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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:
16 January 2015 - 16 January 2015
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
The test is described in OECD 438 and is approved by international regulatory agencies as a replacement for the identification of nonirritant, corrosives/severe irritants in the in vivo Rabbit Eye Assay (OECD 405).

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
Deviations:
no
Qualifier:
according to
Guideline:
EU method B.48 (Isolated chicken eye test method for identifying occular corrosives and severe irritants)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Batch number: 141016
- Appearance: Blue powder
- Purity: ≥85.5%
- Expiry date: 19 Nov 2018
- Storage conditions: Controlled Room Temperature (15-25oC, below 70 RH%)

Test animals / tissue source

Species:
chicken
Strain:
other: ROSS 308
Details on test animals or tissues and environmental conditions:
SOURCE OF COLLECTED EYES
- Source: TARAVIS KFT. (Address: 9600 Sárvár, Rábasömjéni út. 129., Hungary)

- Number of animals: 7 treatements (3 test item, 3 positive and 1 negative control)

- Characteristics of donor animals: age - : approx. 7 weeks old

- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): heads transported to the lab at ambient temperature at the earliest convenience. The heads were inspected for appropriate quality and wrapped with tissue paper moistened with saline, then placed in a plastic box which was closed. The heads were received at the lab. and processed within approximately 2 hours of collection.

- Time interval prior to initiating testing: acclimatization for approximately 45 to 60 minutes

- indication of any existing defects or lesions in ocular tissue samples: If the cornea was in good condition, the eyeball was carefully removed from the orbit for testing. Eyes with a high baseline fluorescein staining (i.e., > 0.5) or corneal opacity score (i.e., > 0.5) were rejected. any eye with cornea thickness deviating more than 10 % from the mean value for all eyes, or eyes that showed any other signs of damage, were rejected and replaced.

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): 30mg
Duration of treatment / exposure:
10s
Observation period (in vivo):
Test eyes were evaluated at approximately 30, 75, 120, 180 and 240 minutes after the post-treatment rinse
Number of animals or in vitro replicates:
7 (3 test item treated, 3 positive control and 1 negative control)
Details on study design:
SELECTION AND PREPARATION OF ISOLATED EYES

NUMBER OF REPLICATES

NEGATIVE CONTROL USED
30 μL of physiological saline (Salsol solution, 0.9% (w/v) NaCl solution)

POSITIVE CONTROL USED
30 mg Imidazole

APPLICATION DOSE AND EXPOSURE TIME
10s

OBSERVATION PERIOD
evaluated at approximately 30, 75, 120, 180 and 240 minutes

REMOVAL OF TEST SUBSTANCE
- Volume and washing procedure after exposure period: 20 mL physiological saline at ambient temperature
- Indicate any deviation from test procedure in the Guideline

METHODS FOR MEASURED ENDPOINTS:
Corneal thickness and corneal opacity were measured at all time points. Fluorescein retention was measured on two occasions, at base line (t=0) and approximately 30 minutes after the post-treatment rinse. Haag-Streit Bern 900 slit-lamp microscope was used for the measurements.

Results and discussion

In vitro

Resultsopen allclose all
Irritation parameter:
percent corneal swelling
Run / experiment:
mean at up to 75 min
Value:
14.1
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation parameter:
percent corneal swelling
Run / experiment:
mean at up to 240min
Value:
27.2
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation parameter:
cornea opacity score
Run / experiment:
mean
Value:
2.33
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Irritation parameter:
fluorescein retention score
Run / experiment:
mean
Value:
2.33
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid

Applicant's summary and conclusion

Interpretation of results:
other:
Conclusions:
Based on this in vitro eye irritation in the isolated chicken eyes test with test item, the test item is not classified as a severe irritant and not classified as nonirritant. It is concluded that an in vivo study is required for proper classification.