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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:
September 2011
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study performed according to OECD 438 and under GLP conditions for indicating severe eye damage. The test is, however, not fully validated for severe eye irritation but the results are sufficiently adequate to conclude on the eye irritation effect of the substance

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2011

Materials and methods

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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Cashmeran
- Molecular formula: C14H22O
- Molecular weight: 206.33
- Physical state: Liquid
- Storage condition of test material: Ambient

Test animals / tissue source

Species:
other: Chicken
Strain:
other: ROSS, spring chickens
Details on test animals or tissues and environmental conditions:
TEST ANIMALS (eye-donors)
- Source: Poultry slaughterhouse v.d. Bor, Amersfoortseweg 118, Nijkerkerveen, The Netherlands
- Age at study initiation: 7 weeks old
- Weight at study initiation: approx. 1.5-2.5 kg

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: saline (negative control) and 5% benzalkonium chloride (BAC, positive control)
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied: 30 uL
- Concentration: Undiluted (100%)
Duration of treatment / exposure:
10 seconds
Observation period (in vivo):
240 minutes (examination at 0, 30, 75, 120, 180 and 240 minutes after treatment)
Number of animals or in vitro replicates:
Number of eyes used:
- Test group: 3
- Positive control: 3
- Negative control: 1
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing: Rinsing with 20 mL saline
- Time after start of exposure: 10 seconds

SCORING SYSTEM: Corneal thickness and corneal opacity were scored at different time points, while fluorescein retention was scored only at 30 minutes after exposure. These endpoint were related to 4 classes of eye irritation (not; slightly; moderately; severely irritating) and finally reconciled to the classification criteria as applicable in EU-CLP, in accordance with the decision criteria in OECD438. Additionally, morphological and microscopic effects were examined.

TOOL USED TO ASSESS SCORE: Slit lamp microscope.

HISTOPATHOLOGY: After treatment, all eyes were preserved in a neutral aqueous phosphate-buffered 4% solution of formaldehyde and corneas were embedded in parraffin wax, sectioned and stained. Histopathological examination was not performed, but the microscopic slides were filed in the archive.

Results and discussion

In vivo

Results
Irritation parameter:
overall irritation score
Basis:
mean
Time point:
other: Not relevant (see remarks)
Score:
110
Max. score:
240
Reversibility:
other: Not relevant
Remarks on result:
other: Irritation index = maximum mean corneal swelling + maximum mean opacity (x20) + mean fluorescein score (x20)
Irritant / corrosive response data:
Cashmeran caused moderate swelling (20%) of the cornea, moderate or moderate to severe corneal opacity (2.2) and moderate or moderate to severe fluorescein retention (2.3). The negative control eye did not show any corneal effect and demonstrated that the general conditions during the tests were adequate. The positive control eyes showed severe corneal effects and demonstrated the suitability and sensitivity of the ICE to detect severe eye irritants.
Other effects:
Wrinkling of the epithelium followed by erosion/loosening of the top-layer of the epithelium were observed in the test substance group. Severe loosening of the epithelium was observed in the positive control group.

Any other information on results incl. tables

Please find in the table below the summary results.

Test material

Maximum score for:

Irritation categories

Irritation index

Classification (EU-CLP

Swelling %

Opacity

Fluorescein retention

Cashmeran

20

2.2

2.3

III;III;III

110

2

Saline (negative control)

0

0.0

0.0

Not applicable, one eye tested

BAC 5% (w/v) (positive control)

27

3.0

3.0

III;IV;IV

147

1

Applicant's summary and conclusion

Interpretation of results:
irritating
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Based on the observed irritation index for Cashmeran and according to the EU-CLP classification schemes of the Isolated Chicken Eye Test, Cashmeran should be classified as irritating and Category 2: “Irritating to eyes”.
Executive summary:

The eye irritating potential of Cashmeran was determined in an isolated chicken eye test performed according to OECD guideline 438 and under GLP conditions.

Cashmeran caused moderate swelling (20%) of the cornea, moderate or moderate to severe corneal opacity (2.2) and moderate or moderate to severe fluorescein retention (2.3). The negative control eye did not show any corneal effect and demonstrated that the general conditions during the tests were adequate. Furthermore, the positive control eyes showed severe corneal effects and demonstrated the suitability and sensitivity of the ICE to detect severe eye irritants. The irritation index for Cashmeran as defined in the OECD guideline was 110 out of a maximum of 240.

Based on the irritation index for Cashmeran the substance does not need to be classified as severely irritating but according to the EU-CLP classification schemes of the ICE, the substance should be classified as irritating and Category 2: “Irritating to eyes”.