. Also, a persistaltic pump was used to supply saline solution at a flow rate of 0.15 - 0.4 ml/minute into the reat of each chamber of the apparatus in order to irrigate the surface of the cornea.

The removed eyes were allowed acclimate for 30 minutes. Three eyes were treated with test material and two eyes were untreated and were controls. For test eyes, 0.1 mL of undiluted test substance was applied to the cornea and allowed to remain for 30 seconds. Then the test material was washed away using 20 mL of saline solution. Immediately following the saline wash, the treat eye was returned to the chamber and continued to be flushed wth saline.

Endpoints included corneal opacity, condition of corneal epithelium, fluorescein uptake, and percentage change of corneal thickness. Following 4 hours of treatment, all test eyes had scores > or = to 1 for corneal opacity. All other endpoints were increased as compared with that of the control eyes.

Results

After analysis of REET results, it was determined that the test material has the potential to cause severe ocular irritancy ex vivo. Therefore, the acute in vivo eye irritation study is not required.

Conclusion

The test material has the potential to cause severe ocular irritancy ex vivo. In accordance with EU CLP Regulation No. 1272/2008 classification of this substance as Eye Dam. Category 1 is required for eye irritation.