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Toxicological information

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:
2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study was conducted in compliance with OECD GLP regulations.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: Liquid
Details on test material:
- Name of test material (as cited in study report): MTDID 948
- Substance type: Mono-constituent
- Physical state: Liquid
- Analytical purity: 94.4%
- Purity test date: 16 April, 2014
- Lot/batch No.: 20014, Unit 027
- Expiration date of the lot/batch: 11 August, 2016
- Storage condition of test material: Ambient temperature

Test animals / tissue source

Species:
other: in vitro method
Strain:
other: in vitro method
Details on test animals or tissues and environmental conditions:
SOURCE OF COLLECTED EYES
- Source: Slaughterhouse v.d. Bor, Nijkerkerveen, The Netherlands
- Number of animals: 4
- Characteristics of donor animals (e.g. age, sex, weight): approximately 7 weeks old, male and female chickens (ROSS, spring chickens)
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): Heads were placed in small plastic boxes on a bedding of paper tissues moistened with isotonic saline. They were transported to the testing facility at ambient temperature.
- Time interval prior to initiating testing: Within 2 hours.
- indication of any existing defects or lesions in ocular tissue samples: None
- Indication of any antibiotics used: No data

Test system

Vehicle:
unchanged (no vehicle)
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 uL
Duration of treatment / exposure:
10 seconds
Duration of post- treatment incubation (in vitro):
After exposure and rinsing, each eye in its holder was returned to its chamber. The eyes were examined at approximately 0, 30, 75, 120, 180 and 240 minutes after treatment. Fluorescein retention was only scored at approximately 30 minutes after treatment. All examinations were carried out with the slit-lamp microscope.
Number of animals or in vitro replicates:
3 (test article and positive control), 1 (negative control)
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): Following treatment, each eye was rinsed with 20 mL of saline
- Time after start of exposure: 10 seconds

SCORING SYSTEM: The following criteria and scoring systems are applied for the assessment of possible effects:
Corneal swelling: Corneal swelling, expressed as a percentage, is calculated according to the following formula: "Corneal thickness at time t minus corneal thickness at time t=0, divided by corneal thickness at time t=0 and multiplied by 100". A negative swelling up to -5% (not unusual for control eyes) will be presented as 0% swelling. The mean percentage of swelling for the three test eyes will be calculated for each of the observation time points of 30, 75, 120, 180, and 240 minutes. The maximum mean percentage (can be at any of the time points) will be used for classification into one of four categories.
Corneal opacity: opacity degree of density (area most dense taken for scoring)
0= no apacity
0.5= very faint opacity (very slight)
1= scattered or diffuse areas, details of iris clearly visible (slight)
2= easily discernible translucent area, details of iris slightly obsucred (moderate)
3= severe corneal opacity, no specific details of iris visible, size of pupil barely discernible (severe)
4= complete corneal opacity, iris invisible (very severe)

Fluorescein retention:
0= no fluorescein retention
0.5= very minor single cell staining (very slight)
1= single cell staining scattered throughout the treated area of the cornea (slight)
2= focal or convluent dense single cell staining (moderate)
3= confluent large areas of the cornea retaining fluorescein (severe)

TOOL USED TO ASSESS SCORE: hand-slit lamp, fluorescein

Results and discussion

In vitro

Resultsopen allclose all
Irritation parameter:
cornea opacity score
Run / experiment:
All
Value:
ca. 0
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
percent corneal swelling
Run / experiment:
Mean
Value:
ca. 0.03
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
fluorescein retention score
Run / experiment:
All
Value:
ca. 0
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: None

DEMONSTRATION OF TECHNICAL PROFICIENCY: Yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes

In vivo

Irritant / corrosive response data:
The test article caused very slight (3%) corneal swelling, no opacity and no fluorescein retntion. Microscopic examination of the corneas did not reveal any abnormalities.

Applicant's summary and conclusion

Interpretation of results:
not irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based on the results of the study, the test article did not show eye irritation potential and is classified as not irritating.
Executive summary:

The test article was evaluated for eye irritation potential in the Isolated Chicken Eye (ICE) test. The study was conducted in compliance with OECD GLP regulations. The test method was based on OECD 438 (2013). The isolated chicken eyes were exposed to a single, neat application of 30 uL of the test article for 10 seconds followed by a 20 mL saline rinse. Negative (saline solution) and positive (benzalkonium chloride) controls were tested in parallel. Three parameters were measured to determine possible adverse eye effects: corneal thickness (expressed as corneal swelling), corneal opacity, and fluorescein retention of damaged epithelial cells. In addition, histopathology of the corneas was performed after exposure. Positive and negative controls performed as expected, indicating a valid test method. The test article caused very slight corneal swelling (3%), no opacity, and no fluorescein retention. Microscopic examination of test article-treated corneas did not reveal any abnormalities. Based on the results of the study, the test article did not show eye irritation potential and is classified as not irritating.