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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:
2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 437 (Bovine Corneal Opacity and Permeability 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

Test animals / tissue source

Species:
other: in vitro
Strain:
other: in vitro
Details on test animals or tissues and environmental conditions:
Not applicable: in vitro

Test system

Vehicle:
unchanged (no vehicle)
Controls:
other: not applicable: in vitro method
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 750 µL
Duration of treatment / exposure:
Corneas were incubated with the test article or appropriate control for 10 minutes at 32 degrees C.
Duration of post- treatment incubation (in vitro):
After rinsing with Minimal Essential Medium (MEM), the corneas were incubated for 120 minutes in Eagle's Minimum Essential Medium. After the completion of the incubation period, opacity was evaluated using an opacitometer. The cell culture medium was then replaced with Na-fluorescein medium and incubated for approximately 90 minutes at 32 C. Following the 90 minute Na-fluorescein exposure, permeability was measured spectrophotometrically.
Number of animals or in vitro replicates:
0, in vitro method
Details on study design:
REMOVAL OF TEST SUBSTANCE
- Washing (if done): The corneas were rinsed with MEM after a 10 minute exposure to the test article or appropriate control.
- Time after start of exposure: 10 minutes

SCORING SYSTEM: Corneal opacity and permeability were measured and used to calculate an in vitro irritancy score (IVIS)

TOOL USED TO ASSESS SCORE: TECAN Infinite M200 Pro Plate Reader

Results and discussion

In vitro

Results
Irritation parameter:
in vitro irritation score
Remarks:
IVIS
Run / experiment:
Score following a 10-minute exposure
Value:
3.9
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Remarks:
measured as mean opacity value + (15 x mean OD450 value)
Other effects / acceptance of results:
The mean in vitro Irritancy Score (IVIS) score following a 10 minutes exposure was 3.9.

Applicant's summary and conclusion

Interpretation of results:
other: Does not cause severe ocular damage
Conclusions:
Based on the results of the test (IVIS=3.9), the test substance is not Category 1, and it is not “no category.” No conclusion can be reached on ocular irritation potential.
Executive summary:

The corneal irritation and damage potential of the test article (clear and colourless liquid, Purity 99.64%, Lot 12912) was tested in the Bovine Corneal Opacity and Permeability test (BCOP test). The study was performed in compliance with OECD GLP (98) 17 (1997). The test method was based on OECD no. 437 (2009), EC No. 440/2008 B. 47 (2010), OTWG- ICCVAM-NICEATM (2006), INVITTOX 127 (2006), and Gautheron, P. et al. 18: 442-449 (1992).

 

Corneas were prepared in cell culture medium and incubated at 32°C for at least 1 hour prior to exposure. Corneas (3/group) were treated with 0.75 mL of undiluted test material and incubated in a horizontal position for 10 minutes at 32°C. A positive control (10% w/v Benzalkonium Chloride) and a negative control (physiological saline) were tested in parallel with the test material. At the end of the exposure period, the corneas were rinsed and incubated in fresh cell culture medium for 120 ± 10 minutes at 32 C. Opacity was evaluated using an opacitometer after the 2 hour post-exposure incubation. Following opacity readings, the cell culture medium was replaced with Na-fluorescein medium and incubated for approximately 90 minutes at 32°C. Following the 90 minute Na-fluorescein exposure, permeability was measured. The mean in vitro irritation score (IVIS) was calculated and the scores were classified according to protocol defined categories.

 

For the test article, the mean IVIS = 3.9 after 10 minutes. The mean opacity score was 4 (range 3 to 5) and the mean permeability score was 0.-0.004 (range -0.012 to 0.001). Controls performed as expected. No pH effect of the test substance was observed on the rinsing medium. 

 

Based on the results of the test (IVIS=3.9), the test substance is not Category 1, and it is not “no category.”