Registration Dossier

Administrative data

Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
05 Mar 2003 - 10 July 2003
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: non-GLP study, alternative ex vivo screeening study.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2003
Report Date:
2003

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 405 (Acute Eye Irritation / Corrosion)
Deviations:
yes
Remarks:
ex vivo study
Principles of method if other than guideline:
The rabbit enucleated eye test is used at SPL as a first stage in the assessment of ocular irritancy potential. The preferred species of choice is the rabbit. The assay has undergone interlaboratory validation and has been shown to reliably detect test materials that are negligible, or moderate to severe ocular irritants compared to the OECD 405 guideline study.
GLP compliance:
yes (incl. certificate)
Remarks:
Dept. of Health, UK

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Physical state: white powder
-The pH of the test item at the concentration of 10% in purified water was 12.2 (SPL).
- Analytical purity: 99.8%
- Lot/batch No.: 4486
- Storage condition of test material: at room temperature, in the dark and under nitrogen gas.

Test animals / tissue source

Species:
other: ex vivo rabbit eyes
Strain:
not specified

Test system

Vehicle:
water
Controls:
yes, concurrent no treatment
Amount / concentration applied:
A volume of 0.1 ml of the test material, which was found to weigh approximately 55 mg (as measured by gently compacting the required volume into an adapted syringe) was sprinkled as evenly as possible over the surface of the cornea. After ten seconds the test material was washed off the cornea using a minimum of 20 ml of saline solution (approximately 32°C). After washing, some of the test material was found to have adhered to the cornea, this did not affect the assessment of irritation or swelling.
Duration of treatment / exposure:
Immediately following washing of the corneal surface, the treated eye was returned to the superfusion chamber and the saline drip repositioned to irrigate the eye. The untreated eyes were similarly washed and used for control purposes.
Observation period (in vivo):
Assessment of corneal cloudiness was made pre-enucleation, post equilibration and approximately 60, 120, 180 and 240 minutes following treatment, according to the numerical evaluation adopted from Advances in Modern Toxicology: Dermatoxicology, 4th Ed, (F Marzulli and H Maibach, eds) Hemisphere Publishing Corporation, Washington DC, 1991, pp 749-815.
Number of animals or in vitro replicates:
Three eyes were treated with test material, two additional eyes remained untreated for control purposes. The treatment eye was removed from thesuperfusion apparatus whilst still being held in the perspex clamp. The clamp/eye was then placed horizontally into a petri dish.

Results and discussion

In vivo

Resultsopen allclose all
Irritation parameter:
other: Corneal Opacity
Basis:
mean
Time point:
other: 1, 2, 3 and 4 hours
Score:
0
Max. score:
0
Reversibility:
fully reversible
Irritation parameter:
other: Corneal Epithelium Condition
Basis:
mean
Time point:
other: 1, 2, 3 and 4 hours
Score:
0
Max. score:
0
Reversibility:
not specified
Remarks on result:
other: normal compared to control eyes
Irritant / corrosive response data:
Corneal Opacity: No corneal effects were noted in the test eyes or control eyes during the study period.
Corneal thickness and Condition: Corneal swelling of the test eyes during the study period was slightly greater than that observed in the control eyes over the same period. The condition of the corneal epithelium of the test eyes and control eyes appeared normal during the study period.
Fluorescein Uptake: No fluorescein uptake was noted in the test eyes or control eyes 240 minutes after treatment.

Applicant's summary and conclusion

Interpretation of results:
moderately irritating
Remarks:
Migrated information Criteria used for interpretation of results: expert judgment
Conclusions:
Under the conditions of the test and following assessment of data for all end points, the test material is considered to have the potential to cause ocular irritancy in vivo. The test material is also likely to cause severe ocular irritancy in vivo in respect of its extreme alkalinity in aqueous solution.
Executive summary:

A screening study was performed to assess the ocular irritancy potential of the test material in the rabbit following application onto the cornea of the enucleated eye. The results of the study are believed to be of value in predicting the ocular irritation potential of the test material in man.

For that purpose 0.1 ml of the test material, which was found to weigh approximately 55 mg, was applied onto the cornea of each of three enucleated eyes which had been maintained at a temperature of 32°C ± 1.5°C within a superfusion chamber. A further two enucleated eyes were treated, for control purposes, with saline solution (0.9% Sodium Chloride).

 

Maximal ocular irritation observations recorded for the test eyes were as follows:

Corneal Opacity

Fluorescein Uptake

Corneal Swelling (%)

Condition of Corneal Epithelium

Test Eyes a [min]

Control Eyes b [min]

Cldy

Area

Int

Area

60

120

240

60

120

240

0

0

0

0

11.2

12.5

20.0

3.4

3.6

3.7

Normal

a          = For each time point the swelling recorded is the mean of three eyes

b         = For each time point the swelling recorded is the mean of two eyes

Cldy = Corneal cloudiness

Int = Intensity of fluorescein uptake

[min] = Minutes following treatment

It can be concluded that under the conditions of the test, the test material is considered to have the potential to cause ocular irritancy in-vivo. The test material is also likely to cause severe ocular irritancy in-vivo in respect of its extreme alkalinity in aqueous solution.